Comparison of bacterial populations of the pig cecum and colon based upon enumeration with specific energy sources.

Concentrations of bacteria in the ceca and colons of pigs were measured by determinations of colony counts on rumen fluid-based media in anaerobic roll tubes. With our most complete medium (medium CCA), the mean colony count of cecal samples from 20 pigs was 2.37 X 10(10) +/- 1.0 X 10(10) (+/- standard deviation)/g (wet weight). The mean number of bacteria attached to or associated with cecal epithelial tissues from three pigs on medium CCA was 2.67 X 10(7) +/- 0.81 X 10(7)/cm2 of tissue. The proportions of gut bacterial populations able to use various energy substrates were estimated on the basis of relative colony counts. The following substrates are listed in descending order of their capacity to support growth of cecal bacteria: glucose, starch, cellobiose, xylose, Trypticase, gastric mucin from swine, mannitol, glycerol, and lactate. The effect of diet upon this distribution was not examined. The relative proportions of bacteria from a given population that were able to grow on various selective media were used as population profiles. Comparisons of populations in this way indicated that differences could be detected between (i) populations from the cecum of littermate pigs, (ii) populations from the cecum and colon of the same pig, and (iii) populations in the lumen of the cecum as compared with populations associated with cecal mucosa.     

Bacteria isolated from the duodenum, ileum, and cecum of young chicks.

Facultatively anaerobic and strictly anaerobic bacteria colonizing the intestinal tracts of 14-day-old chicks fed a corn-based diet were enumerated, isolated, and identified. Colony counts from anaerobic roll tubes (rumen fluid medium) or aerobic plates (brain heart infusion agar) recovered from homogenates of the duodenum, upper and lower ileum, and cecum varied appreciably among samples from individual birds. Anaerobic and aerobic counts from the duodenum and ileum were similar. Anaerobic counts were highest from the cecum (0.7 X 10(11) to 1.6 X 10(11)/g of dry tissue) and exceeded aerobic plate counts by a factor of at least 10(2). Facultatively anaerobic groups (Streptococcus, Staphylococcus, Lactobacillus, and Escherichia coli) comprised the predominant flora of the duodenum and ileum, although large numbers of anaerobes (9 to 39% of the small intestine isolates), represented by species of Eubacterium, Propionibacterium, Clostridium, Gemmiger, and Fusobacterium, were also recovered. Strict anaerobes (anaerobic gram-positive cocci, Eubacterium, Clostridium Gemmiger, Fusobacterium, and Bacteriodes) made up nearly the entire microbial population of the cecum. Scanning electron microscopy of the intestinal epithelia of chicks revealed populations of microbes on the duodenal, ileal, and cecal mucosal surfaces.     

Characterization of predominant bacteria from the colons of normal and dysenteric pigs.

Bacterial populations adherent to the mucosa of the proximal colons of weaned, healthy pigs were compared with populations from pigs with dysentery induced by inoculation with a culture of Treponema hyodysenteriae. Isolates (136) representative of the predominant flora adherent to colonic epithelia of normal pigs and isolates (162) from pigs with dysentery were cultured anaerobically on a rumen fluid-based medium and characterized. Most (71%) of the isolates from colonic epithelia of normal pigs were gram positive, whereas 88% of the epithelia-associated isolates from pigs with dysentery were gram negative. The geometric mean of colony counts was 5.7 X 10(7)/cm2 of colonic tissue from three normal pigs and 7.7 X 10(8)/cm2 from four pigs with dysentery. A number of isolates obtained from contents of the lumens of normal pigs with dysentery were also characterized. Comparison of isolates from epithelial tissue and from contents of the lumens of the same pig indicated that these populations were different. Our results indicate that physiological changes that occur in the colons of pigs with dysentery are accompanied by marked changes in the microbial populations in the colons. The factors which regulate the population changes are not yet understood.     

Intestinal colonization of laboratory rats with Oxalobacter formigenes.

Six strains of Oxalobacter formigenes (anaerobic oxalate-degrading bacteria) were examined for their ability to colonize the gastrointestinal tracts of adult laboratory rats. These rats did not harbor O. formigenes. Strain OxCR6, isolated from the cecal contents of a laboratory rat that was naturally colonized by oxalate-degrading bacteria, colonized the ceca and colons of adult rats fed a diet that contained 4.5% sodium oxalate. Five days after rats were inoculated intragastrically with 10(9) viable cells of strain OxCR6, oxalate degradation rates in cecal and colonic contents increased by 19 and 40 times, respectively. Viable counts of strain OxCR6 from these rats averaged 10(8)/g (dry weight) of cecal contents. Strain OxCR6 was not detected in the cecal contents of inoculated rats fed diets that contained less than 3.0% sodium oxalate. Strains of O. formigenes isolated from the cecal contents of swine, guinea pigs, and wild rats and from human feces also colonized the ceca of laboratory rats; a ruminal strain failed to colonize the rat cecum.     

Characterization of predominant bacteria from the colons of normal and dysenteric pigs

Bacterial populations adherent to the mucosa of the proximal colons of weaned, healthy pigs were compared with populations from pigs with dysentery induced by inoculation with a culture of Treponema hyodysenteriae. Isolates (136) representative of the predominant flora adherent to colonic epithelia of normal pigs and isolates (162) from pigs with dysentery were cultured anaerobically on a rumen fluid-based medium and characterized. Most (71%) of the isolates from colonic epithelia of normal pigs were gram positive, whereas 88% of the epithelia-associated isolates from pigs with dysentery were gram negative. The geometric mean of colony counts was 5.7 X 10(7)/cm2 of colonic tissue from three normal pigs and 7.7 X 10(8)/cm2 from four pigs with dysentery. A number of isolates obtained from contents of the lumens of normal pigs with dysentery were also characterized. Comparison of isolates from epithelial tissue and from contents of the lumens of the same pig indicated that these populations were different. Our results indicate that physiological changes that occur in the colons of pigs with dysentery are accompanied by marked changes in the microbial populations in the colons. The factors which regulate the population changes are not yet understood.     

Enumeration of anaerobic bacterial microflora of the equine gastrointestinal tract

Samples from the duodenum, jejunum, and ileum, as well as from the cecum and colon, were obtained from 11 mature grass-fed horses. Viable counts of total culturable and proteolytic bacteria were made on habitat-simulating media containing 40% clarified ruminal fluid. The mean pHs in the duodenum, jejunum, and ileum were 6.32, 7.10, and 7.47, respectively; the mean pH decreased to 6.7 in the hindgut. The acetate concentration increased along the length of the small intestine and was the only volatile fatty acid present in this gut segment. Molar proportions of acetate, propionate, and butyrate in the hindgut were 85:10:3. Differences in bacterial counts on habitat-simulating media containing equine cecal fluid or clarified ruminal fluid were negligible. Bacterial counts showed a substantial population in the duodenum (ca. 2.9 x 10(6) per g [wet weight] of sample), and this increased to 29.0 x 10(6) in the jejunum and 38.4 x 10(6) in the ileum. Proteolytic bacteria formed a high proportion of the total culturable bacteria, especially in duodenal samples. Counts of proteolytic bacteria per gram (wet weight) of sample were 3.0 x 10(6), 15.6 x 10(6), and 22.0 x 10(6) in the duodenum, jejunum, and ileum, respectively. There was a close relationship between lumenal and mucosal bacterial counts, although actual values were lower in mucosal samples. The mucosal bacterial population in the duodenum was high relative to the lumenal population. Although the comparison of bacterial populations in the hindgut of the horse and white rhino was limited to a single animal, the results were of interest. Counts were higher in the cecum than in the colon for both the horse and the white rhino.(ABSTRACT TRUNCATED AT 250 WORDS)     

Isolation and identification of fecal bacteria from adult swine.

An examination of the fecal microflora of adult swine was made with regard to the efficiency of several roll tube media in enumeration and recovery of anaerobes, the effects of medium constituents on recovery, and the isolation and identification of the predominant kinds of bacteria. Total number of organisms by microscopic bacterial counts varied among fecal samples from 4.48 X 10(10) to 7.40 X 10(10) bacteria/g (wet weight). Comparison of different nonselective roll tube media indicated that about 30% of the fecal bacteria could be recovered with a rumen fluid (40%, vol/vol) medium (M98-5). Recoveries of 21 and 15%, respectively, were obtained with M10 and rumen fluid-glucose-cellobiose agar (RGCA) media. Rumen fluid, Trypticase, sugars, and CO2 gas phase were important components required for maximum recovery with this medium. Similar high recoveries of anaerobes were also obtained with M98-5 containing swine cecal extract of place in rumen fluid or M10 plus swine cecal extract. Significantly lower recoveries were observed with RCGA, media supplemented with swine fecal extracts, reinforced clostridial medium, brain heart infusion agar, and prereduced blood agar. Ninety percent of the bacteria isolated from roll tube media were gram positive and consisted of facultatively anaerobic streptococci, Eubacterium sp., Clostridium sp., and Propionibacterium acnes. The remainder of the flora (8%) included several other species of anaerobes and Escherichia coli. Rumen fluid (or volatile fatty acids), Trypticase, and yeast extract additions to basal media stimulated the growth of anaerobic strains. Variation in the relative proportions of the predominant fecal microflora was observed. This work indicates that satisfactory enumeration, isolation and cultivation of the predominant microflora in swine feces can be obtained when strict anaerobic culture methods and a rumen fluid medium are used.     

Enumeration of anaerobic bacterial microflora of the equine gastrointestinal tract.

Samples from the duodenum, jejunum, and ileum, as well as from the cecum and colon, were obtained from 11 mature grass-fed horses. Viable counts of total culturable and proteolytic bacteria were made on habitat-simulating media containing 40% clarified ruminal fluid. The mean pHs in the duodenum, jejunum, and ileum were 6.32, 7.10, and 7.47, respectively; the mean pH decreased to 6.7 in the hindgut. The acetate concentration increased along the length of the small intestine and was the only volatile fatty acid present in this gut segment. Molar proportions of acetate, propionate, and butyrate in the hindgut were 85:10:3. Differences in bacterial counts on habitat-simulating media containing equine cecal fluid or clarified ruminal fluid were negligible. Bacterial counts showed a substantial population in the duodenum (ca. 2.9 x 10(6) per g [wet weight] of sample), and this increased to 29.0 x 10(6) in the jejunum and 38.4 x 10(6) in the ileum. Proteolytic bacteria formed a high proportion of the total culturable bacteria, especially in duodenal samples. Counts of proteolytic bacteria per gram (wet weight) of sample were 3.0 x 10(6), 15.6 x 10(6), and 22.0 x 10(6) in the duodenum, jejunum, and ileum, respectively. There was a close relationship between lumenal and mucosal bacterial counts, although actual values were lower in mucosal samples. The mucosal bacterial population in the duodenum was high relative to the lumenal population. Although the comparison of bacterial populations in the hindgut of the horse and white rhino was limited to a single animal, the results were of interest. Counts were higher in the cecum than in the colon for both the horse and the white rhino.(ABSTRACT TRUNCATED AT 250 WORDS)     

Isolation, Culture Characteristics, and Identification of Anaerobic Bacteria from the Chicken Cecum

Studies on the anaerobic cecal microflora of the 5-week-old chicken were made to determine a suitable roll-tube medium for enumeration and isolation of the bacterial population, to determine effects of medium components on recovery of total anaerobes, and to identify the predominant bacterial groups. The total number of microorganisms in cecal contents determined by direct microscope cell counts varied (among six samples) from 3.83 x 10(10) to 7.64 x 10(10) per g. Comparison of different nonselective media indicated that 60% of the direct microscope count could be recovered with a rumen fluid medium (M98-5) and 45% with medium 10. Deletion of rumen fluid from M98-5 reduced the total anaerobic count by half. Colony counts were lower if chicken cecal extract was substituted for rumen fluid in M98-5. Supplementing medium 10 with liver, chicken fecal, or cecal extracts improved recovery of anaerobes slightly. Prereduced blood agar media were inferior to M98-5. At least 11 groups of bacteria were isolated from high dilutions (10(-9)) of cecal material. Data on morphology and physiological and fermentation characteristics of 90% of the 298 isolated strains indicated that these bacteria represented species of anaerobic gram-negative cocci, facultatively anaerobic cocci and streptococci, Peptostreptococcus, Propionibacterium, Eubacterium, Bacteroides, and Clostridium. The growth of many of these strains was enhanced by rumen fluid, yeast extract, and cecal extract additions to basal media. These studies indicate that some of the more numerous anaerobic bacteria present in chicken cecal digesta can be isolated and cultured when media and methods that have been developed for ruminal bacteria are employed.     

Enumeration of selected anaerobic bacterial groups in cecal and colonic contents of growing-finishing pigs.

Selected anaerobic bacterial groups in cecal and colonic contents of clinically healthy pigs fed a corn-soybean meal production diet were determined at sacrifice after 4, 8, and 11 weeks on feed, corresponding to intervals within the growing-finishing growth period. By using ruminal fluid-based media, the densities of the culturable anaerobic population; the cellulolytic, pectin-fermenting, pectin-hydrolyzing, xylan-fermenting; and the xylan-hydrolyzing, sulfate-reducing, and methanogenic bacterial populations were estimated. An analysis of variance was performed on these bacterial group variables to examine the effects of phase (weeks on feed), site (cecum or colon), or the interaction of phase with site. The population of total anaerobic bacteria was twice as dense in the colon as it was in the cecum (2 x 10(10) versus 1 x 10(10)/g [wet weight]; P = 0.001). The proportion of cellulolytic bacteria was lower at 4 weeks on feed than at 8 or 11 weeks (23 versus 32%; P = 0.026), while the proportion of pectin-fermenting bacteria depended on the interaction of phase with site (P = 0.021). The numbers of sulfate-reducing bacteria were significantly higher in the colon than in the cecum (6 x 10(7) versus 3 x 10(7); P = 0.014), as were methanogenic bacteria (19 x 10(7) versus 0.6 x 10(7); P = 0.0002). The remaining bacterial groups were stable with respect to phase and site. The results suggest that except for density differences, the microbial communities of the pig cecum and colon are similar in composition throughout the growing-finishing phase.     

Enumeration of selected anaerobic bacterial groups in cecal and colonic contents of growing-finishing pigs

Selected anaerobic bacterial groups in cecal and colonic contents of clinically healthy pigs fed a corn-soybean meal production diet were determined at sacrifice after 4, 8, and 11 weeks on feed, corresponding to intervals within the growing-finishing growth period. By using ruminal fluid-based media, the densities of the culturable anaerobic population; the cellulolytic, pectin-fermenting, pectin-hydrolyzing, xylan-fermenting; and the xylan-hydrolyzing, sulfate-reducing, and methanogenic bacterial populations were estimated. An analysis of variance was performed on these bacterial group variables to examine the effects of phase (weeks on feed), site (cecum or colon), or the interaction of phase with site. The population of total anaerobic bacteria was twice as dense in the colon as it was in the cecum (2 x 10(10) versus 1 x 10(10)/g [wet weight]; P = 0.001). The proportion of cellulolytic bacteria was lower at 4 weeks on feed than at 8 or 11 weeks (23 versus 32%; P = 0.026), while the proportion of pectin-fermenting bacteria depended on the interaction of phase with site (P = 0.021). The numbers of sulfate-reducing bacteria were significantly higher in the colon than in the cecum (6 x 10(7) versus 3 x 10(7); P = 0.014), as were methanogenic bacteria (19 x 10(7) versus 0.6 x 10(7); P = 0.0002). The remaining bacterial groups were stable with respect to phase and site. The results suggest that except for density differences, the microbial communities of the pig cecum and colon are similar in composition throughout the growing-finishing phase.     

Effects of an abrupt diet change from hay to concentrate on microbial numbers and physical environment in the cecum of the pony

Microbial numbers, pH, fluid volume, and turnover rate in the pony cecum were measured during an abrupt change from an all-forage to an all-concentrate diet, both fed at maintenance energy levels. Concentrate feeding resulted in increased (P less than 0.01) numbers of total viable anaerobic bacteria. The numbers of organisms growing on selective starch medium increased (P less than 0.01) when concentrate was fed, while numbers on xylan and pectin media decreased (P less than 0.025). Seven days after the diet change to concentrate, the number of bacteria growing on lactate medium increased (P less than 0.01), followed by a gradual decline. Cellulolytic bacteria occurred in low numbers, ranging from 1.1 x 10(4) to 4.4 x 10(4) per g of cecal contents. Feeding all concentrate decreased both the number of genera (P less than 0.01) and total protozoan numbers (P less than 0.01) in the cecum. Minimum cecal pH values of 6.4 and 5.8 were obtained when forage and concentrate, respectively, were fed, with the minimum pH occurring 6 h postfeeding. Dry-matter percentage of cecal contents followed a diurnal pattern which was the inverse of the pH curve. During forage feeding, the cecum contained an average of 2.2 liters (1.6 to 3.4 liters), which turned over 3.9 times per day. When concentrate was fed, cecal volume averaged 3.9 liters (0.6 to 8.6 liters), with a mean liquid turnover of 4.2 times per day. Microbial numbers and pH changes in the pony cecum associated with an abrupt change in diet from hay to concentrate resembled those which occur in the rumen under similar feeding conditions.     

Effects of an abrupt diet change from hay to concentrate on microbial numbers and physical environment in the cecum of the pony.

Microbial numbers, pH, fluid volume, and turnover rate in the pony cecum were measured during an abrupt change from an all-forage to an all-concentrate diet, both fed at maintenance energy levels. Concentrate feeding resulted in increased (P less than 0.01) numbers of total viable anaerobic bacteria. The numbers of organisms growing on selective starch medium increased (P less than 0.01) when concentrate was fed, while numbers on xylan and pectin media decreased (P less than 0.025). Seven days after the diet change to concentrate, the number of bacteria growing on lactate medium increased (P less than 0.01), followed by a gradual decline. Cellulolytic bacteria occurred in low numbers, ranging from 1.1 x 10(4) to 4.4 x 10(4) per g of cecal contents. Feeding all concentrate decreased both the number of genera (P less than 0.01) and total protozoan numbers (P less than 0.01) in the cecum. Minimum cecal pH values of 6.4 and 5.8 were obtained when forage and concentrate, respectively, were fed, with the minimum pH occurring 6 h postfeeding. Dry-matter percentage of cecal contents followed a diurnal pattern which was the inverse of the pH curve. During forage feeding, the cecum contained an average of 2.2 liters (1.6 to 3.4 liters), which turned over 3.9 times per day. When concentrate was fed, cecal volume averaged 3.9 liters (0.6 to 8.6 liters), with a mean liquid turnover of 4.2 times per day. Microbial numbers and pH changes in the pony cecum associated with an abrupt change in diet from hay to concentrate resembled those which occur in the rumen under similar feeding conditions.     

Differential carbohydrate media and anaerobic replica plating techniques in delineating carbohydrate-utilizing subgroups in rumen bacterial populations.

A basal (BC) medium devoid of added carbohydrates, a complete (CC) medium containing nine carbohydrates were developed for enumerating rumen bacteria. The colony counts on the BC medium were 85 to 100% of those obtained on the CC medium. These colonies were pinpoint size (less than or equal to mm in diameter) but increased in size (2 to 5 mm in diameter) when carbohydrates were subsequently added. With the CC medium or other media tested, the colony counts were 20 to 50% higher on plates than on roll tubes and were about 35% of the direct cell counts. The lower colony counts on roll tubes were shown to result primarily from the loss of viability due to heat stress. The DC media were found by plating techniques to be suitable for differentiating mixed rumen bacterial populations into subgroups based upon carbohydrate utilization as shown by differences in subgroup profiles found within solid and liquid fractions of rumen contents, within rumen contents from animals fed high-forage and high-grain diets, and by correct colony formations by pure cultures of rumen bacteria on appropriate DC media. With simple modifications and use of an anaerobic glove box, replica plating methods and the CC and DC media were found to be a suitable means of rapidly determining the range of utilizable carbohydrate energy sources of rumen bacteria.     

Differential carbohydrate media and anaerobic replica plating techniques in delineating carbohydrate-utilizing subgroups in rumen bacterial populations

A basal (BC) medium devoid of added carbohydrates, a complete (CC) medium containing nine carbohydrates were developed for enumerating rumen bacteria. The colony counts on the BC medium were 85 to 100% of those obtained on the CC medium. These colonies were pinpoint size (less than or equal to mm in diameter) but increased in size (2 to 5 mm in diameter) when carbohydrates were subsequently added. With the CC medium or other media tested, the colony counts were 20 to 50% higher on plates than on roll tubes and were about 35% of the direct cell counts. The lower colony counts on roll tubes were shown to result primarily from the loss of viability due to heat stress. The DC media were found by plating techniques to be suitable for differentiating mixed rumen bacterial populations into subgroups based upon carbohydrate utilization as shown by differences in subgroup profiles found within solid and liquid fractions of rumen contents, within rumen contents from animals fed high-forage and high-grain diets, and by correct colony formations by pure cultures of rumen bacteria on appropriate DC media. With simple modifications and use of an anaerobic glove box, replica plating methods and the CC and DC media were found to be a suitable means of rapidly determining the range of utilizable carbohydrate energy sources of rumen bacteria.     

Influence of high-fiber diet on bacterial populations in gastrointestinal tracts of obese- and lean-genotype pigs

Bacterial populations from gastrointestinal tracts of genetically lean and obese pigs fed a low- or high-fiber diet (0 or 50% alfalfa meal, respectively) were enumerated with rumen fluid media and specific energy sources. Total culture counts in rectal samples declined 56 (P greater than 0.05) and 63% (P less than 0.05) in lean and obese animals, respectively, 3 weeks after feeding the high-fiber diet. After 8 weeks, culture counts had risen and were similar to those obtained before alfalfa was fed (0 week). At slaughter, 12 to 17 weeks after feeding the high-fiber diet, total counts from rectal samples of lean pigs continued to rise and were 13% greater than the 0-week counts, whereas counts from obese animals declined 37% (P greater than 0.05). The number of cellulolytic bacteria in rectal samples of lean-genotype pigs fed the high-fiber diet increased 80 and 71% from 0 to 3 weeks and 3 to 8 weeks, respectively. This overall increases from 0 to 8 weeks in lean pigs was significant (P less than 0.05); however, these increases were not seen in obese pigs. These data suggest that the microflora is initially suppressed when exposed to a high-fiber diet and that later some adaptation takes place, apparently more so in lean than in obese pigs. When specific energy sources were used to delineate the distribution of different bacterial populations in the cecum, colon, and rectum, trends could be detected between high- and low-fiber diets. These data also support the concept that bacteria populations from different sites in the large bowel differ.     

Influence of high-fiber diet on bacterial populations in gastrointestinal tracts of obese- and lean-genotype pigs.

Bacterial populations from gastrointestinal tracts of genetically lean and obese pigs fed a low- or high-fiber diet (0 or 50% alfalfa meal, respectively) were enumerated with rumen fluid media and specific energy sources. Total culture counts in rectal samples declined 56 (P greater than 0.05) and 63% (P less than 0.05) in lean and obese animals, respectively, 3 weeks after feeding the high-fiber diet. After 8 weeks, culture counts had risen and were similar to those obtained before alfalfa was fed (0 week). At slaughter, 12 to 17 weeks after feeding the high-fiber diet, total counts from rectal samples of lean pigs continued to rise and were 13% greater than the 0-week counts, whereas counts from obese animals declined 37% (P greater than 0.05). The number of cellulolytic bacteria in rectal samples of lean-genotype pigs fed the high-fiber diet increased 80 and 71% from 0 to 3 weeks and 3 to 8 weeks, respectively. This overall increases from 0 to 8 weeks in lean pigs was significant (P less than 0.05); however, these increases were not seen in obese pigs. These data suggest that the microflora is initially suppressed when exposed to a high-fiber diet and that later some adaptation takes place, apparently more so in lean than in obese pigs. When specific energy sources were used to delineate the distribution of different bacterial populations in the cecum, colon, and rectum, trends could be detected between high- and low-fiber diets. These data also support the concept that bacteria populations from different sites in the large bowel differ.     

Enumeration and isolation of anaerobic microbiota of piggery wastes.

Media for enumeration of the microbiota of anaerobically stored piggery wastes were tested. Highest colony counts were obtained with 80 to 100% farm slurry supernatant included in the anaerobic roll tube media. Colony counts with these media numbered 2 X 10(9) to 12 X 10(9)/g (wet weight), which represents about 20% of the microscopic counts. Lower percentages of slurry supernatant in the media gave lower colony counts. Addition of glucose, cellobiose, and starch or of Trypticase to media with 20% slurry supernatant did not increase colony counts. Higher values were obtained when hemicellulose preparations were added to these media. Incubation at 25 degrees C gave the highest numbers. Incubation at 15 to 37 degrees C gave counts of about 70 and 10%, respectively, of those at 25 degrees C. Of the colonies picked for isolation, about 20% were obtained in pure culture. The isolates apparently belonged to the genera Peptococcus, Ruminococcus, Peptococcus, Ruminococcus, Pepostreptococcus, and Bacteroides.     

Enumeration and isolation of anaerobic microbiota of piggery wastes

Media for enumeration of the microbiota of anaerobically stored piggery wastes were tested. Highest colony counts were obtained with 80 to 100% farm slurry supernatant included in the anaerobic roll tube media. Colony counts with these media numbered 2 X 10(9) to 12 X 10(9)/g (wet weight), which represents about 20% of the microscopic counts. Lower percentages of slurry supernatant in the media gave lower colony counts. Addition of glucose, cellobiose, and starch or of Trypticase to media with 20% slurry supernatant did not increase colony counts. Higher values were obtained when hemicellulose preparations were added to these media. Incubation at 25 degrees C gave the highest numbers. Incubation at 15 to 37 degrees C gave counts of about 70 and 10%, respectively, of those at 25 degrees C. Of the colonies picked for isolation, about 20% were obtained in pure culture. The isolates apparently belonged to the genera Peptococcus, Ruminococcus, Peptococcus, Ruminococcus, Pepostreptococcus, and Bacteroides.     

Comparison of media for isolation of mouse anaerobic faecal bacteria

A study was made to determine suitable plate-in-bottle media for enumeration and isolation of the anaerobic faecal bacteria of mice. Comparison of non-selective media indicated that colony counts were higher on M-SM10, M10 and M-M98-5 than on EG, W & E medium and BHI, and those on M-SM10 were always the highest. Bacteroidaceae counts on M-SM10, M10, M-M98-5 were particularly high. On the other hand, in chloroform-treated faeces which contain only Clostridial spores, colony counts were higher on EG and W & E medium than on M-SM10, M10 and M-M98-5. These results indicate that suitable non-selective media vary according to the bacterial species to be enumerated and isolated.