Daily energy expenditure and water turnover in two breeds of laying hens kept in floor housing.

Laying hens are increasingly kept in barn or free-range systems, which not only allows birds to move freely but also potentially entails higher energy expenditures due to higher locomotor activity. Therefore, the aim of our study was to quantify the daily energy expenditure (DEE) and water turnover in freely moving laying hens. For that purpose, 10 Lohmann Selected Leghorn (LSL) and 10 Lohmann Brown (LB) hens were obtained from a conventional breeding company at 17 weeks of age. The trial started when birds reached an age of 34 weeks. All 20 birds were kept together in the same littered floor pen (12.1 m2). The pen was equipped with perches, a nest box, feeding and nipple drinkers. The DEE was determined individually for all experimental birds (n = 20) for a total of nine days using the doubly labelled water (DLW) method. Lohmann Brown hens were heavier than LSL hens, but laying rate did not differ between the two breeds, that is, one egg per hen and day during the study period. Average egg mass was 63.1 ±â€¯0.20 g in LB and 61.7 ±â€¯0.12 g in LSL hens, which converted to an egg energy content of 420 and 410 kJ/egg, respectively. Dilution spaces for oxygen and hydrogen differed between the breeds but not the respective turnover rates. Total body water as a percentage of body mass (LB: 54.4%, LSL: 53.8%; SEM = 0.7, F1,18 = 0.41, P = 0.513) and total water intake (TWI) per day (LB: 275 ml/day, LSL: 276 ml/day; SEM = 20, F1,17 = 0, P = 0.994) did not differ between LB and LSL hens. Individual DEE increased with body mass in LB but not in LSL hens. Average DEE did not differ between the two breeds (LB: 1501 kJ/day; LSL: 1520 kJ/day; SEM = 32.1, F1,17 = 2.54, P = 0.131). However, when comparing the DEE on a metabolic mass basis, LSL hens expended with 984 kJ/kg0.75 on average significantly more energy per day than LB hens (895 kJ/kg0.75; SEM = 20.3, F1,18 = 10.1, P = 0.005). Our results suggest that the DLW technique is a viable method to measure the energy expenditure and water turnover over several days in laying hens. Furthermore, we show that laying hens kept in floor pens fit into the general pattern of DEE among wild birds.

Macrophage secretion of miR-106b-5p causes renin-dependent hypertension.

Myeloid cells are known mediators of hypertension, but their role in initiating renin-induced hypertension has not been studied. Vitamin D deficiency causes pro-inflammatory macrophage infiltration in metabolic tissues and is linked to renin-mediated hypertension. We tested the hypothesis that impaired vitamin D signaling in macrophages causes hypertension using conditional knockout of the myeloid vitamin D receptor in mice (KODMAC). These mice develop renin-dependent hypertension due to macrophage infiltration of the vasculature and direct activation of renal juxtaglomerular (JG) cell renin production. Induction of endoplasmic reticulum stress in knockout macrophages increases miR-106b-5p secretion, which stimulates JG cell renin production via repression of transcription factors E2f1 and Pde3b. Moreover, in wild-type recipient mice of KODMAC/miR106b-/- bone marrow, knockout of miR-106b-5p prevents the hypertension and JG cell renin production induced by KODMAC macrophages, suggesting myeloid-specific, miR-106b-5p-dependent effects. These findings confirm macrophage miR-106b-5p secretion from impaired vitamin D receptor signaling causes inflammation-induced hypertension.

Adaptation capacity of Boer goats to saline drinking water.

Due to global climatic changes, water and soil salinization is an increasing worldwide phenomenon, thus creating new threats for farm animal production. The present study was designed to investigate the adaptation capacity of goats towards sodium chloride (NaCl) in drinking water. Twelve non-pregnant Boer goats with an average body weight of 50.5 ± 9.0 kg were kept in individual pens. The study was conducted in four phases applying a two-choice preference test. In the control phase (phase 1) only fresh water was supplied in two containers. In phase 2, water with different salt concentrations (0.25%, 0.5%, 0.75%, 1.0%, 1.25% and 1.5%) was offered in one container and tap water in the other (sensitivity test). During the third phase (adaptation), goats were stepwise habituated to saline water by offering only saline water with different increasing concentrations (between 0% and 1.5% NaCl) in both containers. Subsequently, in phase 4 (sensitivity re-test) the same treatment as in phase 2 was repeated. Goats had ad libitum access to hay, water and a mineral licking block. Individual water and feed intake were recorded daily, while body weight and body condition score were measured every 2nd week. Body weight was not affected by saline water intake, whereas dry matter intake and body condition scores decreased significantly during the experiment. Water intake was significantly (P<0.001) higher in phase 2 (sensitivity test) and phase 3 (adaptation), compared to phase 1 (control) and phase 4 (sensitivity re-test). Total sodium intake followed the same pattern. In phase 2, when goats had the choice between fresh and saline water for the first time they preferred higher salt concentrations and consumed significantly (P<0.001) higher amounts of saline water (75.4 ± 53.2 g/kg BW0.82 per day) than in the re-test (40.4 ± 34.0 g/kg BW0.82 per day) after the habituation period. Thus, salt discrimination rejection thresholds were lowered to 1.25% in phase 4 compared to 1.5% in phase 2. The results suggest that a stepwise adaptation to saline drinking water in goats is an effective method to habituate the animals to saline water intake when concentrations were below 1.5%. Goats reacted more sensitively to the salinity of drinking water after prolonged exposure to saline water indicating flexible regulation mechanisms depending on the total sodium balance of the animal.

Reactions to saline drinking water in Boer goats in a free-choice system.

Salinization of groundwater and soil is a prevalent global issue with serious consequences on animal health and production. The present study was conducted to investigate the capacity of Boer goats to adjust their salt intake from saline drinking water in a free-choice system. In total, 12 non-pregnant Boer goats aged between 1 and 8 years with an average BW of 46.4±8.3 kg were kept in individual pens for 4 weeks. In the control phase (1 week), only fresh water was supplied in five identical buckets for each pen. During the subsequent treatment phase (3 weeks), fresh water and four different concentrations (0.75, 1.0, 1.25 and 1.5% NaCl) of saline water were offered simultaneously in a free-choice system. The positions of the concentrations were changed daily at random. Cut hay and water were provided ad libitum, and a mineral supplement was allocated. Feed and water intake, mineral supplement intake, ambient temperature and relative humidity were recorded daily, whereas BW and body condition score were measured weekly. Dry matter intake, total water intake and total sodium intake were significantly (P<0.001) higher during the treatment phase. Body weight and body condition were not affected by saline water intake. Across the treatment phase, saline water consumption was significantly (P<0.001) lower in young (19.6±27.1 g/kg BW0.82 per day) than in adult goats (27.9±31.5 g/kg BW0.82 per day), indicating that young goats were more sensitive towards the saline water. All goats had a significant preference for fresh water (0% salt) over saline water. At the first offering of the simultaneous choice situation (week 2), animals did not differentiate between the salt concentration of 0.75% and 1.0%. However, with successive treatment (weeks 3 and 4), animals distinguished between saline water concentrations and preferred the 0.75% salt concentration. Salt concentrations of 1% to 1.5% were avoided. The total sodium intake of the goats ranged between 0.37 and 0.55 g /kg BW0.75 per day during the treatment phase, being 8- to 11-fold higher than the daily requirements of sodium for body maintenance. The results suggest that goats are able to differentiate between saline water concentrations and to adjust their sodium intake by quick adjustments in self-selection in a free-choice system. Compared with two-choice preference tests, the present free-choice situation allows evaluating changes in saline water acceptance with prolonged exposure.

Long-term adaptation capacity of ponies: effect of season and feed restriction on blood and physiological parameters.

Domesticated horses are increasingly kept under semi-natural housing conditions, whereas their adaptation capacity is not fully investigated. In all, 10 Shetland pony mares were held under semi-extensive conditions for 1 year. In winter animals were allocated into two feeding groups (60% and 100% of maintenance energy requirement, respectively). Triiodothyronine, thyroxine, non-esterified fatty acids (NEFA), total bilirubin, total protein, triglyceride, glucose, insulin and hair length were measured at monthly intervals, whereas BW, body condition score, cresty neck score and resting heart rate were recorded every 2 weeks. From summer to winter all Ponies showed a reduction in resting heart rate (P0.05). Refeeding of restrictively fed ponies resulted in a rapid increase in resting heart rate and BW and a return of blood parameters to reference values. Adequately supplied animals adapted without difficulty to varying environmental conditions, whereas feed restriction in ponies during winter resulted in reduced resting heart rates suggesting a reduced basal metabolic rate. The energy restriction was compensated by mobilizing body fat reserves which led to changes in blood parameters. Refeeding in feed restricted animals revealed a remarkably quick recovery of physiological and blood parameters to reference values. We therefore suggest that year round-outdoor housing can be a suitable housing system for robust horse breeds provided that an adequate food supply is available.

Seasonal changes of total body water and water intake in Shetland ponies measured by an isotope dilution technique.

Water is an essential nutrient necessary to support life, and adequate water supply is crucial for animal survival and productivity. The present study was designed to determine seasonal changes in the water metabolism of horses under outdoor conditions. Total body water (TBW) and total water intake (TWI) of 10 adult Shetland pony mares were estimated at monthly intervals for 14 mo by using the deuterium dilution technique. During the last 4 mo, 5 ponies were fed restrictively to simulate natural feed shortage in winter, and 5 ponies served as controls. The TBW (kg) was closely related to body mass [TBW (kg) = -2.86 + 0.67 × body mass (kg); P < 0.001; n = 105] explaining 86% of the variation. In contrast to TBW (kg), TBW (%) remained relatively stable across all measurements (57.8 to 71.2%). The TWI showed an increase in summer and a decrease in winter [TWI (mL·kg(-1)·d(-1)) = 15.07 + 23.69 × month - 1.45 × month(2) (R(2) = 0.64, P < 0.01)]. However, TWI measured at ambient temperatures (Ta) < 0°C did not follow the same trend as TWI at Ta > 0°C. Therefore, removing TWI values measured at Ta < 0°C from the analysis resulted in high correlations with locomotor activity (r = 0.87), Ta (r = 0.86), and resting heart rate (r = 0.88). The multiple regression among TWI, Ta, and heart rate explained 84% of the variation in TWI [TWI (mL·kg(-1)·d(-1)) = -13.38 + 1.77 × heart rate (beats/min) + 2.11 × Ta (°C); P < 0.001]. Feed restriction had no effect on TWI and TBW. The TBW content was unaffected by season and physical activity. The established regression equation for TBW and body mass can be used to predict TBW from body mass in ponies under field conditions. The comparison of TWI with published data on drinking water intake revealed that ponies had 1.7 to 5.1 times greater total water intakes when other sources of water such as feed and metabolic water were included. The TWI was highly influenced by environmental conditions and metabolic rate. Contrary to expectation, water supply during the cold seasons might be more critical than under summer conditions when water content of grass is high to allow for the compensation of limited availability of drinking water.

Short communication: milk output in llamas (Lama glama) in relation to energy intake and water turnover measured by an isotope dilution technique.

Despite the fact that llamas have become increasingly popular as companion and farm animals in both Europe and North America, scientific knowledge on their nutrient requirements is scarce. Compared with other livestock species, relatively little is known especially about the nutrient and energy requirements for lactating llamas. Therefore, we aimed to measure milk output in llama dams using an isotope dilution technique and relate it to energy intakes at different stages of lactation. We also validated the dilution technique by measuring total water turnover (TWT) directly and comparing it with values estimated by the isotope dilution technique. Our study involved 5 lactating llama dams and their suckling young. Milk output and TWT were measured at 4 stages of lactation (wk 3, 10, 18, and 26 postpartum). The method involved the application of the stable hydrogen isotope deuterium ((2)H) to the lactating dam. Drinking water intake and TWT decreased significantly with lactation stage, whether estimated by the isotope dilution technique or calculated from drinking water and water ingested from feeds. In contrast, lactation stage had no effect on dry matter intake, metabolizable energy (ME) intake, or the milk water fraction (i.e., the ratio between milk water excreted and TWT). The ratios between TWT measured and TWT estimated (by isotope dilution) did not differ with lactation stage and were close to 100% in all measurement weeks, indicating that the D(2)O dilution technique estimated TWT with high accuracy and only small variations. Calculating the required ME intakes for lactation from milk output data and gross energy content of milk revealed that, with increasing lactation stage, ME requirements per day for lactation decreased but remained constant per kilogram of milk output. Total measured ME intakes at different stages of lactation were similar to calculated ME intakes from published recommendation models for llamas.

Effect of water restriction on drinking behaviour and water intake in German black-head mutton sheep and Boer goats.

The present study was conducted to evaluate and compare the physiological responses of sheep and goats to water restriction using the deuterium dilution technique (D2O) to predict the total water intake (TWI) in both species. In two consecutive trials, 10 non-lactating Boer goats and 10 non-lactating German black-head mutton ewes were each randomly allocated into a treatment and a control group. In the control groups (n = 5, for each species), water was offered ad libitum, whereas the treatment groups (n = 5, for each species) received water 3 h/day on experimental days 8 to 14 and 6 h every 2 days on experimental days 15 to 22. The respiratory rate, rectal temperature, body mass and drinking behaviour were also recorded. The TWI was estimated by D2O for each animal. Water restriction for 21 h/day or 42 h/2 days had no significant (P > 0.05) effect on water intake (WI), feed intake, WI to dry matter intake ratio or body mass in both species. The absence of differences between species in their WIs was also confirmed using D2O. However, sheep had higher respiratory rates and rectal temperatures than goats in both control and treatment groups. Both species showed the ability to tolerate a moderate water shortage by activating several physiological mechanisms and behavioural strategies.

Effect of shearing on water turnover and thermobiological variables in German Blackhead mutton sheep.

Current equations for estimating water requirements in sheep do not differentiate between shorn and unshorn sheep. Furthermore, the effect of shearing on thermoregulative responses in sheep has not been adequately studied under temperate environmental conditions. Therefore, the present study was conducted to investigate the effect of wool coverage on water turnover in relation to thermoregulation in sheep by using the deuterium dilution technique to predict total water intake before and after shearing. Physiological responses, such as water turnover, surface temperature, and rectal temperature, as well as drinking behavior of sheep were also evaluated. Fourteen nonlactating German Blackhead mutton ewes were randomly allocated into 2 groups: a control group (n = 7) that was already shorn, and a treatment group (n = 7) that was left unshorn (wool length: 10.6 ± 1.2 cm). Individual feed and water intakes were recorded throughout the experiment (d 1 to 71). Two weeks after measurements commenced (d 15), treatment sheep were shorn. Water intake was estimated twice for 2 consecutive weeks by using deuterium dilution techniques (d 1 to 15 and d 57 to 71). Ambient temperature (T(a)), relative humidity, and respiratory rate were measured daily, whereas BW, rectal and animal surface temperatures (using infrared thermography), and wool length were measured weekly. In the first 2 wk, when treatment sheep were unshorn, treatment and control ewes differed (P < 0.05) in DMI (52 ± 4 vs. 59 ± 4 g·kg(-0.75)·d(-1)), water intake (165 ± 17 vs. 134 ± 18 g·kg(-0.75)·d(-1)), respiratory rate (66 ± 5 vs. 31 ± 4 breath/min), rectal temperature (39.3 ± 0.2 vs. 38.8 ± 0.1°C), and surface temperatures (body side: 19.3 ± 0.3 vs. 24.5 ± 0.6°C; leg: 25.8 ± 2.4 vs. 27.4 ± 1.6°C). However, after shearing, these differences partly disappeared. The same trend in water intake between groups was confirmed using the isotope dilution technique. We found a significant relationship between T(a) and water intake, respiratory rate, and body surface temperatures. Even under temperate conditions (T(a) < 28°C), shearing significantly reduced core body temperature, water intake, and respiratory rate in German Blackhead mutton sheep, thus indicating heat stress in fleeced animals, which should be considered when determining the optimal shearing time in sheep as well as when estimating water requirements.

Deuterium oxide dilution accurately predicts water intake in sheep and goats.

The aim of this study was to test whether the deuterium oxide dilution technique accurately predicts water intake in sheep and goats. Two other issues were also studied: (i) a comparison of water intake in sheep and goats and (ii) an assessment of whether observations of drinking behaviour can accurately measure the water intake. In this study, eight dry Boer goats and eight dry German Black Head Mutton ewes were kept under controlled stable conditions. Animals had access to hay and water ad libitum. Diurnal drinking behaviour was recorded by video. Individual daily water intake was measured and estimated for 2 weeks by re-weighing water buckets and from water kinetics using the deuterium oxide dilution technique, respectively. In addition, dry matter intakes were directly measured and were significantly higher in sheep than in goats. The average daily water consumption by drinking differed significantly between the two species, with higher intakes in sheep than in goats. Total body water expressed as a percentage of body mass did not differ between species. Measurement methods of total water intake (TWI) using deuterium oxide dilution and re-weighing water buckets did not differ significantly in both species (P = 0.926). Results obtained for measured and estimated TWI confirm that the isotope dilution technique gives reliable results for estimates of water intake in sheep and goats. The higher amount of water intake in sheep was also reflected by their drinking behaviour. Sheep spent approximately 0.3% per 24-h drinking, while Boer goats spent only 0.1%. However, measured and estimated TWIs were only moderately correlated to the daily time spent drinking. The lower water intake found in Boer goats confirms a superior water management capacity compared with Black Head Mutton sheep even under temperate conditions.

Deuterium for estimating total body water and turnover rates in turkeys exposed to different incubation treatments.

Total water intake (TWI) in poultry can be influenced by various factors. Recommendations for water requirements are usually reported on a flock basis without considering individual variation. In the present study, a total of 18 turkeys were used to measure water intake over a 1-wk period starting at 15 wk of age by applying the deuterium dilution technique. Poults originated from eggs exposed to different incubation treatments, with eggs incubated at normal temperature (37.5 degrees C) and eggs subjected to 38.5 degrees C at embryonic d 9 to 12. Experimental birds were kept in flocks of 22 to 30 birds separated by sex and treatment. Feed and water were provided ad libitum. Incubation treatment had no significant effect on any of the parameters investigated (BW, daily gain, water turnover rate, total body water, TWI), whereas sex exerted a significant effect on nearly all traits. Total body water ranged between 60 and 65% of BW, with significantly (P < 0.05) greater values for toms (63.2%) than for hens (60.9%). Males had approximately 30% greater water influxes than females (1,054 +/- 198 vs. 742 +/- 153 mL/d, mean +/- SD). However, the significant influence of sex was eliminated (P = 0.464) when TWI was expressed as grams per kilogram of BW (76 +/- 18 vs. 70 +/- 12 mL/kg of BW; males vs. females). Water consumed averaged 837 mL in male and 569 mL per day in female birds. The present results suggest that the isotope dilution method offers a viable method to measure individual water intake, which can be used for establishing reference values for water consumption in group-housed turkeys.

Measurements of the bodyweight and other physical characteristics of 11 llamas (Lama glama) from birth to weaning.

The bodyweight (bw), thoracic circumference, length and height of 11 llamas were recorded weekly from birth for 27 weeks. Their mean (sd) birth weight was 11.60 (2.35) kg and their weight increased in curvilinear fashion reaching 61.05 (13.75) kg after 27 weeks. Their average daily gains reached the highest value three weeks after birth and then decreased, reaching the lowest value of 262 g per day at 27 weeks. On a metabolic bw(0.83) basis, the growth of the llamas when their dams were at peak lactation was comparable with that of calves, fawns and lambs. All the measurements were significantly affected by the age, but not by the sex of the young or the parity of the dam. The regression models chosen to predict the bodyweight and weight gain of the llamas yielded R(2) ranging from 0.97 to 0.99. Correlations between bodyweight and other single body measurements were between r=0.68 and 0.77. Single or combined body measurements were good predictors of bodyweight, but additional body measurements added little to the already good fit provided by a single measurement (R(2) ranging from 0.93 to 0.98). The thoracic circumference (R(2)=0.98) and length (R(2)=0.96) were good predictors of the bodyweight of the llamas between birth and 27 weeks old.

Measurement of milk intake in suckling llamas (Lama glama) using deuterium oxide dilution.

The objective of the study was to estimate daily milk intake in llama crias and relate nutrient intakes at peak lactation to growth data. Milk intake in 11 suckling llamas was estimated from water kinetics using deuterium oxide (D2O) at d 17, 66, and 128 postpartum. Daily milk intakes averaged 2.6, 2.3, and 2.0 kg at 17, 66, and 128 d postpartum, respectively. Milk intake decreased with age when expressed as daily amount, percentage of body weight (BW), or per kilogram of metabolic size, but the influence of age was eliminated when expressed per gram of daily gain. Because llamas only have one young per parturition, milk intake was equivalent to the daily milk output of the dam, which ranged from 27.6 to 96.9 g/kg of maternal BW(0.75). Compared with different ruminant species, milk production in llamas appears to lie between wild and domestic ruminants used for meat production. Nutrients (dry matter, fat, protein, and lactose) and energy intakes from the milk calculated by combining milk intake and milk composition data decreased with age when expressed as daily amount or per 100 g of BW, but when expressed per gram of daily gain, no clear trend was observed. Maintenance requirement for suckling llamas at peak lactation (17 d postpartum) was 312 kJ of ME/kg of BW(0.83). Combined with milk composition data, the present milk intake estimations at different stages of the lactation can be used to establish recommendations for nutrient and energy requirements of suckling llamas.

Changes in Llama (Lama glama) milk composition during lactation.

Milk samples were collected weekly from 10 llamas during the first 27 wk after parturition under controlled stable conditions. Mean values for the concentrations of the major milk components across the lactation period were 4.70% fat, 4.23% protein, 5.93% lactose, 15.61% dry matter, and 22.62 mg/dL of milk urea N. All constituents were affected by the stage of lactation. There was an increase in fat to protein ratio as protein concentration declined and fat concentration increased. Fat, protein, and lactose concentrations changed during the transition from colostrum to milk. In the first month postpartum, fat concentration remained constant, protein decreased, and lactose increased. Starting with wk 5 postpartum, fat and protein increased and lactose decreased until the end of lactation. Among the major constituents fat had the highest variation. The mean gross energy concentration of milk was 3.88 kJ/g and showed a similar course as protein. Fat contributed 48.0%, protein 26.3%, and lactose 25.7% to the gross energy in the milk. Milk urea N values were higher than those found in ruminants and increased with stage of lactation, whereas the pH decreased. The analyzed milk components were not affected by the lactation number of the animal, except milk urea N. Somatic cell counts indicated the absence of mastitis and revealed that the average somatic cell count of uninfected llamas is lower than in animals usually used for milk production. The 2 algebraic models fitted by a nonlinear regression procedure to the data resulted in suitable prediction curves for the constituents (R2 = 0.76 to 0.94). The courses of major milk constituents in llamas during lactation are similar to those in domesticated ruminants, although different in their values. The established curves facilitate the composition of milk replacers at different stages of lactation for nursing llamas whose dams died or are agalactic.

A kinetic study of immobilized lipase catalysing the synthesis of isoamyl acetate by transesterification in n-hexane.

Isoamyl acetate was synthesized by lipase-catalyzed transesterification of ethyl acetate in n-hexane. The selectivity and rates of ester formation decreased when water content of the immobilized enzyme exceeded 3% (w/w). Experimental observations clearly indicate that the substrates as well as the product (ethanol) act as dead-end inhibitors. A ping-pong bi-bi mechanism with competitive inhibition by substrates and products is proposed that predicts the experimental observation satisfactorily.