Biosynthetic pathways in Oxalobacter formigenes

Oxalate is the only substrate that supports the growth of the gram negative anaerobe, Oxalobacter formigenes. Oxalate is decarboxylated to formate plus CO[subscript]2. A small amount of acetate (0.5-1 mM) is required for biosynthetic reactions. Oxalate is reduced and assimilated into cell biomass by aerobic oxalate-degrading bacteria using either the glycerate pathway or the serine pathway. Oxalate is reduced to 3P-glycerate and assimilated as a C[subscript]3 unit. We detected the enzymatic activities of glycerate pathway but not those of the serine pathway in cell-free extracts of O. formigenes;Four potential sources of carbon for cell biomass are available to O. formigenes, oxalate, acetate, formate and CO[subscript]2. We grew the organism in [superscript]14 C labeled carbon sources and determined the contribution of each of these sources to cell carbon. O. formigenes derived at least 54% of its cell carbon from oxalate and at least 7% from acetate. The only other carbon source utilized was CO[subscript]3. Formate was not incorporated to a significant extent. Carbon from [superscript]14 C-oxalate and [superscript]14 CO[subscript]3 was detected in amino acids derived from [alpha]-ketoglutarate, oxaloacetate, pyruvate, 3P-glycerate and in the aromatic amino acids. Amino acids derived from [alpha]-ketoglutarate, oxaloacetate and pyruvate contained carbon derived from [superscript]14 C-acetate;When O. formigenes was grown on [superscript]13 C-labeled oxalate, acetate or CO[subscript]3,the labeling patterns of the amino acids were consistent with their formation through common biosynthetic pathways. [superscript]13 C from oxalate was detected in the majority of the carbons from all of the amino acids. Approximately 60% of the acetate was incorporated as a C[subscript]2 unit into four amino acids (glutamate, proline, arginine and leucine). The other 40% of the acetate was split and was detected in amino acids derived from oxaloacetate and pyruvate;Enzymatic activities detected in cell-free extracts included: glutamate dehydrogenase, phosphoenolpyruvate carboxylase, pyruvate carboxylase, citrate synthase and isocitrate dehydrogenase. These findings support the [superscript]14 C and [superscript]13 C data which indicate that O. formigenes assimilates acetate into protein using the first third of the TCA pathway and that C[subscript]4 units are formed from C[subscript]3 units by carboxylation of pyruvate or PEP

Lachnospira pectinoschiza sp. nov., an Anaerobic Pectinophile from the Pig Intestine

Pectinophiles are bacteria that utilize pectin and only a few related compounds as substrates. Obligately anaerobic pectinophiles have been isolated from the intestinal tracts and gingivae of humans and from the rumina of cattle. We isolated three strains of pectinophilic bacteria from colonic contents of pigs but were unable to isolate pectinophiles from the rumen contents of four sheep, even when the animals were fed a high-pectin diet. The pectinophiles isolated from pigs were strictly anaerobic, motile, gram-positive rods (0.36 to 0.56 by 2.4 to 3.1 μm). Pectin, polygalacturonic acid, and gluconate were the only substrates that supported rapid growth. All three strains grew slowly on either lactose or cellobiose and fermented fructose after a lag of several days. Pectin was degraded by means of an extracellular pectin methylesterase and a Ca2+-dependent exopectate lyase. A comparison of the 16S rRNA sequences of these isolates with the 16S rRNA sequences of other gram-positive bacteria revealed a specific relationship with Lachnospira multipara (level of similarity, 94%). The Gram reaction, formation of spore-like structures, and the utilization of lactose and cellobiose differentiated the pig isolates from previously described pectinophiles. The pig isolates represent a previously undescribed species of the genus Lachnospira, for which we propose the name Lachnospira pectinoschiza

Long Polar Fimbriae Contribute to Colonization by \u3ci\u3eEscherichia coli\u3c/i\u3e O157:H7 In Vivo

The contribution of long polar fimbriae to intestinal colonization by Escherichia coli O157:H7 was evaluated in sheep, conventional pigs, and gnotobiotic piglets. E. coli O157:H7 strains with lpfA1 and lpfA2 mutated were recovered in significantly lower numbers and caused fewer attachment and effacement lesions than the parent strain

Transmission and Infectious Dose of Escherichia coli O157:H7 in Swine

Escherichia coli O157:H7 is only occasionally isolated from healthy swine, but some experimentally infected animals will shed the organism in their feces for at least 2 months. Potential explanations for the paucity of naturally occurring infections in swine, as compared to cattle, include a lack of animal-to-animal transmission so that the organism cannot be maintained within a herd, a high infectious dose, or herd management practices that prevent the maintenance of the organism in the gastrointestinal tract. We hypothesized that donor pigs infected with E. coli O157:H7 would transmit the organism to naïve pigs. We also determined the infectious dose and whether housing pigs individually on grated floors would decrease the magnitude or duration of fecal shedding. Infected donor pigs shedding 4 CFU of E. coli O157:H7 per g transmitted the organism to 6 of 12 naïve pigs exposed to them. The infectious dose of E. coli O157:H7 for 3-month-old pigs was approximately 6 × 103 CFU. There was no difference in the magnitude and duration of fecal shedding by pigs housed individually on grates compared to those housed two per pen on cement floors. These results suggest that swine do not have an innate resistance to colonization by E. coli O157:H7 and that they could serve as a reservoir host under suitable conditions.This article is from Applied and Environmental Microbiology 70 (2004): 5331, doi:10.1128/AEM.70.9.5331-5335.2004. Posted with permission.</p

Persistent Colonization of Sheep by Escherichia coli O157:H7 and Other E. coli Pathotypes

Shiga toxin-producing Escherichia coli (STEC) is an important cause of food-borne illness in humans. Ruminants appear to be more frequently colonized by STEC than are other animals, but the reason(s) for this is unknown. We compared the frequency, magnitude, duration, and transmissibility of colonization of sheep by E. coli O157:H7 to that by other pathotypes of E. coli. Young adult sheep were simultaneously inoculated with a cocktail consisting of two strains of E. coliO157:H7, two strains of enterotoxigenic E. coli (ETEC), and one strain of enteropathogenic E. coli. Both STEC strains and ETEC 2041 were given at either 107 or 1010CFU/strain/animal. The other strains were given only at 1010CFU/strain. We found no consistent differences among pathotypes in the frequency, magnitude, and transmissibility of colonization. However, the STEC strains tended to persist to 2 weeks and 2 months postinoculation more frequently than did the other pathotypes. The tendency for persistence of the STEC strains was apparent following an inoculation dose of either 107 or 1010 CFU. One of the ETEC strains also persisted when inoculated at 1010 CFU. However, in contrast to the STEC strains, it did not persist when inoculated at 107 CFU. These results support the hypothesis that STEC is better adapted to persist in the alimentary tracts of sheep than are other pathotypes ofE. coli.This article is from Applied and Environmental Microbiology 66 (2000): 4926, doi:10.1128/AEM.66.11.4926-4934.2000.</p

Persistent Colonization of Sheep by Escherichia coli O157:H7 and Other E. coli Pathotypes

Shiga toxin-producing Escherichia coli (STEC) is an important cause of food-borne illness in humans. Ruminants appear to be more frequently colonized by STEC than are other animals, but the reason(s) for this is unknown. We compared the frequency, magnitude, duration, and transmissibility of colonization of sheep by E. coli O157:H7 to that by other pathotypes of E. coli. Young adult sheep were simultaneously inoculated with a cocktail consisting of two strains of E. coli O157:H7, two strains of enterotoxigenic E. coli (ETEC), and one strain of enteropathogenic E. coli. Both STEC strains and ETEC 2041 were given at either 10(7) or 10(10) CFU/strain/animal. The other strains were given only at 10(10) CFU/strain. We found no consistent differences among pathotypes in the frequency, magnitude, and transmissibility of colonization. However, the STEC strains tended to persist to 2 weeks and 2 months postinoculation more frequently than did the other pathotypes. The tendency for persistence of the STEC strains was apparent following an inoculation dose of either 10(7) or 10(10) CFU. One of the ETEC strains also persisted when inoculated at 10(10) CFU. However, in contrast to the STEC strains, it did not persist when inoculated at 10(7) CFU. These results support the hypothesis that STEC is better adapted to persist in the alimentary tracts of sheep than are other pathotypes of E. coli

Indirect Transmission of Escherichia coli O157:H7 Occurs Readily among Swine but Not among Sheep▿

Transmission of Escherichia coli O157:H7 among reservoir animals is generally thought to occur either by direct contact between a naïve animal and an infected animal or by consumption of food or water containing the organism. Although ruminants are considered the major reservoir, there are two reports of human infections caused by E. coli O157:H7 linked to the consumption of pork products or to the contamination of fresh produce by swine manure. The objective of this study was to determine whether E. coli O157:H7 could be transmitted to naïve animals, both sheep and swine, that did not have any direct contact with an infected donor animal. We recovered E. coli O157:H7 from 10/10 pigs with nose-to-nose contact with the infected donor or animals adjacent to the donor and from 5/6 naïve pigs that were penned in the same room as the donor pig but 10 to 20 ft away. In contrast, when the experiment was repeated with sheep, E. coli O157:H7 was recovered from 4/6 animals that had nose-to-nose contact with the infected donor or adjacent animals and from 0/6 naïve animals penned 10 to 20 ft away from the donor. These results suggest that E. coli O157:H7 is readily transmitted among swine and that transmission can occur by the creation of contaminated aerosols