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Biomedical subjects

H W Moon

Publications and source records attributed to H W Moon.

At least 91 records · Page 5Linked to original sources

Colonic compensation in transmissible gastroenteritis of swine.

Absorption of water and electrolytes by the small and large intestine was examined using a nonabsorbable marker technique in 3-day-old and 3-wk-old pigs. One-half of the pigs in each group were orally infected with transmissible gastroenteritis virus; the remaining pigs served as controls. Three-day-old control pigs concentrated the nonabsorbable fluid marker twelve fold along the small and large intestine, indicating an efficiency of about 95% in absorption of the exogenous daily fluid load presented to the intestine. In contrast, the marker concentration in infected pigs showed no change whatsoever along either the small or large intestine, indicating a complete absence of net fluid absorption or secretion in these animals. Three-week-old control pigs concentrated the marker similarly to the 3-day-old group, with the bulk of the fluid absorption occurring in the small intestine. Infected pigs in the 3-wk-old group had marked net fluid secretion in the proximal small intestine, so that about twice the fluid load was presented to the large intestine of the 3-wk-old infected pigs as compared to the 3-day-old infected group. However, in contrast to the 3-day-old infected group, the large intestine of the 3-wk-old infected pigs increased fluid absorption some six times over the control, and this compensatory response prevented diarrhea in these older animals. Analysis of luminal contents indicated that in the older pigs, unabsorbed carbohydrate was almost completely fermented to short-chain fatty acids in the colon, whereas in the younger pigs the carbohydrate passed through the colon unchanged. These results demonstrate that development of microbial digestion, together with rapid short-chain fatty acid absorption, is a primary feature responsible for the colonic compensation in the older pigs with transmissible gastroenteritis.

Age Factors↗

Plasmid-mediated adhesion in enteropathogenic Escherichia coli.

A survey of classical serotype enteropathogenic Escherichia coli has been made with respect to their plasmid profile and ability to adhere to HEp-2 cells. Thirty-one of the 32 strains examined contained a 50-70 Md plasmid, and many exhibited HEp-2 adherence. Strain E2348 (0127:H6), which causes diarrhea in volunteers and is HEp-2-adhesive, was chosen for further study. The large 55 Md plasmid in E2348, pMAR2, has been marked with a transposon coding for ampicillin resistance. E2348 that has been cured of pMAR2 loses the ability to adhere to HEp-2 cells, while HB101, a nonadherent E. coli K12, acquires HEp-2 adhesiveness after gaining this plasmid. Plasmid presence was also shown to correlate with in vivo adhesion to intestine, using the colostrum-deprived piglet model.

Animals↗

Immunization of swine with heat-stable Escherichia coli enterotoxin coupled to a carrier protein does not protect suckling pigs against an Escherichia coli strain that produces heat-stable enterotoxin.

Pregnant swine were immunized parenterally with purified heat-stable Escherichia coli enterotoxin that was made antigenic by coupling it to bovine immunoglobulin G. Immunized swine had high titers of antitoxin in serum and colostrum as measured by radioimmunoassay. However, the heat-stable enterotoxin neutralizing titers of the serum and colostrum from immunized swine were comparatively low. Newborn pigs suckling their immunized dams were not protected against challenge with porcine enterotoxigenic E. coli that produce heat-stable toxin but do not produce heat-labile toxin.

Animals↗

Attaching and effacing activities of rabbit and human enteropathogenic Escherichia coli in pig and rabbit intestines.

Three strains of enteropathogenic Escherichia coli (EPEC), originally isolated from humans and previously shown to cause diarrhea in human volunteers by unknown mechanisms, and one rabbit EPEC strain were shown to attach intimately to and efface microvilli and cytoplasm from intestinal epithelial cells in both the pig and rabbit intestine. The attaching and effacing activities of these EPEC were demonstrable by light microscopic examination of routine histological sections and by transmission electron microscopy. It was suggested that intact colostrum-deprived newborn pigs and ligated intestinal loops in pigs and rabbits may be useful systems to detect EPEC that have attaching and effacing activities and for studying the pathogenesis of such infections. The lesions (attachment and effacement) produced by EPEC in these systems were multifocal, with considerable animal-to-animal variation in response to the same strain of EPEC. The EPEC strains also varied in the frequency and extent of lesion production. For example, three human EPEC strains usually caused extensive lesions in rabbit intestinal loops, whereas two other human EPEC strains usually did not produce lesions in this system.

Animals↗

Characterization of the gene encoding heat-stable toxin II and preliminary molecular epidemiological studies of enterotoxigenic Escherichia coli heat-stable toxin II producers.

The gene encoding heat-stable toxin II (STII) was cloned into an Escherichia coli K-12 strain, and its nucleotide sequence was determined. The deduced amino acid sequence indicates that STII is synthesized within the cell as a 71-amino-acid protein and that neither the DNA nor amino acid sequence bears any similarity to that of heat-stable toxin I. A DNA fragment containing the STII gene was used to probe enterotoxigenic E. coli clinical isolates with various toxin phenotypes and was shown to be useful in detecting all STII and only STII producers.

Amino Acid Sequence↗

Pilus production, hemagglutination, and adhesion by porcine strains of enterotoxigenic Escherichia coli lacking K88, K99, and 987P antigens.

Three strains of enterotoxigenic Escherichia coli which adhered, colonized intensively, and caused disease in pig intestine, but which did not produce pili of the K88, K99, or 987P antigen types were designated 3P(-) ETEC. The 3P(-) ETEC caused mannose-resistant hemagglutination, adhered to porcine intestinal epithelial cells in vitro, and produced pili. However, most bacteria taken directly from the intestine of pigs infected with 3P(-) ETEC appeared to be nonpiliated. Two preparations were isolated from the 3P(-) ETEC. One (material A) contained pili, caused mannose-sensitive hemagglutination, and did not inhibit adhesion of whole bacteria to epithelial cells in vitro. The other (material B) had no demonstrable pili, caused mannose-resistant hemagglutination, and blocked ahesion of bacteria to epithelial cells in vitro. Antiserum against an acapsular mutant (K(-)) of one 3P(-) ETEC strain was absorbed to remove antibodies directed against somatic (O) antigen. The absorbed antiserum agglutinated all three 3P(-) ETEC strains grown in the K(-) form at 37 degrees C, but not when they were grown at 18 degrees C. The absorbed antiserum blocked the hemagglutinating activity of material B, but not of material A. It also reacted (via indirect immunofluorescence) with all of the 3P(-) ETEC when they were grown in pig intestine. The results were interpreted to indicate that: (i) the epithelial adhesive and mannose-resistant hemagglutinating activities of the 3P(-) ETEC strains may be mediated by an antigen contained in material B; (ii) this antigen either is not pilus associated or is associated with pili that are not demonstrable by the methods used here; (iii) the 3P(-) ETEC strains produce type 1 pili which do not mediate their adhesion to intestinal epithelium of pigs.

Adhesiveness↗

Experimental fecal transmission of human cryptosporidia to pigs, and attempted treatment with an ornithine decarboxylase inhibitor.

Fecal material collected from an immunologically deficient man with persistent cryptosporidia infection was stored in potassium dichromate for two weeks and then fed (inoculated) to newborn pigs. The six inoculated newborn pigs shed the organism in their feces starting four to five days afer inoculation and continuing for as long as 22 days after inoculation. Pigs which were killed and necropsied while shedding had cryptosporidia infection of ileum, cecum, and colon. Infected pigs had atrophied ileal villi and flattened irregular cecal and colonic epithelium. Uninoculated littermate controls remained free to the infection and had histologically normal intestinal tracts at necropsy. Treatment of three of the six inoculated pigs with the ornithine decarboxylase inhibitor, DL-alpha-difluoromethylornithine, orally for ten days had no apparent effect on the infection.

Animals↗

Comparison of enterotoxic activities of heat-stable enterotoxins from class 1 and class 2 Escherichia coli of swine origin.

Pig small intestine develops age-dependent resistance to some (class 2 strains) enterotoxigenic Escherichia coli while remaining susceptible to others (class 1 strains). This study tested the hypothesis that class 1 and class 2 strains produce different subtypes of heat-stable enterotoxin (ST). The dose-response curves of small intestine to crude ST preparations from a class 1 and a class 2 strain were compared in several species. In infant mice, the class 1 ST preparation was less active than the class 2 ST preparation, whereas in rabbits the preparations were equally potent. However, in 1-, 7-, and 14-week-old pigs, the class 1 ST preparation was more active than the class 2 preparation. At low doses, both preparations caused reduced absorption in pigs of all three age groups, and at high doses the class 1 preparation caused secretion in all three age groups. In contrast, at high doses the class 2 preparation caused secretion in 1-week-old pigs but only reduced absorption in older pigs. when class 1 and class 2 ST preparations were fractionated by methanol extraction, in both cases the mouse-negative, pig-positive activity was associated with the methanol-insoluble fraction and mouse-positive, pig-positive activity was associated with the methanol-soluble fraction. The results are consistent with a hypothesis that class 1 and class 2 strains of enterotoxigenic E. coli produce different subtypes of ST and that the response of pig intestine to ST varies with both age and toxin subtype.

Aging↗

Fecal transmission of calf cryptosporidia between calves and pigs.

All of 17 calves 1 to 21 days old and 23 of 24 1-day-old pigs inoculated orally with calf feces containing cryptosporidia oocysts became infected with cryptosporidia. Infectivity was maintained by discontinuous passage in calves, with storage in potassium dichromate solution for up to eight weeks between passages. Cryptosporidia shed in calf feces also were infectious for calves directly after collection, without storage in potassium dichromate (two calves). Infected calves and pigs shed cryptosporidia oocysts in their feces for several days (mean of 10 days). Those necropsied at this stage of infection had many cryptosporidia and partial atrophy of villi in their ilea. All of the pigs and 60% of the calves also had cryptosporidia infections in their large intestines at this stage. It was concluded that calf cryptosporidia can be transmitted via feces to calves and to pigs.

Animals↗

Protection against enteric colibacillosis in pigs suckling orally vaccinated dams: evidence for pili as protective antigens.

Pregnant gilts were vaccinated orally with Escherichia coli that produced pilus antigens K99 or 987P. The vaccines were live or dead enterotoxigenic E coli (ETEC) or a liver rough non-ETEC strain which has little ability to colonize pig intestine. Pigs born to the gilts were challenge exposed orally with K99+ or 987P+ ETEC, which did not produce heat-labile enterotoxin or flagella and which produced somatic and capsular antigens different from those of the vaccine strains. Control gilts had low titers of serum and colostral antibodies against pilus antigens, and their suckling pigs frequently had fatal diarrhea after challenge exposure. Serum antibody titers against pilus antigens of the vaccine strains increased in the gilts after vaccination with liver ETEC, and the colostral antibody titers of these gilts were higher than those of controls. Pigs suckling such vaccinated gilts were more resistant than controls to challenge strains were of different pilus types, and it could not be attributed to enterotoxin neutralization by colostrum. In contrast to the live ETEC vaccines given to the pregnant gilts, the liver rough non-ETEC and dead ETEC vaccines stimulated little or no production of antibody against pilu, and the pigs born of these vaccinated gilts remained highly susceptible to challenge exposure. The results support the hypothesis that pilu can be protective antigens in oral ETEC vaccines. It was indicated that in the system reported, protection depended on living bacteria for the production of pilus antigens in vivo or for the transport of pilus antigens across intestinal epithelium.

Administration, Oral↗

Prevalence of pilus antigens, enterotoxin types, and enteropathogenicity among K88-negative enterotoxigenic Escherichia coli from neonatal pigs.

Enterotoxigenic Escherichia coli (ETEC) that were isolated from neonatal pigs and that did not react in preliminary tests for pilus antigen K88 were subjected to additional tests for K88 and for pilus antigens K99 and 987P. Four such isolates produced K88, 9 isolates produced K99, 55 isolates produced 987P, and the remaining 43 isolates produced none of the three pilus antigens (3P(-)). Immunofluorescence tests of ileal sections from pigs were more sensitive for 987P detection than was serum agglutination of bacteria grown from the ileum. Most ETEC that produced K88, K99, or 987P were enteropathogenic (adhered to ileal villi, colonized intensively, and caused profuse diarrhea) when given to neonatal pigs. In contrast, only 3 of the 43 ETEC that produced none of the pilus antigens were enteropathogenic. The isolates were also tested for the type of enterotoxin produced. The K88(+) isolates all produced heat-labile enterotoxin (LT) detectable in cultured adrenal cells (i.e., were LT(+)). None of the 987P(+), K99(+), or enterpathogenic 3P(-) isolates produced LT. However (except for a single K99(+) isolate), they all produced heat-stable enterotoxin detectable in infant mice (STa(+)). Sixteen isolates produced neither LT nor STa but did produce enterotoxin detectable in ligated intestinal loops of pigs (STb). Most of these LT(-) STa(-) STb(+) isolates were also K88(-), K99(-), and 987P(-) and non-enteropathogenic. One of them was K99(+) and enteropathogenic. Our conclusions are as follows. (i) Most enteropathogenic ETEC from neonatal pigs produce either K88, 987P, or K99; however, there are some that produce none of the three antigens. (ii) Immunofluorescence tests for pilus antigens produced in vivo are recommended for the diagnosis of ETEC infections. (iii) Reports of LT(-) STa(-) STb(+) swine ETEC are confirmed; furthermore, such isolates can be enteropathogenic.

Animals↗

Immunization of suckling pigs against enterotoxigenic Escherichia coli-induced diarrheal disease by vaccinating dams with purified K99 or 987P pili: antibody production in response to vaccination.

Pilus-specific antibody levels measured by enzyme-linked immunosorbent assays in serum and colostrum of pregnant swine (dams) were shown to increase after parenteral vaccination with pili. Pilus-specific antibody levels in dams were correlated with protection of their suckling offspring against fatal diarrhea caused by enterotoxigenic Escherichia coli possessing the same pilus as the vaccine.

Animals↗