Protein malnutrition in utero that induces permanent changes in metabolism has been investigated intensively in various animals in recent years, but to the best of our knowledge, not yet in the mink, a strict carnivore. In the present study, minks were fed either a low-protein (LP) diet, i.e., with a protein:fat:carbohydrate ratio of 14:51:35% of metabolisable energy (ME), or an adequate-protein diet (AP), i.e. 29:56:15% of ME, from when implantation was completed until parturition (17.9 +/- 3.6 days). Respiration and balance experiments were performed during both gestation and lactation. Plasma concentrations of leptin, IGF-1, and insulin were determined by radioimmunoassay; the relative abundances of glucose-6-phosphatase (G-6-Pase), fructose-1,6-bisphosphatase (Fru-1,6-P2ase), phosphoenol-pyruvate carboxykinase (PEPCK), and pyruvate kinase (PKM2) were determined in liver, and abundances of adiponectin and leptin in adipose tissue were determined by real-time quantitative PCR (q PCR). The protein supply only affected quantitative metabolism traits during the period of differentiated feeding. The dietary composition was reflected in the nitrogen metabolism and substrate oxidation, but no effects remained during lactation. The LP dams tended to have a smaller liver mass in relation to body weight than did AP dams (2.5% vs. 2.9%; p = 0.09), significantly less leptin mRNA (p < 0.05), and 30.6% fewer kits per mated female (p = 0.03). Furthermore, F1-generation kits exposed to protein restriction during foetal life (FLP1; 10.3 g) had a lower birth weight (p = 0.004) than did F1-generation kits exposed to adequate protein (FAP1; 11.3 g). Differences remained significant until 21 days of age (120.4 g vs. 127.6 g; p = 0.005). The FLP1 foetuses displayed a lower abundance of Fru-1,6-P2ase mRNA (p = 0.007) and of PKM2 mRNA (p = 0.002) than did FAP1 foetuses. Whether these changes during foetal life cause permanent changes in the glucose homeostasis of the offspring and result in the transmission of epigenetic phenotypic changes, as seen in the rat, needs further investigation.

Foetal life malnutrition has been studied intensively in a number of animal models. Results show that especially foetal life protein malnutrition can lead to metabolic changes later in life. This might be of particular importance for strict carnivores, for example, cat and mink (Neovison vison) because of their higher protein requirement than in other domestic mammals. This study aimed to investigate the effects of low protein provision during foetal life to male mink kits on their protein metabolism during the early postweaning period of rapid growth and to investigate whether foetal life protein deficiency affects the response to adequate or deficient protein provision post weaning. Further, we intended to study whether the changes in the gene expression of key enzymes in foetal hepatic tissue caused by maternal protein deficiency were manifested post-weaning. A total of 32 male mink kits born to mothers fed either a low-protein diet (LP), that is, 14% of metabolizable energy (ME) from protein (foetal low -FL), n 5 16, or an adequate-protein (AP) diet, that is, 29% of ME from protein (foetal adequate -FA), n 5 16) in the last 16.3 6 1.8 days of pregnancy were used. The FL offspring had lower birth weight and lower relative abundance of fructose-1,6-bisphosphatase (Fru-1,6-P2ase) and pyruvate kinase mRNA in foetal hepatic tissue than FA kits. The mothers were fed a diet containing adequate protein until weaning. At weaning (7 weeks of age), half of the kits from each foetal treatment group were fed an AP diet (32% of ME from protein; n 5 8 FA and 8 FL) and the other half were fed a LP diet (18% of ME from protein; n 5 8 FA and 8 FL) until 9.5 weeks of age, yielding four treatment groups (i.e. FA-AP, FA-LP, FL-AP and FL-LP). Low protein provision in foetal life lowered the protein oxidation post-weaning compared with the controls (P 5 0.006), indicating metabolic flexibility and a better ability to conserve protein. This could not, however, be supported by changes in liver mass because of foetal life experience. A lower relative abundance of Fru-1,6-P 2 ase mRNA was observed (P , 0.05), being lower in 9.5-week-old FL than in FA kits. It can be concluded that foetal life protein restriction leads to changes in post-weaning protein metabolism through lower protein oxidation of male mink kits.

Protein restriction in utero may give rise to restricted growth as well as induce metabolically related diseases. In order to determine the suitability of mink as an animal model for metabolism studies, the effects of protein restriction during gestation on dam and kit performance must first be determined. Mink dams were fed an adequate protein (AP; crude protein:fat:carbohydrate ratio of 31:55:14% of metabolizable energy, ME) or a low protein diet (LP; 19%:49%: 32% of ME) during the last 21.2 ± 3.3 days of gestation, followed by an adequate diet during lactation. Respiration and balance experiments were performed during late gestation and twice during lactation. The dietary treatment only affected energy metabolism traits significantly during the treatment period in late gestation, such that LP dams oxidized less protein (12% vs 23% of heat production, HE, P = 0.001) but more carbohydrate (37% vs 26% of HE, P < 0.05), and retained less nitrogen (0.3 vs 0.4 g.kg^-0.75.day^-1, P < 0.05) than AP dams. Reproductive performance and kit organ growth were not affected by diet. Kit losses up to weaning were higher in LP than AP dams (2.0 vs 1.4, P < 0.05). LP fetuses weighed less (8.3 vs 11.6 g, P < 0.001) and were shorter (6.2 vs 7.6 cm, P < 0.001) than AP fetuses, however, differences might have been due to different implantation times. LP kits weighed 8.5% less than AP kits at birth, and remained lighter until 21 days of age. The LP diet caused growth restriction and increased kit mortality indicating that the dietary protein supply during late gestation was suboptimal to re-quirements. The long-term consequences of protein restriction in utero in terms of permanent metabolic changes in adulthood now need to be investigated

Malnutrition during foetal life can induce modifications in the phenotype of an individual. The present study aimed to observe effects of low foetal life protein provision on modifications of the phenotype and changes in the progeny of 1-year-old female mink (F 1 generation) offspring of mothers fed a low-protein diet. Traits studied included reproductive performance, energy and protein metabolism, and key hepatic enzymes associated with glucose homeostasis and metabolic hormones. The F 0 generation offspring were fed either a low-protein (14 % of metabolisable energy (ME) from protein -FLP1) or an adequate-protein (29 % of ME from protein -FAP1) diet for the last 17·9 (SD 3·6) d of gestation. The F 1 dams were studied at birth and at 1 year of age, during their first reproductive cycle, after maintenance on an adequate diet from birth and thereafter. Metabolic traits during gestation and lactation were largely unaffected by foetal life protein provision, but birth weight in the F 2 generation was higher (P¼ 0·003) among FLP2 kits than among FAP2 kits. Furthermore, the relative abundance of pyruvate kinase mRNA was significantly (P¼0·007) lower, and fructose-1,6-bisphosphatase mRNA tended (P¼0·08) to be lower in FLP2 foetuses than in FAP2 foetuses, showing some similar difference in the F 2 generation and F 1 generation foetuses, suggesting an effect on some hepatic enzymes affecting glucose homeostasis being transmitted from the F 1 to the F 2 generation. These findings indicate that even though energy and nitrogen metabolism displayed no effect of protein provision during early life, programming effects still appeared at the molecular level in the following generation.

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