Revealing time's secrets at the National Theatre of Costa Rica via innovative software for cultural heritage research.

Establishing affordable, efficient, accessible, innovative, and multidisciplinary methodologies to the diagnosis of the conservation state of an artwork is key to carry out appropriate strategies of conservation and consequently to the creation of modern public policies on cultural heritage. Limited access to large-format paintings is a challenge to restoration scientists seeking to obtain information quickly, in a non-destructive and non-invasive manner, and identify regions of interest. Therefore, we put forward two unique software tools based on multispectral imaging techniques, with the long-term aim to assess the artist's intentions, creative process, and colour palette. This development paves the way for a comprehensive and multidisciplinary understanding of the mysteries encompassed in each pictorial layer, through the study of their physical and chemical characteristics. We conducted the first ever study on Musas I and Musas II, two large-format paintings by Italian artist Carlo Ferrario, located in the National Theatre of Costa Rica. In this study, we used our novel imaging techniques to choose regions of interest in order to study sample layers; while also assessing the works' state of conservation and possible biodeterioration. We explored the applications of our two versatile software tools, RegionOfInterest and CrystalDistribution, and confirmed paint stratigraphies by means of microscopy and spectroscopy analyses (OM, SEM-EDX, Fluorescent microscopy, FTIR-ATR and micro-Raman). In a pilot study, we identified the artist's main colour palette: zinc white, lead white, chrome yellow, lead read, viridian, along with artificial vermilion and ultramarine pigments. We were able to identify artificial vermilion and ultramarine and distinguish them from the natural pigments using CrystalDistribution to map the average size and diameter of the pigment crystals within the paint layers. This study demonstrated that software-based multidisciplinary imaging techniques are novel in establishing preventive and non-invasive methods for historical painting conservation studies, in addition, this study provides tools with great potential to be used in the future in applications such as virtual restoration.

Ammoniation to reduce the toxicity of endophyte-infected tall fescue seed fed to rats.

To assess the efficacy of ammoniation in the detoxification of endophyte-infected tall fescue (Festuca arundinacea Schreb), 40 male Harlan Sprague-Dawley rats were randomly assigned to the following four treatments for 28 d: endophyte-free (E-), endophyte-infected (E+), ammoniated (2% dry matter basis, 7 d) endophyte-free (AE-), and ammoniated endophyte-infected (AE+) tall fescue seed. Total pyrrolizidine alkaloid (N-acetyl and N-formyl loline) and ergovaline contents of endophyte-infected fescue seed were reduced 24 and 54%, respectively, by ammoniation. Endophyte-infected treatment groups had lower (P < 0.01) daily feed intakes (DFI), daily weight gains (DWG), feed efficiencies, and primary serum hemagglutination titers to sheep red blood cell (SRBC) immunization than endophyte-free treatment groups. Performance parameters were higher (P < 0.01) for ammoniated diets in comparison to non-ammoniated diets; however, anti-SRBC titers were not significantly different. When compared to the E+ diet, the AE+ diet increased (P < 0.01) DFI (24%), DWG (41%) and feed efficiency (13%).

Ammoniation to reduce the toxicity of endophyte-infected tall fescue seed fed to rats.

To assess the efficacy of ammoniation in the detoxification of endophyte-infected tall fescue (Festuca arundinacea Schreb.), 40 male Harlan Sprague-Dawley rats were randomly assigned to the following four treatments for 28 d: endophyte-free (E-), endophyte-infected (E+), ammoniated (2% dry matter basis, 7 d) endophyte-free (AE-), and ammoniated endophyte-infected (AE+) tall fescue seed. Total pyrrolizidine alkaloid (N-acetyl and N-formyl loline) and ergovaline contents of endophyte-infected fescue seed were reduced 24 and 54%, respectively, by ammoniation. Endophyte-infected treatment groups had lower (P < 0.01) daily feed intakes (DFI), daily weight gains (DWG), feed effieiencies, and primary serum hemagglutination titers to sheep red blood cell (SRBC) immunization than endophyte-free treatment groups. Performance parameters were higher (P < 0.01) for ammoniated diets in comparison to non-ammoniated die [s; however, anti-SRBC titers were not significantly different. When compared to the E+ diet, the AIE+ diet increased (P < 0.01) DFI (24%), DWG (41%) and feed efficiency (13%).

In vitro and in vivo ruminal and physiological responses to endophyte-infected tall fescue.

Relationships between alkaloid compounds in endophyte-infected tall fescue and ruminal metabolism were studied in two experiments. In the first experiment, different combinations of the pyrrolizidine alkaloids, N-formyl and N-acetyl loline, were incubated with ruminal fluid for 0, 24, or 48 h. Rate of disappearance of N-formyl and N-acetyl loline increased over time. After 48 h, disappearance of N-formyl loline and combinations of N-formyl and N-acetyl loline was greater than N-acetyl loline. Significant amounts of N-formyl and N-acetyl loline were metabolized and converted to loline. In the second experiment, abomasally cannulated sheep were fed increasing amounts of endophyte-infected feed to compare diet digestibility, alkaloid metabolism, and physiological responses. Total tract DM digestibility was greatest for the endophyte-free diet, as were ruminal and total tract ADF, ruminal NDF, and total tract CP digestibilities. N-Formyl and N-acetyl loline recoveries averaged 5% from abomasal contents and 0% in feces. Sixty-eight percent of the pyrrolizidine alkaloids recovered in abomasal contents had been metabolizable to loline. Ergot alkaloids administered in the diet were recovered (50 to 60%) in the abomasal contents, but recovery was only 5% in fecal collections. No significant differences occurred in the physiological parameters measured. Results indicate that response to endophyte-infected tall fescue may be influenced by ruminal metabolism.

Bovine plasma vitamin A responses to selected nutrients, monensin, and endophytic fescue.

Previous research at this station adapted a maximal dose response (MDR) method of evaluating vitamin A status and utilization for use in beef cattle. This method was used in two experiments. In this first experiment, forty-eight crossbred steers (average weight, 284 kg) were fed diets supplemented with salt, monensin or both, and injected with vitamin E, zinc or selenium. Steers receiving monensin had higher (38.5 micrograms/dl, monensin; 31.0 micrograms/dl, controls) initial plasma concentrations of vitamin A (P = .14). However, monensin did not affect post-dosing (MDR) vitamin A concentrations. None of the other dietary treatments or injections affected either pre- or post-dosing concentrations of vitamin A in the plasma. In a second experiment 23 lactating multiparous beef cows (average weight, 500 kg) grazing either fungal endophyte-infected or endophyte-free tall fescue were used to assess possible influences of infected fescue upon vitamin A metabolism. Fungal endophyte infection did not affect either pre- (44.9 micrograms/dl, end-noninf.; 47.7 micrograms/dl, end-inf.) or post-dosing (57.2 micrograms/dl, end-noninf.; 59.3 micrograms/dl, end-inf.) vitamin A concentrations.

Influence of rumen fermentation on response to endophyte-infected tall fescue seed measured by a rat bioassay.

Possible alteration of toxicity of endophyte-infected tall fescue by ruminal fermentation was studied using 28 Harlan Sprague-Dawley rats (avg. initial wt., 141 g). These were assigned randomly to one of four treatments in a 2*2 factorial consisting of a 14-day growth period with weights and feed consumption data determined on days 0, 5, 10 and 14. Treatments were: endophyte-infected Kentucky-31 tall fescue seed (E+) or endophyte free Johnstone tall fescue seed (E-) that was incubated for 0 (NON) or 24 hours (INC) with rumen fluid collected from a 290-kg cannulated steer fed a diet containing 37% endophyte-infected tall fescue hay. Diets consisted of 50% lab chow, 39% fescue seed and 11% rumen contents (air-dry basis). Alkaloid content (N-acetyl plus N-formyl loline) for the four diets were 2540, 2680, 0, and 0 micrograms/g for E+NON, E+INC, E-NON, and E-INC diets, respectively. E- treatments gained faster, consumed more feed and converted feed more efficiently (P < .05) than did E+ fed groups. No difference in feed intake was observed within E+ treatments, however, the E+INC diet gained faster (P < .05) and converted feed more efficiently (P < .05) than did E+NON fed rats (2.56 vs 1.96 g/d and 5.94 vs 7.51 g of feed/g of gain for gain and feed conversion, respectively). Including endophyte-infected seed in rat diets depressed performance (ie. intake, gain and feed efficiency). This depression was partially alleviated by a 24-hour incubation with rumen fluid contents suggesting that toxicity of endophyte-infected tall fescue is lessened due to rumen microbial action.

Metabolism of narasin in chickens and Japanese quail.

Thirty mature chicken hens and 60 mature Japanese quail hens were used to compare pathways of narasin excretion. Carbon-14-labeled narasin was injected into chickens (.7 microCi) and quail (.113 microCi) via cardiac puncture. Blood, sampled at varying times thereafter, and eggs and excreta collected daily for 28 days, were analyzed for 14C. Groups of six chickens and 12 quail were killed prior to [14C]narasin injection and on Days 1, 7, 14, and 28 postinjection to obtain tissue samples for 14C analysis. Blood rapidly cleared the label in both species. Less than 1% of the dose of [14C]narasin remained in blood plasma after 3 h postinjection in both chickens and quail. Label excretion peaked on Day 1 in both species, and most of the 14C was cleared via the excreta (76.7 and 93.6% of the dose for quail and chickens, respectively). Label appeared in the excreta more rapidly and cleared more quickly in quail than in chickens. After 24 h, 68 and 49% of the dose of [14C]narasin appeared in the excreta of quail and chickens, respectively. More label was recovered in the eggs of quail (4.18% of the dose) than in the eggs of chickens (1.32% of the dose). Liver, heart, fat, and ovarian tissues contained traces of radioactivity 1 day postinjection in both species. Muscle and kidney did not contain detectable amounts of label. By Day 7, all tissue had cleared 14C beyond detectable limits. The results indicate that chickens and quail metabolize [14C]narasin via similar pathways and that excretion in quail may be more rapid than in chickens.

Pharmacokinetics of 4-methylimidazole in sheep.

The pharmacokinetics of 4-methylimidazole (4MI), a toxin found in ammoniated forage, was studied after i.v. infusion or oral administration of a single dose of 20 mg 4MI/kg BW to sheep. A two-compartment open model was used to describe i.v. infusion data. Oral data were described by a one-compartment open model. A rapid distribution phase (t1/2 alpha = 28 min) was observed after i.v. infusion. The biological half-lives obtained after i.v. infusion (t1/2 beta = 9.72 h) and oral dosing (t1/2 beta = 9.37 h) were similar. The bioavailability of oral 4MI was .69, with a relatively rapid absorption phase (t1/2abs = 1.52 h). The relatively large volume of distribution (61.6 and 65.8 liters for i.v. infusion and oral dosage, respectively) indicates that 4MI is distributed in the extravascular compartment. A dose of 20 mg/kg BW did not cause any apparent ill effects to the animals.

High-performance liquid chromatographic determination of 4-methylimidazole in sheep plasma and in ammoniated tall fescue hay.

A method for 4-methylimidazole (4MI) extraction and quantitation in body fluids and forage samples was developed. The procedures involve ion-pair extraction of the compound with the quantitation done by ion-pair liquid chromatography. The results indicate that this high-performance liquid chromatographic method is sensitive, reproducible and more rapid than others that have been previously used. The mean recovery of 4MI from plasma and tall fescue (Festuca arundinacea) hay samples were above 95 and 85%, respectively. The versatility of the procedure makes it suitable for the determination of 4MI in body fluids and in forage samples.

Plasma vitamin A levels in cattle in response to large doses of vitamin A.

A dose response procedure was developed for evaluating vitamin A status and utilization in cattle. This could be useful for evaluating diets, vitamin A stores and homeostatic control of vitamin A. Three experiments were designed to determine: 1) size of vitamin A dose required to increase plasma vitamin A concentrations; 2) time after dosing when plasma vitamin A concentrations peak, and 3) if changes in plasma vitamin A concentrations are useful in assessing dietary vitamin A utilization. Using twenty-four steers and heifers, the first two experiments showed that 30X or 40X daily vitamin A requirement (daily requirement calculated as X = body weight in kilograms times 55 International Units of vitamin A) given orally was sufficient to cause plasma vitamin A concentrations to rise 20 hours after dosing. Plasma vitamin A concentrations increased (P less than .01) from 57 to 81 micrograms/dl and 64 to 84 micrograms/dl after a 30X or 40X treatment, respectively. Twenty-X daily requirement resulted in a slight increase after 20 hours (P greater than .05). Seventy-two feedlot steers were used in experiment three to determine if diet affects vitamin A dose response. Variations in feed intake, monensin in the diet or a single dose of vitamin E did not cause significant changes in plasma vitamin A response to vitamin A dosage. Steers receiving monensin had higher initial plasma vitamin A concentrations, but no differences were observed after dosing. Results indicate that dose responses may be useful in assessing dietary vitamin A utilization.

Porcine immunoglobulin transfer after prepartum treatment with selenium or vitamin E.

Responses to prepartum injection of sows with Se and vitamin E (E) were evaluated by determining immunoglobulin (IgA, IgM, IgG) levels in the colostrum and serum of the sows and the serum of their offspring. Fifty-four sows (40 multiparous, 14 primiparous) receiving diets adequate in E and Se according to current NRC (1988) standards were randomly allotted to four treatment groups in which a single i.m. injection of saline (controls), 5 mg of Se, 1,000 IU of E, or both Se and E were given on d 100 of gestation. Sows were bled prior to and 7 d after injection, at farrowing and on d 14 and 28 of lactation. Colostral samples were collected at the initiation of farrowing. Pigs were bled 20 h postpartum and at 14 and 28 d of age. Major immunoglobulin changes in the serum of the sows due to treatment were not seen prior to parturition. Injections of Se and(or) E resulted in higher colostral IgM levels (8.4, 10.7, 9.8 and 9.6 mg/ml, respectively), but only the response from Se was significant (P less than .05). Concentrations of colostral IgA or IgG were not affected by treatment (P greater than .30). Compared with controls, all three treatments increased (P less than .10) IgM concentrations in serum from pigs at birth (28.3, 33.3, 36.0 and 33.5 mg/ml, respectively), whereas IgA and IgG concentrations were not affected (P greater than .30).(ABSTRACT TRUNCATED AT 250 WORDS)

Metabolism of decoquinate in chickens and Japanese quail.

Thirty mature chicken hens and 40 mature Japanese quail hens were used in an experiment to compare pathways of decoquinate (DQ) excretion. Labelled DQ was injected into chickens (.5 microCi via wing vein puncture) and quail (.25 microCi via cardiac puncture) on Day 0. Blood was sampled at 0, 1.5, 3, 6, 9, 12, 24, and 48 h postinjection. Eggs and excreta of chickens and quail were collected for 28 and 14 days, respectively, and analyzed for 14C. Six chickens and eight quail were sacrificed prior to 14C-DQ injection and also on Days 1, 7, 14, and 34 or 32 postinjection. Samples of liver, heart, kidney, bile, skin, fat, and muscle were analyzed for 14C. Blood rapidly cleared 14C in both species, and the half-time of 14C excretion via excreta was more rapid in quail (.37 day) than in chickens (.92 day). Little 14C was found in the eggs of quail (.32% of dose) and chickens (.17% of dose). Quail appeared to excrete peak amounts of detectable 14C 1 day earlier (Day 4) than chickens (Day 5). Liver contained the greatest concentration of 14C on Day 1 in both species. By the end of the experiment, less than 1% of the dose remained in liver or other organs. Results indicate that chickens and quail metabolize 14C-DQ at comparable rates and by similar pathways.

Comparative drug depletion in domestic animals and birds.

Decoquinate (Rhone-Poulenc Inc) and Narasin (Eli Lilly and Co) were selected as model drugs for a comparison of metabolism between major (cattle and chickens) and minor (sheep and quail) species. Decoquinate has been studied in all four species. Narasin studies are in progress in chickens and quail. More than 96% of injected 14C-decoquinate (DQ) was eliminated from blood of all species within 1 hr. Disappearance of the remaining 1 to 4% from blood was rapid for all species. Half-times for DQ appearance in excreta were all less than one day. Cumulative excretion of DQ in eggs of chickens and quail was about 1% for both species. Disappearance of DQ from tissues was essentially complete in 14 days. More than 80% of injected 14C-narasin was eliminated from blood within 1/2 hr. Disappearance of the remainder was rapid for both chickens and quail.

Comparative metabolism of 14C-decoquinate in chickens, quail and sheep.

Removal of pulse doses of 14C-decoquinate from blood was studied in chicken hens, quail hens, and ewe lambs. Estimated clearance before the first post-injection sample at 1 1/2 hr was 96% for sheep, 99% for chickens and over 99% for quail. Half-times for removal of the remaining radioactivity from the blood were 26.8 hr for sheep, 144.7 hr for chickens and 27.2 hr for quail. Treating the chicken data as biphasic yielded a 3.4 hr half-time for an initial fast phase and 210 hr for a later slow phase. Radioactivity disappeared from sheep blood between 2 and 7 d, from chicken blood between 7 and 14 d, and from quail blood between 1 and 2 d. Urine accounted for 35% of the radioactivity administered to sheep. It did not contain detectable activity after the third day.

Nitrogen utilization and digestibility by growing steers and goats of diets that contain monensin and low crude protein.

Two digestion and metabolism experiments were conducted to determine effects of monensin in low-protein diets. Monensin supplementation (27 mg/kg of diet dry matter) of steers (303 kg) fed 8.7% crude protein increased (P less than .01) apparent N digestibility and N retention and decreased (P less than .01) percentage of N apparently absorbed lost in urine. Apparent digestibilities of dry matter, gross energy and acid detergent lignin were increased (P less than .05). Digestibilities of neutral detergent and acid detergent fibers were not affected by monensin. In growing wether goats (15 kg) fed 8.5% crude protein, monensin (23 mg/kg of diet dry matter) improved (P less than .01) apparent N digestibility and apparent N absorption. However, urinary N excretion also increased (P less than .05), resulting in no difference in N retention. Monensin did not affect digestibilities of dry matter or gross energy. Efficiency of feed conversion and average daily gain were improved with monensin supplementation of growing goats fed a low-protein, high-roughage diet. Monensin resulted in typical shift of acetate-to-propionate ratio in both experiments. Results suggest that improved N utilization may account for some benefits of feeding monensin.

Pancreatic amylase, plasma glucose, and insulin responses to propionate or monensin in sheep.

Yearling wethers fitted with reentrant bile-pancreatic duct cannulae were in a two-part study of effects of duodenal propionate infusions or increased ruminal propionate caused by dietary monensin on pancreatic alpha-amylase secretion and glucose and insulin in blood plasma. Continuous duodenal infusion of propionate increased concentrations of glucose and insulin in blood plasma of wethers fed alfalfa. Results supported a direct response of insulin secretion to propionate. Amylase secretion was not affected. Addition of monensin (22 ppm) to an 80% corn diet reduced the ratio of acetate:propionate in rumen, but bile-pancreatic flow and amylase activity were unaffected. Monensin supplementation had little influence on glucose and insulin in blood plasma. Pancreatic alpha-amylase secretion of ruminants seems to be a complex phenomenon that is not regulated strictly by fluctuations of glucose or insulin.

Glucagon influence on gluconeogenesis and oxidation of propionic acid and threonine by perfused ovine liver.

An in situ ovine liver perfusion technique was developed and used to study glucagon effects on utilization of simultaneously infused propionic acid and amino acids. Physiological amounts of propionic acid and amino acids (hydrolyzed casein) were infused into livers along with carbon-14 propionic acid or carbon-14 threonine with and without glucagon. Glucagon (5 mg) caused a 75% increase of glucose synthesis and a 19% increase of labeled carbon dioxide production from carbon-14 propionic acid. There also was a decrease of perfusate urea nitrogen when glucagon was present. Glucagon caused a 76% decrease of carbon-14 threonine utilization by ovine livers, and labeled carbon dioxide production from carbon-14 threonine was only 38% of control when glucagon was infused. From these results, glucagon caused an increase of use of propionic acid and a decrease of use of threonine for energetic pathways in sheep liver. Therefore, glucagon directly or indirectly may mediate amino acid sparing by ruminant liver.

Utilization by growing goats of diets that contain monensin and low or excess crude protein: comparative slaughter experiment.

Monensin was added at 0 or 23 mg/kg dry matter to low (8.3%) or high (17.5%) crude protein diets (2 X 2 factorial arrangement) that were fed to 28 growing wether goats for 194 to 256 d. Average daily gain increased 24% with monensin and 20% with higher crude protein (P less than .05). Feed conversion was improved by both monensin and higher crude protein (P less than .001). During the experiment, digestion and metabolism trials were conducted twice (Periods I and II) with each goat. Apparent dry matter and gross energy digestibility coefficients were not affected by monensin in either period. Digestibility of dietary N was enhanced 7% by monensin in Period II (P less than .01) and tended to be improved in Period I. Higher dietary protein increased all digestibility coefficients (P less than .001). Nitrogen retention was not affected by monensin but was greater (P less than .001) for high-protein-fed goats. Potential changes in empty body composition were assessed by comparative slaughter balance. No effects of dietary treatments on percentage composition of empty body were noted, except a trend for higher protein in monensin-fed goats compared with controls (P less than .1). Empty body protein gain as a percentage of protein intake improved 41% with monensin and decreased 37% with higher dietary protein content (P less than .05). Monensin increased conversion of dietary energy to body ether extract energy (12%, P less than .05), to protein energy (35%, P less than .001) and to total body energy (23%, P less than .001). Magnitude of improvement tended to be greater with low than high dietary protein treatments, suggesting a protein-sparing effect of monensin as determined by comparative slaughter method. However, N retention, measured by intake-excreta balances, was not affected by monensin, suggesting that these point estimates are less than adequate to evaluate true body protein gain.

Essential fatty acid status and characteristics associated with colostrum-deprived gnotobiotic and conventional lambs. Growth, organ development, cell membrane integrity and factors associated with lower bowel function.

A factorial experiment involving gnotobiotic (GN) and conventional (CV) colostrum-deprived lambs and diets formulated to be adequate or deficient in linoleic acid was conducted to determine the effect(s) of the intestinal microflora on the essential fatty acid (EFA) status of the host and subsequent physiological consequences, i.e., growth, organ development, cell membrane integrity and lower bowel function. Lambs were obtained by sterile surgical procedures and housed in sterile isolators or in conventional metabolism stalls for 60 d. Skimmed cow's milk with 6% hydrogenated coconut oil and vitamins A, D and E added with and without .32% of the total calories as linoleic acid was homogenized, bottled and autoclaved, then fed to appetite three to four times daily. The GN lambs supplemented with linoleic acid gained significantly faster between 13 and 41 d of age and more efficiently between 27 and 41 d than the other treatment groups. The absence of dietary linoleic acid decreased liver and spleen weights and, in general, suppressed development of organs except the brain. Red blood cell hemolysis was not affected by treatment. Although showing signs of chronic mild diarrhea, the GN neonatal ruminant differed in Cl- concentration and dry matter percentage of its lower bowel contents from the "classic rodent model." The results indicate that neonatal colostrum-deprived lambs have an EFA requirement, as evidenced by decreased growth and performance characteristics in the GN linoleic deficient vs GN supplemented group, and suggests that the required level is in excess of .32% of the total caloric intake as linoleic acid.(ABSTRACT TRUNCATED AT 250 WORDS)

Acetate and propionate production in the cecum and proximal colon of lambs.

A modified single-injection isotope dilution technique measured acetate and propionate production rates in the cecum and proximal colon of 14 lambs fed a forage (hay) or high corn (concentrate) diet. The modified isotope dilution appeared to have merit. Volatile fatty acid pool sizes were stable and apparently absorption of fermentation products occurred to prevent end product inhibition. Propionate pool sizes were greater (P less than .01) while acetate pool sizes were not significantly greater when lambs received the concentrate diet. Apparent acetate production was higher (P less than .05) in lambs fed the concentrate diet than in forage-fed lambs (239.4 vs 189.5 mmol/d, respectively). Apparent propionate production was also greater (P less than .01) when lambs were fed the concentrate diet rather than the forage diet (431.4 vs 180.7 mmol/d, respectively). These results indicate that acetate and propionate produced in the cecum and proximal colon may be a considerable source of energy and glucose precursors for the growing lamb.