Effects of lactoferrin supplementation on ileal and total tract nutrient digestibility,gastrointestinal microbial populations,and immune characteristics of ileal cannulated,healthy, adult dogs

Abstract

Orally supplemented lactoferrin derived from bovine milk is purported to have beneficial effects on gut health of animals. Bovine lactoferrin (0, 60, or 120 mg/d) was fed to ileal cannulated, adult dogs in a replicated 3 × 3 Latin square design with 14 d periods. Control dogs tended (p = 0.06) to have higher fecal DM concentrations compared with dogs supplemented with 120 mg/d lactoferrin (34.5 vs. 32.9%). Fecal scores ranged from 3.0 – 3.3, suggesting that feces of all dogs was near the desired consistency, with dogs supplemented with 120 mg/d lactoferrin tending (p = 0.08) to have higher fecal scores. Ileal azoreductase activity tended (p < 0.10) to be higher in dogs supplemented with 60 or 120 mg/d lactoferrin (609 vs. 592 nmol/h per g ileal DM, respectively) as compared with unsupplemented dogs (272 nmol/h per g ileal DM). The following bacterial groups were measured: bifidobacteria, Campylobacter spp., Clostridium spp., eubacteria, Escherichia coli, Lactobacillus spp., Staphylococcus spp., and Streptococcus spp. Fecal streptococci concentrations were lower (p < 0.05) for dogs receiving 60 mg/d lactoferrin (8.60 log₁₀ cfu/g fecal DM) as compared with unsupplemented dogs (9.19 log₁₀ cfu/g fecal DM) or dogs receiving 120 mg lactoferrin/d (9.43 log₁₀ cfu/g fecal DM). Dogs supplemented with 120 mg/d lactoferrin tended (p = 0.08) to have higher fecal indole concentrations as compared to unsupplemented dogs (1.80 vs. 1.46 μmol/g fecal DM). Because most bacterial groups measured were unaffected, it appears that lactoferrin did not exhibit prebiotic activity, and based on the data collected, lactoferrin also did not appear to have major effects on indices of health in the dog.     

Influence of supplemental high molecular weight pullulan or gamma-cyclodextrin on ileal and total tract nutrient digestibility, fecal characteristics, and microbial populations in the dog

This study was conducted to determine if supplemental pullulan and gamma-cyclodextrin affect canine nutrient digestibility, microbial populations, and fecal characteristics. Ileal cannulated dogs were fed a commercial diet, and treatments were administered daily in a 5 x 5 Latin square design: (i) no supplement; (ii) 2 g pullulan; (iii) 4 g pullulan; (iv) 2 g gamma-cyclodextrin; (v) 4 g gamma-cyclodextrin. Ileal and fecal samples were collected the last 4 d of each 14-d period. Increasing pullulan tended (p < 0.10) to linearly increase ileal bifidobacteria and lactobacilli and quadratically increase fecal lactobacilli. A similar response was noted in ileal bifidobacteria and lactobacilli with gamma-cyclodextrin. Gamma-Cyclodextrin resulted in a quadratic decrease (p < 0.05) in fecal Clostridium perfringens. Increasing pullulan linearly increased (p < 0.05) fecal score, while gamma-cyclodextrin resulted in a linear decrease (p < 0.05). Pullulan and gamma-cyclodextrin supplementation may have beneficial effects on the microbial ecology of dogs.     

Effects of Supplemental Fructooligosaccharides Plus Mannanoligosaccharides on Immune Function and Ileal and Fecal Microbial Populations in Adult Dogs

The goal of this study was to examine whether supplemental fructooligosaccharides (FOS) plus mannanoligosaccharides (MOS) influenced immune function and ileal and fecal microbial populations of adult dogs. Eight adult dogs surgically fitted with ileal cannulas were used in a crossover design. Dogs were fed 200g of a dry, extruded, kibble diet twice daily. At each feeding, dogs were dosed with either 1g sucrose (placebo) or 2g FOS plus 1g MOS orally via gelatin capsule. Fecal, ileal, and blood samples were collected at the end of each 14-d period to measure microbial populations and immune characteristics. Treatment least squares means were compared using the GLM procedure of SAS. Supplementation of FOS plus MOS increased fecal bifidobacteria and fecal and ileal lactobacilli concentrations. Dogs fed FOS plus MOS also tended to have lower blood neutrophils and greater blood lymphocytes vs placebo. Serum, fecal, and ileal immunoglobulin concentrations were unchanged by treatment. Supplementation of FOS plus MOS beneficially altered indices of gut health by improving ileal and fecal microbial ecology. Supplementation of FOS plus MOS also altered immune function by causing a shift in blood immune cells.     

Selected Gelling Agents in Canned Dog Food Affect Nutrient Digestibilities and Fecal Characteristics of Ileal Cannulated Dogs

Little is known about the effects of gelling agents in canned dog food on nutrient digestibilities and fecal characteristics. Dogs were fed canned diets with either no gelling agent (control) or one of three gelling agents, wheat starch, a guar gum/carrageenan mixture (50:50), and a locust bean meal (LBM)/ carrageenan mixture (50:50) incorporated at two levels (0.2 and 0.5% of the diet on a wet weight basis). Six dogs were fed each diet in a 6 ‐ 7 Youden square design. Dogs fed diets containing gelling agents had higher ileal digestibilities of OM (P = 0.05), fat (P < 0.01), GE (P = 0.02), and total amino acids (P = 0.04) and lower (P < 0.01) total tract DM digestibilities when compared to dogs fed the control diet. Fecal output by dogs fed the control diet, expressed on both an as-is (73.4g/d) and DM (45.6g/d) basis, was lower (P < 0.01) than for dogs fed gelling agent-containing diets (mean 102.3g/d as-is and 57.6g/d DM). Dogs fed the control diet also had lower ( P < 0.01) fecal DM percentages and higher (P = 0.02) fecal scores than dogs fed gelling agent-containing diets.     

Effects of bovine lactoferrin on the immune system and the intestinal microflora of adult dogs

Eighteen Beagle dogs were used to evaluate the effects of bovine lactoferrin (bLF) on immune function and faecal microbial populations. The study comprised three feeding periods, each lasting four weeks. After an initial control Period 1, six dogs per group were supplemented with 0, 120 and 1800 mg bLF/kg dry diet, respectively (Period 2). In Period 3 dogs received again control diets. Peripheral blood mononuclear cell subsets, lymphocyte proliferative response to concanavalin A, phytohaemagglutinin and pokeweed mitogen and plasma IgA and IgG concentrations were analysed. The faecal concentrations of aerobic and anaerobic bacteria, Escherichia coli, Clostridium perfringens, Lactobacillus spp. and Bifidobacterium spp. were determined by cultural methods. Supplementation of bLF increased the number of monocytes, T cells and cytotoxic T cells in the blood and the proliferative response of peripheral blood mononuclear cells. The leukocyte counts were not affected, except monocytes that increased after the supplementation with bLF. Plasma immunoglobulin concentrations were unchanged by treatment. Dogs supplemented with bLF tended to have lower faecal concentrations of E. coli and Clostridium perfringens. In conclusion, bLF seems to alter indices of the cellular immune response and faecal microbial populations of healthy adult dogs.     

Effects of bovine lactoferrin on the immune system and the intestinal microflora of adult dogs

Eighteen Beagle dogs were used to evaluate the effects of bovine lactoferrin (bLF) on immune function and faecal microbial populations. The study comprised three feeding periods, each lasting four weeks. After an initial control Period 1, six dogs per group were supplemented with 0, 120 and 1800 mg bLF/kg dry diet, respectively (Period 2). In Period 3 dogs received again control diets. Peripheral blood mononuclear cell subsets, lymphocyte proliferative response to concanavalin A, phytohaemagglutinin and pokeweed mitogen and plasma IgA and IgG concentrations were analysed. The faecal concentrations of aerobic and anaerobic bacteria, Escherichia coli, Clostridium perfringens, Lactobacillus spp. and Bifidobacterium spp. were determined by cultural methods. Supplementation of bLF increased the number of monocytes, T cells and cytotoxic T cells in the blood and the proliferative response of peripheral blood mononuclear cells. The leukocyte counts were not affected, except monocytes that increased after the supplementation with bLF. Plasma immunoglobulin concentrations were unchanged by treatment. Dogs supplemented with bLF tended to have lower faecal concentrations of E. coli and Clostridium perfringens. In conclusion, bLF seems to alter indices of the cellular immune response and faecal microbial populations of healthy adult dogs.     

Oral administration of Lactobacillus plantarum strain Lq80 to weaning piglets stimulates the growth of indigenous lactobacilli to modify the lactobacillal population

The composition and the number of fecal or intestinal lactobacilli were determined in weaned piglets before (pre-administration and day 0) and during (days 4, 7, and 14) the administration of Lactobacillus plantarum strain Lq80 (daily dose=10(10) cells). Fecal or intestinal lactobacilli were isolated, and the isolates were grouped by RAPD-PCR and further identified by 16S rDNA partial sequences. During administration, the number of lactobacilli in feces and intestinal contents (log cfu/g) increased from day 0 to day 4 (7.96 vs. 10.07, p<0.05), to day 7 (7.96 vs. 10.18, p<0.05), and to day 14 (7.96 vs. 9.02, p=0.07) in the administered group, although L. plantarum was isolated from the feces of piglets in the administered group at 5.0x10(6 cfu/g. The level of culturable lactobacilli was significantly higher on day 7 in the administered group than in the control group (8.93 vs. 10.18, p)=0.0002). Furthermore, the minor species in the lactobacillal population under the control condition become detectable with the administration of strain Lq80. The oral administration of L. plantarum strain Lq80 was effective to stimulate the development of the lactobacillal population in the intestine of post-weaning piglets.     

Effect of microbial phytase on apparent ileal amino acid digestibility of phosphorus-adequate diets in growing pigs

Six ileally cannulated pigs (mean initial body weight 34.8 kg) were used to study the effect of microbial phytase on apparent ileal digestibility of P, total N and amino acids. Three P-adequate diets (digestible P concentration 2.3 g kg(- )l) containing barley (B), soyabean meal (S) or a mixture of the two (BS) with or without phytase supplement (1000 FTU x kg(-1)) were fed to pigs using a 6 x 6 Latin square design. The addition of phytase increased (p < 0.05) apparent ileal P digestibility of diets B, S and BS by 16.5, 19.2 and 19.2%, respectively. There was no effect of phytase on the ileal digestibility of total N. Apparent ileal digestibility of amino acids tended to increase in the BS diet supplemented with phytase (mean improvement of 2.2%); but no significant difference was found for any amino acid as compared with the unsupplemented diet. To asses the additivity of apparent amino acid digestibility, the determined values for the BS diet were compared to those calculated from digestibilities found in diets B and S. There were no significant differences between the determined and calculated values. It is concluded that the addition of microbial phytase to P-adequate diets does not affect ileal amino acid digestibility in growing pigs and that the apparent amino acid digestibility values determined in single ingredients may be additive when included into a complex diet.     

Gastrointestinal and immunological responses of senior dogs to chicory and mannan-oligosaccharides

Thirty-four senior dogs (pointers 8-11 years, beagles 9-11 years) were used to evaluate the effects of oligosaccharides on nutritional and immunological characteristics. Dogs were randomly allotted to treatments [1% chicory (CH), 1% mannan-oligosaccharide (MOS), 1% chicory + 1% MOS (CM), or no supplementation (control, CON)] in a parallel design with a 4 week baseline period followed by a 4 week treatment period. Dietary supplementation with MOS or CM tended (P = 0.07) to increase food intake due, in part, to an increase in fermentable fibre and a decrease in energy content of the diet. Although wet faecal output increased (P < 0.05) for dogs supplemented with MOS or CM, when corrected for food intake, no differences were noted. The CM treatment increased (P < 0.05) faecal score (1 = hard and dry, 5 = watery liquid), although these scores remained in a desirable range (3 to 3.5). Chicory supplementation increased (P = 0.07) fat digestibility. Chicory or MOS increased (P < or = 0.05) faecal bifidobacteria concentrations 0.4 and 0.5 log10 cfu/g DM, respectively, compared to the CON, while MOS decreased (P < 0.05) faecal E. coli concentrations. Oligosaccharides did not affect white blood cell (WBC) concentrations, but CH and CM tended to increase (P = 0.10) neutrophil concentrations compared to control dogs. Peripheral lymphocyte concentrations were decreased in dogs supplemented with MOS (P = 0.06) and CM (P < 0.05). Chicory and MOS alter faecal microbial populations and certain indices of the immune system of senior dogs.     

Effects of a probiotic Lactobacillus acidophilus strain on feed tolerance in dogs with non-specific dietary sensitivity

This study investigated the effects of the probiotic Lactobacillus acidophilus strain DSM 13241 in dogs with non-specific dietary sensitivity (NSS). Six adult German Shorthair Pointers with NSS consecutively received a control dry diet and the same diet supplemented with the probiotic (6 x 10(6) cfu/g) for 12 weeks each, followed by another control period of four weeks. Frequency of defecations, faecal quality and nutrient digestibility were determined. Faeces were cultured for Clostridium perfringens, Escherichia spp., lactobacilli and bifidobacteria and quantitative fluorescence in situ hybridisation (FISH) was performed. Feeding the probiotic improved faecal consistency, faecal dry matter and defecation frequency (p < 0.05). Faecal concentrations of culturable lactobacilli and bifidobacteria increased numerically, but not significantly, also the decrease of the numbers of C. perfringens and Escherichia spp. did not reach the level of significance. Results from FISH showed more constant bacterial populations. It can be concluded that L. acidophilus DSM 13241 can stabilise the digestive processes in dogs with NSS.     

Calcium supplementation effect on calcium balance in endurance-trained athletes during prolonged hypokinesia and ambulatory conditions

Calcium (Ca) supplements may be used to normalize Ca-balance changes but little is known about the effect of Ca supplements on Ca balance during hypokinesia (decreased kilometers per day). The aim of this study was to evaluate the effect of daily intakes of Ca supplements on Ca balance during hypokinesia (HK). Studies were done during 30 d of a pre-HK period and during 364 d of a HK period. Forty male athletes aged 23–26 yr were chosen as subjects. They were divided equally into four groups: unsupplemented ambulatory control subjects (UACS), unsupplemented hypokinetic subjects (UHKS), supplemented hypokinetic subjects (SHKS), and supplemented ambulatory control subjects (SACS). The SHKS and UHKS groups were kept under an average running distance of 0.7 km/d. In the SHKS and SACS groups supplemented with 35.0 mg Ca lactate/kg body weight. Fecal Ca loss, urinary excretion of Ca and phosphate (P), serum concentrations of ionized calcium (Ca_I) total Ca, P, and Ca balance, intact parathyroid hormone (iPTH) and 1,25 dihydroxyvitamin D (1,25(OH)2D), anthropometric characteristics and peak oxygen uptake were measured. Fecal Ca excretion, urinary Ca and P excretion, serum Ca_I, total Ca, and P concentration, and negative Ca balanced increased significantly (p ≤ 0.01) in the SHKS and UHKS groups when compared with the SACS and UACS groups. Serum, urinary, and fecal Ca changes were much greater and appeared much faster in the SHKS group than in the UHKS group. Serum iPTH and 1,25 (OH)2 D, body weight, and peak oxygen uptake decreased significantly (p ≤ 0.01) in the SHKS and UHKS groups when compared with the SACS and UACS groups. In contrast, the corresponding parameters remained stable in the SACS and UACS groups when compared with the baseline control values. It was concluded that during prolonged HK, urinary and fecal Ca excretion and serum Ca concentration increased significantly despite the presence of a negative Ca balance; thus, Ca supplements cannot be used to normalize negative Ca balance during prolonged HK.     

Reducing the potential copper toxicity of concentrates to sheep by the use of molybdenum and sulphur supplements

A high copper (Cu) diet (45.3 μg Cu/g DM) was given to three groups of animals, ♂ or ♀ Scottish Blackface and ♂ Finnish Landrace lambs, without added molybdenum (Mo), or with 2, 4, 8 or 16 mg Mo/kg DM added in a 3 × 5 factorial experiment lasting 18–27 weeks. Sodium sulphate, providing 2 g S/kg, was added with each Mo supplement.Six of the nine lambs not given supplementary Mo + S died of Cu poisoning but those given Mo + S survived. Histological evidence of subclinical hepato-toxicity was found in Mo + S supplemented lambs but it decreased in severity as the level of added Mo increased. Plasma aspartate amino-transferase (PAAT) concentrations were elevated in unsupplemented lambs from week 9 and in lambs given 2 mg Mo/kg from week 12 but they remained normal in lambs given 4–16 mg Mo/kg DM. Successive increments in dietary Mo reduced the increase in liver Cu after 18–20 weeks from 1450 to 735, 483, 445 and 131 μg/g DM. The proportion of ingested Cu (y%) retained in the liver was related to dietary Mo (x, mg/kg DM) by the equation y = 2.6 ? 1.66 log x ± 0.21 (r = 0.98; 2 d.f.).Finnish Landrace lambs retained 50% less Cu in their livers, had lower PAAT levels and showed less histological evidence of liver damage than ♂ Scottish Blackface lambs. The latter had higher PAAT levels and a higher mortality from Cu poisoning than ♀ Scottish Blackface lambs although the two sexes retained similar proportions of ingested Cu in their livers.The results are discussed in relation to the practical use of Mo + S to prevent Cu poisoning in sheep.     

Effects of dietary supplementation of DL-methionine or 2-hydroxy-4-(methylthio)-butanoic acid on food intake, nutrient digestibility, nitrogen balance, and urinary and blood metabolites in healthy, growing dogs

The aim of this study was to evaluate effects on nutritional responses of supplemental DL-methionine and 2-hydroxy-4-(methylthio) butanoic acid (HMTBA) in a commercial-type diet in growing dogs. A nitrogen balance study was conducted as a randomized complete block design using 30 Pointer puppies (72-d-old; 5.5 kg). A corn and poultry byproduct meal based diet was supplemented with 0.1 or 0.2% DL-methionine or HMTBA on an equimolar basis. Organic matter and gross energy tended (p < 0.10) to be less digestible by dogs fed the 0.1% HMTBA diet compared with the 0.2% DL-methionine diet, but other nutrients were unaffected. Postprandial urinary calcium tended (p < 0.10) to be lower for the basal and HMTBA treatments. Fecal ammonia tended (p < 0.10) to be lower for the 0.1% HMTBA diet than for the 0.2% DL-methionine diet. At the levels tested, DL-methionine and HMTBA appear to act similarly when included in a corn and poultry by-product meal diet fed to young dogs.     

Influence of supplemental high molecular weight pullulan or γ-cyclodextrin on ileal and total tract nutrient digestibility,fecal characteristics,and microbial populations in the dog

Abstract

This study was conducted to determine if supplemental pullulan and γ-cyclodextrin affect canine nutrient digestibility, microbial populations, and fecal characteristics. Ileal cannulated dogs were fed a commercial diet, and treatments were administered daily in a 5×5 Latin square design: (i) no supplement; (ii) 2 g pullulan; (iii) 4 g pullulan; (iv) 2 g γ-cyclodextrin; (v) 4 g γ-cyclodextrin. Ileal and fecal samples were collected the last 4 d of each 14-d period. Increasing pullulan tended (p < 0.10) to linearly increase ileal bifidobacteria and lactobacilli and quadratically increase fecal lactobacilli. A similar response was noted in ileal bifidobacteria and lactobacilli with γ-cyclodextrin. γ-Cyclodextrin resulted in a quadratic decrease (p < 0.05) in fecal Clostridium perfringens. Increasing pullulan linearly increased (p < 0.05) fecal score, while γ-cyclodextrin resulted in a linear decrease (p < 0.05). Pullulan and γ-cyclodextrin supplementation may have beneficial effects on the microbial ecology of dogs.     

Novel encapsulation improves recovery of probiotic strains in fecal samples of human volunteers

Probiotic supplements can contribute to maintaining health and ameliorating various disease symptoms. Probiotics can be delivered in many forms with crucial differences in their survival during gastrointestinal (GI) passage. Previously, a novel encapsulation, Probiotic Pearls™ Acidophilus, Integrative Therapeutics, LLC, USA (Pearls), was shown to increase survival in vitro after exposure to gastric conditions. Here, we compare fecal recovery in human volunteers consuming Pearls or a conventional hard-shelled gelatin capsule. We performed a randomized double-blinded, two-armed trial, with six healthy subjects in each 12-day study arm. In fecal samples collected at baseline, twice during the intervention period, and after washout, we compared colony counts between the two encapsulation methods. The identity of the colonies was confirmed by colony morphology, strain-specific PCR, and 16S rRNA gene sequencing. We further performed a comprehensive 16S rRNA gene sequencing-based analysis to identify differential effects on overall microbiota composition. We detected an average log increase in bifidobacteria of 0.152 cfu/g with gelatin and 0.651 cfu/g with Pearls capsules (p > 0.05). Total lactobacilli counts increased in both groups with no difference between the groups. However, the supplemented Lactobacillus acidophilus NCFM decreased to baseline levels within 7 days after end of supplementation with gelatin capsules while 3.11 log cfu/g higher counts compared to baseline (p = 0.05) remained for Pearls. Targeted qPCR largely confirmed the trends observed by viable plate counts. Protecting the probiotic strains by Pearls encapsulation results in higher recovery rates of the supplemented lactobacilli and bifidobacteria in fecal samples and increased persistence, suggesting an improved survival and viability that might increase efficacy towards achieving desired health benefits.

     

The influence of an increased cobalt supply on ruminal parameters and microbial vitamin B12 synthesis in the rumen of dairy cows

The aim of the study was to examine the effects of an elevated dietary cobalt supply to dairy cows on rumen fermentation parameters and microbial vitamin B12 synthesis in the rumen. Five lactating dairy cows fitted with a ruminal and a duodenal cannula were subsequently fed either a ration containing only the native cobalt content (0.17 mg Co/ kg DM) or a ration supplemented with cobalt sulphate (0.29 mg Co/kg DM). The pH-value, the ammonia concentration as well as the concentration and the molar proportions of short chain fatty acids in the rumen were not significantly influenced by feeding the ration with the higher cobalt content. While there was no difference in microbial protein flow, the cobalamin flow at the duodenum was significantly elevated in supplemented animals (3.67 +/- 0.69 vs. 8.63 +/- 2.22 mg B12/d). The efficiency of cobalt utilisation for ruminal vitamin B12 synthesis was calculated to be 7.1 +/- 1.3% for the unsupplemented and 9.5 +/- 2.4% for the supplemented ration. Further investigation has to prove if there are any benefits for cows resulting from the elevated cobalamin synthesis measured, caused by feeding higher amounts of dietary cobalt.     

Magnesium deposition and depletion in magnesium supplemented rats during and after hypokinesia and vivarium control

Hypokinesia (diminished movement) induces significant magnesium (Mg) changes; however, little is known about Mg deposition and Mg depletion during HK. Measuring the Mg level in some tissues during HK and post-HK and Mg supplement, we aimed to establish Mg deposition and Mg depletion during prolonged HK. Studies were done on 408, 13-wk-old male Wistar rats (370-390 g) for a 15-d pre-HK period, a 98-d HK period, and a 15-d post-HK period. Rats were equally divided into four groups: unsupplemented vivarium control rats (UVCR), unsupplemented hypokinetic rats (UHKR), supplemented vivarium control rats (SVCR), and supplemented hypokinetic rats (SHKR). Both UHKR and SHKR were kept in small individual cages. The SVCR and SHKR took 53 mg Mg/d. During the HK period, plasma, urinary, and fecal Mg levels increased significantly (p < or = 0.05), whereas during the post-HK period Mg deposition, muscle and bone Mg content decreased significantly (p < or = 0.05) in UHKR and SHKR when compared with their pre-HK values and their respective vivarium controls (UVCR and SVCR). During the initial days of the post-HK period, plasma, urinary, and fecal Mg levels decreased significantly (p < or = 0.05), whereas during the post-HK period Mg deposition, muscle and bone Mg content remained significantly (p < or = 0.05) depressed in UHKR and SHKR when compared with UVCR and SVCR, respectively. However, during the HK period and post-HK period Mg deposition, bone, muscle, plasma, urinary, and fecal Mg levels changed significantly (p < or = 0.05) more in SHKR than UHKR. By contrast, during the HK period and post-HK period. Mg deposition, muscle, bone, plasma, urinary, and fecal Mg values change insignificantly (p > 0.05) in UVCR and SVCR when compared with their pre-HK values. It was concluded that reduced muscle, bone, plasma, urinary, and fecal Mg during post-HK and Mg supplement may demonstrate Mg depletion, whereas higher Mg loss during HK despite reduced muscle and bone Mg and Mg depletion might demonstrate Mg deposition incapacity during HK.     

Ileal bile acid transport regulates bile acid pool, synthesis, and plasma cholesterol levels differently in cholesterol-fed rats and rabbits

We investigated the effect of ileal bile acid transport on the regulation of classic and alternative bile acid synthesis in cholesterol-fed rats and rabbits. Bile acid pool sizes, fecal bile acid outputs (synthesis rates), and the activities of cholesterol 7alpha-hydroxylase (classic bile acid synthesis) and cholesterol 27-hydroxylase (alternative bile acid synthesis) were related to ileal bile acid transporter expression (ileal apical sodium-dependent bile acid transporter, ASBT). Plasma cholesterol levels rose 2.1-times in rats (98 +/- 19 mg/dl) and 31-times (986 +/- 188 mg/dl) in rabbits. The bile acid pool size remained constant (55 +/- 17 mg vs. 61 +/- 18 mg) in rats but doubled (254 +/- 46 to 533 +/- 53 mg) in rabbits. ASBT protein expression did not change in rats but rose 31% (P < 0.05) in rabbits. Fecal bile acid outputs that reflected bile acid synthesis increased 2- and 2.4-times (P < 0.05) in cholesterol-fed rats and rabbits, respectively. Cholesterol 7alpha-hydroxylase activity rose 33% (24 +/- 2.4 vs. 18 +/- 1.6 pmol/mg/min, P < 0.01) and mRNA levels increased 50% (P < 0.01) in rats but decreased 68% and 79%, respectively, in cholesterol-fed rabbits. Cholesterol 27-hydroxylase activity remained unchanged in rats but rose 62% (P < 0.05) in rabbits. Classic bile acid synthesis (cholesterol 7alpha-hydroxylase) was inhibited in rabbits because an enlarged bile acid pool developed from enhanced ileal bile acid transport. In contrast, in rats, cholesterol 7alpha-hydroxylase was stimulated but the bile acid pool did not enlarge because ASBT did not change. Therefore, although bile acid synthesis was increased via different pathways in rats and rabbits, enhanced ileal bile acid transport was critical for enlarging the bile acid pool size that exerted feedback regulation on cholesterol 7alpha-hydroxylase in rabbits.     

Control of foodborne pathogens during sufu fermentation and aging

Control of the foodborne pathogens Escherichia coli O157:H7, Salmonella typhimurium, Staphylococcus aureus, and Listeria monocytogenes during sufu fermentation was evaluated. Before fermentation, pathogens were inoculated onto tofu (substrate for sufu) at 5 log cfu/g or 3 log cfu/g, and starter culture (Actinomucor elegans) was inoculated at 3 log cfu/g. After 2 days of fermentation at 30 degrees C, the four pathogens reached 7 to 9 log cfu/g, and the mold count reached 6 to 7 log cfu/g. After fermentation, sufu samples were aged in a solution of 10% alcohol + 12% NaCl. After 1 month of aging, the total bacterial count was 6 to 7 log cfu/g, but all foodborne pathogens and mold were reduced to nondetectable levels. The total bacterial count decreased after aging for 2 months and 3 months, but the differences were not significant (P > 0.05) compared with the count after 1 month. Microorganism in experimental sufu from different aging periods and in commercial sufu were compared. A total of 270 isolates were purified and identified by the BBL Crystal Identification System. From the experimental sufu samples, 49 Bacillus spp. (20.4%), 167 Enterococcus spp. (69.6%), 6 Shewanella putrefaciens (2.4%), and 18 miscellaneous gram-negative bacilli (7.5%) were identified. From commercial sufu samples, 17 Bacillus spp. (56.7%), 2 Enterococcus durans (6.7%), 5 miscellaneous gram-negative bacilli (16.7%), 5 Corynbacterium aquaticum (16.7%), and 1 Shewanella putrefaciens (3.3%) were obtained. Although the longer aging period did not significantly decrease the total bacterial count, it may help in the development of sufu flavor. This study showed that sufu fermentation and aging can control common foodborne pathogens, so sufu is a safe product even though its preparation does not include pasteurization.     

Effects of iron,zinc, calcium,and vitamins on the activity and contents of human placental copper/zinc and manganese superoxide dismutases

One hundred seventy-nine pregnant women, ages 15–45 yr, were divided into three groups. Group A was orally given one spansule per day containing 150 mg dried ferrous sulfate, 61.8 mg zinc sulfate, and 500 μg folic acid, starting from the first 4 wk of pregnancy and ending at the day of delivery. Similarly, group B was given one tablet containing 625 mg calcium carbonate, 1000 mg vitamin C, 300 IU Vitamin D, 1350 mg citric acid, and 15 mg Vitamin B6. Group C was without any supplements and served as a control. Mothers who received iron/zinc supplements (group A) during pregnancy had significantly higher copper/zinc superoxide dismutase activity in their placentae than calcium/vitamin-supplemented mothers (group B) or unsupplemented mothers (group C). The enzyme activity increased with age of the mothers from 15 to 40 yr, then decreased after in both supplemented groups, whereas this increase and decrease occurred at early age in the unsupplemented group. Immunochemical quantitation of the enzyme contents showed no significant difference between the supplemented and unsupplemented groups, suggesting that the observed increase in the enzyme activity might arise from posttranslational processing of the enzyme. The placental manganese superoxide dismutase activity and contents, however, were similar in the supplemented groups, whereas they were slightly higher in the unsupplemented group; the overall superoxide dismutase-like activities in the placentae were the highest in iron-zinc supplemented group and the lowest in the unsupplemented group.