Acceptability of flavoured pharmaceutically non-active mini-tablets in pet cats tested with a rapid 3-portal acceptance test with and without food.

Palatable oral pharmaceuticals are crucial for feline medication. The pharmaceutical industry prefers synthetic flavours over organic ones because of hygiene and regulatory issues. The aim of this study was to find a palatable synthetic flavour for future taste-masking of feline pharmaceuticals. The hypothesis was that synthetic meat aromas and free amino acids would be palatable to cats. The palatability of 18 synthetically flavoured mini-tablets was screened with 10-19 pet cats using a rapid 3-portal acceptance test with and without food. The tested flavours were synthetic amino acids (L-carnitine, l-glutamic acid monosodium salt hydrate, l-leucine, l-methionine, l-phenylalanine, l-proline, and taurine), d-(+)-Maltose monohydrate and thiamine hydrochloride. Furthermore, thiamine hydrochloride was combined with amino acids (l-cysteine, l-leucine, l-methionine and l-proline) and synthetic meat flavours (2-acetylpyridine, 2-acetylthiazole, 2-pentylpyridine and 4-hydroxy-5-methyl-3(2H)-furanone). The negative control was a non-flavoured placebo mini-tablet, while positive controls were an organic yeast-flavoured mini-tablet and a yeast- and fish-based commercial vitamin tablet in mini-tablet form. No significant differences were detected between palatable synthetic flavours and the placebo, nor between the synthetic flavours and the yeast flavour. In general, the mini-tablet seemed to be small enough to be accepted inside a food item. These results differ from the earlier literature about the taste preferences of cats for amino acids, and hence free amino acids should not be considered palatable to cats based purely on previous findings.

Difficulties in administration of oral medication formulations to pet cats: an e-survey of cat owners.

The purpose here was to determine the problems cat owners encounter in medicating their cats with orally administered drugs at home. The study was carried out as an open e-questionnaire survey addressed to cat owners in which the authors focused on the oral administration route. A total of 46 completed questionnaires were included in the survey. In the study, 46 cats received 67 orally administered drugs. Approximately half of the drugs were registered for use in cats by the European Medicines Agency (54 per cent), and there were also off-label drugs registered for human (36 per cent) and canine medication (7.4 per cent) and an ex tempore drug (3.0 per cent). The owners were unable to give the doses as prescribed for their cats for one-fourth of the medications (16/67). Drugs that were registered for feline medication were significantly more palatable than drugs registered for other species (odds ratio (OR) 4.9), and liquid formulations were significantly more palatable than solid formulations (OR 4.8). However, most of the owners (22/38) preferred a solid dosage form, while few (4/38) chose a liquid formulation. The results indicate that there is still a need for more palatable and easily administered oral drugs for cats.

Quality and production trait genetics of farmed European whitefish, Coregonus lavaretus.

We present here phenotypic and genetic parameters for the major quality and production traits of farmed European whitefish. A total of 70 families were produced by mating each of 45 sires to an average of 1.6 dams and each of the 52 dams to an average of 1.3 sires. A total of 2,100 individuals were recorded for survival, and 507 individuals for growth and quality-related traits. The 4 major results were as follows: first, all traits exhibited nonzero heritabilities except for fillet gaping and fillet protein%. The heritabilities for the production traits were harvest weight (0.42 ± 0.10), gutted weight (0.40 ± 0.10), fillet weight (0.36 ± 0.09), maturity score (0.27 ± 0.11, on liability scale), survival (0.19 ± 0.05, on liability scale), carcass% (0.14 ± 0.07), and fillet% (0.11 ± 0.06). The heritabilities for the quality traits were condition factor (0.49 ± 0.10), fillet lipid% (0.37 ± 0.10), muscle texture (0.30 ± 0.09), Distell lipid reading (0.26 ± 0.09), fillet lightness (0.16 ± 0.07), fillet gaping (0.04 ± 0.06), and fillet protein% (0.04 ± 0.06). Second, the quality traits that were significantly genetically correlated with each other were all related to lipid deposition. Increasing fillet lipid% (an undesired change in whitefish) was genetically related to desired lighter fillet color [genetic correlation (r(G)) = 0.70 ± 0.22] and to undesired greater condition factor (0.39 ± 0.17). None of the other genetic correlations between condition factor, fillet lipid%, muscle texture, fillet lightness, fillet gaping, and fillet protein% were significant. Third, BW and gutted weight were genetically related to the quality traits that were genetically related to lipid deposition. Increasing harvest weight was genetically related to high fillet lipid% (r(G) = 0.59 ± 0.14), lighter fillet color (0.61 ± 0.25), and to greater condition factor (0.60 ± 0.12). All other genetic correlations of harvest weights with the quality traits were nonsignificant, indicating that rapid growth was not genetically related to gaping and softer flesh. Fourth, none of the genetic correlations of carcass%, fillet%, maturity, and survival with the quality traits were significant, implying weak genetic integration between the traits. Yet, marginally significant genetic correlations were found for fillet lipid% with maturity score (r(G) = -0.46 ± 0.24) and survival (0.36 ± 0.19). These results provide the genetic basis for assessing the potential to improve product quality via selective breeding.

New insights into segregation during tabletting.

The aim of this study was to evaluate how different granule size distributions affect the tablet compression process. The emphasis was on developing new analytic methods for compression data for entire batch. In all, 18 batches of granules containing theophylline and lactose were tabletted, using an instrumented eccentric tabletting machine. During tablet compression, upper and lower punch forces were recorded. Mathematical methods were developed for analysing the compression data during tabletting. The results suggested two types of undulation in the tabletting data: (1) short-time scale variation or tablet-to-tablet changes in force data and (2) long-time scale undulation describing the changes occurring throughout the tabletting process, such as segregation. These undulation phenomena were analysed, using various mathematical methods. In addition the results suggest that smaller particles have better tabletting properties, to a certain limit. However particle size alone cannot explain the tabletability of granules.

Contamination of surface run-off water and soil in two horse paddocks.

Increased stabling of horses near to cities has led to interest in the environmental effects of paddocks. In this study, the contamination of horse paddocks was examined by determining the nutrient and micro-organism contents in the surface run-off waters and the electrical conductivity, pH and phosphorus, potassium and nitrate contents of top soils. Two open-stable paddocks were studied, one cleaned and the other left uncleaned, with a stocking density of 37.5 animalsha(-1) in both. The feeding and drinking places were the most contaminated areas of both paddocks. In spring, after seven months of use, the nutrient concentrations in the surface run-off water from paddocks were 3.4-18.8mg/l for total phosphorus, 3.0-15.0mg/l for phosphate and 18.3-140.0mg/l for total nitrogen, indicating a risk to surface waters. Summer rain generated surface run-off, especially from the feeding area of the stock-free uncleaned paddock.

Process analysis of fluidized bed granulation.

This study assesses the fluidized bed granulation process for the optimization of a model formulation using in-line near-infrared (NIR) spectroscopy for moisture determination. The granulation process was analyzed using an automated granulator and optimization of the verapamil hydrochloride formulation was performed using a mixture design. The NIR setup with a fixed wavelength detector was applied for moisture measurement. Information from other process measurements, temperature difference between process inlet air and granules (T(diff)), and water content of process air (AH), was also analyzed. The application of in-line NIR provided information related to the amount of water throughout the whole granulation process. This information combined with trend charts of T(diff) and AH enabled the analysis of the different process phases. By this means, we can obtain in-line documentation from all the steps of the processing. The choice of the excipient affected the nature of the solid-water interactions; this resulted in varying process times. NIR moisture measurement combined with temperature and humidity measurements provides a tool for the control of water during fluid bed granulation.

Heat-shock protein expression is absent in the antarctic fish Trematomus bernacchii (family Nototheniidae).

The heat-shock response, the enhanced expression of one or more classes of molecular chaperones termed heat-shock proteins (hsps) in response to stress induced by high temperatures, is commonly viewed as a 'universal' characteristic of organisms. We examined the occurrence of the heat-shock response in a highly cold-adapted, stenothermal Antarctic teleost fish, Trematomus bernacchii, to determine whether this response has persisted in a lineage that has encountered very low and stable temperatures for at least the past 14-25 million years. The patterns of protein synthesis observed in in vivo metabolic labelling experiments that involved injection of (35)S-labelled methionine and cysteine into whole fish previously subjected to a heat stress of 10 degrees C yielded no evidence for synthesis of any size class of heat-shock protein. Parallel in vivo labelling experiments with isolated hepatocytes similarly showed significant amounts of protein synthesis, but no indication of enhanced expression of any class of hsp. The heavy metal cadmium, which is known to induce synthesis of hsps, also failed to alter the pattern of proteins synthesized in hepatocytes. Although stress-induced chaperones could not be detected under any of the experimental condition used, solid-phase antibody (western) analysis revealed that a constitutively expressed 70 kDa chaperone was present in this species, as predicted on the basis of requirements for chaperoning during protein synthesis. Amounts of the constitutively expressed 70 kDa chaperone increased in brain, but not in gill, during 22 days of acclimation to 5 degrees C. The apparent absence of a heat-shock response in this highly stenothermal species is interpreted as an indication that a physiological capacity observed in almost all other organisms has been lost as a result of the absence of positive selection during evolution at stable sub-zero temperatures. Whether the loss of the heat-shock response is due to dysfunctional genes for inducible hsps (loss of open reading frames or functional regulatory regions), unstable messenger RNAs, the absence of a functional heat-shock factor or some other lesion remains to be determined.

Tissue-specific expression of zebrafish (Danio rerio) heat shock factor 1 mRNAs in response to heat stress.

All organisms respond to environmental, chemical and physiological stresses by enhanced synthesis of an evolutionarily conserved family of proteins known as heat shock proteins (HSPs) or stress proteins. Certain HSPs are also expressed constitutively during cell growth and development, and they function as molecular chaperones. The transcriptional regulation of hsp genes is mediated by the heat shock transcription factor (HSF). The stress response has been studied mostly in mammalian cell lines or organisms normally maintained under constant laboratory conditions. There is much less information on the regulation of the stress response of animals, such as fish, that have to tolerate large fluctuations in environmental and internal conditions. To characterize the regulation of the heat shock response in fish, we have cloned the first heat shock transcription factor from fish, zebrafish Danio rerio. Phylogenetic analysis confirms that the isolated zebrafish HSF belongs to the HSF1 family and is therefore designated zHSF1. Analysis by reverse transcriptase polymerase chain reaction (RT-PCR) shows the presence of two zHSF1 mRNA forms that are expressed in a tissue-specific fashion upon exposure to heat stress. Both forms are expressed in gonads under all conditions; in liver and to a lesser extent in the gills, the longer splice form of zHSF1 disappears upon heat shock. We present evidence for a unique tissue-specific regulation of HSF1 upon exposure to elevated temperature.

Effects of heat shock and hypoxia on protein synthesis in rainbow trout (Oncorhynchus mykiss) cells.

We examined the effects of heat stress (from 18 degreesC to 26 degreesC) and low oxygen tension (1% O2=1 kPa) on protein synthesis in primary cultures of hepatocytes, gill epithelial cells and fibroblast-like RTG-2 cells of rainbow trout Oncorhynchus mykiss. All these cell types displayed elevated levels of 67, 69 and 92 kDa proteins, whereas a 104 kDa protein was induced only in RTG-2 cells. Hypoxia induced a cell-type-specific response, increasing the synthesis of 36, 39 and 51 kDa proteins in the gill epithelial cells. The regulation of the heat-shock response in fish hepatocytes showed that an HSF1-like factor is involved in the transcriptional induction of the hsp70 gene. Consequently, there was a pronounced accumulation of hsp70 mRNA. Furthermore, the kinetics of activation of DNA binding and the increase in hsp70 gene expression showed a remarkable correlation, indicating that hsp70 expression is regulated at the transcriptional level in these trout cells.

Haemoglobin function in intact lamprey erythrocytes: interactions with membrane function in the regulation of gas transport and acid-base balance.

Haemoglobin function within lamprey erythrocytes offers a unique solution to gas transport among vertebrates. Lamprey haemoglobin within intact erythrocytes is in oligomer/monomer equilibrium and has an oxygen affinity similar to that of haemoglobin in other active fishes. The cooperativity of oxygen binding, which is reduced at low pH values, the effect of protons and the effect of the concentration of haemoglobin on its oxygen affinity are all due to dissociation/association reactions of the haemoglobin molecules. The permeability of the lamprey red cell membrane to acid and base equivalents is very low, and plasma bicarbonate cannot therefore be dehydrated to carbon dioxide to any significant extent during the residence time of blood in the gills. This potential limitation on carbon dioxide excretion is overcome, however, by the high intraerythrocytic pH and the marked oxygenation-linked pH changes in the erythrocyte, which are due to the large Haldane effect of the haemoglobin. Owing to the relative impermeability of the erythrocyte membrane to acid equivalents, intraerythrocytic haemoglobin cannot take part in the acid-base buffering of the extracellular compartment. As a consequence, extracellular acid loads cause marked fluctuations in plasma pH.

Effect of haemoglobin concentration on the oxygen affinity of intact lamprey erythrocytes

We investigated whether the oxygen affinity of lamprey haemoglobin decreases with increasing oxygen concentration at the high (10­25 mmol l-1 monomeric) haemoglobin concentrations prevailing within the erythrocytes. The intracellular concentration of haemoglobin was experimentally adjusted by shrinking the cells osmotically: the osmolality of the equilibration medium was increased from approximately 250 mosmol kg-1 by 90 mosmol kg-1 to 340 mosmol kg-1 or by 180 mosmol kg-1 to 430 mosmol kg-1 by adding sucrose in the medium. This increased the mean cellular haemoglobin concentration from 16.9±0.23 mmol l-1 (monomeric haemoglobin) to 20.0±0.20 mmol l-1 (monomeric haemoglobin) and to 23.0±0.36 mmol l-1 (monomeric haemoglobin), respectively (means ± s.e.m., N=35­40; all the samples from 7­8 different pools of blood at each osmolality combined). The oxygen equilibrium curves at each osmolality were determined by Tucker's method. An increase in haemoglobin concentration shifted the oxygen equilibrium curve to the right as indicated by the P50 values, which were 4.26±0.07 kPa at the lowest, 4.64±0.13 kPa at the intermediate and 5.64±0.40 kPa (means ± s.e.m., N=7­8) at the highest haemoglobin concentrations. The decrease in haemoglobin oxygen-affinity was attributed to the volume changes, since the intracellular pH did not decrease with increasing mean cellular haemoglobin concentration. Thus, the variations in red blood cell volume commonly observed during hypoxia may play a role in the regulation of haemoglobin function.