Relative contribution of foetal and post-natal nutritional periods on feeding regulation in adult rats.

AIM: The aim of this study was to assess the contribution of both foetal and/or post-natal nutritional periods on feeding regulation in adult rats. METHODS: Body weight gain, adipose tissue development, food preferences and feeding pattern under regular chow or Western diets were characterized on four experimental groups of rats: pups born from protein-restricted dams (R) and weaned by control (RC) or R dams (RR) and pups born from control dams weaned by C (CC) or R dams (CR). RESULTS: Rats born with intrauterine growth restriction (IUGR) and fed a Western diet at adulthood appeared predisposed to body weight gain and more fat accretion, whereas CR rats, despite their preference for high-fat diet and their hyperphagia for Western diet, did not show significant increase in fat tissue. Daytime food intakes, as well as their speed of ingestion, were found modified in RC and RR. Alterations in the hypothalamic appetite regulatory mechanisms were investigated through neuropeptide expression analysis. IUGR rats showed altered expression of key elements of leptin and NPY signalling, while CR rats exhibited lesser expression of enterostatin, MC4r and HT-1Br mRNA. CONCLUSION: Altogether, these results indicate that peri-natal nutrition has different lasting effects on feeding pattern and hypothalamic appetite regulation, depending on the time window insult.

Quantification of Penicillium camemberti and P. roqueforti mycelium by real-time PCR to assess their growth dynamics during ripening cheese.

Real-time PCR has been applied to quantify mycelium of Penicillium camemberti and Penicillium roqueforti during ripening of model cheese curd and surface mould-ripened cheeses. Total fungal DNA was first validated as an indicator of mycelial biomass in pure liquid culture and then in model curds at different stages of ripening. To imitate cheese matrix effects, DNA was extracted from curd mixed with known amounts of fresh mycelium of P. camemberti or P. roqueforti and was used as biomass standards for further quantitative real-time PCR. Mycelial mass per cheese (mg/g) was then directly obtained from fluorescence data. In model cheese curd, mycelial mass of P. camemberti increased from 2.8 at d4 to 596 mg/g at d11 whereas P. roqueforti increased from 0.3 to 6.3 mg/g during the same period. P. camemberti showed a fast development in Coulommiers from d2 to d9 (66 to 119 mg/g) and a 100-fold increase in Carré (0.85 to 85 mg/g). While mycelial biomass reached a maximum at d9 in Coulommiers, it still developed in Carré until d45. For the first time, cheese manufacturers have a powerful technique to monitor mycelial growth dynamics of their fungal cultures, which represents an important step for controlling cheese making.

Isolation and analysis of differentially expressed genes in Penicillium glabrum subjected to thermal stress.

Penicillium glabrum is a filamentous fungus frequently involved in food contamination. Numerous environmental factors (temperature, humidity, atmospheric composition, etc.) or food characteristics (water activity, pH, preservatives, etc.) could represent potential sources of stress for micro-organisms. These factors can directly affect the physiology of these spoilage micro-organisms: growth, conidiation, synthesis of secondary metabolites, etc. This study investigated the transcriptional response to temperature in P. glabrum, since this factor is one of the most important for fungal growth. Gene expression was first analysed by using suppression subtractive hybridization to generate two libraries containing 445 different up- and downregulated expressed sequence tags (ESTs). Expression of these ESTs was then assessed for different thermal stress conditions, with cDNA microarrays, resulting in the identification of 35 and 49 significantly up- and downregulated ESTs, respectively. These ESTs encode heat-shock proteins, ribosomal proteins, superoxide dismutase, trehalose-6-phosphate synthase and a large variety of identified or unknown proteins. Some of these may be molecular markers for thermal stress response in P. glabrum. To our knowledge, this work represents the first study of the transcriptional response of a food spoilage filamentous fungus under thermal stress conditions.

Myelotoxicity of trichothecenes and apoptosis: an in vitro study on human cord blood CD34+ hematopoietic progenitor.

Previous studies have revealed that hematological disorders associated with trichothecenes intoxication in humans could result from hematopoiesis inhibition. The most frequent and potent trichothecene mycotoxins are T-2 toxin and deoxynivalenol (DON), respectively. Apoptosis induction by these two toxins was investigated in vitro on human hematopoietic progenitors (CD34+ cells). Hoechst coloration, DNA fragmentation and annexin-V/PI labeling in flow cytometry showed that T-2 toxin, in contrast to DON, induced apoptosis in CD34+ cells. T-2 toxin effect was dose- and time-dependent with a significant increase of apoptotic cells as early as 3h after incubation at 10(-7) M and a maximum reached at 12 h. This observation evidenced the high sensitivity of hematopoietic progenitors to T-2 toxin. The inhibition of T-2 toxin-induced apoptosis by a pan-caspase inhibitor (Z-VAD-fmk) suggested the involvement of caspases. The proportional increase of caspase-3 specific activity (DEVDase) with T-2 toxin concentration confirmed its role in the process. After incubation of CD34+ cells with T-2 toxin, in conditions that induced apoptosis, clonal expansion of granulo-monocytes, erythrocytes and megakaryocytes precursors was dose-dependently inhibited. The hematological effects observed in T-2 toxin mycotoxicosis could then be assigned to hematopoiesis inhibition by apoptosis. Different mechanisms that need to be further elucidated are involved in DON myelotoxicity.

Effect of Ochratoxin A on human haematopoietic progenitors proliferation and differentiation: an in vitro study.

Ochratoxin A (OTA) is a mycotoxin food and feed-contaminant known to induce nephro and hepatotoxicity in human and animal, and related to human Balkan Endemic Nephropathy. However, haematological troubles are also observed in case of acute OTA intoxication. These disorders observed in animals emphasise the necessity to determine if OTA exposure induce damage to haematopoietic system in human. The effect on haematopoiesis has been evaluated using in vitro clonogenic assays of the three lineages i.e., platelet, red and white blood cell progenitors. Human erythroblastic and granulomonocytic progenitor proliferation is decreased in the presence of 10(2) microM OTA. Platelet progenitors were destroyed at 10(2) microM OTA. For the lowest concentrations haematopoietic progenitor proliferation is not affected by OTA. Comparison with other mycotoxins known to be myelotoxic shows that OTA is less myelotoxic than trichothecenes.

Pancreatic secretory response to feeding in the calf: CCK-A receptors, but not CCK-B/gastrin receptors are involved.

In bovine species, as in human, the pancreas predominantly expresses cholecystokinin-B (CCK-B)/gastrin receptors. However, the role of this receptor in the regulation of meal-stimulated pancreatic enzyme release has not been determined. In milk-fed calves, we previously described prandial patterns of exocrine pancreatic secretion and a long prefeeding phase was observed. The present study was aimed at determining both the role of external stimuli in the outset of the prefeeding phase and the implication of pancreatic CCK-A and CCK-B/gastrin receptors in the mediation of pancreatic response to feeding. The first objective was studied by suppressing external stimuli associated with food intake (unexpected meal) and the second by infusing highly specific and potent antagonists of CCK-A (SR 27897) and CCK-B/gastrin (PD 135158) receptors during the prandial period. When calves were given an unexpected meal, the long prefeeding increase in pancreatic secretion was absent. SR 27897 (but not PD 135158) inhibited the preprandial phase and greatly reduced postprandial pancreatic juice and enzyme outflows. The expectancy of a meal seemed to elicit an increased pancreatic response right before a meal and CCK-A receptors may mediate this information via neural pathways. The implication of CCK and CCK-A receptors in mediating the postfeeding pancreatic response was also demonstrated. The participation of CCK-B/gastrin receptors in this regulation was not demonstrated.

Effects of melatonin on liver estrogen receptor and vitellogenin expression in rainbow trout: an in vitro and in vivo study.

Although melatonin is believed to mediate many seasonal and circadian effects of photoperiod on reproduction in salmonids, the precise mechanisms underlying such effects are still largely unknown. Recent data of the literature indicate a relationship between melatonin and expression of estrogen receptors (ER) in various tissues. In this study, the effects of melatonin on estrogen receptor and/or vitellogenin expression were studied by a combination of in vivo and in vitro experiments. In yeast stably expressing ER and transfected with an estrogen-responsive element-beta-galactosidase reporter gene, melatonin had no effect on basal or E2-stimulated ER expression. Incubation of hepatocyte aggregates with melatonin (10(-8) to 10(-4)) for 16 or 48 h did not modify the E2-stimulated ER and vitellogenin mRNA, as measured by dot blots. Finally, neither pinealectomy nor melatonin implants caused any effect on basal or E2-stimulated ER and vitellogenin mRNA contents in the liver. Altogether, these results suggest that, although we cannot exclude potential effects at the brain or pituitary levels, melatonin has no or little effects on estrogen receptor in the liver.

Expression of clock gene in the brain of rainbow trout: comparison with the distribution of melatonin receptors.

To identify brain structures potentially acting as biological clocks in rainbow trout (Oncorhynchus mykiss), the expression sites of a trout homolog of the mouse clock gene were studied and compared with that of melatonin receptors (Mel-R). For this purpose, a partial sequence of the trout clock gene, including a PAS domain, was obtained by reverse transcription-polymerase chain reaction and used to perform in situ hybridization. The highest density of clock transcripts was observed in the periventricular layer (SPV) of the optic tectum, but a weaker expression was detected in some pretectal nuclei, such as the posterior pretectal nucleus (PO) and the periventricular regions of the diencephalon. Comparison of the hybridization signal in fish sacrificed at 08:00 and 17:00 did not indicate major changes in clock expression levels. Comparison of adjacent sections alternatively treated with clock and Mel-R probes suggests that both messengers are probably expressed in the same cells in the SPV and PO. In addition, in situ hybridization with a glutamate decarboxylase 65 probe, demonstrates that cells expressing clock and Mel-R in the optic tectum are gamma-aminobutyric acid neurons. The tight overlapping between the expression of Mel-R and clock transcripts in cells of the PO and SPV suggests a functional link between these two factors. These results indicate that the optic tectum and the pretectal area of the rainbow trout are major sites of integration of the melatonin signal, express the clock gene, and may act as biological clocks to influence behavioral and endocrine responses in trout.

Exogenous CCK and gastrin stimulate pancreatic exocrine secretion via CCK-A but also via CCK-B/gastrin receptors in the calf.

A predominance of the pancreatic cholecystokinin (CCK) receptor of the B/gastrin subtype (CCK-B/G) was reported in calves older than 1 month. Specific CCK-A and CCK-B/G receptor antagonists (SR 27897 and PD 135158, respectively) were used to identify the CCK receptor subtype involved in exogenous CCK- and gastrin-induced exocrine pancreatic responses. Conscious calves (2 months old) with catheterized pancreas, jugular vein and duodenum were used; the pancreatic juice was continuously reinfused. CCK (30 pmol kg-1 min-1, 40 min) evoked an increase in pancreatic juice flow and enzyme secretion, while the same dose of gastrin increased enzyme secretion alone. CCK-induced pancreatic secretion was abolished by SR 27897 (15 nmol kg-1 min-1, 55 min) and reduced by PD 135158 (0.15 nmol kg-1 min-1, 55 min). Gastrin-induced enzyme secretion was reduced by PD 135158 (50% to 90%) and to a lesser extent by SR 27897 (50% to 60%). These results demonstrate that CCK and gastrin in the physiological range stimulate pancreatic exocrine secretion in calves and that these effects are partly mediated by CCK-B/G receptors. Although CCK-A receptors are not predominantly expressed, they seem to play a major role in the response of pancreatic exocrine secretion to CCK.

Glucocorticoid receptor immunoreactivity in neurons and pituitary cells implicated in reproductive functions in rainbow trout: a double immunohistochemical study.

In order to identify the nature of the glucocorticoid receptor (GR)-expressing neurons and pituitary cells that potentially mediate the negative effects of stress on reproductive performance, double immunohistochemical stainings were performed in the brain and pituitary of the rainbow trout (Oncorhynchus mykiss). To avoid possible cross-reactions during the double staining studies, combinations of primary antibodies raised in different species were used, and we report here the generation of an antibody raised in guinea pig against the rainbow trout glucocorticoid receptor (rtGR). The results obtained in vitellogenic females showed that GnRH-positive neurons in the caudal telencephalon/anterior preoptic region consistently exhibited rtGR immunoreactivity. Similarly, in the anterior ventral preoptic region, a group of tyrosine hydroxylase-positive neurons, known for inhibiting gonadotropin (GTH)-2 secretion during vitellogenesis, was consistently shown to strongly express GR. Finally, we show that a large majority of the GTH-1 (FSH-like) and GTH-2 (LH-like) cells of the pituitary exhibit rtGR immunoreactivity. These results indicate that cortisol may affect the neuroendocrine control of the reproductive process of the rainbow trout at multiple sites.

Differential tissular expression of the CCK(A) and CCK(B) gastrin receptor genes during postnatal development in the calf.

Local and temporal expression of CCK(A) and CCK(B)/gastrin receptor genes was studied in the calf with a quantitative Reverse Transcription-Polymerase Chain Reaction (RT-PCR) method. Cerebral cortex, antrum, fundus, gall bladder, pancreas and liver were analyzed in calves at 0, 2, 7, 21, 28 and 150 days of age. Cerebral cortex and pancreas expressed both receptor genes with a ratio between CCK(A) and CCK(B)/gastrin receptor transcripts varying according to the age. Gall bladder and fundus showed an exclusive expression of CCK(A) and CCK(B)/gastrin receptor mRNAs, respectively, with the highest levels of transcripts in newborn and 28-day-old calves. The rank order for CCK(A) receptor mRNA expression was gall bladder > pancreas > cerebral cortex >>> antrum and that for CCK(B)/gastrin receptor mRNA expression was cerebral cortex / pancreas / fundus >> antrum. No CCK(A) and CCK(B)/gastrin receptor mRNA was detected in liver, regardless of the age of calves. The present data represent a basis for a better understanding of the ontogeny of physiological functions linked to the CCK(A) and CCK(B)/gastrin receptors.

Gut regulatory peptide levels in bovine fetuses and their dams between the 3rd and 9th months of gestation.

Several gut regulatory peptides were measured by radioimmunoassay between 3 and 9 months of gestation in the plasma of 91 bovine fetuses and their dams, in fetal gastric content and in amniotic fluid. During gestation, plasma peptide concentrations did not change in cows. Likewise, fetal plasma concentrations of cholecystokinin, somatostatin, secretin and vasoactive intestinal polypeptide showed no variation while those of gastrin, pancreatic polypeptide and gastric inhibitory polypeptide increased during the last 6 months. Peptide levels in the fetus were higher than or equal to maternal concentrations. At 8-9 months of gestation, gastrin, CCK, secretin and somatostatin concentrations in amniotic fluid were lower than those measured in fetal gastric content and in maternal and fetal plasma. Therefore, a substantial endogenous endocrine production of regulatory peptides by the fetus probably exists as early as the third month of gestation, accompanied by a release into the lumen of the gut.

Source of dietary protein influences kinetics of plasma gut regulatory peptide concentration in response to feeding in preruminant calves.

The kinetics of the peripheral plasma concentrations of eight gut regulatory peptides were examined in response to feeding in preruminant calves. Two experiments were carried out in animals fed milk substitutes either based on milk protein (control diet) or in which casein had been replaced by hydrolyzed fish (fish diet in experiment 1) or whey (whey diet in experiment 2) protein concentrate. In contrast to the control diet, the latter two did not coagulate within the abomasum. No variation was observed in plasma concentrations of gut regulatory peptides during 1-1.4 hr before the morning meal regardless of the nature of the dietary protein. With the control diet, the meal was followed by an increase in cholecystokinin, gastrin and gastric inhibitory polypeptide and a fall in secretin, vasoactive intestinal polypeptide and motilin, whereas no significant change was observed for somatostatin and pancreatic polypeptide. The replacement of casein by protein substitutes did not greatly modify the pattern of plasma responses to feeding, but the prefeeding and postfeeding levels were highly affected. We conclude that the most important characteristic influencing plasma gut peptide concentrations is the ability of dietary protein to clot in the abomasum, consequently determining the pattern of gastric emptying, and that variations appear depending on the origin of protein substitutes in relation to the duodenal content and mainly to the digesta pH.

Comparison of the kinetics of pancreatic secretion and gut regulatory peptides in the plasma of preruminant calves fed milk or soybean protein.

Exocrine secretion from the pancreas and concentrations of cholecystokinin, gastrin, secretin, and somatostatin in plasma were measured in relation to feeding in 70- to 120-d-old preruminant calves fed either a milk diet or a soybean diet. Pancreatic fluid was continuously collected, measured, and reintroduced in catheterized calves. Blood samples were withdrawn for measurements of gut regulatory peptide concentrations in plasma. A slight increase in outflow of pancreatic fluid was observed 30 min before the milk diet was introduced but not before the soybean diet was fed. In contrast, concentrations and outflows of protein and trypsin immediately after feeding were higher when calves were fed the soybean diet. Overall, during the first 5 h postfeeding, the outflow of pancreatic fluid was 40% higher when the milk diet was fed than when the soybean diet was fed. No difference in outflow of protein was observed, but that of trypsin was 82% higher when the soybean diet was fed. This enhanced enzyme secretion could have been related to the increased plasma concentrations of gastrin and cholecystokinin after the soybean diet was fed. Secretin release was less in calves fed the milk diet that in calves fed the soybean diet during the first 2 h postfeeding, suggesting that this gut peptide along with gastrin and cholecystokinin, contributed to the stimulation of enzyme secretion. Plasma gut regulatory peptides could be influenced by the soybean diet, which does not coagulate in the stomach, inducing faster gastric emptying of protein and fat, and by the chemical form of protein from the soybean diet and the lower susceptibility of these proteins to protease compared with casein. However, the resulting enhancement of pancreatic trypsin secretion and activity seemed to be insufficient to increase the digestibility of soybean protein up to a level similar to that of milk.

Diet modifies elastase I and II activities and mRNA levels during postnatal development and weaning in piglets.

Little information is available on the expression of pancreatic elastase I and II, despite their role in protein milk digestion. We studied the developmental changes and the effects of diet composition on both elastase I and II expression in suckled and weaned piglets. We measured their activities and levels of their corresponding mRNA. Forty-two piglets were assigned to seven groups according to age and diet. Piglets were slaughtered at birth (Group 1), or suckled up to 13 d (Group 2) or 21 d (Group 3), fed a milk substitute from 14 to 21 d (Group 4) or to 56 d (Group 7), suckled up to 21 d and then fed a dry starter up to 56 d (Group 5), or fed a milk substitute from 14 to 21 d and then a dry starter up to 56 d (Group 6). At 21 d pancreatic function was not modified by the source and the form of milk consumed. The specific activity of elastase II was maximum at birth and declined sharply thereafter, whereas that of elastase I markedly increased after weaning. The presence of milk protein in the diet did not prevent the sharp decrease in elastase II activity observed with age. During the 13 d period of suckling sow's milk, the mRNA patterns indicated that the regulation was at the mRNA and post-transcriptional levels, whereas after weaning and depending on the source of dietary protein, it was essentially translational and/or post-translational. Taken together, our results provide evidence of the early expression of elastase I and II genes that could enhance protein digestion. It seems that elastase II might be a predominant pancreatic protease during the milk-feeding period, whereas elastase I might be related to weaning.

Specific regulation of pancreatic elastase I and II mRNA expression during postnatal development in the calf: reverse transcriptase-polymerase chain reaction analysis.

The specific regulation of pancreatic elastase I and II mRNA expression as well as of the protein, RNA, and DNA contents were determined during ontogeny in the calf. Specific activities and mRNA concentrations were quantified by spectrophotometry and reverse transcriptase-polymerase chain reaction, respectively. Calves were either milk-fed or weaned until slaughter at different ages. The biosynthesis of elastases I and II was modulated by postnatal development and weaning, leading to specific gene expression profiles. The levels of elastase I activity and of the corresponding mRNAs were found to evolve in a roughly similar way. On the contrary, elastase II activity level decreased sharply during postnatal development, while no changes were observed in the corresponding mRNA levels. After weaning, elastase I activity and mRNA levels, as well as elastase II mRNA levels, increased. However, the magnitudes of elastase I activity and mRNA inductions were different. Therefore, the expression of each gene in the calf pancreas is more or less independently regulated and the regulation is mainly pretranslational (elastase I) or translational (elastase II) during postnatal development and both pretranslational and translational at weaning. The translational efficiency of elastase I and II mRNAs might be influenced by the nature of dietary proteins.

Method of measurement of pancreatic elastase II activity and postnatal development of proteases in human duodenal juice and bovine and porcine pancreatic tissue.

A specific method for pancreatic elastase II activity analysis was developed. True elastase II activity could be discriminated from that of elastase I and chymotrypsin. The postnatal development of four pancreatic proteases in the duodenal juice of children and in the pancreatic homogenates of calves and piglets was measured. The study was carried out on patients without (14 children) and with (5 children) pancreatic insufficiency. Calves and piglets were either milk-fed or weaned until slaughter at different ages. Profiles of enzyme development were globally similar in milk-fed piglets and calves, while in children without pancreatic insufficiency, no significant change was observed between 4 and 168 months. In children with pancreatic insufficiency, enzyme activity was low. In animals, elastase II and chymotrypsin activities were maximal at birth, decreased with age, and probably were associated with the digestion of milk protein. In contrast, elastase I and trypsin activities increased markedly after weaning in connection with the intake of solid food.

Kinetics of pancreatic exocrine secretion and plasma gut regulatory peptide release in response to feeding in preruminant and ruminant calves.

Pancreatic exocrine secretion and plasma cholecystokinin, gastrin, secretin, and somatostatin concentrations were examined in relation to feeding in 70- to 120-day-old preruminant and ruminant calves. The apparatus used was designed to immediately re-infuse the animal's own pancreatic juice and to carry out accurate measurements of the juice flow in real time and to take samples. In the preruminants, pancreatic juice, protein, and trypsin flows increased from 45 min before and until 15 min after the meal and decreased sharply thereafter over a period of 30 min. while protein and trypsin concentrations peaked after feeding. A significant increase in plasma gastrin and cholecystokinin (CCK), a fall in secretin and no change in somatostatin were observed after milk ingestion. By contrast, in the ruminants, feeding had no effect on the pancreatic secretion and on the plasma concentrations of these peptides. Similar and simultaneous patterns of juice flow and secretin, as well as of protein and trypsin concentrations, CCK and gastrin, could support the hypothesis that these gut regulatory peptides play a significant role in the regulation of the pancreatic function. In preruminant calves, the existence of cephalic, gastric and intestinal phases is discussed. In the ruminants, that of the ruminal phase is questionable.

Bovine pancreatic preproelastases I and II: comparison of nucleotide and amino acid sequences and tissue specific expression.

Clones encoding bovine preproelastases I and II were isolated from a pancreatic cDNA library and were sequenced in order to define the structural characteristics of these enzymes. The bovine 947- and 884-nucleotide preproelastase I and II cDNAs encode proteins containing a signal peptide of the same length (16 amino acids), but with a slightly different number of amino acids for the activation peptide (10 and 12, respectively) and the mature enzyme (240 and 241, respectively). Considering amino acid sequences, each enzyme shares a high degree of identity (76-86%) within species. In contrast, only 55.3% identity is found between bovine elastases I and II. This difference could explain partly their own specificity. Analysis of the expression of the elastases in various bovine tissues demonstrated that they are specifically expressed in high levels in the pancreatic gland. These two approaches (structure and expression) allowed us to characterize the bovine pancreatic elastases I and II.

Response of the calf pancreas to differently processed soya bean and pea diets.

The purpose of this study was to determine the effect of replacing skim-milk powder by differently treated soya bean or pea products on growth, pancreas size and pancreatic enzyme activities in calves. Three separate experiments have been performed. In experiments 1 and 2, 28 and 21 male Holstein calves were divided into 4 or 3 groups, respectively, and fed either dairy products or milk substitutes in which protein was mainly provided by soya bean products differing in their protein concentration due to the technological processing applied. In experiment 3, 45 male Holstein calves were divided into 3 groups and were fed either dairy products, or raw or flaked pea flour as a protein source. After an experimental period of 99 +/- 4 days in experiments 1 and 2, and of 88 days in experiment 3, animal growth rate was significantly lower with raw pea flour (16%) and with the soya bean diet, which was highly concentrated in carbohydrates and allergenic proteins (13-27%). Pancreas weight decreased significantly (16-18%) with pea diets and tended to be lower (NS) with the water extracted, concentrated and heated flour (soya bean). Amylase-specific activity increased significantly (43%) with pea diets but showed opposite tendencies with the most refined soya bean products. Proteolytic enzyme activities were slightly influenced by dietary protein source, but this was not as obvious as in the literature reviewed. Specific messenger RNAs corresponding to amylase, trypsin and chymotrypsin seemed to increase (NS) with the soya bean diets, particularly with the less elaborated one. However, further investigations are required before any conclusions may be drawn concerning regulation levels of pancreatic adaptation to dietary protein. According to this study and the literature, results concerning pancreatic response to diets were different suggesting that the origin of soya bean, pea seeds and technological treatments applied to them were of great importance. Also, the level of incorporation into milk substitute and the presence of more or less antinutritional factors could influence pancreatic enzyme variations by complex mechanisms.