A Quantitative Comparison of the Faecal Microflora of Baboons Fed a Natural Diet or a Synthetic Diet Complete or Deficient in Pyridoxine or Riboflavin

S ummary . Quantitative methods were used to study the microflora of faeces produced by a group of baboons receiving fresh fruit and vegetables (natural diet), and 3 groups fed a synthetic diet deficient in either pyridoxine or riboflavin, or which contained both vitamins (pair fed). There was no significant difference between the microbial counts of any of the groups of organisms studied in the pyridoxine or riboflavin deficient and pair fed baboons. Compared with baboons on natural diet, faecal counts from baboons fed the synthetic diet showed highly significant increases in Clostridium welchii and in lactose-fermenting enterobacteria, together with a considerable decrease in the number of lactobacilli. Small increases in the numbers of micrococci, staphylococci and faecal streptococci were noted, whilst the numbers of yeasts were reduced slightly. Whilst the change from the natural to the synthetic diet caused a great alteration in the flora, the synthetic diet flora appeared to be extremely stable and generally unaffected by drastic modifications of the synthetic diet. However, marked alteration did occur on re-feeding with the natural diet. Data on the faecal microflora of baboons are compared briefly with published data for man and other animals, and the effects of dietary changes on the composition of the faecal flora are discussed.     

Vitamin B12 Production by the Gastro-intestinal Microflora of Baboons Fed either a Vitamin B12 Deficient Diet or a Diet Supplemented with Vitamin B12

The vitamin B₁₂-producing capacity of micro-organisms isolated from baboon faeces and gastric contents was measured using Lactobacillus leichmannii . The animals were fed either a diet deficient in or supplemented with vitamin B₁₂ (controls). Samples of gastric and small intestinal contents, obtained at laparotomy from two young vitamin B₁₂-deprived baboons, contained varying quantities of vitamin B₁₂. Many of the organisms isolated from these aspirates produced vitamin B₁₂ in vitro . The highest levels of vitamin B₁₂ were produced by anaerobic organisms. Gastric juice samples from vitamin B₁₂ deprived and control baboons contained similar types of organisms with like vitamin B₁₂-producing capacity. The vitamin B₁₂ content, pH and total bacterial counts of gastric juice samples aspirated after a 6 h fast from vitamin B₁₂ deprived baboons were not significantly different from those of the control animals. The pH values of gastric juice samples aspirated 18 h after feeding, however, were significantly lower than those of 6 h fasting samples in both groups. The mean vitamin B₁₂ levels in the total volumes of gastric juice aspirated after each fasting period were similar. The possible involvement of the gastrointestinal flora in the vitamin B₁₂ status of the baboon is discussed.     

Effects of dietary lactose and purified diet on intestinal microflora of rats

In the rats fed on lactose-containing purified diet (PDL), bifidobacteria remarkably increased in number in both the jejunum and cecum as compared with those fed on control diet (PD) or conventional diet (NMF). Besides, lactobacilli, streptococci, staphylococci, and Escherichia coli increased in number in the jejuna and/or ceca of rats fed on PDL as compared with those fed on PD. The incidence and numbers of Lactobacillus species were remarkably different between the rats fed on PDL and those on any of the other diets. Indole-producing Bacteroides spp. were rarely isolated from the PDL-fed rats. The cecum was markedly enlarged in the PDL rats and reduced in the PD rats as compared with that in the NMF animals.     

Effect of fermented feed on the microbial population of the gastrointestinal tracts of pigs.

An in vivo experiment was performed with pigs to study the inhibitory effect of fermented feed on the bacterial population of the gastrointestinal tract. Results demonstrated a significant positive correlation between pH and lactobacilli in the stomach contents of pigs in dry feed as well as in the stomach contents of pigs fed fermented feed. Furthermore, a significant positive correlation between the pH and the numbers of bacteria in the family Enterobacteriaceae in the contents of the stomach of pigs fed dry feed was found. In the stomach contents of pigs fed fermented feed, a significant negative correlation was found between the concentration of the undissociated form of lactic acid and the numbers of Enterobacteriaceae. The numbers of Enterobacteriaceae in the contents of the stomach, ileum, cecum, colon, and rectum of pigs fed fermented feed were significantly lower compared with the contents of the stomach, ileum, caecum, colon, and rectum of pigs fed dry feed. The numbers of total lactobacilli were significantly higher in the stomach contents of pigs fed fermented feed and in the ileum contents of one pig group fed fermented feed compared with the contents of pigs fed dry feed. However, the influence of lactobacilli on numbers of Enterobacteriaceae could not be demonstrated. It was concluded that fermented feed influences the bacterial ecology of the gastrointestinal tract and reduces the levels of Enterobacteriaceae in the different parts of the gastrointestinal tract.     

THE SELECTIVE ACTION OF THALLOUS ACETATE FOR LACTOBACILLI

SUMMARY: The effect of thallous acetate has been examined on the growth of 23 strains of lactobacilli representing 13 different species. All strains grew satisfactorily in both broth and agar media containing 0.1% of thallous acetate. Of the 18 other organisms also examined the growth of the streptococci was unsuppressed, staphylococci, a micrococcus, B. licheniformis and Pr. mirabilis were partially inhibited and strains of Bact. coli, B. cereus, Chr. prodigiosum and Ps. fluorescens were almost completely inhibited by that concentration. By its use a valuable selective medium for isolating lactobacilli from natural habitats of mixed flora was obtained, other organisms, with the exception of streptococci, being almost completely eliminated.     

Effect of Fermented Feed on the Microbial Population of the Gastrointestinal Tracts of Pigs

An in vivo experiment was performed with pigs to study the inhibitory effect of fermented feed on the bacterial population of the gastrointestinal tract. Results demonstrated a significant positive correlation between pH and lactobacilli in the stomach contents of pigs in dry feed as well as in the stomach contents of pigs fed fermented feed. Furthermore, a significant positive correlation between the pH and the numbers of bacteria in the family Enterobacteriaceae in the contents of the stomach of pigs fed dry feed was found. In the stomach contents of pigs fed fermented feed, a significant negative correlation was found between the concentration of the undissociated form of lactic acid and the numbers of Enterobacteriaceae. The numbers of Enterobacteriaceae in the contents of the stomach, ileum, cecum, colon, and rectum of pigs fed fermented feed were significantly lower compared with the contents of the stomach, ileum, caecum, colon, and rectum of pigs fed dry feed. The numbers of total lactobacilli were significantly higher in the stomach contents of pigs fed fermented feed and in the ileum contents of one pig group fed fermented feed compared with the contents of pigs fed dry feed. However, the influence of lactobacilli on numbers of Enterobacteriaceae could not be demonstrated. It was concluded that fermented feed influences the bacterial ecology of the gastrointestinal tract and reduces the levels of Enterobacteriaceae in the different parts of the gastrointestinal tract.     

Endogenous infection in mice with streptozotocin-induced diabetes. A feature of bacterial translocation

Slc:ddY mice that received a single intraperitoneal injection of 200 mg/kg streptozotocin (STZ) were examined for persistency of diabetes (changes of indigenous bacterial floras, and bacterial translocation. Significant diabetes (increase in plasma glucose and decrease in insulin) was recognized 2 weeks after the injection, and persisted for 12 weeks. The numbers of aerobic gram-negative bacilli, staphylococci (including micrococci), and streptococci in caecal and oral floras were significantly increased, but the numbers of anaerobic bacteria in caecal flora were hardly changed. Bacterial translocation of indigenous bacteria to the mesenteric lymph node, lung, or kidney was detectable in some mice 2 weeks after the injection. The incidence of bacterial translocation in these STZ-treated mice then increased; infection caused by several organisms, e.g., Klebsiella pneumoniae, Staphylococcus epidermidis, streptococci, or Lactobacillus sp., occurred in lung, liver, spleen, kidneys, and mesenteric lymph node. No indigenous bacteria were cultured from these organs of control mice. This endogenous infection may have been due to the over population of several bacterial strains caused by disruption of indigenous floras along with depression of immunological function.     

The Gastrointestinal Bacteria of Mink (Mustela vison L): Influence of Age and Diet

Total numbers of aerotolerant and anaerobic bacteria, and densities of Enterobacteriaceae, lactobacilli, staphylococci, salmonella and shigella, and campylobacteria were enumerated in the contents of the stomach, small intestine (and the associated mucosa), and colon of mink beginning at 2 weeks of age to adulthood, and in adults that were fed diets with different levels and types of fiber or food deprived. Highest densities of all bacterial groups were found in the colon at all ages (up to 108 cfu per g for total anaerobes), but were 2–4 orders of magnitude lower than those of other mammals. When all regions were pooled, significant age-related increases (p<0.05) were detected for anaerobes, aerobes, and staphylococci, and these coincided with the dietary shift at weaning. Enterobacteriaceae did not vary with age. Lactobacilli were never common isolates, but were detected more often after weaning, particularly in adults fed diets containing the 2 sources of fiber. Campylobacteria were detected only at 2 weeks of age, and salmonella and shigella were not isolated from any of the experimental mink. Total bacterial densities, the relative proportions of the bacterial groups, and age- and diet-related effects differ from those known for other mammals, which may be related to the carnivorous diet and rapid movement of digesta through the GIT.     

The normal microbial flora of the baboon vagina.

The microbial flora of the upper vagina and cervix was examined in 38 adult baboons at various stages of the menstrual cycle. The mean number of different species isolated from each baboon was 9.5, with species of Bacteroides, Corynebacterium and group D streptococci predominating. Lactobacilli and mycoplasmas were found in 47.4 and 44.7% of the animals, respectively. No ureaplasms were isolated. Cyclical variations in the microbial flora were minimal.     

Characteristic faecal flora of NC mice

The composition of faecal flora of NC mice was compared with that of CF #1 mice. NC- and CF #1-germfree (GF) mice were cage-mated with NC- or CF #1-conventional (CV) mice in an isolator. The faecal flora of these ex-GF mice was dependent on the recipient mouse strain modifying colonization by the donor mouse bacteria. Although NC- and CF #1-pups removed by hysterectomy were fostered to different strains, almost all these mice at 8 weeks old had a strain characteristic pattern of faecal flora regardless of the foster strains. In GF mice mono-associated with a Lactobacillus strain or a Bifidobacterium strain isolated from faeces of CV mice, the numbers of these bacteria in the stomach and small intestine of NC mice were lower than those of CF #1 mice. In GF mice associated with chloroform-treated faeces of CV mice, and a Lactobacillus strain or a Bifidobacterium strain, the numbers of these bacteria in the stomach and all parts of the intestine of NC mice were considerably lower than those of CF #1 mice. These results suggested that the composition of faecal flora of NC mice were characteristic, i.e. the fact that the numbers of lactobacilli were low compared with CF #1 mice with ordinary faecal flora and the colonization of bifidobacteria, peptococcaceae and eubacteria on ES agar in NC mice intestine differed, was due to genetic factors.     

Microbial Penicillinase in Soft Types of Cheese

Penicillinase activity was found in commercial samples of Limburger, Liederkranz, and Brick, as well as in Roquefort and Danish Blue cheeses. The numbers of penicillinase-producing bacteria in the cheese flora were estimated by use of the most probable numbers technique coupled with penicillinase determinations on the individual cultures. Although streptococci constituted the predominant flora in most of the samples, significant numbers of penicillinase-producing bacteria were also observed. Six of seven penicillinase-producing isolates that were studied in detail were micrococci; the other was an unidentified gram-positive, catalase-positive rod. The results suggest the widespread occurrence of penicillinase-forming micrococci on the surfaces of soft types of cheese.     

Effect of Streptococcus faecalis BIO-4R on intestinal flora of weanling piglets and calves

The effect of oral administration of Streptococcus faecalis BIO-4R, an antibiotic-resistant lactic acid bacterium, on the intestinal flora of weanling piglets and cows reared on antibiotic-containing diet was investigated. Fourteen days after administration of the bacteria, the intestinal flora of the piglets was examined. Animals of the administered group had stabilized lactic flora such as bifidobacteria, streptococci, and lactobacilli, whereas most animals of control group had reduced lactic flora. On the other hand, abundant yeasts were detected from the cecum, colon, and feces of the control animals, but the levels were significantly lower in the animals given strain BIO-4R. The density of Salmonella in the intestine appeared to be reduced after the administration of strain BIO-4R. The number of BIO-4R cells was shown to be 10 times lower in the duodenum and jejunum than in the ileum, suggesting that strain BIO-4R might have grown transiently in the ileum. The similar trend toward stabilization of the lactic flora was also observed in cows after administration of BIO-4R. In addition, an antagonistic effect of the strain against yeasts and Salmonella was suggested. These findings indicate that the oral administration of strain BIO-4R is one of the useful methods whereby the potentially deleterious effect of antibiotics on the intestinal flora of farm animals may be minimized.     

Effect of Streptococcus faecalis BIO-4R on intestinal flora of weanling piglets and calves.

The effect of oral administration of Streptococcus faecalis BIO-4R, an antibiotic-resistant lactic acid bacterium, on the intestinal flora of weanling piglets and cows reared on antibiotic-containing diet was investigated. Fourteen days after administration of the bacteria, the intestinal flora of the piglets was examined. Animals of the administered group had stabilized lactic flora such as bifidobacteria, streptococci, and lactobacilli, whereas most animals of control group had reduced lactic flora. On the other hand, abundant yeasts were detected from the cecum, colon, and feces of the control animals, but the levels were significantly lower in the animals given strain BIO-4R. The density of Salmonella in the intestine appeared to be reduced after the administration of strain BIO-4R. The number of BIO-4R cells was shown to be 10 times lower in the duodenum and jejunum than in the ileum, suggesting that strain BIO-4R might have grown transiently in the ileum. The similar trend toward stabilization of the lactic flora was also observed in cows after administration of BIO-4R. In addition, an antagonistic effect of the strain against yeasts and Salmonella was suggested. These findings indicate that the oral administration of strain BIO-4R is one of the useful methods whereby the potentially deleterious effect of antibiotics on the intestinal flora of farm animals may be minimized.     

In vitro fermentability and physicochemical properties of fibre substrates and their effect on bacteriological and morphological characteristics of the gastrointestinal tract of newly weaned piglets

Abstract

Fermentability of fibre has a great impact on the bacterial flora along the gastrointestinal tract of newly weaned piglets. Therefore, this parameter was determined by incubating in vitro different fibre substrates (chicory roots, sugar beet pulp, wheat bran and corn cobs) with contents of jejunum or caecum sampled from slaughtered pigs. Incubating with small intestinal contents, lactic acid was the only fermentation product. Fermentability was highest for chicory roots, followed by wheat bran and sugar beet pulp, while corn cobs were not fermented. Based on SCFA formed in the incubations with caecal contents, ranking of the fermentability of the fibre substrates was in the same order. The effect of adding different fibre substrates to diets of newly weaned piglets on bacteriological and morphological aspects of the gastrointestinal tract was also investigated. In Experiment 1 three groups of five piglets, weaned at four weeks of age, received a control feed (C), C supplemented with corn cobs (50 g/kg) or with chicory roots (20 g/kg). In Experiment 2, diet C was supplemented with sugar beet pulp (120 g/kg) or with wheat bran (75 g/kg). After three weeks animals were euthanized and digesta were sampled from stomach, proximal and distal jejunum, caecum and colon. Furthermore, mucosal scrapings were prepared and tissue samples were taken from jejunum, caecum and colon. Viscosity was determined for jejunal, caecal and colon contents. Corn cobs in the feed increased the number of total bacteria, lactobacilli and bifidobacteria in the stomach and proximal duodenum, while a decreased count of streptococci in distal jejunum contents was noted. Chicory roots increased the counts of Escherichia coli in the distal jejunum and on the mucosa, while sugar beet pulp decreased the number of lactobacilli on the mucosa only. Wheat bran seemed to increase the count of E. coli in jejunal digesta and on the mucosa, and also the number of lactobacilli in the stomach and jejunum. Bifidobacterial numbers were increased but only in the proximal part of the jejunum. Fibre substrates affected the concentration of lactate and SCFA in different parts of the intestinal tract. Feeding corn cobs increased villus length in the proximal jejunum by 13%. The number of intra-epithelial lymphocytes in the villous epithelium of proximal and distal jejunum was decreased by corn cobs and chicory roots supplementation while beet pulp and wheat bran had the opposite effect. In Experiment 1, apoptotic index of the mucosa of the distal jejunum was very low and decreased when corn cobs were fed. Mitotic index in the crypts was only affected by the wheat bran diet and a small decrease was noted. It was concluded that the fermentability of fibre was not an ideal criterion for predicting its effects on the flora. The effect of fibres on viscosity of digesta was negligible probably explaining the lack of clear and consistent influences on the intestinal mucosa.     

In vitro fermentability and physicochemical properties of fibre substrates and their effect on bacteriological and morphological characteristics of the gastrointestinal tract of newly weaned piglets

Fermentability of fibre has a great impact on the bacterial flora along the gastrointestinal tract of newly weaned piglets. Therefore, this parameter was determined by incubating in vitro different fibre substrates (chicory roots, sugar beet pulp, wheat bran and corn cobs) with contents of jejunum or caecum sampled from slaughtered pigs. Incubating with small intestinal contents, lactic acid was the only fermentation product. Fermentability was highest for chicory roots, followed by wheat bran and sugar beet pulp, while corn cobs were not fermented. Based on SCFA formed in the incubations with caecal contents, ranking of the fermentability of the fibre substrates was in the same order. The effect of adding different fibre substrates to diets of newly weaned piglets on bacteriological and morphological aspects of the gastrointestinal tract was also investigated. In Experiment 1 three groups of five piglets, weaned at four weeks of age, received a control feed (C), C supplemented with corn cobs (50 g/kg) or with chicory roots (20 g/kg). In Experiment 2, diet C was supplemented with sugar beet pulp (120 g/kg) or with wheat bran (75 g/kg). After three weeks animals were euthanized and digesta were sampled from stomach, proximal and distal jejunum, caecum and colon. Furthermore, mucosal scrapings were prepared and tissue samples were taken from jejunum, caecum and colon. Viscosity was determined for jejunal, caecal and colon contents. Corn cobs in the feed increased the number of total bacteria, lactobacilli and bifidobacteria in the stomach and proximal duodenum, while a decreased count of streptococci in distal jejunum contents was noted. Chicory roots increased the counts of Escherichia coli in the distal jejunum and on the mucosa, while sugar beet pulp decreased the number of lactobacilli on the mucosa only. Wheat bran seemed to increase the count of E. coli in jejunal digesta and on the mucosa, and also the number of lactobacilli in the stomach and jejunum. Bifidobacterial numbers were increased but only in the proximal part of the jejunum. Fibre substrates affected the concentration of lactate and SCFA in different parts of the intestinal tract. Feeding corn cobs increased villus length in the proximal jejunum by 13%. The number of intra-epithelial lymphocytes in the villous epithelium of proximal and distal jejunum was decreased by corn cobs and chicory roots supplementation while beet pulp and wheat bran had the opposite effect. In Experiment 1, apoptotic index of the mucosa of the distal jejunum was very low and decreased when corn cobs were fed. Mitotic index in the crypts was only affected by the wheat bran diet and a small decrease was noted. It was concluded that the fermentability of fibre was not an ideal criterion for predicting its effects on the flora. The effect of fibres on viscosity of digesta was negligible probably explaining the lack of clear and consistent influences on the intestinal mucosa.     

Detection of Caries-Inducing Microorganisms in Hyposalivated Rats without Infection of Mutans Streptococci

Rampant dental caries was induced in hyposalivated rats fed a high sucrose diet without infection of mutans streptococci, in which increased numbers of lactobacilli and S. aureus were demonstrated in the oral flora. Administration of either penicillin or piperacillin, effective against all isolates of lactobacilli, markedly inhibited the caries induction in these rats, while severe dental caries was induced in hyposalivated rats given vancomycin that is inhibitory against S. aureus. These results suggested that certain lactobacilli might induce dental caries in hyposalivated rats fed a sucrose diet. Three strains of Lactobacillus species isolated from the hyposalivated rats were made resistant to erythromycin. The caries-inducing activity of these erythromycin-resistant lactobacilli was studied in hyposalivated rats giving erythromycin in the drinking water at a concentration of 500 μg/ml. After a 61-day experimental period, severe dental caries was induced in hyposalivated rats infected with L. fermentum TY1R. On the other hand, low caries incidence was found in hyposalivated rats infected with either L. acidophilus TY7R or L. plantarum TY3R. These results indicate that L. fermentum may be one of causative agents of dental caries in hyposalivated rats fed a sucrose diet.     

Changes in the bacterial cecal flora of mice infected with Trichuris muris (Schrank, 1788).

Cecal microorganisms of mice were categorized and enumerated weekly during the developmental cycle of infection with the whipworm, Trichuris muris. The cecal bacterial population consisted of Escherichia coli, Proteus spp, Acinetobacter lwoffi (Mima polymorpha), aerobic lactobacilli, staphylococci, enterococci, and anaerobes (bacteroides, streptococci, and lactobacilli) in control and T. muris-infected mice. The aerobic lactobacilli and the anaerobes constituted the greatest number of organisms in both groups. In week three there was a decrease in the number of these organisms, and in week four fewer of these and of all other organisms in the worm-infected mice when compared to controls. The most significantly reduced bacterial counts were observed during the period of T. muris self-cure.     

Intestinal Microflora Associated with Enteritis of Early-Weaned Pigs

Studies were conducted on the enumeration of 7 groups of fecal microflora including total aerobes, total anaerobes, coliforms, lactobacilli, staphylococci, streptococci and yeasts and molds of 18-day old piglets. These pigs were early weaned (21 days) on different modifications of an early-weaning ration. The above mentioned microflora were enumerated again when some of the pigs in a replicate started scouring. The occurrence of diarrhea was always associated with significant increases in the numbers of coliforms and corresponding decreases in the lactobacilli counts. No other single group of fecal microflora differed in the scouring and non-scouring animals. The composition of the early-weaning ration offered to the animals did not, in itself, influence the fecal microflora to any appreciable extent. In another series of experiments, enumeration of coliforms and lactobacilli was conducted on samples obtained from different segments of the intestinal tracts of scouring and non-scouring pigs. Increased numbers of coliforms and decreased numbers of lactobacilli were observed at all levels of the intestinal tracts of the scouring animals. However, these changes were more marked in the duodenal samples than in those obtained from other parts of the intestine.     

Gastrointestinal flora of cotton rats

The gastrointestinal (GI) flora of cotton rats was examined. No lactobacilli were detected in any part of the GI tract. Anaerobes, including Peptococcaceae, Bacteroidaceae, bifidobacteria and eubacteria, were the predominant bacteria in the stomach, small intestine, caecum and faeces. Aerobes and facultative anaerobes, including Enterobacteriaceae and streptococci, were detected at low numbers and very low frequency of occurrence in all parts of the GI tract. Sixty-one isolates of bifidobacteria were recovered from the stomach, small intestine, caecum and faeces of cotton rats. They were identified as Bifidobacterium animalis, B. pseudolongum biovar a and b. The study showed that the GI flora of cotton rats seem to be very different from the GI flora in other rodents.     

Changes with time in the oral microflora and dental caries induction in hyposalivated rats fed on sucrose diet.

The effects of hyposalivation on the induction of dental caries and the change in oral microflora were examined at weekly intervals in Sprague-Dawley rats fed on diet 2000 containing 56% sucrose. In hyposalivated rats, significant dental caries was induced within one week and its severity increased with the experimental period. Bacteriological examinations demonstrated that the number of total cultivable microorganisms, lactobacilli and Staphylococcus aureus increased significantly shortly after surgical induction of hyposalivation, while the number of streptococci and yeasts did not increase significantly until the 7th week, beyond which time remarkable gross caries developed. A positive correlation was found between the caries score and the recovery of lactobacilli from mandibles of hyposalivated rats, while there was no statistically significant correlation between the caries score and the recoveries of S. aureus. On the other hand, dental caries was not induced in control rats fed on sucrose diet with no surgically-induced hyposalivation. It was also found that the number of lactobacilli increased significantly shortly after diet 2000 was given to control rats, but S. aureus was rarely recovered from the mandibles of control rats throughout the experiments. The roles of lactobacilli and S. aureus in the induction of dental caries under the hyposalivated condition were discussed and it was suggested that lactobacilli may play some significant role in the induction of dental caries in hyposalivated rats.