Prevalence and relationships of sensory taint, 5α-androstenone and skatole in fat and lean tissue from the loin (Longissimus dorsi) of barrows, gilts, sows, and boars from selected abattoirs in the United States.

This study assessed prevalence of boar taint in backfat and lean of barrows, gilts, sows, and boars, collected from abattoirs, without knowledge of the farms of origin, in different regions in the United States. Concentrations of 5α-androstenone (liquid chromatography/mass spectroscopy) and skatole (liquid chromatography with fluorescent detection) in backfat were measured. A trained panel evaluated boar taint aroma in heated samples. Mean 5α-androstenone and skatole levels were low among barrows, gilts, and sows, whereas 55.8% of boars scored above a 1.0 µg/g threshold for 5α-androstenone concentrations and 34.2% were above a 0.2 µg/g threshold for skatole concentrations. Mean aroma scores for backfat and lean from barrows, gilts, and sows were low. In comparison, 59.2% of boars had elevated mean aroma scores from fat samples and 31.7% from lean. Importantly, boar taint aroma was detectable by the trained panel in at least some animals in each of the sex classes.

Subsecond adsorption and desorption of dopamine at carbon-fiber microelectrodes.

High-repetition fast-scan cyclic voltammetry and chronoamperometry were used to quantify and characterize the kinetics of dopamine and dopamine-o-quinone adsorption and desorption at carbon-fiber microelectrodes. A flow injection analysis system was used for the precise introduction and removal of a bolus of electroactive substance on a sub-second time scale to the disk-shaped surface of a microelectrode that was fabricated from a single carbon fiber (Thornel type T650 or P55). Pretreatment of the electrode surfaces consisted of soaking them in purified isopropyl alcohol for a minimum of 10 min, which resulted in S/N increasing by 200-400% for dopamine above that for those that were soaked in reagent grade solvent. Because of adsorption, high scan rates (2,000 V/s) are shown to exhibit equivalent S/N ratios as compared to slower, more traditional scan rates. In addition, the steady-state response to a concentration bolus is shown to occur more rapidly when cyclic voltammetric scans are repeated at short intervals (4 ms). The new methodologies allow for more accurate determinations of the kinetics of neurotransmitter release events (10-500 ms) in biological systems. Brain slice and in vivo experiments using T650 cylinder microelectrodes show that voltammetrically measured uptake kinetics in the caudate are faster using 2,000 V/s and 240 Hz measurements, as compared to 300 V/s and 10 Hz.

Effect of pH and surface functionalities on the cyclic voltammetric responses of carbon-fiber microelectrodes.

Carbon electrodes are useful for the detection of oxidizable species with cyclic voltammetry. In particular, carbon-fiber microelectrodes have been employed for the measurement of several neurotransmitters in brain tissue. However, during cyclic voltammetry with carbon-fiber electrodes the current varies with changes in concentration of some inorganic cations as a result of their interaction with surface functional groups. The electrode's response to the hydronium ion is a particular concern because its voltammetric response occurs over a broad range of potentials that overlap those of neurotransmitters of interest such as dopamine. This is especially a problem in vivo because simultaneous changes of dopamine and pH frequently occur in brain tissue. In this work, voltammetric current changes are shown to arise from pH dependent shifts in the peak potentials of background voltammetric waves that arise from species confined to the carbon-fiber electrode surface. Polishing the electrode with alumina suspended in cyclohexane in an environment containing lowered oxygen, a method previously demonstrated to remove oxides from the carbon surface, leads to a substantial reduction in the sensitivity to pH changes. However, this is accompanied by a loss in signal amplitude for dopamine. The dopamine response can be restored using the cation exchanger Nafion without significantly increasing the pH response. To investigate which oxide functional groups play a direct role in the electrode's current responses to changes in pH, surface-confined carbonyl and alcohol functionalities were chemically modified. In both cases, the modification did not affect the carbon-fiber electrode's responsiveness to changes in pH. Nonetheless, the polishing technique proved to be effective in reducing pH interferences in in vivo applications.

Effects of preweaning exposure to a starter diet on enterotoxigenic Escherichia coli-induced postweaning diarrhea in swine.

Experiments were conducted to evaluate the effect of restricted feeding of a starter diet to suckling pigs (creep feeding) in a model of postweaning colibacillosis. The hypothesis that restricted creep feeding primes an intestinal allergic reaction to starter diet ingested after weaning was tested. Twenty-eight suckling pigs were fed a starter diet for 3 h/d on days 7, 8, and 9 after birth (creep-fed). Twenty-six suckling pigs were not fed the diet until 3 weeks of age (not creep-fed), when all pigs were weaned and given the starter diet. One day after weaning, 24 creep-fed and 22 not creep-fed pigs were inoculated with K88+ enterotoxigenic Escherichia coli, and 4 pigs in each group were kept as noninoculated controls. Among inoculated pigs (principals), 10 creep-fed and 12 not creep-fed pigs were found to be genetically resistant to K88+ E coli and remained healthy during the 6-day postinoculation period, as did the noninoculated controls. Eighteen (10 creep-fed and 8 not creep-fed) of the 24 genetically susceptible principals developed diarrhea after inoculation. There were no significant differences in the incidence and severity of diarrhea, amount of body weight loss, and mortality between creep-fed and not creep-fed susceptible principal pigs. Histologic examination of intestine from control pigs and principals that survived for 6 days after infection did not reveal any substantial morphologic difference between creep-fed and not creep-fed groups. In conclusion, creep feeding was not required for the production of diarrhea in this model. Creep feeding did not induce morphologic changes characteristic of an allergic reaction in the small intestine.

Extravasation of lymphocytes via paracortical venules in sheep lymph nodes: visualization using an intracellular fluorescent label.

In rodents and humans, lymphocytes extravasate into lymph nodes via specialized paracortical venules lined with high endothelium (HEV). Sheep and other ruminants do not have morphologically defined HEV in their lymph nodes. It has been assumed that lymphocyte extravasation in these species proceeds via analogous structures; i.e., paracortical venules lined with low to medium endothelium. In this study, lymphocyte suspensions were prepared from surgically excised lymph nodes of sheep and labeled with an intracellular fluorescent dye, H33342. Labeled cells were infused intravenously back into donors, and sheep were killed at various intervals after infusion. Frozen sections of lymph nodes were examined microscopically for the location of labeled cells. Ten minutes after infusion, labeled cells were seen in the lumen of venules located in the paracortical region of the nodes. At later time points, cells were seen apparently migrating through the venule walls and in the adjacent paracortical tissue. Similar experiments were performed in which H33342-labeled murine lymphocytes were infused into syngeneic mice. When equivalent cell numbers (based on animal size) were infused, no obvious differences were seen between location and kinetics of appearance of labeled cells in lymph nodes of sheep compared to those of mice. These results indicate that lymphocyte extravasation in sheep proceeds via paracortical venules in lymph nodes. The function of these venules appears to be analogous to HEV in nonruminant species.

In vitro adhesion of K88ac+ Escherichia coli to Peyer's patch and peripheral blood lymphocytes, buccal and rectal epithelial cells or intestinal epithelial brush borders of weaned pigs.

Escherichia coli adhesion assays were conducted using isolated porcine peripheral blood lymphocytes, Peyer's patch lymphocytes, rectal epithelial cells or brush borders, buccal epithelial cells and brush borders from small intestinal epithelial cells. The cells and brush borders were tested for their ability to bind K88-piliated enterotoxigenic E. coli Strain M1823B (K88ac) and E. coli Strain 1476 (K-12, K88ac). Comparison of adhesive phenotypes of 37 weaned pigs as determined by the adhesion assay with small intestinal brush borders and the adherence of K88ac+ enterotoxigenic E. coli to peripheral blood lymphocytes, Peyer's patch lymphocytes and rectal epithelial cells or brush borders, revealed no correlation. In vitro adhesion of K88ac-bearing E. coli was always negative with buccal epithelial cells. K88ac strains varied in their ability to adhere to lymphocytes and rectal epithelial cells or brush borders, indicating that the mechanism of adherence is unrelated to K88-mediated adhesion observed in animals that had the receptors on small-intestinal epithelial-cell brush borders. The non-piliated control E. coli Strain 123 adhered to fresh peripheral blood lymphocytes, and less intensively to frozen-thawed peripheral blood lymphocytes or Peyer's patch lymphocytes. It was concluded that none of the cell types or brush borders, except small-intestinal epithelial-cell brush borders, could be used as targets for phenotyping pigs for the presence of the K88 receptors that have been associated with adhesion and colonization of K88+ enterotoxigenic E. coli in the porcine small intestine.

Lymphocyte recirculation in cattle: patterns of localization by mammary and mesenteric lymph node lymphocytes.

We examined patterns of lymphocyte localization in female dairy cattle following infusion of 51Cr-labeled autologous lymphocytes prepared from surgically excised mammary or ileal mesenteric lymph nodes. Labeled lymphocytes prepared from mammary lymph nodes were recovered in proportionally high numbers from mammary and prescapular lymph nodes, and in low numbers from intestinal mesenteric nodes. This pattern was observed in both heifers and lactating cows. In contrast, labeled lymphocytes prepared from ileal mesenteric lymph nodes of lactating cows were recovered in proportionally high numbers from intestinal mesenteric nodes, and in low numbers from mammary and prescapular nodes. These findings, when compared with previous results in sheep and swine, support the hypothesis that lymphocytes do not migrate efficiently between the gut and mammary gland of ruminants.

F41 pili as protective antigens of enterotoxigenic Escherichia coli that produce F41, K99, or both pilus antigens.

Pigs suckling dams that have been vaccinated with pilus antigen are protected against challenge with enterotoxigenic Escherichia coli (ETEC) strains that express the same pilus antigen. However, some ETEC strains express more than one pilus antigen. Pregnant swine were vaccinated either with E. coli HB101 that harbored a recombinant plasmid coding for F41 expression (F41+) or with the HB101 parent strain that carries the pHC79 vector (F41-). Suckling pigs born to vaccinated dams were challenged with ETEC that expressed either K99, F41, or both pilus antigens. Production of F41 in vivo was demonstrated by immunofluorescence assay of sections of ileum and by seroconversion against F41 antigen by pigs challenged with F41+ and K99+ F41+ ETEC strains. The F41+ vaccine protected against challenge with an F41+ ETEC strain. In contrast, F41+ vaccination did not protect against challenge with K99+ or K99+ F41+ ETEC strains. The F41- vaccine did not protect against challenge with any strain used. The results indicate that K99+ F41+ ETEC strains produce F41 antigen in the small intestine during disease and that F41+ vaccination can be a protective antigen if the challenge strain expresses only F41 antigen, but that F41+ vaccination may not protect against strains that produce both K99 and F41 antigens.

Experimental disease in infant goats induced by a Mycobacterium isolated from a patient with Crohn's disease. A preliminary report.

Pilot studies were done to assess the pathogenicity of a Mycobacterium which had been recovered from the diseased ileum of a patient with Crohn's disease. In four separate studies, pairs of infant goats served as subjects. One of each pair received an oral inoculum of freshly harvested Mycobacterium species strain Linda suspended in cream. A littermate or stablemate which received only cream served as control. Necropsies were done at three, five, six, and 10 months postinoculation. Each of the four inoculated animals developed segmental granulomatous disease of the ileum or ileum and more proximal segments of small intestine, and regional lymph nodes. The earliest lesion occurred in Peyer's patches of the ileum and consisted of granulomatous clusters of epithelioid cells and giant cells, without caseation, which often occurred in a mantle of lymphocytes between the germinal centers and the muscularis mucosae. Nine of 10 such granulomas were free of acid-fast bacilli. In more advanced lesions, there was confluence of granulomas and ulceration of the mucosal surface. Two of the four inoculated animals also had lymphocytic lymphangitis in affected segments. Although the Mycobacterium Linda was recovered from intestinal segments of all four animals, acid-fast bacteria were not demonstrable in the intestines in two of them. Control animals remained free of lesions and acid-fast bacilli and were negative by bacteriologic culture. The Mycobacterium species strain Linda represents an enteric pathogen capable of inducing granulomas of the distal small intestine of susceptible species. The lesions produced have distinct similarities to those occurring in Crohn's disease.

Effects of microbial and host variables on the interaction of rotavirus and Escherichia coli infections in gnotobiotic calves.

Naturally occurring mixed infections with Escherichia coli and rotavirus have been associated with fatal diarrhea of calves about 1 week old. Experiments were designed to reproduce this syndrome in gnotobiotic calves. Clinical, microbiological, and pathologic data were used to assess severity of disease and mechanisms of the interaction between the 2 infections. An initial study involved 5- to 8-day-old gnotobiotic calves inoculated with a strain of enterotoxigenic E coli (ETEC) and a strain of rotavirus. Calves were observed for 2 days after they were inoculated; fatal diarrhea was not produced. In later studies, variables were tested to identify those that might contribute to fatal diarrhea. Variables which did not result in fatal or severe diarrhea or which did not cause disease that was more severe in dually inoculated calves than that in monoinoculated calves were increasing feed to 2 times base line, increasing dose of ETEC to 10 times base line, inoculating calves when they were 2 days old, using a strain of E coli that causes colisepticemia, and using a different strain of rotavirus. When the observation period was extended from 2 days to 6 days after calves were inoculated, severe, watery, fatal diarrhea occurred in 6 of 12 calves by 32 to 72 hours after dual inoculation was given. Fatal diarrhea was associated with intensive colonization by the ETEC in the caudal half of the small intestine. Microscopic lesions were similar between dually inoculated and rotavirus-monoinoculated calves, except there was more severe atrophy of ileal villi of dually inoculated calves.(ABSTRACT TRUNCATED AT 250 WORDS)

Capsule reduces adherence of enterotoxigenic Escherichia coli to isolated intestinal epithelial cells of pigs.

Previous reports have demonstrated that heat-stable (A-type) capsule on piliated enterotoxigenic Escherichia coli enhances colonization of enterotoxigenic E. coli in the small intestine and enhances virulence of enterotoxigenic E. coli. In this report, four encapsulated enterotoxigenic E. coli strains and one encapsulated nonenterotoxigenic strain of E. coli and their nonencapsulated mutants were tested for adhesion to isolated intestinal epithelial cells or brush borders from neonatal pigs. The enterotoxigenic E. coli also expressed the K99 pilus antigen. The nonencapsulated mutants of the four enterotoxigenic E. coli adhered in higher numbers than did the encapsulated parental strains. Both the encapsulated and nonencapsulated forms of enterotoxigenic E. coli 431 grown at 18 degrees C (K99 production suppressed) adhered poorly to the isolated cells. The nonenterotoxigenic E. coli 1793 which does not express K99 antigen also adhered poorly in both encapsulated and nonencapsulated forms. Fab fragments of anticapsular immunoglobulin G failed to block the effect of capsule on adherence of strain 431. The results indicated that K99 was the principal mediator of in vitro adhesion of the enterotoxigenic E. coli strains and that capsule impedes the in vitro adhesion. They also suggested that the capsular enhancement of colonization by such strains in vivo probably is by some mechanism other than enhanced adhesion to epithelium.

Type 1 pili (F1) of porcine enterotoxigenic Escherichia coli: vaccine trial and tests for production in the small intestine during disease.

This study was performed to determine whether the F1 (type 1) pili of a porcine strain of enterotoxigenic Escherichia coli are protective antigens and whether they are produced in the pig small intestine during disease caused by an enterotoxigenic E. coli. Reciprocal cross-absorption experiments with antisera prepared against F1 pili purified from enterotoxigenic E. coli 431 (O101:K30,99:H-:F1) and P14 (O149:K91,88ac:8H+:F1) demonstrated that the F1 antigens of the two strains were closely related or identical. Pregnant swine vaccinated with a vaccine prepared from strain P14 (F1+) responded with a significant increase in antibody against F1 in their serum and colostrum. However, the vaccinated dams did not significantly protect their suckling pigs against fatal challenge with strain 431. There was no evidence of F1 pilus production in the strain 431-infected pigs, as determined by immunofluorescent staining of ileal sections, direct electron microscopic examination of bacteria from ilea, and titration of serum agglutinins in convalescent pigs. It was concluded that strain 431 did not produce F1 in the small intestine during disease and that F1 was not a protective antigen in this system.

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.

Studies with an unclassified virus isolated from diarrheic calves.

A transmissible agent (Breda agent) was isolated from a calf with diarrhea and shown to be infectious by inoculation orally into gnotobiotic and conventionally reared calves. The "Breda" agent had the morphology of a virus and possessed a hemagglutinin. Antigenic studies showed the virus to be antigenically different from bovine coronavirus, parainfluenza 3 virus, bovine rotavirus, bovine parvovirus and bovine pestivirus (BVD). Attempts to culture the virus in cell or organ cultures or in embryonated eggs, were unsuccessful. The virus was either spherical or kidney shaped, with 7-9 nm peplomers on the surface. A few particles possessed coronavirus processes of 17-20 nm, but these were arranged irregularly and were thought to be tissue debris. Three out of eight experimental calves developed severe diarrhea and the lesions in the small and large intestines were similar to those reported for coronavirus. The virus replicated in the jejunal and ileal regions of the small intestine and in the spiral colon, as judged by immunofluorescence. The virus multiplied in all experimental calves and was excreted in the feces; excretion correlating with the onset of diarrhea or a change in the appearance of the feces. There was little or no malabsorption measured by the uptake of D-xylose and the fact that infection of both the crypt and villus epithelial cells was observed, suggests that the pathogenesis may be different from rotavirus and coronavirus. Fourteen of forty seven calves in the outbreak were infected with the virus, virus was not identified in other farm outbreaks of the disease.

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.

Development of resistance with host age to adhesion of K99+ Escherichia coli to isolated intestinal epithelial cells.

When isolated intestinal epithelial cells from neonatal and older pigs, calves, and mice were tested for adhesion by K99+ enterotoxigenic Escherichia coli, cells from older animals were resistant to adhesion.