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H W Moon

Publications and source records attributed to H W Moon.

At least 109 records · Page 6Linked to original sources

Effects of age, ambient temperature, and heat-stable Escherichia coli enterotoxin on intestinal transit in infant mice.

Some interrelationships among age, ambient temperature, intestinal transit, and enterotoxigenic Escherichia coli infection were studied in an infant mouse model. The transit of dye in the small intestine was accelerated during the response to heat-stable E. coli enterotoxin. Transit in the small intestine of normal mice accelerated with increased age (from less than 17 h to 8 days old) and accelerated with increased ambient temperature (from 25 to 37 degrees C). Transit was more rapid in the jejunum than in the ileum throughout the range of experimental conditions studied. E. coli strains that do not produce any of the pili known facilitate intestinal colonization were cleared from the small intestine more rapidly at 37 degrees C than at 25 degrees C. This clearance was thought to be due to accelerated transit at the higher temperature. In contrast, a strain of E. coli that produces K99 (pili previously shown to facilitate intestinal colonization in other species) was not cleared from the small intestine and colonized more intensively at 37 degrees C than at 25 degrees C. Intensified colonization by this strain was thought to be due to increased production of K99 at the higher temperature. It was suggested that sluggish intestinal transit may also be characteristic of the neonates of other species and be one of the factors predisposing them to intestinal colonization by enteropathogens. It was speculated that this predisposition may be enhanced if the neonates are chilled. However, the effect of ambient temperature on intestinal transit in homeothermic neonates such as pigs, calves, and humans may be different from that in mice because neonatal mice are poikilothermic.

Aging↗

Localization and activities of lysosomal enzymes in jejunal and ileal epithelial cells of the young pig.

The localization and the specific activities of lysosomal enzymes in jejunal and ileal epithelium of suckling pigs were determined. Acid phosphatase was found in pinocytotic vesicles and vacoles at both intestinal sites in pigs examined during (less than or equal to 1 day old) or shortly after (2 days old) closure. These pinocytolically active vesicles and vacuoles were determined to be phagolysosomes. In the ileum, phagolysosomes persisted well after closure, at an age (10 days) when ileal epithelium retains the capacity for pinocytotic uptake of immunoglobulin, but does not transport it to blood. Neither jejunum nor ileum contained phagolysosomes at an age (30 days) when the intestine had lost both uptake and transport capacities. The specific activities of acid phosphatase and cathepsin B-1 in jejunum were similar for all age groups. However, the activities of these enzymes in ileum of 10-day-old pigs were three and five times greater than at any other age or site. The results are consistent with phagolysosomes in the ileum of 10-day-old pigs functioning as a "barrier" which prevents transport of the macromolecules which are taken up pinocytotically by this epithelium. The results do not permit conclusions whether (or not) the phagolysosomes which appear in jejunum and ileum of pigs less than or equal to 1 day old contribute to closure, because the transcellular route for immunoglobulin absorption in pigs has not been precisely defined.

Acid Phosphatase↗

Pathogenic relationships of rotavirus, Escherichia coli, and other agents in mixed infections in calves.

Infection with agents interpreted as causing or contributing to diarrhea (rotavirus, coronavirus, enterotoxigenic Escherichia coli, and cryptosporidia) were demonstrated in 24 of 32 newborn calves that had naturally occurring diarrheal disease. The calves were from 12 herds in Iowa. Infections as well as enteric lesions and hypoglobulinemia occurred more frequently among diarrheal calves than among nondiarrheal calves from these same herds. In most calves, infections were mixed; ie, both viruses, one or both viruses plus cryptosporidia, or rotavirus plus enterotoxigenic E coli.

Animals↗

Effects of age and ambient temperature on the responses of infant mice to heat-stable enterotoxin of Escherichia coli: assay modifications.

The response of infant mice to heat-stable enterotoxin from Escherichia coli was affected by the age of the mice (2, 4, 6, and 8 days) and by the ambient temperature (25, 30, and 37 degrees C) after exposure to the enterotoxin. The younger mice and/or mice held at lower temperatures tended to accumulate intestinal fluid (high gut weight/body weight ratios), but older mice and/or mice held at higher temperatures tended to respond with diarrhea and low gut weight/body weight ratios. The standard infant mouse assay forheat-stable E. coli enterotoxin can be simplified, without loss of sensitivity or reliability, by holding the mice at 37 degrees C after exposure and using diarrhea as the index of response. Diarrhea can be detected easily by incorporating dye in the inocula and (at the end of the assay) checking for dye mixed with feces on the rear quarters of the mice or on a sheet of white paper placed under them during incubation.

Aging↗

Immunization of suckling pigs against enteric enterotoxigenic Escherichia coli infection by vaccinating dams with purified pili.

Pregnant swine (gilts) were vaccinated parenterally with a suspension of purified pili from the porcine enterotoxigenic Escherichia coli strain 987 (09:K103::NM). Gilts injected with placebo served as controls. Suckling pigs born to gilts in both groups were challenged intragastrically with virulent strain 987. The percentage of deaths, incidence and duration of diarrhea, numbers of E. coli in the ilea, and E. coli attachment to the villous epithelia were significantly less in suckling pigs of vaccinated gilts than in those of controls. These results are consistent with the hypothesis that pili of some enterotoxigenic E. coli facilitate adhesion to intestinal epithelia. Vaccination of dams with pili appears to be a means of immunizing against diarrheal disease caused by enterotoxigenic E. coli in suckling neonates. This work confirms the role of somatic pili as colonization and virulence factors and provides another example of safe and effective purified pilus vaccines.

Animals↗

In vitro adhesion of Escherichia coli to porcine small intestinal epithelial cells: pili as adhesive factors.

Escherichia coli strains with pili (K99 or 987P) known to facilitate intestinal colonization adhered in vitro to porcine intestinal epithelial cells. These strains adhered equally to both ileal and jejunal epithelial cells. A laboratory E. coli strain that has type 1 pili also adhered to porcine intestinal epithelial cells. When nonpiliated cells derived from 987P+, K99+, or type 1 pilus+ strains were used for in vitro adhesion assays, they failed to adhere. The attachment of piliated bacteria to epithelial cells was a saturable process that plateaued at 30 to 40 bacterial cells attached per epithelial cell. Competitive inhibition of bacterial cell attachment to epithelial cells with purified pili showed that only purified 987P competed against the 987P+ strain and only purified type 1 pili competed against the type 1 pilus+ strain. Competition between a K99+ strain and K99 was not consistently achieved. K99+, 987P+, and type 1 pilus+ bacteria could be prevented from adhering to epithelial cells by Fab fragments specific for K99, 987P, or type 1 pili, respectively. Fab fragments specific for non-K99 bacterial surface antigens did not inhibit adhesion of the K99+ strain. It is concluded that adhesion of E. coli to porcine intestinal epithelial cells in vitro is mediated by pili and that the epithelial cells used apparently had different receptors for different pili.

Animals↗

Immunization of suckling pigs against enterotoxigenic Escherichia coli-induced diarrheal disease by vaccinating dams with purified 987 or K99 pili: protection correlates with pilus homology of vaccine and challenge.

Pregnant gilts were vaccinated with purified strain 987 pili (987P), strain K99 pili, or a saline-formaldehyde control. Suckling pigs born to vaccinated gilts were allowed to consume colostrum and were then challenged intragastrically with one of three enterotoxigenic Escherichia coli strains: 987 (O9:K103, 987P:NM), 74-5208 (02O:K101, 987P:NM) or 431 (O101:K30, 99:NM). In litters where the dam was vaccinated with the same pilus as that possessed by the challenge organism, the incidence and duration of diarrhea and the degree of intestinal colonization (either duration or extent) were less than those of the other vaccine groups. Surviving pigs in the homologous vaccine groups also had better weight gains than pigs in the other vaccine groups. The experiments extend and confirm previous reports that vaccination of the dam with purified pili confers protection to neonatal suckling pigs against diarrheal disease caused by enterotoxigenic E. coli strains that possess the same pili. Protection did not extend to enterotoxigenic strains possessing different pili.

Animals↗

Bovine cryptosporidiosis: a transmission and scanning electron microscopic study of some stages in the life cycle and of the host-parasite relationship.

Transmission and scanning electron microscopy of the ileal mucosae from 12 calves infected with Cryptosporidium sp. showed cryptosporidia free in the lumen and attached to epithelium. The attached parasites were interpreted to be extracellular and adherent to the microvillous border of epithelial cells. Stages of the organism included crescentic-free merozoites, trophozoites, schizonts, gametes and oocysts. Attached parasites were detected chiefly at villous tips and all stages were present on a single villus. Attachment sites were characterized by absence or disintegration of microvilli, disorganization of the terminal web and development of a specialized attachment zone. There were increased numbers of lysosomes and irregularities in the nuclear membrane of parasitized epithelial cells. It was concluded that cryptosporidia exist in bovine intestine as extracellular parasites and cause epithelial changes that in turn probably cause or contribute to diarrheal disease.

Animals↗

Colonization of porcine small intestine by Escherichia coli: colonization and adhesion factors of pig enteropathogens that lack K88.

The colonizing and adhesive attributes of enterotoxigenic acapsular and/or nonpiliated mutants from K88-negative enteropathogenic Escherichia coli strains were compared with their capsulated and piliated parents (parents were piliated when grown in vitro and in vivo). Acapsular, nonpiliated mutants from three different colonizing strains of enteropathogenic E. coli lost their ability to colonize the ileum of newborn pigs. Acapsular, piliated and capsular, nonpiliated mutants were derived from one of the parental strains (987), and both mutants lacked the ability to colonize the ileum of pigs. The only mutants available from a fourth strain (431) were acapsular and piliated, and they colonized as well as their parents. These data indicate that both capsule and pili are involved in colonization by strain 987. In contrast, capsule is not required for colonization by strain 431, but pili may be.

Animals↗

Intestinal colonization and adhesion by enteroxigenic Escherichia coli: ultrastructural observations on adherence to ileal epithelium of the pig.

Colonization of pig ileum by enterotoxigenic Escherichia coli that were enteropathogenic for pigs but that lacked K88 antigen (K88-) resulted in morphological characteristics similar to those reported for K88+ strains. Strains of enterotoxigenic E. coli from three different K88-serotypes adhered to the villous epithelium. In sections examined by transmission electron microscopy, adherent bacteria were separated from each other and from epithelial microvilli by peribacterial electron-lucent regions. The enterotoxigenic E. coli had appendages that extended into these regions. The appendages were morphologically characteristic for each strain. It is possible that these appendages were pili, polysaccharide K antigens, or structures resulting from some interaction between pili and polysaccharide. Certain pili or pilus-like structures may be virulence attributes that facilitate adhesion of enterotoxigenic E. coli to the intestinal epithelium.

Animals↗

Occurrence of K99 antigen on Escherichia coli isolated from pigs and colonization of pig ileum by K99+ enterotoxigenic E. coli from calves and pigs.

Several strains of enterotoxigenic Escherichia coli (ETEC) isolated from pigs were found to have an antigen (K99) previously reported only on strains of calf and lamb origin and which facilitates intestinal colonization in the latter two species. Several human ETEC were also tested for K99; however, none were positive. Each of four K99-positive ETEC strains of calf origin and one of pig origin produced K99 in pig ileum in vivo, adhered to villous epithelium in pig ileum, colonized pig ileum, and caused profuse diarrhea in newborn pigs. In contrast to the K99-positive strains above, four K99-negative ETEC from humans and chickens and one K99-positive ETEC from a calf either did not colonize pig ileum or did so inconsistently. When the K99-negative strains did colonize, they had little or no tendency to adhere to intestinal villi. These results are consistent with the hypothesis that K99 facilitates adhesion to and colonization of pig ileum by some ETEC.

Animals↗

Colonization of porcine intestine by enterotoxigenic Escherichia coli: selection of piliated forms in vivo, adhesion of piliated forms to epithelial cells in vitro, and incidence of a pilus antigen among porcine enteropathogenic E. coli.

In contrast to K88-positive porcine enterotoxigenic Escherichia coli (ETEC), K88-negative porcine ETEC strains did not adhere to isolated intestinal epithelial cells in vitro. However, they did adhere to intestinal epithelium in vivo. Growth of one such ETEC (strain 987) in pig small intestine consistently yielded a greater percentage of piliated cells than did growth in vitro. This increase was demonstrable by electron microscopy, by change in colonial morphology, and by agglutination in specific antisera against the pili of strain 987. In contrast to the stored stock culture (which contained very few piliated cells), richly piliated forms of strain 987 did adhere to isolated intestinal epithelial cells in vitro. A series of porcine E. coli strains was tested for agglutinability in antiserum against the pili of strain 987, and several K88-negative ETEC strains were agglutinated. These data are consistent with the hypothesis that pili facilitate intestinal adhesion and colonization by K88-negative ETEC strains.

Antigens, Bacterial↗

Colonization of porcine small intestine by Escherichia coli: ileal colonization and adhesion by pig enteropathogens that lack K88 antigen and by some acapsular mutants.

Seven K88-negative porcine enteropathogenic Escherichia coli, representing three different serogroups, caused severe diarrhea and characteristically colonized the ileum, but not the jejunum, of intragastrically exposed newborn pigs. Bacterial counts of intestinal contents and wall, fluorescence, and scanning electron microscopy all suggested that these strains colonized the ileum by adhesion to the villous epithelium. However, in ligated intestinal loops, these enteropathogenic E. coli strains adhered to jejunal epithelium as well as to ileal epithelium. Acapsular (K-) mutants, derived from one of the principal strains, retained their colonizing and adhesive abilities, whereas K- mutants from three other enteropathogenic E. coli strains did not. It is suggested that: (i) these K88-negative enteropathogenic E. coli colonize the ileum by adhesion, and (ii) the adhesion of some K-88-negative strains is mediated by surface factors other than, or in addition to, the polysaccharide K antigen.

Animals↗

Etiologic diagnosis of diarrheal disease of calves: frequency and methods for detecting enterotoxin and K99 antigen production by Escherichia cola.

Escherichia coli isolated from calves in Minnesota and Montana were tested for enterotoxigenicity via bio-assay of cell-free broth culture fluid and for K99 antigen via a serum agglutination test. Infant mice were used to assay for heat-stable enterotoxin (ST), and adrenal cells in culture were used to assay for heat-labile enterotoxin (LT). Forty-six of the 345 E coli isolates produced ST enterotoxin, but none produced LT enterotoxin. Thirty-five of the 46 enterotoxigenic isolates had K99 antigen, and only 9 of 66 nonenterotoxigenic isolates so tested had this antigen. The enterotoxigenicity of 28 additional E coli isolates known or suspected to be calf enteropathogens and provided by investigators from 3 different laboratories was also tested. All isolates from 2 laboratories produced ST but not LT. All isolates from the 3rd laboratory produced LT but not ST. Escherichia coli organisms that were positive in the infant mouse assay also caused positive ligated, jejunal-loop responses in calves and in 9-day-old (but not in 5-week-old) pigs. It was concluded that the infant mouse and adrenal cell tests for ST and LT, combined with the agglutination test for K99, would be useful in the diagnosis of enteric enterotoxic colibacillosis of calves.

Agglutination Tests↗

Heat-stable Escherichia coli enterotoxin production in vivo.

Hysterectomy-derived, colostrum-deprived piglets were infected with enterotoxigenic Escherichia coli on day 4 of life. Samples of feces and intestinal contents were collected and tested in infant mice for enterotoxic activity. Positive enterotoxic responses were observed in mice given filtrates of feces and intestinal contents from piglets infected withe enterotoxigenic E. coli known to produce heat-stable enterotoxin but not heat-liabile enterotoxin in vitro. It is concluded that heat-stable enterotoxigenic E. coli induce diarrhea by production of heat-stable enterotoxin in vivo.

Animals↗