The antimicrobial activity of fatty acids, monolaurin, citric, succinic, fumaric, malic and lactic acid was determined in cultures of two strains of Escherichia coli, three strains of Salmonella sp. and two strains of Clostridium perfringens. Antimicrobial activity was expressed as minimum inhibitory concentration (MIC) that prevented growth and glucose utilization in treated cultures. Caprylic acid was the only acid inhibiting glucose utilization in all cultures. Its MIC varied from 1 to 3 mg/ml. Strains CCM 3954 and CCM 4225 of E. coli were inhibited also by capric acid at 5 mg/ml. Strains CCM 4435<sup>T </sup>and CNCTC 5459 of Cl. perfringens were inhibited by medium-chain fatty acids (C<sub>8</sub> to C<sub>14</sub>), oleic acid and one strain also by linoleic acid. The minimum MICs were those of lauric and myristic acid (between 0.1 and 0.2 mg/ml). Growth of Cl. perfringens, but not other bacteria, was inhibited also by monoglyceride of lauric acid (MIC = 3 mg/ml), and by citric acid (MIC = 4 mg/ml). Inhibitory effects of other acids were not observed at 5 mg/ml. Caprylic and lauric acid did not influence the K<sup>+ </sup>permeability of the cytoplasmic membrane in cells of E. coli CCM 4225 and Cl. perfringens CCM 4435<sup>T</sup>, respectively. In cultures of both strains of E. coli treated with caprylic acid at 5 mg/ml, and in those of Cl. perfringens CCM 4435<sup>T </sup>treated with lauric acid at 1 mg/ml, or with its monoglyceride at 5 mg/ml, the transmission electron microscopy revealed damage of cytoplasmatic structures. In cells of Cl. perfringens the separation of inner and outer membranes was apparent, the integrity of the outer membrane, however, was maintained. It can be concluded that medium-chain fatty acids are more efficient antimicrobials than other, more polar organic acids tested.
Two hundred and thirty colonies from the caecal contents of six rabbits were picked up and, after a 2-d incubation, were microscopically characterized using Gram staining. Large Gram-negative (34%) and small Gram-negative (30%) irregular rods, Gram-negative (27%) and Gram-positive (8%) cocci were found. Eleven isolates (Bacteroides ovatus (6 strains), B. thetaiotamicron, B. caccae, B. stercoris, B. capillosus and Capnocytophaga ochracea) were identified using commercial tests for measuring their catalase activity, metabolite production, etc., and testing their growth in 20% bile. Bacteria belonging to the genus Bacteroides were demonstrated to be the principal pectinolytic organisms in the rabbit caecum.
Sirotek K., E. Santos, V. Benda, M. Marounek: Isolation, Identification and Characterization of Rabbit Caecal Mucinolytic Bacteria. Acta Vet. Brno 2003, 72: 365-370.The aim of our study was to isolate, identify and characterize mucinolytic bacteria from the rabbit caecum. Two hundred and thirty mucin-grown colonies from the caecal contents of 7 rabbits were examined microscopically after the Gram staining. Gram-negative irregular rods were the most numerous mucinolytic isolates (34.1%), followed by gram-negative cocci and short rods (22.5%), gram-positive rods (17.3%), gram-positive cocci (16.6%) and gram-positive sporeforming rods (9.5%). An attempt was done to identify 31 typical isolates on basis of their morphology, biochemical characteristics and production of metabolites. In addition, bacterial cells were hybridized with fluorescently labelled probes for Bacteroides/Prevotella and Clostridium genus. Four isolates were identified at the species level as Mitsuokella multiacidus, three isolates as Bacteroides capillosus and one isolate as Actinomyces izraeli. All isolates except the last one belong to the Bacteroidaceae family. One strain could be assigned to the Bacteroides/Prevotella genus on basis of the hybridization test. Other mucinolytic isolates were not identified as their characteristics did not correspond to any previously described bacterial species. No Clostridium sp. strain was detected. In two M. multiacidus strains high activities of extracellular mucin lyases were found. The pH-optimum of lyases was 6.2. Calcium cations were necessary for their optimal function. This work extends general knowledge about fermentation of carbohydrate and nitrogen substrates in rabbit caecum. Rabbit, caecum, mucin, bacteriaMucin is a glycoprotein present in the mucosal layer lining mammalian gastrointestinal, respiratory and reproductive tract. The carbohydrate moiety can be either an acidic mucopolysaccharide containing uronic acids and hexosamines or an oligosaccharide containing L-fucose and sialic acid (S alyers and Leedle 1983). Mucin composition is tissue specific and differs in different animal species. Mucin is a part of the defense mechanism as it prevents bacteria, viruses and dietary lectins from attaching to the intestinal or other epithelium. Another mucin function is the protection of the mucosal epithelium against acidic and proteolytic damage in the stomach and intestine. In the digestive tract, mucin insures a smooth passage of the foodpulp. Holmes et al. (1974) observed in pigs that large amounts of mucin were not digested in the small intestine, but fermented in the hindgut. Microorganisms responsible for mucin fermentation in the human colon are partially known. Salyers et al. (1977) isolated from the human colon several Bacteroides strains able to degrade mucins and plant polysaccharides. B ayliss and Houston (1984) identified some mucin fermenters as members of the Bacteroides genus, other resembled Bifidobacterium sp., Clostridium septicum, and Eubacterium sp. Crociani et al. (199...
Two strains of Bifidobacterium globosum were isolated from caecal contents of rabbits in a search for potential probiotics. Both strains fermented glucose, galactose, pentoses, maltose, raffinose and starch. Common coccidiostats (monensin, salinomycin) and antimicrobial growth promotors (avoparcin, bacitracin, nitrovin, virginiamycin) supplied at 10 mg/L inhibited their growth in cultures with glucose. Fermentation parameters of bifidobacteria on glucose and starch differed. More formate and ethanol and less lactate were produced during growth on glucose than during growth on starch. When growing on starch, the two strains of bifidobacteria produced 1 mol lactate per 5.6 and 5.7 mol acetate, respectively. Corresponding values during growth on glucose were 17.3 and 8.4 mol of acetate per mol of lactate. Starch-grown cells accumulated more saccharides than cells grown on glucose (1.48 vs. 0.41 and 3.12 vs. 1.18 mmol glucose units per 1 g of dry matter, respectively).
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