Increasing the dietary supply of carotenoids through forage supplementation: Effect on nitrogen and mineral retention in captive Golden pheasants (Chrysolophus pictus).

This experiment was conducted to evaluate the effects of feeding different levels of green forages on feed consumption, nutrient and mineral utilization in Golden pheasants (GP). Twenty-seven female GP (BW 617-635 g) were randomly distributed into three groups of nine birds each in an experiment based on completely randomized design (CRD). Birds in group T1 were fed a conventional zoo diet containing 1.4% green forages; however, the diets of the birds in groups T2 and T3 contained 2.7% and 5.0% of green forages, respectively. Intake of total carotenoids increased with increased level of green forages in the diet. Apparent retention of N, Ca, and Zn was higher in GP laying hens fed diet containing 5.0% green forages as compared to those fed conventional diet containing 1.4% green forages. Results of the present study indicate that inclusion of 5% green forage in the diet of GP would improve the utilization of N, Ca, and Zn without any adverse effect on intake and utilization of other nutrients. Data related to nutrient intake, basal endogenous losses (BEL) and coefficient of retention (COR) of N, Ca, P, Mg, Fe, Cu, Mn, and Zn are novel and may be of use for future research. Zoo Biol. 35:522-532, 2016. © 2016 Wiley Periodicals, Inc.

Monoterpenes as inhibitors of digestive enzymes and counter-adaptations in a specialist avian herbivore.

Many plants produce plant secondary metabolites (PSM) that inhibit digestive enzymes of herbivores, thus limiting nutrient availability. In response, some specialist herbivores have evolved digestive enzymes that are resistant to inhibition. Monoterpenes, a class of PSMs, have not been investigated with respect to the interference of specific digestive enzymes, nor have such interactions been studied in avian herbivores. We investigated this interaction in the Greater Sage-Grouse (Phasianidae: Centrocercus urophasianus), which specializes on monoterpene-rich sagebrush species (Artemisia spp.). We first measured the monoterpene concentrations in gut contents of free-ranging sage-grouse. Next, we compared the ability of seven individual monoterpenes present in sagebrush to inhibit a protein-digesting enzyme, aminopeptidase-N. We also measured the inhibitory effects of PSM extracts from two sagebrush species. Inhibition of aminopeptidase-N in sage-grouse was compared to inhibition in chickens (Gallus gallus). We predicted that sage-grouse enzymes would retain higher activity when incubated with isolated monoterpenes or sagebrush extracts than chicken enzymes. We detected unchanged monoterpenes in the gut contents of free-ranging sage-grouse. We found that three isolated oxygenated monoterpenes (borneol, camphor, and 1,8-cineole) inhibited digestive enzymes of both bird species. Camphor and 1,8-cineole inhibited enzymes from chickens more than from sage-grouse. Extracts from both species of sagebrush had similar inhibition of chicken enzymes, but did not inhibit sage-grouse enzymes. These results suggest that specific monoterpenes may limit the protein digestibility of plant material by avian herbivores. Further, this work presents additional evidence that adaptations of digestive enzymes to plant defensive compounds may be a trait of specialist herbivores.

Melatonin regulates splenocytes proliferation via IP3-dependent intracellular Ca2+ release in seasonally breeding bird, Perdicula asiatica.

Melatonin plays an important role in the immune regulation of birds. Both endogenous and exogenous melatonin modulates lymphocyte proliferation via its specific membrane receptors, Mel(1a), Mel(1b) and Mel(1c), though the mechanisms behind this process are poorly understood. We investigated the differences in melatonin membrane receptor Mel(1a), Mel(1b) and Mel(1c) expression by western blot and reverse transcription reaction and the in vitro effect of melatonin on the intracellular Ca(2+) concentration ([Ca2+]i) in splenocytes of the Indian Jungle Bush Quail, Perdicula asiatica. We used a non-selective melatonin receptor antagonist for Mel(1a) and Mel(1b), luzindole, and the selective Mel(1b) blocker, 4P-PDOT to check the specific role of melatonin receptor on ([Ca2+]i). The expression of Mel(1a), Mel(1b) and Mel(1c) receptors mRNA and protein was upregulated by melatonin (10(-7) M) with a significant high rise in ([Ca2+]i), which was differentially blocked by supplementation of antagonist, luzindole (10(-7) M) and 4P-PDOT (10(-7) M). Furthermore, we noted in vitro effect of melatonin and 2-aminoethoxydiphenyl borate (2-APB), a cell-permeable antagonist of inositol 1, 4, 5-trisphosphate (IP3) receptor to check the rise in ([Ca2+]i) through the IP3 pathway. Significantly low ([Ca2+]i) was noted in melatonin and 2-APB pretreated splenocytes when compared with splenocytes where 2-APB was absent. Thus, our data suggest that melatonin through its membrane receptor induced the elevation of ([Ca2+]i) via IP(3)-dependent pathway for splenocyte proliferation in P. asiatica.

Effect of ginger (Zingiber officinale Roscoe) on growth performance, nutrient digestibility, serum metabolites, gut morphology, and microflora of growing guinea fowl.

A 56-day feeding trial was conducted to investigate the effect of dietary supplementation of dried ginger meal (DGM) on the growth performance, nutrient digestibility, serum parameters, gut morphology, and microflora of growing helmeted guinea fowl (Numidia meleagris). One hundred sixty 28-day-old male keets were assigned to four dietary treatments. There were 40 birds per treatment replicated four times with 10 keets each. The experimental diets consisted of the basal diet (control), ginger-supplemented diets containing 20, 40, and 60 g/kg DGM, respectively. Guinea fowls fed diet supplemented with DGM had higher (P <0.05) final body weight, body weight gain and lower (P <0.05) feed intake. Optimum weight gain was obtained with supplementation level of 36.15-g DGM (R (2) = 0.923). Guinea fowls fed diet supplemented with 40 g/kg DGM recorded the highest (P <0.05) coefficient of total tract apparent digestibility of dry matter, ether extract and longest (P <0.05) duodenal and ileal villus heights. The crypt depth of the duodenum and ileum reduced (P <0.05) with increasing level of dietary supplementation of DGM. Dietary supplementation of DGM resulted in increased (P <0.05) total serum protein, serum albumin and low (P <0.05) serum cholesterol concentration. Ileum content of birds fed diet supplemented with 40 g/kg DGM recorded the highest (P <0.05) lactobacillus count. In conclusion, dietary inclusion of 40-g DGM per kilogram diet is hereby recommended for improved growth, apparent nutrient digestibility, gut morphology, serum chemistry, and stimulation of balanced intestinal microflora in growing guinea fowl.

Effects of dietary selenium supplementation on tissue selenium distribution and glutathione peroxidase activity in Chinese Ring necked Pheasants.

The objective of this study was to determine the concentration of total selenium (Se) and the proportions of total Se comprised as selenomethionine (SeMet) and selenocysteine (SeCys) in the postmortem tissues of female pheasants (Phasianus Colchicus Torquator) offered diets that contained graded additions of selenised-enriched yeast (SY) or a single comparative dose of sodium selenite (SS). Thiobarbituric acid reactive substances (TBARS) and tissue glutathione peroxidase (GSH-Px) activity of breast (Pectoralis Major) were assessed at 0 and 5 days postmortem. A total of 216 female pheasant chicks were enrolled into the study. Twenty-four birds were euthanased at the start of the study, and samples of blood, breast muscle, leg muscle (M. Peroneus Longus and M. Gastrocnemius), heart, liver, kidney and gizzard were collected for determination of total Se. Remaining birds were blocked by live weight and randomly allocated to one of four dietary treatments (n = 48 birds/treatment) that either differed in Se source (SY v. SS) or dose (control (0.17 mg total Se/kg), SY-L and SS-L (0.3 mg/kg total Se as SY and SS, respectively) and SY-H (0.45 mg total Se/kg)). Following 42 and 91 days of treatment, 24 birds per treatment were euthanased, and samples of blood, breast muscle, leg muscle, heart, liver, kidney and gizzard were retained for determination of total Se and the proportion of total Se comprised as SeMet or SeCys. Whole blood GSH-Px activity was determined at each time point. Tissue GSH-Px activity and TBARS were determined in breast tissue at the end of the study. There were increases in both blood and tissues to the graded addition of SY to the diet (P < 0.001), but the same responses were not apparent with the blood and tissues of selenite-supplemented birds receiving a comparable dose (SY-L v. SS-L). Although there were differences between tissue types in the distribution of SeMet and SeCys, there were few differences between treatments. There were effects of treatment on erythrocyte GSH-Px activity (P = 0.012) with values being higher in treatments SY-H and SS-L when compared with the negative control and treatment SY-L. There were no effects of treatment on tissue GSH-Px activity, which is reflected in the overall lack of any treatment effects on TBARS.

Identification of carotenoid pigments and their fatty acid esters in an avian integument combining HPLC-DAD and LC-MS analyses.

Yellow-orange-red ornaments present in the integuments (feathers, bare parts) of birds are often produced by carotenoid pigments and may serve to signal the quality of the bearer. Although carotenoid esterification in tissues is a common phenomenon, most of the work on avian carotenoids has been focused on the identification of free forms or have been done after sample saponification. Here we determined free and esterified carotenoid composition in a bird species with red ornaments: the red-legged partridge (Alectoris rufa). Carotenoids from leg integument were extracted and processed by TLC to separate three major carotenoid groups (free form, mono- and diesters with fatty acids), whereas saponified extracts gave only free forms of carotenoids. TLC fractions were then analyzed by HPLC-DAD with C18 phase column for a preliminary identification of carotenoid groups. The final characterization of free carotenoids and its esters with fatty acids was performed with direct extracts analyzed by LC-MS and LC-MS/MS with a C30 phase, always with a system coupled to DAD. The main carotenoid (λ(max) 478 nm and [M+H](+) at m/z 597.2) was identified as astaxanthin by comparison with standards. A second carotenoid (λ(max) between 440 and 480 nm and [M+H](+) at m/z 581.3) was not identified among any of the commercially available carotenoid standards, although it could correspond to pectenolone according to its fragmentation pattern. Both the unidentified carotenoid and astaxanthin formed monoesters with fatty acids, but only astaxanthin was in its diesterified form. Monoesters were mainly formed with palmitic, stearic, oleic and linoleic acids. Complementary analyses of fatty acid composition in partridge integument by GC-MS revealed high amounts of these and other fatty acids, such as myristic, arachidic and docosanoic acids. The combination of HPLC-DAD and LC-MS/MS spectra was especially useful to identify the carotenoids present in the esterified forms and the probable masses of the fatty acids included in them, respectively.

Selective fatty acid mobilization from adipose tissues of the pheasant (Phasianus colchicus mongolicus) during food deprivation.

Avian response to fasting has been examined intensively in penguins (Aptenodytes spp.) adapted to long-term food deprivation but less in species experiencing shorter fasts. Thus, the selectivity in (i) incorporating different fatty acids (FA) from diet into total lipids of white adipose tissue (WAT) and liver and (ii) mobilizing FA from these tissues was examined in pheasants Phasianus colchicus mongolicus fed or fasted for 4 d. Dietary FA were selectively incorporated into intra-abdominal and subcutaneous WAT having a similar composition. The WAT lipids contained higher proportions of saturated and monounsaturated FA and less polyunsaturated FA (PUFA) than the dietary profile. However, the isomers of 20:1 and 22:1 were incorporated inefficiently into the WAT lipids. The essential C18 PUFA precursors having smaller percentages in the pheasant tissues than in the diet were likely converted into longer-chain derivatives probably utilized to a great extent for structural lipids of muscles and organs. During food deprivation, the pheasants preferentially utilized 16:1n-7, 18:3n-3, 18:1n-9, and 16:0 but preserved long-chain saturated and unsaturated FA. Mobilization was more efficient for shorter-chain FA and increased with Delta9-desaturation. The hepatic FA profile was resistant to the 4-d period of food deprivation. The results demonstrate that the incorporation of FA into WAT and their mobilization from lipid stores are selective not only in mammals but also in birds.

Seasonal variations in the crop contents of scavenging Helmeted Guinea Fowls (Numida meleagris, L.) in Parakou (Benin).

1. An experiment was carried out with 120 helmeted guinea fowls during one year in Parakou (Benin). Feed intake, ingredient and chemical composition, along with the nutritional adequacy of scavenging diets were measured during the rainy season (November-February) and dry season (March-October) in order to propose supplementation strategies. Ingredients found in crops were identified and allocated into 6 main categories (supplemental feed, seeds, green forages, animal materials, mineral matter and unidentified materials). 2. Mean dry weights of crop contents were significantly higher in the rainy than in the dry season. Amounts and proportions of supplemental feed and seeds were not significantly different between seasons, whereas those of green forage, animal materials and mineral matter were higher in rainy season. Supplemental feed, especially maize and sorghum, was the largest component of the crop content in both seasons. The most represented grass seeds were Panicum maximum (rainy season) and Rottboellia cochinchinensis (dry season). 3. Dietary concentrations of organic matter, non-nitrogen extract and metabolisable energy were higher in the dry season, while mineral concentrations were higher in the rainy season. There were no significant differences between the two seasons in dry matter, crude protein or crude fibre. 4. Scavenging provided insufficient nutrients and energy to allow guinea fowls to be productive. Therefore, more nutritionally balanced supplementary feed would be required during both seasons.

Effect of enzyme supplementation on the metabolisable energy content of solvent-extracted rapeseed and sunflower seed meals for chicken, guinea fowl and quail.

(1) The nitrogen-corrected apparent metabolisable energy (AME(N)) content of solvent-extracted rapeseed and sunflower seed (un-decorticated) meals in relation to species (chicken, guinea fowl and quail) and dietary addition of feed enzymes (0 or 0.5 g/kg diet) was evaluated by a diet replacement method in a 3 x 2 factorial design. (2) The metabolism trial was conducted at two substitution levels (200 and 400 g/kg diet) of each meal with or without supplementation of commercial enzyme preparation in 6 individuals or 6 groups of cockerels, guinea fowls and quails. (3) The experimental diets were fed for a period of 12 d followed by a 3-d collection period during which total feed consumed and droppings output were quantitatively recorded. (4) The AME(N) values of rapeseed meal for cockerels, guinea fowls and quails were 8.4, 8.7 and 8.8 MJ/kg, respectively, while the corresponding values for sunflower seed meal were 6.1, 6.1 and 6.2 MJ/kg dry matter, without enzyme supplementation. (5) The AME(N) value of rapeseed meal did not improve with enzyme supplementation. However, AME(N) values of sunflower seed meal significantly increased with enzyme supplementation, from 6.1 to 6.5 MJ/kg dry matter. (6) Since AME(N) values of rapeseed meal and sunflower seed meal were similar in chicken, guinea fowl and quail, values reported for chicken could, therefore, be used for guinea fowl and Japanese quail.

The effects of fasting and refeeding on biochemical parameters in the red-legged partridge (Alectoris rufa).

Every year, about three million farm-reared red-legged partridges are released in Spain for hunting purposes, most of them for restocking where the success is very low. Some factors such as the sudden shift from abundant commercial energy-rich diet to natural food and absence of experience at looking for natural food resources could cause a rapid physical deterioration, being more vulnerable to predation and disease. To determine fasting capacity in red-legged partridges, we fasted 32 birds during 4 days and then we refed them, collecting blood samples and body masses during the experiment. Red-legged partridges entered in the third critic phase of fasting, in which body proteins became the main energetic source, after 48 h of fasting. The fasting period caused an average loss of 17% of their initial body mass and only attained a 93.6% of their initial body mass after 24 days of refeeding. The limited capacity to resist fasting in the red-legged partridge could be one of the reasons for the low success in restocking, mainly in those where neither supplementary feeding nor acclimation period is allowed for birds before they are finally released.