A Comparative Study of the Rabbit and Pig Gut Loop Systems for the Assay of Escherichia coli Enterotoxin

A comparison was made between segments of pig and rabbit small intestine in their response to heat-labile (LT) and heat-stable (ST) preparations from porcine enteropathogenic Escherichia coli. Either whole cell lysates or dialysed broth culture supernatants were used as sources of LT and soft agar culture fluids as a source of ST. Whole cell lysates of all thirteen LT-producing E. coli strains tested regularly elicited fluid accumulation in rabbit gut loops. Whole cell lysates of certain E. coli strains considered to be nonenteropathogenic in pigs could also elicit a positive response in rabbit gut loops. When graded doses of LT were tested in pig and rabbit gut loops, the rabbit was more sensitive and is therefore considered preferable to the pig for quantitation of LT. In the rabbit, upper (jejunal) and lower (ileal) small intestine were compared for their response to LT and it was found that ileal loops were twice as sensitive but more prone to false positive reactions. When soft agar culture fluids of several enteropathogenic E. coli strains were tested in the rabbit, the response was inconsistent, and it was concluded that the rabbit is unsuitable for the assay of the heat-stable enterotoxin.     

Limitations of the Infant Mouse Test for Escherichia coli Heat Stable Enterotoxin

A study was undertaken to evaluate the response of different test systems to preparations of heat-stable enterotoxin (ST) derived from Eschericihia coli strains recovered from diarrheal disease of humans, pigs and calves. Sterile broth culture supernatants of enterotoxigenic strains of E. coli were heated at 65°C for 30 minutes and tested for the presence of heat-stable enterotoxin. Three test systems, namely, ligated intestine of weaned pigs, ligated intestine of rabbits and the infant mouse test were used in attempts to detect ST in the culture supernatants. Two patterns of reaction were observed in response to ST-containing preparations: either the preparation elicited a response in the three tests or the preparation elicited a reaction only in the ligated pig intestine. A response in all three tests were observed for 5/5 human ST-producing E. coli, 5/5 bovine enterotoxigenic E. coli, 5/5 “atypical” porcine enterotoxigenic E. coli, 3/3 St⁺LT^- porcine E. coli of serogroup O138:K81 and 4/24 LT⁺ST⁺ porcine E. coli. A response only in the ligated pig intestine was obtained with 5/5 ST⁺LT^- porcine E. coli belonging to serogroups other than O138:K81 and to 20/24 ST⁺LT⁺ E. coli from pigs. The results are consistent with the view that there are two kinds of ST, one of which (ST1) reacts in all three tests and the other (ST2) which reacts only in the ligated pig intestine. The findings underscore the limitations of the infant mouse test as a means of detecting ST in porcine isolates of E. coli, since the test fails to detect ST produced by a large number of these E. coli strains. There appeared to be a relationship between kind(s) of ST produced and the animal species from which the producing organism was recovered.     

Adrenergic regulation of vascular smooth muscle tone in calf digital artery

Radioligand binding studies and functional assays on isolated smooth muscle preparations were performed in order to obtain a biochemical and functional characterization of the beta-adrenoceptor (beta-AR) subtypes involved in regulation of the smooth muscle relaxation of the calf's common digital artery. The results indicate that the common digital artery possesses two beta-AR populations (40% beta(1) and 60% beta(2)) and the beta(2)-subtype appears to predominate as far as function is concerned. Only the beta(2)-AR agonists clenbuterol and fenoterol caused dose-related relaxant effects, antagonized by propranolol, when tested in preparations precontracted both with PGF(2alpha) (1.4 x 10(-5) m) and noradrenaline (1.2 x 10(-6) m). In noradrenaline precontracted preparations the beta(1)-AR selective agonists dobutamine and xamoterol caused vasodilation which was not antagonized by (+/-)propranolol. While the functional relaxant effects of dobutamine may be attributed to its potent competitive alpha-AR blocking activity, further investigations are required to explain the effect of xamoterol. The vasodilator effect of (+/-)isoproterenol was irregular. The recorded contractile effects, mainly at dosages greater than 10(-6) m, suggest the loss of drug selectivity for beta-AR and alpha-AR activation. Indirect evidence indicates that the alpha-adrenoceptor (alpha-AR) population in this tissue which produces a strong contraction is functionally dominant over the beta-AR, suggesting limited therapeutic benefit for beta-AR drugs to control blood flow disorders in the calf's distal limb.     

Hemagglutinating and hydrophobic surface properties of salmonellae producing enterotoxin neutralized by cholera anti-toxin

Eleven Salmonella strains known to produce enterotoxin under aerobic culture conditions in deferrated (DF) medium at 37°C were shown to produce enterotoxin with and without aeration at 22, 28, 37 and 42°C. Heat-labile enterotoxin was generally produced with growth temperatures up to 37°C irrespective of aeration. Heat-stable enterotoxin was produced up to 42°C, mainly aerobically, as indicated by infant mouse assay (IMA), by six of the eleven strains tested. Nine strains produced heat-stable rapid permeability factor (RPF) in rabbit skin.Cholera anti-toxin neutralized reactivities of Salmonella heat-labile enterotoxin in four different biological assays. Mixed gangliosides also neutralized this activity in the cell—test systems.With guinea-pig erythrocytes, all strains underwent mannose-resistant hemagglutination (MRHA) irrespective of growth temperatures, i.e. 22 and 37°C or medium, i.e., DF, tryptose soy broth (TSB) and colonization factor antigen (CFA) agar. At both growth temperatures, CFA agar-grown cells of each strain caused MRHA of bovine erythrocytes. Excepting three Salmonella typhimurium strains, DF broth-grown cells gave MRHA of bovine, chicken and human group A erythrocytes, CFA agar-grown cells caused MRHA of chicken and human blood, whereas TSB-grown cells caused few MRHA reactions.Salmonellae producing both heat-stable, (ST) and heat-labile, (LT) enterotoxins adsorbed to Phenyl Sepharose whereas salmonellae that produced only LT enterotoxin did not.The presence of MRHA adhesions did not correlate with cell-surface hydrophobicity. However, mannose-resistant hemagglutinins may occur more commonly among salmonellae than has been previously recognized.     

Immunity to Escherichia coli in Pigs: Anti-enterotoxins in Colostrum and Serum from Vaccinated and Non-vaccinated Sows

Colostrum from non-vaccinated sows did not contain naturally occurring antibodies to heat labile Escherichia coli enterotoxin. Vaccination of sows by either the intramuscular or intramammary routes with a live formalinized E. coli vaccine resulted in the production of colostrum capable of neutralizing the heat labile toxin. Intramammary vaccination resulted in the production of colostrum which significantly reduced the enterotoxigenic effects of the vaccine strain of E. coli organisms but not that of a heterologous strain.

Vaccination of the sows resulted in the production of serum antibodies to heat labile enterotoxin. Antibodies to heat stable enterotoxin were not demonstrable in the colostrum of either non-vaccinated or vaccinated sows.

     

Virulence factors of Escherichia coli isolated from cows with acute mastitis

A total of 184 Escherichia coli isolates recovered from cows with acute mastitis were examined for recognized pathogenic mechanisms and virulence factors commonly found in pathogenic groups of E coli. A modification of the Eng procedure (for detecting complement deficiencies in serum) was used to test for resistance to different animal sera. The Sereny test (for invasiveness), infant mouse test (for heat-stable enterotoxin), and Y-1 adrenal tumor cell assay (for heat-labile enterotoxin) were used. Hemagglutination tests, using rabbit, sheep, and guinea pig RBC, were done with and without added mannose. All of the 184 isolates were serum resistant in all tested sera. None of the isolates was invasive. Only 1 isolate was positive for heat-stable enterotoxin and 2 cultures were positive for heat-labile enterotoxin. Multiple patterns of hemagglutination were observed. The majority of the isolates exhibited both mannose-sensitive and mannose-resistant hemagglutinins with guinea pig and rabbit RBC. A few strains were positive only in mannose-sensitive or mannose-resistant hemagglutination tests. A few strains were negative in all hemagglutination tests. Based on our results, E. coli from cows with acute mastitis lack the virulence factors commonly observed in other E coli groups associated with disease. Serum resistance was the only characteristic that could be related to virulence.     

Virulence profiles of enterotoxigenic, shiga toxin and enteroaggregative Escherichia coli in South African pigs

Enterotoxigenic Escherichia coli (ETEC) and shiga toxin E. coli (STEC) are important causes of colibacillosis in piglets. Recently, enteroaggregative E. coli heat-stable enterotoxin 1 (EAST-1) has been implicated in pig diarrhoea. This study investigated the prevalence of enterotoxin [heat-labile toxins (LT), heat-stable toxin a (STa), heat-stable toxin b (STb)], shiga toxins (Stx1, Stx2, Stx2e), enteroaggregative heat-stable E. coli (EAST-1), associated fimbriae (F4, F5, F6, F41, F18ab, F18ac) and non-fimbrial adhesins [adhesin involved in diffuse adherence 1 (AIDA-1), attaching and effacing factor, porcine attaching- and effacing-associated factor] in South African pigs. A total of 263 E. coli strains were isolated from Landrace (n?=?24), Large White (n?=?126), Duroc (n?=?28) and indigenous (n?=?85) breeds of piglets aged between 9 and 136 days. PCR was used in the analysis. Virulent genes were detected in 40.3 % of the isolates, of which 18.6, 0.4 and 17.5 % were classified as ETEC, STEC and enteroaggregative E. coli (EAEC), respectively. Individual genes were found in the following proportions: STb (19.01 %), LT (0.4 %), STa (3.4 %), St2xe (1.1 %) and EAST-1 (20.2 %) toxins. None of the tested fimbriae were detected in ETEC and STEC isolates. About one third of the ETEC and STEC isolates was tested negative for both fimbrial and non-fimbrial adhesins. Twenty-five pathotypes from ETEC-, EAEC- and STEC-positive strains were identified. Pathotypes EAST-1 (30.2 %), STb (13.2 %) and STb/AIDA-1 (10.4 %) were most prevalent. The study provided insight on possible causes of colibacillosis in South African pigs.     

Enterotoxigenic Escherichia coli in the jejunum of piglets with neonatal diarrhea

Thirty-two Escherichia coli colonies were taken from the primary step of cultivation of the jejunal contents of each of 10 dead piglets which had suffered from diarrhea. The organisms of each colony were examined for the presence of adhesion fimbria (F4 (K88) and F5 (K99)), production of heat-stable and heat-labile enterotoxin and of colicins.The presence of heat-labile enterotoxin in the intestinal content of the necropsied pigs was also tested, and results correlated with enterotoxin production of the isolated E. coli strains. In all but 3 pigs, 50–80 % of the E. coli strains were found to produce one or both of the enterotoxins and to possess the F4 of the F5 antigen. All bacteria producing both heat-labile and heat-stable enterotoxin proved to belong toi O group 149 and to possess the F4 antigen. Strains from 1 pig belonged to O group 64 and possessed the F5 antigen; these bacteria produced heat-stable enterotoxin only. Most of the enterotoxin-producing E. coli also produced colicins.After each subcultivation, the strains produced less heat-labile enterotoxin, some becoming negative when assayed.     

Effects of adrenergic agonists and antagonists on the blood pressure and heart rate of the pig fetus

The effects of adrenergic agonists and antagonists on blood pressure and heart rate were investigated in 18 chronically catheterised pig fetuses aged between 102 and 109 days of gestation (term is 114 days). One fetus had been decapitated in utero at 42 days of gestation. The alpha adrenergic agonist methoxamine produced a small but dose dependent hypertension and a dose related slowing in heart rate. The beta adrenergic agonist isoprenaline decreased mean arterial pressure in a dose related manner and produced tachycardia. Propranolol, a beta adrenergic blocker, increased mean arterial pressure and decreased heart rate. The response to subsequent alpha adrenergic blockade with phentolamine was hypotension and a slight bradycardia. Decapitation at 42 days of gestation did not seem to change the fetal responsiveness to adrenergic agonists but removed the blood pressure response to beta blockade. These observations indicate that the heart and circulation of the pig fetus are under adrenergic vasomotor control during late gestation.     

Autonomic innervation of the airways

Summary

A review is given of the literature concerning the autonomic innervation of airway smooth muscle. The cholinergic, adrenergic and non‐cholinergic non‐adrenergic (NANC) systems in humans and several animal species are discussed. The diagnostic and therapeutic possibilities and limitations of new receptor specific agonists and antagonists are also discussed.     

Evaluation of different cell cultures for the detection of heat‐labile enterotoxins of Escherichia coli strains isolated from pigs

The sensitivity of various cell cultures to heat‐labile enterotoxins (LT) and Verocytotoxin (VT) of fifteen E. coli strains isolated from cases of pig colibacillosis in Poland was estimated and compared with the effect of enterotoxins of four standard E. coli strains. Often tested cell cultures, only the following were susceptible: CHO, Vero, GMK, and HeLa.

Eight strains showed CTE in HeLa and CHO cells and five of these reacted in Vero cells. The results appear to suggest that some of the tested E. coli strains isolated from pigs produced VT enterotoxin. Morphological changes caused by the above mentioned E. coli toxins in Vero and GMK cells took the form of cell rounding, followed by cell dissolution.     

Enterotoxin plasmids in bovine and porcine enterotoxigenic Escherichia coli of O groups 9, 20, 64 and 101.

Fourteen strains of Escherichia coli of serogroups characteristic of porcine class 2 enterotoxigenic E. coli isolated from pigs or calves were selected for genetic studies. The strains were examined for their ability to cotransfer a number of plasmid-mediated properties during conjugation with E. coli K-12. These properties were antibiotic resistance, and the production of heat-stable enterotoxin, the K99 antigen and colicin and the ability to ferment raffinose. Distinction was made between the two types of heat-stable enterotoxin, STa and STb. All 14 strains were antibiotic resistant and 11 of them cotransferred antibiotic resistance and heat-stable enterotoxin. One strain which transferred heat-stable enterotoxin also transferred the raffinose gene. Among six K99-positive strains which transferred heat-stable enterotoxin, five always cotransferred K99. Three strains had 100% cotransfer of colicin as well as heat-stable enterotoxin and K99. Drug resistance determinants were cotransferred at high frequency with heat-stable enterotoxin for six of eight multiple drug resistant enterotoxigenic E. coli. A 100% cotransfer of combinations of heat-stable enterotoxin, K99, colicin and antibiotic resistance was often associated with a single plasmid band on agarose gel electrophoresis. For some strains, the genes for STa and STb were on the same plasmid and for others they were on separate plasmids. The enterotoxin plasmids ranged in size from 5.2 to 85 Mdal. Heterogeneity in molecular size occurred among enterotoxin plasmids in E. coli of the same serogroup and recovered from the same animal host species.     

Influence of catecholamines, prostaglandins and thyroid hormones on growth hormone secretion by chicken pituitary cells in vitro

In young chickens plasma concentrations of growth hormone (GH) are depressed by prostaglandins (PG) E1 and E2, epinephrine, norepinephrine, alpha 2 and beta agonists or thyroid hormones. A primary culture of chicken adenohypophyseal cells was used to examine the direct effects of these agents at the level of the pituitary as evaluated by GH release in the presence and absence of growth hormone releasing factor (GRF). Following collagenase dispersion and culture (preincubation, 48 hr) cells were exposed (incubation, 2 hr) to test agents, except for thyroid hormones which were added during the preincubation, and incubation period. Growth hormone release was increased (P less than .05) in the presence of PGE1 (10(-8)M by 34%; 10(-7)M by 54%), PGE2 (10(-8)M by 29%; 10(-7)M by 29%), PGF2 alpha (10(-8)M by 28%), and the beta agonist isoproterenol (10(-7)M by 46%). Basal GH release from chicken pituitary cells was not affected by dopamine, norepinephrine, epinephrine, thyroxine (T4), triiodothyronine (T3), or alpha adrenergic agonists. Growth hormone releasing factor stimulated GH release was not affected by the presence of prostaglandins E1, E2 or F2 alpha in the incubation media. However, GRF stimulated GH release was reduced by high doses of catecholamines: dopamine (10(-6)M by 34%), norepinephrine (10(-6)M by 74%), epinephrine (10(-8)M by 47%; 10(-7)M by 41%; 10(-6)M by 89%), and by the alpha 1 adrenergic agonist, phenylephrine (10(-7)M by 52%), the alpha 2 agonist, clonidine (10(-8)M by 34%; 10(-7)M by 83%) and the beta agonist, isoproterenol (10(-7)M by 64%).(ABSTRACT TRUNCATED AT 250 WORDS)     

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.

     

Enterotoxigenicity,hemagglutination and cell-surface hydrophobicity in Aeromonas hydrophila,A. Sobria and A. Salmonicida

Thirty-one Aeromonas hydrophila, 13 A. sobria and two A. salmonicida strains of diverse sources were tested for enterotoxigenicity, hemagglutination and cell surface hydrophobicity. Although 93% of the culture supernatant fluids of the Aeromonas strains exhibited cytotoxic effects on Y1 adrenal and Chinese hamster ovary (CHO) cells, typical rounding of Y1 adrenal cells was reproducibly observed before cytotoxicity for 80% of the isolates within 1 h of exposure.Twenty-eight strains were positive for delayed permeability factor (DPF) activity in rabbit skin. Culture filtrates of 16 of 20 strains that were positive both in the Y1 adrenal cell test and for DPF activity elicited fluid accumulation in rabbit ileal loops. The DPF and ileal loop activities were neutralizable by cholera antitoxin. All, except two strains each of A. sobria and A. hydrophila, produced a heat-stable, rapid permeability factor (RPF) detected in rabbit skin. Heat-treated culture supernatant fluids of two A. hydrophila and one A. sobria isolate gave positive responses in the infant mouse assay. Nine other strains gave borderline reactions.When A. hydrophila and A. sobria isolates were grown in broth, approximately 90% agglutinated bovine, chicken, human group A and guinea-pig erythrocytes in the presence of mannose at 4°C and/or 20°C. The two A. salmonicida isolates produced mannose resistant hemagglutination (MRHA) of these four blood types.Hydrophobic interaction chromatography indicated adhesive potential in 61% A. hydrophila and 100% A. sobria strains expressing weak to strong hydrophobic cell surface properties. The results of these investigations strongly imply that the Aeromonas strains produce a cytotonic enterotoxin immunologically related to cholera toxin. Adhesive characteristics were commonly found in both clinical and routine isolates.     

Affinity of isoxsuprine for adrenoreceptors in equine digital artery and implications for vasodilatory action

We used isolated equine digital arteries to study the vasodilatory mechanism of isoxsuprine, and fowl caecum preparations to investigate the affinity of the drug for beta-adrenoceptors. Isoxsuprine is a potent vasodilator of arterial smooth muscle that has been precontracted by an alpha-adrenoceptor agonist such as noradrenaline (log EC50 = -6.33 [-5.98; -6.68]). The present study indicates that its effect is due to alpha-adrenoceptor blockade since: (1) after a long lasting exposure to cumulative doses of isoxsuprine the vasoconstricting action of noradrenaline cannot be restored; (2) isoxsuprine does not promote relaxation on preparations precontracted by PGF2alpha; (3) isoxsuprine shifts the dose-response curve of noradrenaline to the right; and (4) its affinity (pK(B) = 6.90 [6.60; 7.20]) in this experiment is comparable to that in noradrenaline-precontracted preparations and is 14 times lower than that of the selective alpha1-adrenergic antagonist prazosin [pK(B) = 8.04 (7.40; 8.68]). The affinity of isoxsuprine for beta-adrenoceptors was 100 times lower than that of isoprenaline when tested on fowl caecum. This preparation has a large beta-adrenoceptor and negligible alpha-adrenoceptor population concerned with the control of smooth muscle motility. Our data suggest that the alpha-mediated effect of isoxsuprine on horse arterial smooth muscle is due to higher affinity of the drug for alpha- than beta-adrenoceptors rather than low concentration or functionality of beta-sites at this site. According to these data, pure beta2-agonists seem to be more profitable tools to determine vasodilation of the arterial bed in horses legs.     

Detection of Escherichia coli O157 and Escherichia coli O157:H7 by the immunomagnetic separation technique and stx1 and stx2 genes by multiplex PCR in slaughtered cattle in Samsun Province, Turkey

This study was conducted to investigate the presence of Escherichia (E.) coli O157 and E. coli O157:H7 and stx1 and stx2 genes on cattle carcasses and in rectal samples collected from Samsun Province of Turkey. A total of 200 samples collected from cattle carcasses and the rectal contents of 100 slaughtered cattle from two commercial abattoirs were tested using the immunomagnetic separation technique and multiplex PCR methods. E. coli O157 and E. coli O157:H7 were detected in 52 of the 200 samples (26%) tested. Of the positive samples, 49 were E. coli O157 and three were E. coli O157:H7. The E. coli O157 strain was isolated from 24 carcasses and 25 rectal samples, while E. coli O157:H7 was isolated from two carcasses and one rectal sample. Of the 49 samples positive for E. coli O157, 32 were from the rectal and carcass samples of the same animal, while two E. coli O157:H7 isolates were obtained from rectal swabs and carcasses of the same animal. The stx1 and stx2 genes were both detected in 35 E. coli O157 isolates and one E. coli O157:H7 isolate, but the stx2 gene was only detected alone in two E. coli O157 isolates. Overall, 16 carcasses tested positive for E. coli O157 and one carcass tested positive for E. coli O157:H7 based on both carcass and rectal samples. Overall, the results of this study indicate that cattle carcasses pose a potential risk to human health due to contamination by E. coli O157 and E. coli O157:H7 in the feces.     

Observational study on antimicrobial resistance in Escherichia coli and Salmonella isolates from Ontario calf samples submitted to a diagnostic laboratory from 2007 to 2020

The objectives of this study were to i) describe Escherichia coli and Salmonella isolates; ii) investigate the temporal trends in antimicrobial resistance (AMR) profiles; and iii) evaluate the impact of season and age on these AMR profiles from diagnostic and post-mortem samples in Ontario calves ≤ 2-months-old submitted from 2007 to 2020 to the Animal Health Laboratory in Guelph, Ontario, Canada. Antimicrobial susceptibility testing results were measured by the Kirby-Bauer disk diffusion method. A total of 1291 isolates with AMR profiles were obtained from calves, with E. coli (n = 434) and Salmonella (n = 378) being the most common bacteria characterized for AMR. For E. coli, 79% of isolates tested showed a positive result in F5/K99, whereas for Salmonella isolates, S. Typhimurium (33%) and S. Dublin (22%) were the 2 most common serotypes identified. Multivariable logistic regression models were built to evaluate AMR profiles for E. coli (n = 414) and Salmonella (n = 357) to each antimicrobial tested. Most E. coli isolates (91%) and Salmonella isolates (97%) were resistant to at least one of the antimicrobials tested. In general, E. coli and Salmonella had higher odds of resistance in calves aged ≥ 2 wk compared to 1-week-old calves, and little difference was seen in the level of resistance over the years observed or between seasons in most of the antimicrobials tested. Prospective research should investigate potential risk factors for the development of AMR in calves examples being antimicrobial use and farm management practices.     

Detection of the enteroaggregative Escherichia coli heat-stable enterotoxin 1 (EAST1) gene and its relationship with fimbrial and enterotoxin markers in E. coli isolates from pigs with diarrhoea

The presence of the astA gene responsible for production of enteroaggregative Escherichia coli heat-stable enterotoxin 1 (EAST1) was examined in E. coli strains isolated from pigs with postweaning diarrhoea. Two hundred and seven isolates were tested using PCR for the astA marker and for heat-labile I (LTI), heat-stable I (STI), and heat-stable II (STII) enterotoxin genes. Moreover, the isolates were also analysed for their serotypes (O and K antigens) as well as for fimbrial adhesins using agglutination methods. It was shown that 96 (46.4%) of the isolates possessed the astA genetic determinant. The most common EAST1-positive E. coli serotype was O149:K91 and these strains were mostly LTI/STII-positive. A close correlation between the presence of F4 fimbriae and the EAST1 gene was also observed: 88 of 96 (91.7%) astA(+) isolates tested possessed the F4 antigen. Thus, EAST1 enterotoxin may represent an additional virulence determinant playing a role in the pathogenesis of porcine colibacillosis.     

Comparative prevalence of four enterotoxin genes among Escherichia coli isolated from swine

Presence of Escherichia coli enterotoxin genes LT (heat-labile enterotoxin), STaP (heat-stable enterotoxin a, porcine genotype), STaH (heat-stable enterotoxin a, human genotype), and STb (heat-stable enterotoxin b) among 874 swine isolates of E coli was determined, using DNA probes and the DNA colony hybridization technique. Of the 874 isolates evaluated, 45% hybridized with at least one of the enterotoxin gene probes and were designated as enterotoxigenic E coli (ETEC). Eighty-five percent of the ETEC were from pigs with enteric colibacillosis. The remaining 15% were from pigs with edema disease or various other diseases, and from healthy swine. Seventy-four percent of the ETEC hybridized with the STb probe, 52% with STaP, and 31% with LT; ETEC did not hybridize with the STaH probe. Most of the ETEC hybridized with more than one enterotoxin gene probe. Isolates that hybridized with the LT probe also hybridized with STb. The most prevalent gene combination was LT-STb. However, 35% of the ETEC from neonatal (less than or equal to 1 week old) swine with enteric colibacillosis were of the STaP-only genotype, and 33% of the ETEC from older swine with enteric colibacillosis were of the STb-only genotype.