Response of Gnotobiotic Pigs to Escherichia coli

In a study of the response of gnotobiotic pigs to coliform infections, 45 one-week-old germfree pigs were divided into five groups and each group was inoculated orally with a different strain of Escherichia coli. Three of these were enteropathogenic swine strains, P307[08:K87(B), K88 a,b (L):H19]; P570 [0138:K81]; P568[0141:K85a,b(B), K88a,b(L):H4], one was a virulent human strain, H224, [026:K60(B6)], and one was a non-enteropathogenic swine strain, P581[OX13:K68]. It was attempted to protect a portion of the pigs with orally administered specific antisera and sera from non-immunized specific pathogenfree (SPF) pigs. Observations were made on the clinical response, bacterial counts of feces and intestinal contents, gross pathological changes, distribution of the organisms in organs and serum hemagglutinin titers.

Infection with E. coli P307 resulted in diarrhea, dehydration and death, unless the pig was protected with specific antiserum. The pigs infected with E. coli P570 had a transient diarrhea but retained their appetites and recovered. Those infected with the other three strains remained healthy throughout. No circulating hemagglutinating antibody against the test strains of E. coli could be detected in any of the pigs seven days or earlier post-inoculation.

Relationship could not be established between the numbers of viable E. coli in the feces and the presence of clinical colibacillosis. Orally administered specific antiserum afforded protection against strain P307, but did not reduce the number of E. coli in the gut or alter their distribution in the internal organs. This suggested that the protective effect of specific antibody in the intestine was due to its action on a metabolite (enterotoxin) produced by E. coli P307 rather than the organism itself.

     

Studies of Escherichia coli in Gnotobiotic Pigs: I. Experimental Reproduction of Colibacillosis*

The successful experimental production of colibacillosis in young gnotobiotic pigs by the oral administration of E. COLI (08:K.:H21 was accomplished. Colibacillosis was not produced in gnotobiotic pigs infected with either of the 2 other strains of E.COLI employed.

Necropsy observations were presented and discussed. Emphasis was placed on the absence of grossly visible enteritis in pigs that died in the acute phase of the disease. The gross appearance of the intestinal tract changed some hours after death to resemble that associated with enteritis.

Bacteremia was detected in the gnotobiotic pigs orally infected with each of the 3 strains of E. COLI, and the special characteristics of the colostrum-deprived pig were stressed as important factors in these findings.

     

Polyerositis and Arthritis Due to Escherichia coli in Gnotobiotic Pigs

Forty gnotobiotic pigs from six litters were exposed orally to Escherichia coli 083:K·:NM at 69 to 148 hours of age, while 17 pigs from the same litters served as unexposed controls. Clinical signs of infection included fever, anorexia, diarrhea, lameness, and reluctance to move.

Eighty-four percent of the exposed pigs in four litters died, while only 13% in two litters died. Gross and microscopic lesions included serofibrinous to fibrinopurulent polyserositis in 96% of the exposed pigs in four litters and 33% of the exposed pigs in two litters. A few pigs had gross and/or microscopic lesions of arthritis. Escherichia coli was routinely isolated from the serous and synovial cavities of infected pigs.

Anti-hog cholera serum administered orally as a colostrum substitute gave partial protection against E. coli infection.

     

Studies of Escherichia coli in Gnotobiotic Pigs: III. Evaluation of Orally Administered Specific Antisera*

The protective effect of orally administered immune (E. COLI 08:K.:H21) serum was demonstrated in experimentally infected gnotobiotic pigs. The temporary protective effect of the immune serum and the correlation of protection with IN VITRO antibody determinations were discussed. The results indicated that the protective action of the immune serum is apparently not dependent upon the complement-antibody bactericidal system. It was suggested that the most plausible mechanism of action of the immune serum was the inactivation of endotoxin in the lumen of the intestine.     

Studies of Escherichia Coli in Gnotobiotic Pigs: II. The Immune Response*

The immune response of orally infected and parenterally vaccinated (E. COLI 08:K.:H21) gnotobiotic pigs was studied by bactericidal and indirect hemagglutination test methods. The bacterial agglutination test proved to be of no value in this study.

The results demonstrated that antibody was detectable within 8 days following oral infection, but the titers remained very low until after the pigs were vaccinated intravenously. The titers of the sera were markedly increased following intravenous vaccination.

There was no detectable absorption of antibody from the gut of gnotobiotic pigs fed immune serum at 4 to 6 days of age and no detectable difference between the immune response of these pigs and those not fed serum.

     

Studies of Escherichia Coli in Gnotobiotic Pigs: IV. Comparison of Enteropathogenic and Nonenteropathogenic Strains

Four gnotobiotic pigs were infected with an enteropathogenic strain of Escherichia coli, and 4 were infected with a nonenteropathogenic strain of E. coli. Pigs killed in pairs at 6, 12, 24, and 48 hours PI. Four pigs were maintained as germfree controls. The discussions were based on the results of 1) clinical observations, 2) necropsy observations, 3) counts of viable E. coli in segments of the small intestine, 4) attempts to isolate E. coli from the heart, liver, and bile, 5) microscopic examination of fixed intestinal sections to determine the location of E. coli and morphologic evidence of the host response, and (6) determinations of the pH of the contents of the various portions of the gastrointestinal tract.

No diarrhea, fluid accumulation, or impairment of the digestive capacity were noted in the pigs infected with the nonenteropathogenic strain of E. coli. The number of viable E. coli detected in the respective segments of the homogenized small intestine was similar in pigs infected with either strain.

Diarrhea occurred continuously starting 18 hours PI in the pigs infected with the enteropathogenic strain and killed 24 or 48 hours PI. The pH of the contents of the cecum and colon became markedly more alkaline simultaneously with the increase in the heterogeneity and fluid content of the cecum and colon and thus appeared to correlate well with the onset of the clinical diarrhea. No enteritis was detected grossly or microscopically.

The characteristics that determine the enteropathogenicity of a strain of E. coli could not be defined from the results, but it was noted that the host response appeared to be quite similar to that of infant rabbits experimentally infected with Vibrio cholera.

     

The Significance of Proliferation and Enterotoxin Production by Escherichia coli in the Intestine of Gnotobiotic Pigs

The significance of enterotoxin production and proliferative ability of Escherichia coli in the intestinal tract as related to porcine enteric colibacillosis was studied in 68 gnotobiotic pigs.

The animals were monocontaminated at seven to ten days of age with eight selected strains of E. coli. The strains were two naturally occurring porcine enteropathogens — P155 (0149:K91;K88a,c:H10) and P307 (08:K87;K88a,b:H19), two nonenteropathogenic strains — P104 (0139:K82:H1) and F11 (018-ab:K?:H14), and four enterotoxigenic derivatives of the above strains — P104(P155), P104(P307), F11(P155) and F11(P307). The response of the animals was evaluated on the basis of clinical observations and necropsy lesions 22 hours after exposure to the organisms. E.coli counts were determined at seven different levels of the intestinal tract. Cell free extracts of the intestinal contents were examined for enterotoxic activity by the ligated pig intestine loop test.

All of the strains possessing the enterotoxin plasmid produced enterotoxin in the pig's intestine and were capable of causing diarrhea. The nonenteropathogenic E. coli failed to do so. The strains possessing the P155 enterotoxin plasmid were more virulent than the corresponding derivatives with the P307 enterotoxin plasmid. Strains P155, P307 and P104(P155) proliferated in the upper small intestine at a greater rate and were more virulent than the other strains. The numbers attained in the upper small intestine by the other enterotoxigenic derivatives were comparable to those of their nonenteropathogenic parent strains.

It was considered that enterotoxin produced by E. coli was the essential factor for causing a diarrheic response in gnotobiotic pigs. The virulence of each of the tested strains appeared to be governed by the degree of enterotoxicity associated with a particular enterotoxin plasmid, the numbers attained by these organisms in the upper small intestine, (but not in the lower small intestine or in the colon), and by other undetermined factors.

     

Experimental Colibacillosis in Gnotobiotic Baby Pigs II. Pathology

Sixty-two neonatal gnotobiotic pigs were used in three experiments to determine the lesions produced by two closely related strains of Escherichia coli O138:K81:NM (of Michigan origin) and O138:K81 (of Minnesota origin). Exposure was by subcutaneous injection of bacterial culture into the umbilical stump or by oral inoculation.

Gross signs common to monocontaminated pigs included distention of the flaccid small and large intestines with fluid contents. Edema was prominent in various tissues of most pigs exposed via the umbilical stump but not in those exposed orally.

Histological lesions were predominantly in the gastrointestinal tract and were variable. At one extreme acute hemorrhagic enteritis was present in two pigs, while at the other extreme in a few pigs it was difficult to distinguish tissues of infected pigs from those of noninfected germfree pigs. Significant histological lesions common to monocontaminates included mild inflammatory reaction, hydropic degeneration of the intestinal epithelium, evidence of interference with normal function of the villus-draining mechanisms, and vascular changes generally indicated by edema.

The findings suggest that interference with normal absorption of nutrients plays at least some role in the pathogenesis of colibacillosis in young gnotobiotic pigs.

     

Studies of Escherichia Coli in Gnotobiotic Pigs: V. Evaluation of the Effects of Orål and Parenteral Administration of Immune Serum

The effects of orally and/or parenterally administered immune serums were evaluated in 36 gnotobiotic pigs infected with an enteropathogenic strain of Escherichia coli. Pigs were euthanatized at predetermined time intervals between 6 and 48 hours prostinfection. The results were evaluated on the basis of: 1) clinical observations, 2) necropsy observations, 3) counts of viable E. coli in segments of the small intestine, 4) attempts to isolate E. coli from the heart, liver, and bile, 5) microscopic examination of fixed intestinal sections to determine the location of E. coli and morphologic evidence of the host response, and 6) determination of the pH of the various portions portions of the gastrointestinal tract.     

Duration of Mycoplasma hyopneumoniae Infection in Gnotobiotic Pigs

Sixteen gnotobiotic pigs raised in flexible plastic isolators (four pigs per isolator) were inoculated with a culture of Mycoplasma hyopneumoniae. One pig was killed and underwent necropsy at weekly intervals for the following 16 weeks. Macroscopic lesions were observed in the lungs of 13 of 16 pigs and microscopic lesions were found in 14 of 16 pigs. Mycoplasma hyopneumoniae was cultured from the trachea or lungs from 10 of the 16 pigs. Scanning electron microscope studies showed areas of damage to the cilia, collections, of leucocytes and mucus, and mycoplasma in the trachea as well as the bronchi. These conditions were found in all the pigs seen at necropsy from nine to 16 weeks postinoculation and there was no evidence of noticeable regression or recovery during this 16 week period.     

载铜蒙脱石对大肠杆菌和仔猪腹泻的影响

对通过阳离子交换反应合成的载铜蒙脱石 (MMT- Cu) ,应用 XRD衍射分析显示 ,载铜反应前后 ,蒙脱石的(0 0 1)面网间距从原先的 1.5 4 4 nm增加到了 1.5 88nm ,表明铜是以水合阳离子或复合阳离子形式进入蒙脱石晶格层间位置。体外试验表明 ,MMT- Cu对大肠杆菌 K88具有较强的抗菌能力。MMT- Cu对仔猪的试验表明 :与对照组相比 ,日粮中添加 0 .2 % MMT- Cu使仔猪日增重提高了 13.89% (P<0 .0 1) ,料重比下降了 8.38% (P<0 .0 5 ) ,平均腹泻率降低了 71.80 % (P<0 .0 1) ,结肠内容物中大肠杆菌数显著下降 (P<0 .0 1) ,空肠绒毛高度及绒毛高度与腺窝深度的比率分别提高了 19.2 1% (P<0 .0 5 )和 37.10 % (P<0 .0 1)。     

Experimental Colibacillosis in Gnotobiotic Baby Pigs I. Microbiological and Clinical Aspects

Sixty-two gnotobiotic pigs were used in three experiments to determine the means whereby two related strains of Escherichia coli colonized the intestinal tract. Pigs were exposed by a method simulating neonatal contamination of the umbilical stump. Bacteremia was produced within one and one half hours, and by 24 hours the infection was generally well established in the gastrointestinal tract. By 48 hours after exposure, the bacteremia had subsided so that only an occasional isolation from organs other than the gastrointestinal tract was made.

Oral exposure of one litter of germfree pigs produced heavy colonization of the entire gastrointestinal tract within four hours. Evidence of intermittent bacteremia was present in pigs of this litter. Diarrhea appeared earlier when pigs were exposed orally than when they were exposed by way of the umbilical stump.

     

沼液喂猪效果分析

先后用21头大约克×沙乌头和27头大约克×梅山猪作了两次在日粮中添加沼液的试验。试验Ⅱ因雨水冲淡了沼液而收效不明显。在试验Ⅰ中,添加15—30％沼液组和添加20％—40％沼液组的平均日增重分别比对照组高出10.99％和13.26％;每kg增重消耗饲料量分别比对照组减少7.36％和10.34％;经济收益分别比对照组多17.22元和23.66元。试验猪贪睡少动、皮肤红润、被毛光亮。试验证明,沼液是一种优良添加剂,用来喂猪,不仅提高了养猪的经济效益,而且促进了生态系统内的物质循环。