Influence of dietary protein content and source on colonic fermentative activity in dogs differing in body size and digestive tolerance.

Low-consistency, high-moisture feces have been observed in large dogs (Canis lupus familiaris), compared with small dogs, and particularly in sensitive breeds (e.g., German Shepherd dogs). The aim of this work was to determine if greater colonic protein fermentation is responsible for poorer fecal quality in large sensitive dogs. Twenty-seven bitches were allotted to 4 groups based on size and digestive sensitivity: small, medium, large tolerant, and large sensitive. Five experimental diets varying in protein source [highly digestible wheat gluten (WG) vs. medium digestible poultry meal (PM), and protein concentration from 21.4 to 21.6 (LP) to 38.2 to 39.2% CP (HP)] were tested. Diets were fed for 14 d and followed by a 12-d transition period. Digestive fermentation by-products were investigated in fresh stools [ammonia, phenol, indole, and short chain fatty acids including acetate, propionate, and butyrate (C2 to C4 SCFA), branched-chain fatty acids (BCFA), and valerate] and in urine (phenol and indole). Bacterial populations in feces were identified. The PM diets resulted in greater fecal concentrations of ammonia, BCFA, valerate, indole, and C2 to C4 SCFA than WG diets (P = 0.002, P < 0.001, P = 0.039, P = 0.003, and P = 0.012, respectively). Greater concentrations of ammonia, BCFA, and valerate were found in the feces of dogs fed HP compared with LP diets (P < 0.001, P < 0.001, and P = 0.012, respectively). The concentrations of ammonia, valerate, phenol, and indole in feces of large sensitive dogs were greater (P < 0.001, P < 0.001, P = 0.002, and P = 0.019, respectively) compared with the other groups. The Enterococcus populations were greater in feces of dogs fed with PMHP rather than WGLP diets (P = 0.006). Urinary phenol and indole excretion was greater when dogs were fed PM than WG diets (P < 0.001 and P = 0.038, respectively) and HP than LP diets (P = 0.001 and P = 0.087, respectively). Large sensitive dogs were prone to excrete a greater quantity of phenol in urine (P < 0.001). A diet formulated with highly digestible protein, such as WG, led to reduced concentrations of protein-based fermentation products in feces together with improved fecal quality in dogs, especially in large sensitive ones. Poor fecal quality in large sensitive dogs could be partly related to the pattern of protein fermentation in the hindgut.

Variations of plasma leptin in show horses during a work season.

Leptin is an adipocytokine mainly expressed by adipose tissue. Secretion of leptin in healthy animals is closely related to fat mass and metabolic activity. The aim of this study was to investigate plasma leptin variations, in relation to nutritional and exercise parameters in adult show horses during a work season. EDTA-blood samples were taken at rest from 37 Iberian horses. Body weight, body condition score and fat percentage determined by ultrasonic measurement of rump fat thickness were measured. Plasma leptin was determined with a multi-species RIA kit. Linear mixed effects model was used to assess relationship between plasma leptin and other biological parameters. Plasma leptin concentration was <12.6 ng/ml (mean = 2.8 ± 1.6 ng/ml) and was significantly higher during training periods (p < 0.0001) (4.5 ± 1.7 ng/ml) than in show periods (2.0 ± 1.1 ng/ml), despite a significant increase (p < 0.0001) in energy intake. The body weight remained almost constant. The plasma leptin concentrations were significantly affected by exercise (p < 0.0001), body weight (p = 0.04) and BCS (p < 0.0001), but were not affected by percentage of fat. In conclusion, the marked decrease in leptin values observed during a period of intense (i.e. excessive) exercise could result from an adaptation to cumulative alterations in energy balance, to exercise per se or to a combination of both.

[Non allergic disorders associated with domestic moulds]. / Pathologie non allergique et exposition aux moisissures domestiques.

INTRODUCTION: Mouldy surfaces are encountered in up to 20 % of dwellings. Because this indoor air contamination is so widespread, respiratory physicians should be aware of its effects on health and especially of its impact on respiratory diseases. BACKGROUND: The air contaminants within mouldy dwellings are very diverse. Therefore, a given heath effect cannot be attributed specifically to an individual contaminant. In the field of respiratory diseases, excluding asthma and allergy, long-term exposure to indoor moulds has been recognized as a risk factor for both ENT and bronchial symptoms. Hydrophilic moulds seem to have a larger health impact than other mould species. Among respiratory diseases, inhalation fever and, to a lesser extent, childhood respiratory infections are linked to exposure to moulds. In contrast, the relationship between exposure to indoor moulds and diseases such as sinusitis, mucous irritation syndrome, recurrent respiratory infections in adults, COPD and pulmonary haemorrhage has not been clearly established. VIEWPOINT: There are still many scientific uncertainties in this field. However, the authorities are becoming more active in dealing with unhealthy buildings and encouraging research. CONCLUSION: The health impact of mouldy dwellings represents a major public health issue. It needs incentives from institutions and financial support as well as the involvement of many specialists.

Influence of dietary protein content and source on fecal quality, electrolyte concentrations, and osmolarity, and digestibility in dogs differing in body size.

When fed the same diet, large-breed dogs tend to produce feces of poorer quality compared with small-breed dogs. Moreover, German shepherds, although having a BW similar to Giant Schnauzers, are particularly prone to digestive intolerance, producing feces of poor consistency and increased moisture. Digestive tolerance reflects the reaction of the animal to the diet, and it can be assessed by determining fecal quality (consistency, moisture, volume, odor, and color). This study was conducted to assess the effect of protein source and content on fecal quality, and to determine whether greater digestibility and lesser fecal osmolarity and electrolyte concentrations are associated with improved fecal quality in dogs differing in body size and digestive tolerance. Twenty-seven healthy female dogs were divided into 4 groups according to BW and digestive tolerance: small, medium, large tolerant, and large sensitive. Five diets, varying in protein source (wheat gluten, poultry meal, and a 50:50 mixture of both sources) and concentration (22, 29, and 39% CP on a DM basis for low, medium, and high, respectively) were tested. The present study was divided in 2 phases: 2 diets were studied in a crossover design in phase I, and 3 diets were studied in a Latin square design in phase II. Diets were fed for 14 d, followed by a 12-d transition period. Fecal score (1 = dry and hard feces, to 5 = liquid diarrhea), moisture, electrolytes (Na and K), and osmolarity, and digestibility of DM, energy, fat, CP, and ash were determined. Fecal score and moisture (P < 0.001) were less and overall digestibility (P < 0.001 for DM, CP, fat, ash, and energy) was greater for wheat gluten than for poultry meal diets. Large dogs had the greatest fecal score and moisture (P < 0.001), together with the greatest overall digestibility (P < 0.001 for DM, P = 0.054 for CP, P = 0.005 for ash, and P = 0.003 for energy). Osmolarity was less for wheat gluten-based diets (P < 0.001), and was not affected by dog size. Fecal electrolyte concentration varied mainly with dog group (P = 0.005 for Na, and P < 0.001 for K), being greater in large sensitive dogs compared with small dogs. Wheat gluten was proved to be a suitable protein source for modulating fecal quality in dogs, particularly in sensitive breeds. Poorer fecal quality in large sensitive dogs can be related to greater digestibility and greater fecal electrolyte concentrations, but not to fecal osmolarity.

Evaluation of nested and real-time PCR assays in the diagnosis of candidaemia.

Polymerase chain reaction (PCR) assays have a very low theoretical detection threshold and are therefore advocated for the diagnosis of fungaemia. However, their effectiveness in this respect remains to be assessed. This study compared real-time PCR (Can-G) and nested PCR assays with blood culture for the diagnosis of Candida spp. bloodstream infections. A total of 200 clinical blood samples obtained from 110 patients at risk for developing a systemic fungal infection, hospitalized in the University Hospital of Sfax (Tunisia), were submitted to testing by culture, nested PCR and real-time PCR. Blood culture was positive in 36 patients. When compared with culture, the Can-G assay (81% sensitivity, 96% specificity) performed better than the nested PCR assay (86% sensitivity, 54% specificity). The real-time PCR assay, which avoids both the contamination hazard with amplicons that may cause false-positive results and the use of time-consuming post-PCR steps, appears more suitable than the nested PCR assay for the laboratory diagnosis of Candida spp. bloodstream infections. In this study, real-time PCR did not enhance the diagnostic sensitivity for Candida spp. bloodstream infections compared with conventional blood culture; however, it may lead to earlier implementation of an adequately targeted antifungal treatment.

Colonic permeability is higher in Great Danes compared with smaller breed-dogs.

Fed the same dry diet, large dogs show poorer fecal quality than small ones. A high colonic permeability could explain a low water and electrolyte net balance leading to high fecal water content. This experiment was conducted to evaluate colonic permeability in dogs varying in body size and to determine whether colonic permeability is related to fecal sodium concentration and fecal quality. Four breeds of dogs were used: six Miniature Poodles (MP), six Standard Schnauzers (SS), six Giant Schnauzers (GS) and six Great Danes (GD). Colonic permeability was evaluated using the ratio of urinary lactulose to sucralose (L:S) after oral administration. Fecal sodium concentration was measured by flame photometry. The urinary L:S ratio was significantly lower in GD, indicating a higher colonic permeability, than in the three other breeds (0.35 ± 0.12 for GD and 0.51 ± 0.05 for MP). GD also presented the higher fecal sodium concentrations and the poorest fecal quality. The higher fecal sodium concentration observed in GD could be explained by the higher colonic permeability and both these variables could be important explanations for higher fecal moisture in large dogs.

[Housing and health counselling services in the management of allergic respiratory disease]. / Le Conseil Habitat-Santé dans la prise en charge des maladies allergiques respiratoires.

INTRODUCTION: Input from Housing and health counselling services is advisable when a patient's health seems to be impaired by their housing conditions. METHODS: 650 home visits have been performed by our organisation since 2002. Each visit includes a questionnaire to assess respiratory as well as non-respiratory indoor risk factors, Acarex test to assess mite-allergen content in mattress dust, mould sampling and, when appropriate, air sampling for measurement of volatile organic compounds and aldehydes. RESULTS: The dwellings studied were mostly flats located in the downtown and occupied by a tenant. In most instances, several health hazards were identified. These hazards, in decreasing occurrence included: mold (74.4%), mite infestation in mattress dust (56.3%), cleaning products accessible to children's'hands (47.8%), dangerous electrical circuits (21.1%), exposure to chemical air pollutants (9.0%), exposure to an electromagnetic field (2.8%). Numerous fungal species were identified often occurring in association. CONCLUSION: This service allowed the identification of numerous and various health hazards. Its efficacy and effectiveness remains to be evaluated.

Liver lipid metabolism.

The liver plays a key role in lipid metabolism. Depending on species it is, more or less, the hub of fatty acid synthesis and lipid circulation through lipoprotein synthesis. Eventually the accumulation of lipid droplets into the hepatocytes results in hepatic steatosis, which may develop as a consequence of multiple dysfunctions such as alterations in beta-oxidation, very low density lipoprotein secretion, and pathways involved in the synthesis of fatty acids. In addition an increased circulating pool of non-esterified fatty acid may also to be a major determinant in the pathogenesis fatty liver disease. This review also focuses on transcription factors such as sterol-regulatory-element-binding protein-1c and peroxisome proliferator-activated receptor alpha, which promote either hepatic fatty acid synthesis or oxidation.

Adipose tissue gene expression in obese dogs after weight loss.

Body weight (BW) mainly depends on a balance between fat storage (lipogenesis) and fat mobilization (lipolysis) in adipocytes. BW changes play a role in insulin resistance (IR), the inability of insulin target tissue to respond to physiological levels of insulin. This results in inhibition of lipogenesis and stimulation of lipolysis. Weight gain leads to IR whereas, weight loss improves insulin sensitivity (IS). The aim of this study was to evaluate the effect of weight loss and recovery of IS on the expression of genes involved in lipogenesis and lipolysis in weight losing dogs. Gene expression was studied in both subcutaneous and visceral adipose tissue. Obese dogs received a hypoenergetic low fat high protein diet (0.6 x NRC recommendation). Before and after weight loss, IS was assessed using the euglycaemic hyperinsulinaemic clamp. Gene expression of IRS-2, SREBP, intracellular insulin effectors, ACC, FAS, FABP, ADRP, PEPCK, lipogenesis key proteins, perilipin and HSL, lipolysis key proteins were quantified using real-time RT-PCR in subcutaneous and visceral fat. BW decreased from 15.2 +/- 0.5 to 11.4 +/- 0.4 kg (p < 0.05) over 78 +/- 8 days. When obese, dogs were insulin resistant. After weight loss, IS was improved. In the subcutaneous adipose tissue, the expression of only the IRS-2 was increased. In the visceral adipose tissue, the expression of the genes involved in the lipogenesis was decreased whereas one of the genes implied in the lipolysis did not change. The expression profile of genes involved in lipid metabolism, as measured after weight loss, is indicative for a lower lipogenesis after weight loss than in obese dogs. Our results also confirm dramatic differences in the lipid metabolism of visceral and subcutaneous fat. They should be completed by comparing gene expression during weight losing and normal weight steady state.

Hormonal disturbances associated with obesity in dogs.

Obesity is associated with multiple endocrine alterations and changes in the concentration of circulating hormones. However, few studies have explored such alterations in dogs with naturally acquired excess weight. In the present study, we investigated the effect of naturally acquired obesity on cortisol, insulin-like growth factor (IGF)-1 and prolactin secretion in dogs. Thirty-one overweight dogs were enrolled in the trial. Blood samples were collected before and after adrenocorticotrophic hormone (ACTH) injection. Free thyroxine (fT4), cortisol, thyroid-stimulating hormone (TSH), IGF-1, prolactin and fructosamine were assayed. Body weight excess increased significantly with age and neutered dogs were more obese than entire ones. The ACTH stimulation test was within the normal range for 26 of 31 dogs. Prolactinaemia was increased in seven dogs and IGF-1 in six dogs. Twenty dogs had a fructosamine concentration >340 microm. Interestingly, 18 of 31 dogs showed disturbances of thyroid function based on high TSH and/or low fT4 baseline concentration, with 11 dogs showing both. According to these parameters only six of 31 dogs were free of hormonal disturbances. These results revealed the high incidence of such disturbances, especially thyroid dysfunction, in obese, but otherwise apparently healthy dogs. They demonstrate the importance of examining endocrine function during the initial evaluation of obese dogs to avoid failure of any nutritional treatment.

Colonic fermentation of inulin increases whole-body acetate turnover in dogs.

Metabolism of acetate from colonic fermentation was investigated in dogs. Beagle dogs (n = 9) were fed a control diet for 17 d followed by a 3% inulin-enriched diet (from chicory) for 4 and 21 d. On 3 occasions, the dogs were administered simultaneously infusions of [1-(13)C]acetate i.v. and [1,2-(13)C(2)]acetate intrarectally. Peripheral acetate concentration and turnover did not change over time after consumption of an inulin-enriched diet for 4 d. After 21 d of consuming the inulin-enriched diet, the whole-body acetate turnover increased significantly by 31% from (mean +/- SEM) 15.6 +/- 2.2 to 20.4 +/- 2.9 micromol/(kg . min) without a change in concentration. The rate of colonic acetate production that reached the peripheral circulation was 4.8 +/- 1.8 micromol/(kg . min). However, no [1,2-(13)C(2)]acetate tracer was recovered in the peripheral circulation. The fraction of oxidized tracer was higher in the gut (64 +/- 3%) than in peripheral circulation (46 +/- 3%) in dogs fed an inulin-enriched diet for 21 d. In conclusion, colonic fermentation of inulin occurred and indirectly stimulated whole-body acetate turnover in dogs fed an inulin-enriched diet for 21 d.

Relationship between total transit time and faecal quality in adult dogs differing in body size.

Fed the same diet, large and giant-breed dogs have higher faecal moisture and increased frequency of soft stools than small ones. This could be the result of physiological differences, such as a different gastrointestinal transit time. In this study, we have correlated mean total transit time (MTT) with body size and faecal consistency in dogs varying in body size. Fifty dogs from 13 different breeds were used, from a Dachshund to a Great Dane. The MTT was determined using coloured plastic beads [Cummings and Wiggins, Gut, Vol. 17 (1976), p. 219], and faecal consistency was scored daily during the study. We confirmed the strong correlation between height at the shoulder (body size) and faecal score (r = 0.76; p < 0.0001). The MTT increased with body size, from 22 h for a Miniature Poodle to 59 h for a Giant Schnauzer. We found significant positive correlations (p < 0.0001) between MTT and body size as well as faecal scores (r = 0.71 and 0.70 respectively). In the present study, we observed an effect of body size on MTT. In our 50 healthy dogs a longer MTT was related to a poorer faecal quality. Previous studies reported no relationship between body size and the upper gastrointestinal transit time in healthy dogs. So, we hypothesized that body size would mainly affect colonic transit time and that a longer colonic residence time would be related to a poorer faecal quality by promoting fermentation activity.

Effect of size on electrolyte apparent absorption rates and fermentative activity in dogs.

The objectives of the study were to assess overall electrolyte absorption capacity as well as fermentative activity in dogs varying in body size in order to verify whether these parameters can explain the poor faecal quality in large dogs. Four breeds of dogs were used: six Miniature Poodles (MP), six Standard Schnauzers (SS), six Giant Schnauzers (GS) and six Great Danes (GD). Net absorption rates of electrolytes were assessed by calculating the apparent digestibility of sodium and potassium and measuring their content in stool. Fermentative activity was calculated by measuring faecal concentrations of lactic acid and short-chain fatty acids. The results showed higher faecal concentrations of fermentation products and electrolytes as well as poorer digestibility of electrolytes in Giant Schnauzers and Great Danes. Strong positive correlations were found between these different parameters and stool quality (scores and moisture). Lower overall electrolyte absorption and higher fermentative activity could thus be factors capable of explaining poorer faecal quality in these two large dog breeds.

Insulin resistance and changes in plasma concentration of TNFalpha, IGF1, and NEFA in dogs during weight gain and obesity.

Obesity-induced insulin resistance (IR) is a common problem in humans as well as domestic dogs. It is well-known that this syndrome is associated with many modifications but it is still unclear if the changes are alterations or adaptations. The purpose of this study was to develop obesity-induced IR in dogs, through a long-term overfeeding period, and to explore hormonal and metabolic disturbances associated with the development of this syndrome. Dogs were overfed for 7 months. Body weight increased by 43 +/- 5%, and insulin sensitivity decreased by 44 +/- 5%. Plasma insulin-like growth factor 1 (IGF1), tumour necrosis factor alpha (TNFalpha), and non-esterified fatty acids (NEFA) concentrations progressively increased during the overfeeding period (IGF1: 111 +/- 13 to 266 +/- 32 ng/ml, p < 0.001; TNFalpha: 5 +/- 5 to 134 +/- 41 pg/ml; NEFA: 0.974 +/- 0.094 to 1.590 +/- 0.127 mmol/l, p < 0.05). These metabolic and hormonal impairments are associated with IR, in obese dogs, and could explain, at least in part, the outbreak of this syndrome.

Wall relative humidity: a simple and reliable index for predicting Stachybotrys chartarum infestation in dwellings.

UNLABELLED: Because the indoor mold Stachybotrys chartarum has been considered as potentially responsible for serious health effects, its identification in dwellings with water damages is of utmost importance. As such dwellings are many, it would be of great value to have a simple and reliable index for predicting its presence. The aim of the study was to compare measurements of wall relative humidity (RH) to mold identification in 458 samples from 100 dwellings. Mold identification was performed by direct microscopic examination of a sample collected on the wall by the gummed paper technique. Mean (+/- s.d.) wall RH (%) was much higher (97.0 +/- 6.1) on the 30 samples where S. chartarum was identified compared with the 291 samples where other molds were identified (41.8 +/- 36.9) and to the 137 samples where no molds were identified (38.9 +/- 34.8). There was no straightforward relationship between wall and room RH. In conclusion, this study clearly demonstrate that the simple measurement of wall RH can be used as a reliable index for discarding and suspecting S. chartarum infestation in dwellings. PRACTICAL IMPLICATIONS: This paper suggests that very high relative humidity (RH) within walls is a strong risk factor for their infestation with the 'toxic mold' Stachybotrys chartarum. Besides, data from the literature demonstrate that other molds are able to produce mycotoxins when RH is very high. Thus, measurement of wall RH, which is easy to perform and very cheap, could be used as a screening tool to select those dwellings where mold identification should be performed and remediation should be promptly carried out.

Echinococcus granulosus strain typing in North Africa: comparison of eight nuclear and mitochondrial DNA fragments.

Recent studies of Echinococcus granulosus molecular strain typing have enabled a better understanding of the transmission cycle of cystic echinococcosis. There have been many publications in this area but there is a need for the evaluation of these tools. We have attempted to respond to this need in our study, which assessed 8 DNA fragments of 40 E. granulosus cysts from North Africa. Parasitological material was collected from 5 types of intermediate hosts, in 5 different countries. The primers chosen to amplify DNA targets were defined either in nuclear DNA, or in mitochondrial DNA. After amplification, PCR products were sequenced. The sequences obtained were aligned and comparisons were made within the group and with GenBank sequences. Whether the target was nuclear or mitochondrial, the same 2 main groups of genotypes were found. The first one, the 'sheep' strain, was found in the human, sheep and cattle samples collected in North Africa. The second one, the 'camel' strain, was found in the camel cysts and cattle and human cysts from Mauritania. These findings further confirm the congruence of the data given by the nuclear and the mitochondrial genome.

Cystic echinococcosis in Algeria: cattle act as reservoirs of a sheep strain and may contribute to human contamination.

In Algeria, cystic echinococcosis (CE) is a serious economic and public health problem. The common sheep/dog cycle is usually considered as the major source of human contamination. But to date the main strain of Echinococcus granulosus involved in the human contamination and the role of other hosts are still unknown. This paper reports an original work performed in northern Algeria combining field observations and molecular analysis. In a first step, examination of 6237 carcasses in slaughterhouses showed high infection and fertility rates in cattle and dromedaries. Then, in a second step, we used a molecular biology approach to identify the E. granulosus strain(s) involved. Forty-six samples from various origins were collected. They were analysed using comparison of PCR-amplified DNA sequences with one genomic (BG 1/3) and two mitochondrial (COI and NDI) targets. Results show the presence of a "sheep" strain of E. granulosus in North Algeria circulating between cattle and ovines and infectious to humans, whereas in South Algeria, a "camel" strain and a "sheep" strain were found to circulate in camels and in sheep, respectively. This study also reports an ambiguous genotype which resembled the "sheep" strain genotype (Gl) on the basis of the partial COI gene sequence, whereas on the basis of the partial NDI gene sequence, it was similar either to the "sheep" strain (Gl) or to the "camel" strain (G6). Besides its basic interest, our study confirms the role of other hosts (mainly cattle) in leading to transmission to humans and suggests that control measures should not only target sheep.

Influence of age and body size on the digestibility of a dry expanded diet in dogs.

The objectives of the study were (i) to evaluate the effects of age and body size on the apparent digestibility of a dry expanded diet and (ii) to determine whether poor faecal quality could be related to lower nutrient utilization. Four breeds of dogs were used: six Miniature Poodles (MP), six Medium Schnauzers (MS), six Giant Schnauzers (GS) and six Great Danes (GD). Digestive trials were performed at 11, 21, 35 and 60 weeks of age. Faecal moisture and scoring were recorded at the same periods. Digestibility coefficients of all macronutrients increased significantly (p < 0.05) with age in all four breeds. Organic matter digestibility increased from 80.2 +/- 1.5 to 85.5 +/- 1.5% in MP; from 80.2 +/- 0.9 to 86.3 +/- 0.5% in MS; from 79.6 +/- 4.2 to 88.9 +/- 0.5% in GS and from 85.7 +/- 1.5 to 88.4 +/- 0.8% in GD. Crude protein digestibility increased from 74.5 +/- 4.0 to 81.5 +/- 1.9% in MP; from 74.3 +/- 3.9 to 82.6 +/- 0.9% in MS; from 71.8 +/- 6.0 to 85.9 +/- 0.9% in GS and from 80.8 +/- 4.6 to 84.8 +/- 1.3% in GD between 11 and 60 weeks of age, respectively. Mean faecal moisture also increased over the growth, except in GS. Nutrient digestibility was significantly higher in large dogs at all age periods. However, larger dogs showed lower faecal scores and increased faecal moisture. In conclusion, these results show (i) an effect of age and body size on nutrients digestibility and (ii) that the lower faecal quality observed in healthy large breed dogs does not appear related to lower nutrient utilization.

Simultaneous measurement of plasma concentrations and 13C-enrichment of short-chain fatty acids, lactic acid and ketone bodies by gas chromatography coupled to mass spectrometry.

A new method has been developed for the simultaneous measurement, in a reduced plasma sample, of concentration and 13C-isotopic enrichment of acetic, propionic, butyric, lactic, acetoacetic and beta-hydroxybutyric acids by gas chromatography coupled to mass spectrometry. After plasma deproteinisation, a diethylic extraction and a N-tert.-butyldimethylsilyl-N-methyltrifluoroacetamide derivatisation were performed. Both diethyl extraction and derivatisation procedures were optimised using the central composite designs methodology. The optimised method provides good linearity, intra-day and within-day repeatability. Except for beta-hydroxybutyric (49 microM) and acetoacetic acid (5 microM), detection limits were ranging between 0.2 and 0.7 microM allowing uses of this method for colonic metabolism studies.