Maturation of the intestinal digestion and of microbial activity in the young rabbit: impact of the dietary fibre:starch ratio.

The developmental changes of intestinal digestive potential and caecal microbial activity were described in suckling and weaned rabbits according to two feeding programmes. Two groups of thirteen litters were fed from 18 to 42 days old a "High" or a "Medium" NDF:starch ratio diet (resp. 2.7 vs 2.0, groups HL and ML) with similar protein and lipid levels, and from 42 to 70 days old the two groups were fed a "Low" NDF:starch ratio diet (1.7). From 25 to 32 days (weaning), the milk and solid feed intake were 22% and 41% higher in ML group (P<0.05), and the mortality by diarrhoea was 4 units lower (P<0.01). The whole tract digestive efficiency increased by 10% before weaning, and remained steady (organic matter) or decreased (lipids, protein) after weaning. Energy digestibility was 0.623 and 0.686 for High and Medium diets respectively. From 25 to 42 days, total enzymatic activity in intestinal content increased for chymotrypsin (5-fold, P<0.001), lipase (10-fold, P<0.001), amylase (17-fold, P<0.01) and maltase (11-fold, P<0.001), while trypsin doubled after weaning. The feeding programme only affected the amylase and maltase activities, that were higher in HL group (P<0.05). The volatile fatty acids concentration in the caecum was not significantly different among the groups, but it increased by 44% 10 days after weaning. The bacterial fibrolytic enzymes, increased by 30% after weaning and were similar among the two groups. The study revealed that the intestinal digestive maturation and the caecal microbial activity of the rabbit evolved markedly between 3 and 5 weeks of age, and was weakly affected when the NDF:starch ratio decreased from 2.7 to 2.0.     

Male sexual behavior contributes to the maintenance of high LH pulsatility in anestrous female goats

The objective of this study was to determine the importance of male sexual behavior in stimulating LH secretion in anovulatory female goats. Two groups of females (n = 10 per group) were each exposed to a buck in sexual rest and submitted to natural daylength. In one group, the buck was awake, whereas in the other group, it was sedated to prevent its sexual behavior. Two other groups of goats (n = 10 per group) were exposed to sexually active bucks that had been exposed to 2.5 months of long days. In one group, the buck was awake, and in the other group, it was sedated. LH secretion was determined every 15 min from 4 h before introducing the bucks to 8 h after, then every 15 min again from 20 to 24 h after introducing the bucks. The bucks submitted to natural daylength did not stimulate LH secretion (P > 0.05), whether they were sedated or not. In contrast, both the awake and the sedated light-treated bucks induced an increase (P < 0.05) of LH pulsatility in the first 4 h following their introduction. However, pulsatility remained elevated until 24 h in the females exposed to the light-treated awake buck, whereas in the group with the light-treated sedated buck, pulsatility diminished (P < 0.05) after the first 4 h of stimulation by the buck. In conclusion, the sexual behavior of males contributes to the maintenance of a high LH pulsatility up to 24 h after introduction into a group of anovulatory goats.     

Adaptability of the digestive function according to age at weaning in the rabbit: I. Effect on feed intake and digestive functionality

The functional adaptability of the digestive system to the level of feed intake was investigated in the young rabbits by comparing two groups of 12 litters each, weaned at 21 (W21) or 35 (W35) days of age. From 14 days onwards, rabbits were fed a pelleted feed (NDF: 332 g/kg, CP: 177 g/kg, starch: 98 g/kg, as-fed basis). Until 49 days of age, the profile of digestive enzymes was weekly determined in the small intestinal content and mucosa, as well as caecal fermentation traits and fibrolytic activities. In the W21 group, the solid feed intake was increased by 57% between 21 and 35 days (P < 0.01), while the daily body growth was lower from 21 till 42 days (-17%, P < 0.05) when compared with the W35 group. Activities of enzymes of pancreatic origin were only scarcely influenced by the weaning age. In the W21 group, amylase activity tended to be lower at 28 days of age (-36%, P = 0.064), and trypsin activity was decreased by 31% at 49 days of age (P < 0.01). Lipase activity was similar in both weaning groups. Duodenal and jejunal activities of maltase and aminopeptidase N (APN) were higher on day 28 in the W21 group as compared with the W35 group (×1.4 to ×2.4, respectively, P < 0.05). On day 35, duodenal APN activity was twice as higher in the W21 group than in the W35 group (P < 0.01). In caecum, major differences between both weaning groups were observed at 28 days of age with a decrease in ammonia concentration (-43%, P < 0.01) in W21 compared with W35 rabbits. Conversely, the acetate proportion was 5% higher in the W21 group (P < 0.01) on day 28. In conclusion, the digestive tract of early-weaned rabbits showed some adaptative properties in response to nutritional environment changes, but they were insufficient to maintain their growth rate.     

A note on the effectiveness of behavioural rehabilitation for reducing inter-dog aggression in shelter dogs

The effectiveness of a rehabilitation program for reducing inter-dog aggression was evaluated at the municipal animal shelter. Sixteen dogs (of 60 examined) met the study criteria of medium inter-dog aggression as determined by an inter-dog aggression test. These dogs received a 10-day treatment of daily rehabilitation for 30 min (rehabilitation group, n = 9) or daily release into an outdoor enclosure for 30 min (control group, n = 7). Rehabilitation consisted of desensitising and counter-conditioning dogs to the approach of other “stimulus” dogs. Most dogs in the rehabilitation group showed a decline in aggression scores when re-tested after the last treatment (day 11), and differed significantly from the control dogs which showed either an increase or no change in aggression scores (U = 8.5, P < 0.01). Rehabilitation dogs also showed lower frequencies of aggressive body postures (“facing the stimulus dog”, P < 0.05, and “stiff posture”, P < 0.10) and higher frequencies of less assertive postures (“ears back”, P < 0.05, and “lowered neck”, P < 0.10) on day 11. The differences between groups were no longer significant when a reduced sample of dogs was tested 1 week after rehabilitation ended (day 18). The study shows short-term reduction of inter-dog aggression through rehabilitation, but further work is needed on effective ways of maintaining the behavioural change.     

l-arginine supplementation of milk liquid or dry diets fed to pigs after weaning has a positive effect on production in the first three weeks after weaning at 21 days of age

A total of 48, 21-day-old weaned pigs, was used in a 2 × 2 factorial arrangement of treatments with the factors being diet type (milk liquid replacer vs. dry feed) and l-arginine (ARG) supplementation (0 vs. 6 g ARG/kg) to test the hypothesis that dietary supplementation with ARG would increase performance of pigs after weaning. Pigs were fed the experimental diets for 10 d (days) after weaning and then transitioned over a 3-d period to a dry Phase II diet fed in meal form devoid of supplemental ARG. The study ended at d 21. There were five replicates (pens) per treatment (a total of 12 pigs per treatment). Blood samples were collected from two pigs per replicate on d 7 and 16 of the experiment, and free amino acids (AA) and plasma urea nitrogen (PUN) levels analysed. Milk-fed pigs outperformed (P<0.001) dry-fed pigs for the first 10 d of the experiment as well as for the total 21-d period. At d 7, milk-fed pigs had higher (P<0.05) levels of most free indispensable and dispensable amino acids in their plasma. In both the milk-fed and dry-fed pigs supplemented with ARG, average daily feed intake (ADFI, P<0.05) and average daily gain (ADG, P<0.05) were increased during the dietary transition period (d 11–14), when pigs were being changed to the Phase II diet. The difference in production in the transition period caused a tendency for ARG-supplemented pigs to eat more feed (P<0.1) and grow faster (P<0.5) over the 21-d experimental period. Pigs supplemented with ARG had higher plasma ARG levels (P<0.05) at d 7 after weaning and lower plasma urea levels (P<0.05) at both d 7 and 16 after weaning. These data show the benefits of feeding a milk liquid diet as well as of ARG supplementation after weaning on production indices.     

Both substrate availability and utilisation contribute to the defence of core temperature in response to acute cold

Acute cooling significantly increases energy demand in non-hibernators for the defence of core temperature but the contribution of the liver to thermogenesis is poorly understood. A two-tracer method to estimate lipid metabolism in cold-naïve control (CON) and cold-acclimated (CA) rats was employed to quantify hepatic rates of fat metabolism. Both fenofibrate, to increase liver mass and fat oxidation and dichloroacetate (DCA) to inhibit fat oxidation were used to alter lipid metabolism in CON animals. Following acute cooling, CA led to a doubling of the time to reach a core temperature 25 °C (P < 0.001), whereas DCA treatment decreased time of cooling (P < 0.01). DCA-treatment increased the gradient of Arrhenius-transformed rate–pressure product (P < 0.01). CA increased both palmitate uptake (P < 0.001) and β-oxidation (P < 0.01) whilst DCA treatment decreased uptake (P < 0.01) and β-oxidation (P < 0.05). Tissue-specific estimates of metabolism revealed that CA led to a 12-fold increase in β-oxidation for brown adipose tissue (P < 0.001) whilst fenofibrate halved β-oxidation in the liver (P < 0.01) despite doubling the liver mass (P < 0.001) and DCA decreased hepatic β-oxidation to 15% of control levels. Taken together, these results suggest that the liver has minimal contribution to thermogenesis in the rat, with brown adipose tissue significantly increasing both fat uptake and oxidation in response to CA.     

Effect of substitution of wheat starch by potato starch on the performance,digestive physiology and health of growing rabbits

The goal of this research was to study the effect of the substitution of wheat starch by potato starch (PS) on the performance, health and digestion of growing rabbits. Three experimental diets were formulated with 0%, 7% and 14% PS (PS0, PS7 and PS14, respectively) and similar starch contents (22% dry matter basis), proteins and fibre. The three diets were administered to three groups of 48 rabbits from weaning (28 days) to slaughter (70 days), and growth and health measurements were made. Another 10 rabbits per diet (30 rabbits at each age), reared under similar conditions, were slaughtered at 6 to 10 weeks of age, and the digesta were collected to analyse the caecal microbial activity (pH, volatile fatty acids (VFA) levels, fibrolytic activity) and the starch concentration in the ileal digesta. At the same ages, the whole tract digestibility coefficients were measured in 10 other rabbits for each treatment (30 rabbits). The feed intake between 28 and 42 days of age (days) increased by 11% (P < 0.05) in PS0 v. PS14. Over the whole growth period (28 to 70 days), weight gain was similar among diets (40.5 g/day), whereas the feed intake and feed conversion increased (8.5% and 5.2%, respectively; P < 0.05) with the PS14 diet. Mortality and morbidity were not affected by the diets. The starch concentration of the ileal contents increased (P < 0.01) with the addition of PS to the diet (0.39%, 0.77% and 1.08% for diets PS0, PS7 and PS14, respectively). Starch digestibility was 0.8 percentage units higher (99.8% v. 99.0%) with the PS0 diet than the PS14 diet (P = 0.04). The bacterial cellulolytic activity in the caecum tended to be higher with the PS14 diet (P = 0.07). The total VFA caecal concentration increased (P < 0.01) only in 6-week-old rabbits with PS7 compared with PS0 (54.7 v. 74.5 mmol/l). Protein digestibility and ileal starch concentration decreased (P < 0.05) with age (6 v. 10 weeks), and hemicelluloses digestibility increased (P < 0.05). At 10 weeks of age, rabbits showed a higher VFA pool (6.25 mol) and proportion of butyrate (15.9%) and a lower proportion of acetate (79.3%), ammonia level (7.5 mmol/l) and C3/C4 ratio (0.31) than at 6 weeks of age. The intake of potato starch had no effect on the performance, caecal microbial activity or digestive health of growing rabbits.     

Effects of dietary zinc on performance, nutrient digestibility and plasma zinc status in Cashmere goats

Thirty-six 1.0-year-old Liao Ning Cashmere goat wethers (BW = 22.01 ± 0.59 kg) were used to determine the effects of dietary zinc (Zn) level on the performance, nutrient digestibility and plasma Zn status during the cashmere fiber growing period. The goats were randomly divided into four groups that were fed a basal diet containing 22.3 mg Zn/kg dry matter (DM) with 0, 15, 30 or 45 mg Zn/kg DM as reagent grade ZnSO₄·7H₂O. The experiment lasted 60 days including a 7-day metabolism trial. Both average daily gain (ADG) and feed efficiency were improved (P < 0.05) by Zn supplementation and were higher (P < 0.05) for the treatment groups supplemented with 30 and 45 mg Zn/kg DM compared with 15 mg Zn/kg DM. Zn supplementation had no influence on the length and diameter of cashmere fiber (P > 0.05). Digestibility of DM, crude protein (CP), neutral detergent fiber (NDF) and acid detergent fiber (ADF) did not differ among treatments (P > 0.05). Plasma Zn concentrations were increased (P < 0.01) by Zn supplementation and were higher (P < 0.05) for the treatment groups supplemented with 30 and 45 mg Zn/kg DM compared with 15 mg Zn/kg DM. Zn apparent absorption rate and apparent retention rate were decreased (P < 0.05) by Zn supplementation, but did not differ among Zn supplemented treatments (P > 0.05). In conclusion, a control diet containing 22.3 mg Zn/kg DM was inadequate for achieving optimal growth performance in Cashmere goats, and the recommended level of dietary Zn for such goats is 52.3 mg/kg DM during the cashmere fiber growing period.     

Effects of early contact between non-littermate piglets and of the complexity of farrowing conditions on social behaviour and weight gain

In this study we tested if contact possibilities between non-littermate piglets and complexity of farrowing conditions affect the pre- and post-weaning behaviour, weight gain and skin lesions of piglets. Suckling sows were either kept in a group housing system (GH), in a single pen loose housing system (LH), or in conventional farrowing crates (FC). In the single pen systems a piglet door to the adjacent pen was opened on d 10 after farrowing in half of the pens so that piglets were able to enter the neighbouring pen (LH+ and FC+). For control, in the other half of single pens no piglet doors were opened (LH− and FC−). In the group housing system piglets also were allowed to freely move within the whole system on d 10 after farrowing. After weaning on d 28 piglets were kept in littered rearing pens in an open stable holding 20 piglets each. Piglets from contact pens were mixed with those they previously had contact to whereas piglets from control pens were mixed with unfamiliar litters. Data were obtained from 230 litters (113 sows with 1935 farrowed piglets). All piglets were scored for skin lesions immediately before and 4 days after opening the piglet doors, as well as immediately before and 4 days after moving into rearing pens. Behaviour (biting, fighting, drinking and laying) of piglets was recorded in the rearing pens in a 48-h period after weaning for 2 × 4 h. Treatments did not affect the level of skin lesions in the rearing period (H = 8.72, df 4, ns) nor daily weight gain until weaning (F_4,216 = 1.21, ns). In the 48 h after moving to rearing pens, less intensive agonistic behaviour (fighting and biting) was observed in contact piglets (H = 53.36, df 4, P < 0.0001). Four days after weaning control piglets showed significantly higher numbers and more severe skin lesions than contact piglets and, in addition, lesion scores of piglets from the larger single farrowing pens with straw bedding were significantly lower compared to the single farrowing crate (H = 33.86, df 4, P < 0.0001). The latency for lying in the new rearing pen was decreasing (F_4,93 = 25.76, P < 0.001) and the latency for drinking (F_4,81 = 3.43, P = 0.01) was increasing with decreasing complexity and space allotment of the housing system but were not related to whether the piglets have had contact to other litters before weaning. Five weeks after weaning weight gain (F_4,204 = 7.01, P < 0.0001) and BW (F_4,207 = 5.34, P < 0.001) were higher in treatments offering contact. Our results show that familiarising piglets from different litters 10 day post partum by establishing contact possibilities through a piglet door reduces social stress at weaning and increases weight gain after weaning. Farrowing pens with straw bedding and enlarged space as offered in the farrowing pens and the group housing system can further decrease the level of harmful agonistic interactions after mixing unacquainted litters at weaning and can improve the adaptation of piglets towards the new environment of the rearing pen.     

Effects of dietary copper on growth performance, nutrient digestibility and fiber characteristics in cashmere goats during the cashmere slow-growing period

Thirty-six 2.5-year-old wether Inner Mongolian White Cashmere Goats (IMWG) (BW = 42.7 ± 3.44 kg) were used to determine the effects of dietary copper (Cu) concentration on growth performance, nutrient digestibility and fiber characteristics during the cashmere slow-growing period. Wethers were stratified by weight and randomly assigned to four dietary treatments, which included a control diet containing 5.60 mg Cu/kg DM, the control diet supplied, respectively, with 10, 20 and 30 mg Cu/kg DM (total dietary Cu level of 5.60, 15.6, 25.6 and 35.6 mg/kg DM). The experiment lasted 50 days including a 10-day preliminary trial and 10-day metabolism trial. Average daily feed intake (ADFI) did not differ among treatment groups (P > 0.05), except that the supplement providing 30 mg Cu/kg DM decreased average daily gain and gain efficiency (P < 0.05). Copper supplementation had no influence on digestibility of DM, CP and ADF (P > 0.05), however, NDF digestibility of the treatment group supplemented with 30 mg Cu/kg DM was lower compared with that of other groups (P < 0.05). Length and growth rate of cashmere fiber were higher in the treatment group supplemented with 20 mg Cu/kg DM compared with other groups (P < 0.05), but cashmere diameter was not affected by Cu supplementation (P > 0.05). In conclusion, supplementation of Cu at the levels of 10, 20 and 30 mg/kg DM to the basal diet containing 5.60 mg Cu/kg DM had no influence on ADFI or nutrient digestibility of DM, CP and ADF in cashmere goats, while 30 mg Cu/kg DM supplementation had a negative effect on growth performance and NDF digestibility. However, 20 mg Cu/kg DM supplementation of the basal diet enhanced cashmere growth. Hence, the appropriate supplemental level during the cashmere slow-growing period is deemed to be 20 mg Cu/kg DM (total dietary Cu level of 25.6 mg/kg DM).     

Cloning and overexpression of a maltase gene from Schizosaccharomyces pombe in Escherichia coli and characterization of the recombinant maltase

The Schizosaccharomyces pombe maltase structural gene (SPMAL1⁺) was amplified from genomic DNA of S. pombe by PCR. An open reading frame of 1740 bp, encoding a putative 579 amino-acid protein with a calculated molecular mass of 67.7 kDa was characterized in the genomic DNA insert of plasmid pQE30. The specific maltase activity in the induced transformants was 21 times higher than that in wild-type. However, the estimated molecular mass of the purified recombinant maltase was 44.3 kDa by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). The optimal temperature and pH of the purified recombinant maltase were 40 °C and 6, respectively. The recombinant maltase was weakly activated by Mg²⁺, Ca²⁺, Na⁺, and Ba²⁺, but was strongly inhibited by Hg²⁺, Ag⁺ and Cu²⁺, EDTA, and PMSF. The purified maltase could actively hydrolyse ρ-nitrophenyl glucoside (PNPG), maltose, dextrin, and soluble starch. The results demonstrate that maltase from S. pombe was different from that from other yeasts, and might be usefully exploited in the future by the biotechnology industry or lead to the development of new molecular genetic tools.     

Effect of level of fibre on performance and digestion traits in growing rabbits

An experiment was conducted to determine the effects of level of fibre on production performance, gastrointestinal tract development, caecum fermentation and fibrolytic activity in weaner to 2-month-old New Zealand rabbits. One hundred weaner rabbits were allocated in individual cages for five treatments in which they were fed each diet with ADF at 130 (NDF, 274.1; ADL, 39.7; CP, 158.4; EE, 26.8; starch, 252.5 g/kg), 160 (NDF, 292.9; ADL, 41.9; CP, 160.3; EE, 25.6; starch, 249.4 g/kg), 190 (NDF, 312.4; ADL, 50.5; CP, 157.6; EE, 24.2; starch, 218.2 g/kg), 220 (NDF, 343.9; ADL, 51.2; CP, 158.5; EE, 22.2; starch, 217.6 g/kg), and 250 g/kg (NDF, 375.7; ADL, 66.0; CP, 156.9; EE, 22.4; starch, 220.8 g/kg) original matter basis, respectively. The results were: ADG and F/G deteriorated with increasing fibre (P<0.05); ADI increased with increasing dietary ADF (P=0.002). The stomach relative weight (g/kg LW), small intestine relative weight (g/kg LW), caecum weight, caecum content weight and caecum relative weight (g/kg LW) all increased with increasing ADF (P=0.003, 0.007, 0.002, 0.010 and <0.001, respectively). Quadratic effects of level of ADF on the villus height, villus height/crypt depth of the duodenum and the villus height/crypt depth of the jejunum were obtained; cubic effects of level of ADF on the crypt depth of the jejunum and the villus height/crypt depth of the ileum were obtained. Cubic effects of level of ADF on the pH, the butyric acid (% total VFA) and C2/C4 were obtained. The NH₃–N concentration dropped and the acetic acid (% total VFA) increased when the dietary ADF increased (P<0.001). The fibrolytic activity measured in the caecal contents increased when the dietary ADF increased (P<0.001). Providing a ration consisting mainly of clover meal, wheat straw and peanut hulls, the optimum dietary ADF content for weaner to 2-month-old growing meat rabbits was shown to be 190 g/kg feed.     

Effect of a disinfection strategy on the methicillin‐resistant Staphylococcus aureus CC398 prevalence of sows,their piglets and the barn environment

Aims

To assess, in a cleaned and disinfected barn environment, the efficacy of an animal disinfection strategy to reduce the livestock‐associated methicillin‐resistant Staphylococcus aureus (LA‐MRSA) prevalence in sows, their offspring and the barn environment.

Methods and Results

On each farm, six sow rounds were sampled; sows were divided into either a test or control group. Per round, 20 sows and 40 of their piglets were sampled at different time points together with the barn environment. The disinfection strategy of the test groups consisted of washing the sows with a shampoo followed by disinfection of the skin with a solution containing chlorhexidine digluconate and isopropanol. On the first day of disinfection and 6 days after stopping the disinfection, a significant decrease (P < 0·01) of on average 68 and 66% in sow MRSA prevalence was observed on both farms, whereas no decrease was seen in the control groups. Just before weaning, 21–28 days after the end of the disinfection strategy, the difference in MRSA prevalence between both groups was reduced to 4% and no longer significant (P = 0·20). The MRSA prevalence of the piglets in the test groups was significantly lower (26%; P < 0·01) 6 days after the end of disinfection. Just before weaning, this difference was reduced to 5% but still significant (P < 0·01). In the swine nursery unit, no significant difference (P = 0·99) was seen between both groups. Based on semi‐quantitative counts, a relationship (r² > 0·6; P < 0·01) was seen between MRSA contamination in the barn environment and the MRSA prevalence in pigs.

Conclusion

Results show that the tested disinfection strategy reduces temporarily the sow and piglet MRSA status, but does not result in a final reduction in MRSA at weaning or in the nursery unit.

Significance and Impact of the Study

First report on the efficacy of an animal disinfection strategy to reduce LA‐MRSA prevalence in sows, their offspring and the barn environment.     

Influence of diets supplemented with vitamins C and E on pirarucu (Arapaima gigas) blood parameters

This study evaluated the influence of diets supplemented with 500, 800, 1200 mg kg^− 1 of vitamin C (ascorbic acid or AA) and vitamin E (α-tocopherol or α-T) on the physiological responses of pirarucu fed for 2 months. Weight and mortality were not affected by dietary vitamin type or their concentrations. Significant increase (p < 0.05) on the red blood cells count was obtained on treatments with 800 and 1200 mg AA kg^− 1 and on the hemoglobin concentration on treatment with 500 mg α-T kg^− 1 relatively to control. Mean corpuscular volume presented a significant decrease (p < 0.05) on treatment with 800 and 1200 mg AA kg⁻¹ when compared to control. Mean corpuscular hemoglobin concentration was significantly high (p < 0.05) on treatment with 500 mg α-T kg^− 1. Only in vitamin C treatments, we noticed a significant increase (p < 0.05) in the number of leucocytes relative to control. All fish in the vitamin-supplemented treatments, except 500 mg AA kg^− 1, had high total protein values compared to control. Fish treated with 800 or 1200 mg α-T kg^− 1 also showed increases in plasma glucose concentrations. Our results suggest that 800 and 1200 mg AA kg^− 1 are probably the most suitable concentrations for pirarucu diets, although high vitamin E diets are not necessary for quantitative leucocyte increases for this species.     

Isolation and partial characterization of three isoamylases of Rhyzopertha dominica F. (Coleoptera: Bostrichidae)

Three isoamylases of Rhyzopertha dominica (termed RdA70, RdA79, and RdA90 according to their relative mobility in gel electrophoresis) were isolated by ammonium sulfate fractionation and hydrophobic interaction chromatography. RdA70 and RdA79 showed an optimal pH of 7.0, whereas for RdA90 the optimal pH was 6.5. The three isoamylases remained stable at 50 °C for 1 h, but at 60 °C, all lost 50% of their activity in 20 min and were completely inactivated in 1 h. RdA70 and RdA79 were inhibited by albumin extracts from wheat samples varying widely in amylase inhibitory activity; however, RdA90 was highly resistant to inhibition. β-Mercaptoethanol up to 30 mM increased the activity of the three isoamylases by 2.5-fold. The action pattern of the three isoamylases was typical of endoamylases; however, differences were observed on the hydrolytic efficiency rates measured as V_max/K_m ratio on starch, amylopectin, and amylose. The hydrolyzing action of RdA90 on starch and amylopectin (V_max/K_m = 90.4 ± 2.3 and 78.9 ± 6.6, respectively) was less efficient than that on amylose (V_max/K_m = 214 ± 23.2). RdA79 efficiently hydrolyzed both amylopectin and amylose (V_max/K_m = 260.6 ± 12.9 and 326.5 ± 9.4, respectively). RdA70 hydrolyzed starch and amylose at similar rates (V_max/K_m = 202.9 ± 5.5 and 215.9 ± 6.2, respectively), but amylopectin was a poor substrate (V_max/K_m = 124.2 ± 7.4). The overall results suggest that RdA70 and RdA79 appear to belong to a group of saccharifying isoamylases that breaks down long fragments of oligosaccharide chains produced by the hydrolytic action of RdA90. The simultaneous action of the three isoamylases on starch, aside from the high resistance of RdA90 to wheat amylase inhibitors, might allow R. dominica to feed and reproduce successfully on the wheat kernel.     

Trehalose accumulation in Baudoinia compniacensis following abiotic stress

Baudoinia compniacensis is a microfungus recently described as the principal agent of fouling known as “warehouse staining”, affecting building exteriors, fixtures and vegetation surfaces in areas proximate to distillery aging warehouses, commercial bakeries and other areas subject to low-level ethanol vapour exposure. The surfaces most affected tend to be highly exposed and undergo extreme diurnal temperature fluctuations. In previous work, we have demonstrated the existence of heat-inducible putative chaperone proteins that may also be induced by low-level exposures to ethanol vapour (e.g., <10 ppm). The present study investigated the cellular accumulation of trehalose, a disaccharide identified in some microorganisms to be important in the protection of cell components during adverse stress conditions, such as thermal stress. Following heat shock at 45 °C, we observed a 2.5-fold accumulation of trehalose relative to unheated controls maintained at 26 °C. Peak trehalose concentrations of 10 mg g⁻¹ dry wt were seen at 90 min after heat treatment, followed by a gradual return to post-treatment by 150 min. Exposure of B. compniacensis cells to ethanol resulted in a similar increased accumulation of trehalose compared to unexposed controls. These findings imply that trehalose may be important in the tolerance of this fungus to abiotic stresses, such as heat and solvent exposure, and suggest future research directions for the control and prevention of warehouse staining.     

Effects of inclusion of neutral detergent soluble fibre sources in diets varying in forage particle size on feed intake, digestive processes, and performance of mid-lactation Holstein cows

The aim of this study was to evaluate effects of partial replacement of neutral detergent soluble fibre (NDSF) for starch in diets varying in particle size (PS) of alfalfa hay on chewing activities, ruminal fermentation, nutrient digestibility and performance in mid-lactation dairy cows. Eight multiparous Holstein cows (146 ± 6.0 d in milk; 36.7 ± 2.57 kg milk/d) were used in a replicated 4 × 4 Latin square design with four 21 d periods with the last 7 d for data collection. The experiment was a 2 × 2 factorial arrangement with 2 levels of NDSF (low = 85 g/kg or high = 130 g/kg diet dry matter) each combined with 2 PS (short = 20 mm or long = 40 mm) of alfalfa hay. Results show that forage PS alone, or in combination with NDSF inclusion, had no effect on dry matter (DM) intake. Although total chewing, eating and ruminating times were not affected by treatments, eating time per kg of neutral detergent fibre (NDF) ingested was higher in long versus short alfalfa hay-based diets (P<0.05). Feeding long forage PS increased sorting of the diet against particles >19 mm, and in favor of those <8 mm (P<0.05). Feeding diets high in NDSF lowered DM intake (P<0.05), but increased apparent digestibility of all nutrients including NDF (P<0.05) independent of forage PS. Ruminal pH and concentrations of total volatile fatty acids were unaffected by dietary treatments, however the proportion of butyrate was higher in ruminal fluid of cows fed high NDSF diets (P<0.05). Changes in milk composition included lower milk crude protein content in high NDSF diets and higher lactose content for short hay-based diets (P<0.05). That milk yield and milk energy output were similar in low versus high NDSF diets suggests that high NDSF-fed cows had higher energy efficiency due to lower DM intake. Results suggest that, independent of forage PS, NDSF sources can be successfully included to partly replace starchy grains in diets exceeding minimum fibre recommendations.     

Effect of renoguanylin on hydrogen/bicarbonate ion transport in rat renal tubules

Renoguanylin (REN) is a recently described member of the guanylin family, which was first isolated from eels and is expressed in intestinal and specially kidney tissues. In the present work we evaluate the effects of REN on the mechanisms of hydrogen transport in rat renal tubules by the stationary microperfusion method. We evaluated the effect of 1 μM and 10 μM of renoguanylin (REN) on the reabsorption of bicarbonate in proximal and distal segments and found that there was a significant reduction in bicarbonate reabsorption. In proximal segments, REN promoted a significant effect at both 1 and 10 μM concentrations. Comparing control and REN concentration of 1 μM, JHCO₃⁻, nmol cm^− 2 s^− 1 − 1,76 ± 0,11_control × 1,29 ± 0,08_{REN 10 μM}; P < 0.05, was obtained. In distal segments the effect of both concentrations of REN was also effective, being significant e.g. at a concentration of 1 μM (JHCO₃⁻, nmol cm^− 2 s⁻ 1 − 0.80 ± 0.07_control × 0.60 ± 0.06_{REN 1 μM}; P < 0.05), although at a lower level than in the proximal tubule. Our results suggest that the action of REN on hydrogen transport involves the inhibition of Na⁺/H⁺exchanger and H⁺-ATPase in the luminal membrane of the perfused tubules by a PKG dependent pathway.