Colonisation phenotype and colonisation potential differences in Campylobacter jejuni strains in chickens before and after passage in vivo

Campylobacter jejuni strains isolated from chicken faeces and from humans suffering from gastroenteritis were used to determine the colonisation phenotypes and colonisation potential of these strains in chickens. Five different colonisation types were observed ranging from immediate and sustained colonisation to completely non-colonising. Phenotype one showed immediate colonisation with prolonged excretion of viable C. jejuni bacteria. Phenotype two showed delayed colonisation with prolonged excretion of viable C. jejuni bacteria. Phenotype three showed immediate colonisation and delayed clearing of viable C. jejuni bacteria. Phenotype four showed delayed colonisation and delayed clearing of viable C. jejuni bacteria. Strains of phenotype five could not colonise chickens. Inoculum levels to obtain maximum caecal colonisation for each phenotype for strains cultured in vitro and in vivo was also determined. Following passage in vivo through a chicken, the minimum inoculum required for sustained colonisation dropped approximately 1000-fold, however, the colonisation phenotype remained unchanged before and after a passage in vivo.     

Enteric bacteria: Friend or foe?

The normal gastrointestinal tract contains an enormous number of aerobic and anaerobic bacteria which normally enjoy a symbiotic relationship with the host but can have adverse effects with local and systemic consequences. The small intestine constitutes a zone of transition between the sparsely populated stomach and the luxuriant bacterial flora of the colon. Regulation of the intestinal flora depends on complex interactions between many factors including secretion of gastric acid, intestinal motility, biliary and pancreatic secretions, local immunity, the surface glycocalyx and mucus layer, and diet. Microbial interactions are also important, and can involve alterations in redox potential, substrate depletion and production of substances such as bacteri-ocins that inhibit bacterial growth. The beneficial effect of the normal enteric flora include the competitive exclusion of potentially pathogenic organisms, and the production of nutrients such as short-chain fatty acids (which represent an important energy source for the colonic mucosa) and vitamins. Detrimental effects of the enteric flora include competition for calories and essential nutrients, particularly by bacteria located in the small intestine, and a capacity to damage the mucosa, in some circumstances causing or contributing to inflammatory bowel disease. These problems can be accentuated by interference with the physiological regulation of intraluminal bacteria allowing overgrowth by a normal resident, or colonisation by transient pathogens. The pathophysiological consequences may involve direct damage to the intestinal mucosa, and bacterial metabolism of intraluminal constituents, for example forming deconjugated bile acids and hydroxylated fatty acids which stimulate fluid secretion. Additional problems arise if there is interference with the mucosal barrier since this can result in increased passage of bacteria and bacterial products stimulating mucosal inflammation, while bacterial translocation can result in bacteraemia and septicaeniia. Problems associated with bacterial pathogens are illustrated by the properties of the spectrum of pathogenic Escherichia coli, some of which facilitate long-term colonisation by adherence to the surface or invasion of enterocytes.     

Dietary carbohydrates affect caecal fermentation and modify nitrogen excretion patterns in rats. I. Studies with protein-free diets.

In a two-factorial experiment on 96 young male rats, the effects of substituting 10% raw potato starch (PS), pectins (PEC), or cellulose (CEL) for corn starch (CS) were studied using an unsupplemented protein-free (PF) diet or a PF diet supplemented either with DL-methionine or urea. The pH and the short chain fatty acids (SCFA) content in caecal digesta, as well as caecal digesta and tissue weights were determined and used as the criteria of caecal fermentation intensity. Blood urea level, amount of N excreted via faeces and urine, DAPA content, and amino acid composition of faecal protein were analyzed as indices of protein metabolism. A 10-day adaptation period to the carbohydrates fed with the casein diet preceded the experimental period of feeding the respective carbohydrates with protein-free diets. Dietary carbohydrates significantly influenced total and individual SCFA content in caecal digesta, as well as other parameters related to the intensity of fermentation. Potato starch and pectins were more intensively fermented than cellulose. Supplementation of the PF diet with methionine and urea affected only caecal isobutyric and valeric acid content in a way dependent on the carbohydrates present in the diet. Carbohydrates significantly altered the routes of N excretion. Faecal excretion was increased by all carbohydrates studied compared to corn starch, pectins had the most marked effect. Urinary excretion was significantly increased by cellulose (as compared with the PEC and PS groups) and decreased by pectins as compared with all other groups. There was an interaction between the effects of carbohydrates and type of protein-free diet on faecal and urinary excretion. The sum of amino acids in faecal protein was the lowest on the PEC diet, but the amino acid composition expressed as a per cent of total amino acid content was similar in all groups. It can be concluded that dietary carbohydrates alter the excretion patterns of endogenous nitrogen in rats in different ways and that this effect is related to the intensity of their fermentation in the hind gut.     

Reflux of bile acids from the duodenum into the rumen of cows with a reduced intestinal passage

The purpose of this study was to find out whether the concentration of bile acids in the rumen of healthy cows differs from that of cows with a reduced gastrointestinal passage. The investigations were performed on 277 heifers and cows of the Swiss Braunvieh, Simmental and Swiss Holstein breeds. Forty of these had traumatic reticuloperitonitis, 30 had right or left abomasal displacement, 14 had an ileus of the duodenum, 36 had an ileus of the jejunum and 54 had a caecal dilatation. The other 103 animals had disorders which did not affect the digestive system and were used as controls. The mean (+/- se) bile acid concentration in the rumen fluid of the control animals was 1.0 +/- 0.30 mumol/litre. The bile acid concentrations in the rumen fluid of the cows with a duodenal or jejunal ileus were significantly higher than those of the other groups, demonstrating that bile acids may reflux from the duodenum into the rumen of cows with an ileus in the small intestine.     

Effect of thermal treatment of potato products on nutrient decomposition in the digestive tract of swine. 1. Passage and digestibility of nutrients in the various portions of the intestine

In addition to a basic ration (barley, dried green fodder, fishmeal) pigs (castrated males, 100 ... 130 kg live weight) received a supplement of raw potato starch, raw ensiled potatoes, cooked potato starch or steamed ensiled potatoes; nutrient and amino acid passage and digestibility (absorption) were measured in the small intestine, the upper large intestine and the total digestive tract with the help of simple cannulae at the distal ileum and the middle of the hind gut and the estimation of the HCl insoluble ash as an indicator. In addition, the amounts of bacteria in faeces were determined and the influence of raw and cooked potato starch on N retention was ascertained. When raw potato products were fed, distinctly higher amounts of dry matter, organic matter and starch pass the ileum, which get into the colon undigested, i.e. the digestibility of these substances is low at the end of the ileum (raw potato starch 24%, starch of raw ensiled potatoes 36%). Up to the middle of the colon there is intensive bacterial starch degradation so that in this place the difference between raw and thermically treated potatoes is only small. No matter what the previous treatment of the potatoes was, there is hardly any starch in faeces, i.e. it was nearly 100% digested up to the end of the digestive tract. The passage and digestibility resp. of ash and crude cellulose from the basic ration is not influenced by the supplement of raw or thermically treated potato products in the individual sections of the digestive tract. At the end of the ileum there are no differences between the apparent digestibility of the crude protein and absorption of amino acids of supplemented raw or cooked potato starch or steamed ensiled potatoes; after the use of raw ensiled potatoes they are, however, significantly more reduced than the comparison of analyses of faeces shows. In the large intestine the high amount of potato starch stimulates the metabolism and the reproduction of faecal bacteria as is shown by an increase by 20-30% in bacteria N excretion in faeces. Apparent crude protein digestibility and amino acid absorption are diminished by that. The increased N excretion in faeces after the supplement of raw potato starch to the basic ration is-in comparison with the supplement of heated starch-compensated by a commensurately lower N excretion in urine so that N retention practically remains uninfluenced by the supplement of variously treated potato starch.(ABSTRACT TRUNCATED AT 400 WORDS)     

Observations on avian intestinal spirochaetosis

Three SPF-laying hens were inoculated into the crop with avian intestinal spirochaetes which previously had been passaged in broiler chicks by oral inoculation (isolate 1380). Mild persisting gastrointestinal disorder developed; at nine months post inoculation spirochaetes were readily demonstrated in caecal faeces. Histologic examination of the caecal mucosa revealed many spirochaetes covering the mucosal surface and filling up the crypts lumina. Spirochaetes were found in intra- and subepithelial locations and in gaps running through the epithelium. These gaps often opened into subepithelial cavities crowded with spirochaetes ('gaplike lesions'). These lesions were seen mostly on the tips of the villi and in the deeper parts of the crypts. Massive erosion or desquamation of epithelium heavily infested by spirochaetes occurred. These findings indicate colonisation of the mucosal surface and of the crypts, penetration of the mucosa and colonisation of subepithelial compartments with spirochasetes in poultry suffering from intestinal spirochaetosis.     

Lowering of the incidence of puerperal mastitis in the sow by protection of the mammae from contamination

Twelve sows each farrowed in an experimental pen designed especially for this study or in a crate. Viable counts of enterobacteriaceae were performed in samples taken from the laying area and from the teats. Secretion from every mammary complex was examined repeatedly for bacteria and for somatic cells. The sows in the experimental pen did not lay down in their own faeces. The viable counts in samples from the laying area and the teats were much lower than with the sows kept in farrowing crates. Infection with E. coli was observed in 3 mammary complexes of the sows in the experimental pen as compared to 27 complexes of the sows in the crate. More than half of the infections was detected in the samples taken before farrowing began. In the average the bacteria persisted for 1.3 days. On the first 4 days of life piglets sucking teats with cytologically defined mastitis had an average daily gain of 105 g as compared to 125 g with piglets sucking healthy teats. In conclusion puerperal mastitis is a consequence of faecal contamination of the mammary gland. Soiling of the laying area with faeces and urine can be reduced by improvements in the farrowing accommodations.     

Nitrogen metabolism in the large intestine of ruminants. 7. Metabolism of intracecally-infused 15N urea with supplement of starch in bulls

Three bulls with an average live weight of 228 kg were fitted with ileo-caecal reentrant cannulas for the experiment. The rations were composed of 3 kg maize silage and 3 kg wheat straw pellets per animal and day. In a previous period 50% of the digesta was collected over 12 hours and stored deep-frozen. In the main period the digesta flow was interrupted for 30 hours. The digesta flow was collected quantitatively. In the caecal part of the re-entrant cannula previously collected digesta and starch (over 30 hours) as well as 15N urea (over 24 hours) were supplemented. The amount of starch corresponded to about 10% of the DM of the digesta. Analyses of the urine, faeces, ileum digesta and blood plasma were carried out. The quota of starch clearly stimulates bacterial processing in the large intestine so that 20.5% of the supplemented 15N was excreted in faeces within 24 hours. 91.2% of the 15N in the faeces was localised in the bacteria fraction. Individual differences of the animals distinctly show the connection between the excretion of the 15N in faeces and urine. A decreased isotope excretion in faeces of 17.2% for animal 3 in contrast to the 23% for animals 1 and 2 showed an increased elimination of the 15N through the kidney with 32.7% instead of 25.2%. The largest proportion of the ileum digesta, i.e. 46%, can be localised in the 15N urea fraction; the NH3-fraction is also distinctly labelled. With time progressing, the 15N quota flowing from the rumen to the small intestine increases.     

Quantitative observations on experimental reo-like virus (rotavirus) infection in colostrum-deprived calves.

Ten calves were challenged with one of two strains of reo-like virus (rotavirus). Changes in the daily faecal and urinary outputs were monitored and packed cell volume, plasma sodium, potassium and urea levels were measured. Faeces were examined for the presence of rotavirus by direct electron microscopy and immunofluorescence in cultures of PK(15) cells. All calves excreted rotavirus in the faeces for several days. Two calves remained clinically normal throughout the experiment, but in the remaining calves, faeces became mucoid in consistency and yellow-white in colour. In only two calves did the daily faecal output exceed 500 g with a fall in the dry matter content to less than 10 per cent. Slightly elevated blood urea levels and hyperkalaemia were the only changes observed in blood chemistry and these quickly returned to normal. Virus antigen was observed in the epithelial cells by immunofluorescence in the proximal and middle small intestine of calves. Pathological lesions occurred predominantly in the proximal small intestine of nine calves examined.     

Absorption and use of amino acids infused into the cecum of growing pigs. 4. Comparative studies on oral or cecal administration of 15N-lysine and 15N-urea, respectively

12 pigs were divided into 4 groups. All animals received an identical basal diet deficient in lysine and additional isonitrogenous amounts of 66.4 mmol N in the form of 15N-lysine and 15N-urea resp. orally or caecally. Caecal application was carried out as permanent infusion through caecal cannulae. N- and 15N-balances were ascertained of all animals and the remain of the labelled nitrogen was determined. From the comparison of the N-balances the conclusion can be drawn that though caecally applied N-compounds, whether they were infused as amino acids or as non-amino acid-N, disappear in the large intestine, i.e. are digested, do not, however, improve the N-balance but are excreted as additional urine-N. Subsequent to oral application, lysine or urea are almost quantitatively absorbed in the small intestine. Absorbed lysine is used in the synthesis of body protein, absorbed urea, however, is almost completely excreted in urine (83% of the 15N-amount absorbed). 15N-excretion in faeces after the oral application of 15N-lysine and 15N-urea resp. was less than 1% of the 15N-amount applied, after caecal infusion, however, it was approximately 6%, the biggest part of which (70-77%) was incorporated in bacteria protein. After caecal infusion the main quota of the infused 15N-amount (greater than or equal to 80%) was excreted in urine, most of it in the form of urea. After the oral application of 15N-lysine this could be detected in both the TCA-soluble fraction of the serum and the serum protein. After caecal infusion 15N in the TCA-soluble fraction of the serum could mainly be found as NPN, absorption and incorporation of intact 15N-lysine were considerably lower. An calculation showed that the maximum of the absorbed 15N-amount in the form of lysine was 3% and that of the infused amount was 1.8%. It can generally be doubted that the absorption of lysine in the large intestine is significant in the protein metabolism. The absorption of utilizable lysine is practically completed at the end of the ileum. In the large intestine mainly the ammonia by the catabolic activity of the intestinal flora is absorbed and subsequently excreted through the intestines.     

Experimental study of some factors limiting 'competitive exclusion' of salmonella in chickens

Some factors affecting the efficiency of competitive exclusion of Salmonella typhimurium var copenhagen in chicks were studied experimentally under gnotobiotic conditions. Axenic chickens were given dilute suspensions of adult faeces and exposed to the salmonellae. Prevention of colonisation of the gut by salmonellae ('competitive exclusion') was variable and depended possibly in part on the source of the adult faeces used to protect the chicks. Exclusion was also dose-dependent, a large inoculum of the salmonellae (10(7) viable organisms per animal) leading to colonisation in treated chicks. An inapparent carrier state was sometimes produced by lower doses of the salmonellae (10(2) viable organisms per animal), but bursts of excretion still appeared after inoculation of the salmonellae. In inapparent healthy carriers, the inoculation of a dose of Eimeria tenella oocysts that was known to produce subclinical caecal coccidiosis led to the shedding of large numbers of salmonellae for over two weeks, and the use of 'competitive exclusion' in poultry as a preventive measure for salmonella infections might thus be limited by the frequent occurrence of subclinical coccidiosis in the field.     

Comparison between dialysis methods and centrifugation/filtration for collecting fecal fluid from calves

Faecal samples of different consistency from 15 calves were subjected to two methods to separate faecal fluid. Osmolality as well as the concentrations of electrolytes and short chain fatty acids were determined for each of the fluid samples obtained and the results compared. For dialysis the following procedure was used: small dialysis bags filled with dextran 10% were placed in the cooled (4 degrees C) faecal sample to which gentamycin was added to inhibit fermentation. Equilibrium between dialysate and surrounding fluid was reached after 24 hours. However, depending on the consistency of the faeces, after 6 hours 90 to 95% of the final osmolality at 24 hours was reached. In regard to osmolality, sodium, potassium and chloride concentrations as well as to concentrations of short chain fatty acids no significant differences could be observed between the faecal fluid obtained by dialysis or by centrifugation (3500 g, 20 min, 4 degrees C) and subsequent filtration. Distinctly higher concentrations of calcium and inorganic phosphate were measured in the filtrates; the probable explanation is that these ions in faeces are largely present as poorly diffusible compounds (in particular with faeces of a more solid consistency) or bound to the cell membranes.     

Detection of Mycobacterium avium subsp. paratuberculosis by buoyant density centrifugation, sequence capture PCR and dot blot hybridisation

Detection of Mycobacterium avium subsp. paratuberculosis (M. paratuberculosis) by polymerase chain reaction (PCR) is often hampered by the lack of efficient methods for sample treatment. We report a protocol for analysis of faecal samples based on buoyant density centrifugation in Percoll and IS900 sequence capture PCR combined with a dot blot assay for detection of low-grade infection of M. paratuberculosis. Serial dilutions of M. paratuberculosis genomic DNA and M. paratuberculosis bacteria were used to assess the sensitivity of the method. The final evaluation was performed with spiked faecal samples, which also were analysed by culture. The presence of PCR inhibitory substances in processed faecal samples was evaluated by including a PCR internal control. By using buoyant density centrifugation, sequence capture PCR, and dot blot hybridisation, we achieved a sensitivity of 10(3)CFU (colony forming units)/g of faeces. The detection limit by culture was assessed to 10(2)CFU/g of faeces. We conclude that the described protocol is a fast and sensitive alternative to bacterial culture of faecal samples.     

Some aspects of the gastrointestinal microflora of veal calves fed different rations: a pilot study

The gastrointestinal microflora of veal calves reared on different diets was studied because the nature of this microflora affects the quality of veal as a result of contamination of carcass surfaces with intestinal contents during slaughter. Diet A consisted of a milk substitute, diet B of milk substitute + straw pellets and diet C of milk substitute + straw pellets + concentrates. In the rumen fluid of calves reared on diet A significantly higher counts of Gram-negative bacteria but lower counts of thermotrophic enterobacteriaceae were found than in calves reared on diets B or C. As for the faecal flora, diets B and C seem to result in significantly lower counts of Gram-negative bacteria and thermotrophic enterobacteriaceae. In 46% of the faecal specimens and 62% of the specimens of rumen fluid from calves fed on milk substitute only, Pseudomonas aeruginosa was isolated in mean counts of 4.1 log cfu/g and 5.2 log cfu/g respectively. P. aeruginosa could not be isolated from any specimen from calves receiving straw pellets. These results indicate that the inclusion of straw pellets in the diet of veal calves may increase the bacteriological safety and keeping quality of veal.     

Acute in vivo interactions of Helicobacter equorum with its equine host

REASONS FOR PERFORMING STUDY: A novel urease-negative Helicobacter species has been isolated from faecal samples of clinically healthy horses, but no information is available about the main sites of colonisation in the equine gastrointestinal tract nor is the pathogenic potential of this microorganism known. An experimental infection in horses was therefore carried out. METHODS: Four horses were infected with H. equorum strain CCUG 52199T and subjected to euthanasia at 10 (n = 2) and 30 days (n = 2) post inoculation. A fifth animal was inoculated with phosphate buffered saline and used as control. Gastrointestinal samples were examined histologically and bacteriologically. These samples, as well as faecal material collected at regular intervals, were also subjected to PCR analysis. RESULTS: All horses remained clinically healthy and no specific macroscopic lesions were identified, nor were there any microscopic changes. H. equorum-DNA was detected in the faeces during the whole experiment in all infected animals but not in the negative control. Sites of colonisation were caecum, colon and rectum. CONCLUSIONS: H. equorum is able to colonise the equine lower bowel and is excreted in faeces without apparent pathology. No association between the presence of the organism and gastrointestinal disease was demonstrated.     

Coccidial infection with Eimeria tenella and caecal defaecation in chicks.

1. The periodicity of caecal defaecation was investigated in chicks infected with Eimeria tenella. The birds were illuminated for 12 h/d (07.00-19.00) and faeces were collected at 3-h intervals for up to 21 d. 2. In uninfected birds caecal faeces were never collected during the dark period. During the periods 07.00-10.00 h, 10.00-13.00 h, 13.00-16.00 h and 16.00-19.00 h, caecal defaecation occurred on approximately 44, 19, 55 and 91% of the total number of days respectively. 3. In infected birds production of haemorrhagic caecal faeces began from between 90 and 156 h after infection and continued during both light and dark periods for between 12 and 48 h. After this period, no caecal faeces were observed for between 39 and 120 h. Normal caecal defaecation resumed from between 198 and 264 h.     

Relationship of retinoid and carotenoid metabolism with caecotrophy in rabbits

The relationship of retinoid and carotenoid metabolism with caecotrophy was studied in adult female New Zealand White rabbits kept in individual metabolic cages. Caecotrophy was prevented by the use of plastic collars. The dry matter, crude protein, fibre, fat and ash contents of hard and soft faecal samples were determined. The retinoid (retinol and retinyl palmitate) and carotenoid (canthaxanthin, beta-carotene, beta-cryptoxanthin, lutein and zeaxanthin) levels of the blood, liver, kidney, caecal content and faeces were also measured by high-performance liquid chromatography (HPLC). The prevention of caecotrophy resulted in a significant decrease of blood retinol (P < 0.001) and retinyl palmitate (P < 0.01) concentration but it did not cause any significant change in the retinol and retinyl palmitate contents of the liver and kidney. The caecal content (25.78 +/- 6.87 microg/g) and the soft faeces (34.52 +/- 10.48 microg/g) contained the retinoids in similar amounts. Various carotenoids were found in considerable amounts in different types of faeces, while in the tissues (blood, liver and kidney) these pigments did not occur in substantial amounts. Total carotenoid concentration was similar in the caecal content (11.23 microg/g) and in the caecotroph (13.85 microg/g). On the basis of the results it can be assumed that the retinoid content of rabbit feed could be lowered in the presence of adequate caecal function and caecotrophy.     

Effects of probiotics: alone and in a mixture of Biosacc plus Zinc Bacitracin on the caecal microflora of Japanese quail

The aim of this study was to investigate the effects of two commercially available probiotics, alone and in combination with an antibiotic, on the caecal flora of Japanese quail (Coturnix coturnix japonica) reared under unstressed conditions. Thirty-four 90-day-old Japanese quail were selected for this study. The birds were divided into four groups, two groups of nine birds and two groups of eight birds. The animals in these groups were given feed containing 0.5 kg per tonne of probiotics alone (Protexin or Biosacc), a mixture of probiotic plus antibiotic (Biosacc plus Zinc Bacitracin at 0.5 and 0.0525 ppm, respectively), and a group where no supplement was added to be used as controls. The total count of aerobic bacteria, lactobacilli, enterobacteriaceae, coliforms, enteroccoci, salmonellae, sulphite-reducing anaerobic bacteria (clostridia), and pH values in the caecal content of the birds were examined. No significant differences were detected among the four groups for pH values and bacterial number (p>0.05), except for sulphite-reducing anaerobic bacteria (p<0.001). These results suggest that the use of probiotics alone and/or a mixture of a probiotic plus antibiotic as a feed supplement does not have a major suppressing effect on the majority of bacterial groups in the caecal flora of mature, healthy Japanese quail reared in unstressed conditions.     

In vitro and in vivo binding of trimethoprim and sulphachlorpyridazine to equine food and digesta and their stability in caecal contents

Binding of antibiotics to food has received little attention in equine medicine, although such binding could potentially reduce the bioavailability and clinical efficacy. In the present study, binding of trimethoprim (TMP) and sulphachlorpyridazine (SCP) to hay, grass silage and concentrate was investigated in vitro in buffer at pH 6.8 at different concentrations. The binding of TMP and SCP to caecal contents was also studied. In addition, the degradation of TMP and SCP by the caecal microflora was investigated by incubating sterilized and non-sterilized caecal contents for 3 h at 37° under anaerobic conditions and comparing the TMP and SCP contents. Further, a TMP/SCP powder formulation was adminstered orally with concentrate at a dose rate of 5 mg/kg TMP and 25 mg/kg SCP to three ponies with a caecum fistula; the animals were deprived of food for 8 h before administration. Blood samples, caecal contents samples and faecal samples were collected and analysed for TMP and SCP concentrations by means of high performance liquid chromatography (HPLC). Three non-fistulated ponies, acting as control animals, were fed the same dose of TMP/SCP with concentrate after 8 h of food deprivation and blood samples were taken. The percentage of in vitro binding of TMP as well as SCP to hay, grass silage and concentrate at concentrations of 4 μg/mL to 10 μg/mL was high (60-90%). TMP and SCP were also extensively bound to caecal contents (50-70%). At spiking concentrations above 10 μg/mL the percentage of binding decreased. There was no evidence of biodegradation of TMP or SCP in caecal contents. In vivo, both drugs could be detected in the caecal contents and in the faeces of three fistulated ponies. However, the fistulated ponies differed from the control ponies in that their TMP and SCP plasma concentrations were higher, and two fistulated ponies did not show double peaks in their plasma concentration-time curves. Therefore, the fistulated ponies did not provide an optimal model for in vivo binding studies. Despite this limitation, it can be concluded that binding of TMP and SCP to food is a major cause of the limited bioavailability of these drugs in the horse. It is hypothesized that the binding is reversible, and that a second absorption phase occurs in the large intestine, but part of the administered dose remains bound as both drugs were found in the faeces.     

The measurement of glucocorticoid concentrations in the serum and faeces of captive African elephants (Loxodonta africana) after ACTH stimulation

Conventionally, the assessment of adrenal responses to stress relies on blood sample collection. However, blood collection from animals is impossible without restraint or immobilisation that influences results. This study was undertaken to validate recently established enzyme immunoassays that measure faecal glucocorticoid metabolites in elephants, and to perform a preliminary investigation into the biological relevance of this non-invasive method for use in assessing the degree of stress in this species. Four juvenile African elephants were injected i.m. with 2.15 mg synthetic adrenocorticotrophic hormone (Synacthen, Novartis, Switzerland). Blood and faecal samples were collected over 4 h and 7 d respectively. Concentrations of serum cortisol and faecal cortisol metabolites were determined using immunoassay. Variability of basal and peak values in blood and faeces was observed among the elephants. After ACTH injection, serum cortisol concentrations increased by 400-700%. An 11-oxoaetiocholanolone enzyme immunoassay (EIA) proved best suited to measure cortisol metabolites (11,17-dioxoandrostanes) when compared to a cortisol and corticosterone EIA in faecal samples. Concentrations of faecal 11,17-dioxoandrostanes increased by 570-1070%, reaching peak levels after 20.0-25.5 h. Greater levels of glucocorticoid metabolites were measured in faecal samples from elephants kept in small enclosures compared to levels in the faeces of animals ranging over a larger area. The results of this preliminary study suggest that non-invasive faecal monitoring of glucocorticoid metabolites is useful in investigating adrenal activity in African elephants.