Food and water intake, growth, and adiposity of Sprague-Dawley rats with diet board for 24 months.

Ad libitum (AL) feeding of rats leads to obesity and increased result variability, as well as premature morbidity and mortality. It may also alter metabolism and responses to foreign compounds. Moderate dietary restriction (DR) reduces these untoward effects without compromising the sensitivity of rodent bioassays. The diet board (DB) is a novel method for achieving moderate DR in group housing. Food pellets are firmly attached into grooves in an aspen board, and rats have to gnaw the wood in order to eat. Food is available continuously, but due to the effort involved rats eat less. This study simulated a chronic safety test to assess the long-term effects of DB feeding. A total of 146 male and female outbred Sprague-Dawley rats, nine weeks old at onset, were housed in groups of three and fed either AL or with DBs for two years. Food and water consumption were measured at six time points. The rats were weighed every one to two weeks. Body and tibial lengths and epididymal fat weight were measured at necropsy. Modified body mass index was calculated at five time points after one year of age. DB feeding reduced body weight and fat tissue moderately, more so in males. DB males ate less than AL males, but no differences were seen in the total food consumption in the females. There was no consistent difference in the within-group variations of the measured parameters. DB is a workable DR method, albeit some modification could enhance and standardize its DR effects, especially in female rats.

Refining cage change routines: comparison of cardiovascular responses to three different ways of cage change in rats.

Cage change is one of the unavoidable routines in laboratory rodent care. However, cage change disrupts the rodents' olfactory environment and can evoke stress reactions. In this study, the short-term cardiovascular responses to three different cage change procedures were compared with telemetric monitoring. These procedures were: placing the rats into a new, clean cage (NEW), transferring the old cage lid into the clean cage (LID) and transferring an enrichment object into the clean cage (ENR) with the animals. Seven outbred rats (four Hsd:Sprague-Dawley and three HsdBrlHan:WIST) were instrumented with telemetric transmitters. The reactions were recorded during the 24 h following the cage change procedures. All cage change procedures (and also simple handling) caused elevated heart rate and mean arterial pressure levels for up to 5 h after the procedure, with the largest effect seen during the first hour. The reactions observed after cage change were significantly (P < 0.05) greater than those observed after simple handling. The reactions after NEW were significantly higher than the reactions after ENR or LID, though the results were dependent on the stock. In Wistar rats the LID procedure resulted in smaller reactions than ENR. In Sprague-Dawley rats, the differences between ENR and LID were not so clear, but the transfer of scent-marked material into the new cage decreased the reactions compared with the NEW procedure also in this stock. Based on these results, using the old cage lid on the new cage could reduce the disturbance of cage change in rats.

Comparison of ear tattoo, ear notching and microtattoo in rats undergoing cardiovascular telemetry.

Individual and permanent identification of experimental animals is a common and often essential research practice. There is little information available on the short-term effects of these procedures on the animals. In this study, seven rats were implanted with telemetric devices. The effects of three different identification methods (ear tattoo, ear notching and microtattoo) were compared. Cardiovascular data were collected for 24 h after the procedures. Time periods of 0-1, 1-4, 4-16 h (dark) and 16-24 h after the procedure were analysed separately. The most pronounced differences in measured parameters were observed during the first hour after the procedures were performed. Mean arterial pressure (MAP) was significantly higher (P < 0.012) following the ear tattoo than the microtattoo procedure by a difference of approximately 5 mmHg. Heart rate (HR) was significantly elevated (P < 0.001) after ear tattoo compared with both ear notching (Δ = 31 beats per minute [bpm]) and microtattoo (Δ = 44 bpm). During the 1-4 h period and the following dark period, the MAP was highest in the ear notching group, but no differences were observed in the HRs. During the following dark period (4-16 h) and the next day (16-24 h) differences in MAP and HR were minor. In conclusion, microtattoo appears to cause the mildest changes in HR and blood pressure. Based on these results, ear tattoo and ear notching should be replaced by microtattoo whenever possible.

The effects of intra-cage aspen tube on cardiac morphology and gene expression.

According to the European recommendations rodents should be provided with a nest box if there is insufficient nesting material to build a complete, covered nest. Rats are generally poor nest builders; hence an additional structure is needed. Optimally, housing refinement may be combined with better science; at least it should not detract from the scientific integrity. In order to evaluate these options, there is a need to assess the items used in individual research projects. Studies investigating molecular mechanisms of cardiac hypertrophy and heart failure are typically long-lasting studies; therefore, refinement of the housing of rats in these studies is important. The aim of this study was to evaluate in rats whether a wooden tube has any impact on cardiac morphology or on basal gene expression of atrial natriuretic peptide (ANP) and brain natriuretic peptide (BNP); known markers of cardiac overload, hypertrophy and heart failure. The experimental protocol simulated cardiovascular studies, but without any surgical operations. A total of 42 male Hsd:SD rats were used in an eight-week experiment. After weaning, the experimental group was provided with a rectangular aspen tube and nesting material, and the control group with only nesting material. ANP and BNP gene expression were measured from the left ventricles with Northern blot analysis postmortem along with the absolute weights of the whole heart, left and right atria and left and right chambers. The weights of the whole heart and left chamber were also analysed in relation to body weight. No statistically significant differences were observed in any of these variables. The inter-individual variation was also unchanged by the cage item. In conclusion, the aspen tube does not disrupt research results or alter the number of animals needed and can therefore be recommended for enrichment purposes in cardiovascular studies.

Role of cage material, working style and hearing sensitivity in perception of animal care noise.

During daily care, laboratory animals are exposed to a variety of sounds which may have effects on welfare and also cause physiological and behavioural changes. So far, almost no attention has been paid to individual sounds or the sound level caused by animal care or the sound level inside the animal cage. In this study, sounds from selected rat care procedures were recorded: pulling cage out of the rack, placing it onto a table and replacing the cage back into the rack; with measurements made inside the rat cage and in the adjacent cage. Diet was poured into the food hopper and sounds were recorded inside the cage and also the adjacent cage. The work was repeated in a calm and also in a hurried style, using stainless steel and polycarbonate cages. Finally, the sounds produced by running tap water were recorded. Differences between rat and human hearing were compared using novel species-specific sound level weightings: R-weighting for rats dB(R) and H-weighting for human dB(H). Hurried work with steel caused sound exposure levels exceeding 90 dB(R) when the cages were placed into the rack and about 80 dB(R) when pulling them out of the rack or placing onto a table. With polycarbonate, the levels were 10-15 dB(R) lower. Unhurried calm working produced lower sound exposure levels than hurried working in many procedures. When the procedures were repeated with measurements in the adjacent cage, the sound exposure levels were lower, but the results were similar. Pouring food pellets into a hopper above the rat's head caused 15 dB(R) higher sound exposure levels than pouring food to an adjacent cage. In general, humans hear these sounds about 10-15 dB louder than rats. In conclusion, cage material, working style and hearing sensitivity all have an impact on the sound exposure level in the rodent cage. With correct working methods, high sound levels can be efficiently avoided in most cases.

R-weighting provides better estimation for rat hearing sensitivity.

Since sounds may induce physiological and behavioural changes in animals, it is necessary to assess and define the acoustic environment in laboratory animal facilities. Sound studies usually express sound levels as unweighted linear sound pressure levels. However, because a linear scale does not take account of hearing sensitivity-which may differ widely both between and within species at various frequencies-the results may be spurious. In this study a novel sound pressure level weighting for rats, R-weighting, was calculated according to a rat's hearing sensitivity. The sound level of a white noise signal was assessed using R-weighting, with H-weighting tailored for humans, A-weighting and linear sound pressure level combined with the response curves of two different loudspeakers. The sound signal resulted in different sound levels depending on the weighting and the type of loudspeaker. With a tweeter speaker reproducing sounds at high frequencies audible to a rat, R- and A-weightings gave similar results, but the H-weighted sound levels were lower. With a middle-range loudspeaker, unable to reproduce high frequencies, R-weighted sound showed the lowest sound levels. In conclusion, without a correct weighting system and proper equipment, the final sound level of an exposure stimulus can differ by several decibels from that intended. To achieve reliable and comparable results, standardization of sound experiments and assessment of the environment in animal facilities is a necessity. Hence, the use of appropriate species-specific sound pressure level weighting is essential. R-weighting for rats in sound studies is recommended.

Osseointegration of dental implants in bone irradiated with 40, 50 or 60 gy doses. An experimental study with beagle dogs.

Construction of different kinds of prostheses on irradiated bone tissues after tumour surgery is a complicated part of modern implantology. In irradiated regions of the jaws where the bone layers are thin and the blood supply also is minimal, knowledge of the impact of each dose of radiation is very important. The aim of this study was to compare the bone response around titanium implants loaded with fixed bridges in dog mandibles irradiated with total doses of 40, 50 or 60 Gy. The results suggest that after irradiation with 40-50 Gy, when the dose is fractioned in order to achieve higher tolerance of the tissues compared with single dose irradiation, titanium implants may become osseointegrated in the mandibles. For high success rates, however, careful planning of radiotherapy and selection of implantation site with an adequate blood supply are essential.

Transgenic mice aberrantly expressing human ornithine decarboxylase gene.

We have generated transgenic mice carrying human ornithine decarboxylase gene. Two different transgene constructs were used: (i) a 5'-truncated human ornithine decarboxylase gene and (ii) an intact human ornithine decarboxylase gene. Transgenic mice carrying the 5'-truncated gene did not express human ornithine decarboxylase-specific mRNA. Transgenic mice carrying the intact human ornithine decarboxylase gene expressed human-specific ornithine decarboxylase mRNA in all tissues studied. However, as indicated by actual enzyme assays, the expression pattern was highly unusual. In comparison with their wild-type littermates, the transgenic mice exhibited greatly elevated enzyme activity in almost every tissue studied. Ornithine decarboxylase activity was moderately elevated in parenchymal organs such as liver, kidney, and spleen. Tissues like heart, muscle, lung, thymus, testis, and brain displayed an enzyme activity that was 20 to 80 times higher than that in the respective tissues of nontransgenic animals. The offspring of the first transgenic male founder animal did not show any overt abnormalities, yet their reproductive performance was reduced. The second transgenic founder animal, showing similar aberrant expression of ornithine decarboxylase in all tissues studied, including an extremely high activity in testis, was found to be infertile. Histological examination of the tissues of the latter animal revealed marked changes in testicular morphology. The germinal epithelium was hypoplastic, and the spermatogenesis was virtually totally shut off. Similar examination of male members of the first transgenic mouse line revealed comparable, yet less severe, histological changes in testis.

Osseo-integration of dental implants in radiated mandibles: an experimental study with beagle dogs.

This investigation was planned to detect the ability of radiated bone to integrate with dental implants and to study the load-bearing capacity of these implants. In five beagle dogs, the premolars were extracted, after which one side of the mandible was radiated with 40 Gy. Three months later, two hollow-cylinder implants were placed on each side of the mandible. After a three-month period of osseo-integration, bridges were constructed on the implants, and they were loaded for six months. Both clinical and x-ray examinations showed that all implants had integrated well and bore load without change in their host sites for a six-month period, at which time the dogs were killed.

Evaluation of anaesthetic potency of medetomidine-ketamine combination in rats, guinea-pigs and rabbits.

In many animal species ketamine hydrochloride is combined with xylazine, an alpha 2-adrenoceptor agonist, to achieve anaesthesia. The purpose of this study was to evaluate the anaesthetic efficacy of various dose combinations of another alpha-2-adrenoceptor agonist medetomidine and ketamine in rats, guinea pigs and rabbits. Both sexes of all three species in groups of five were used. In rats, dose combinations medetomidine/ketamine, both in mg/kg, used were 0.25/60, 1.0/60 and 0.5/70; in guinea pigs 0.5/40 and in rabbits 0.5/25 and 0.5/60. In rats and rabbits the righting reflex was lost within 2-3 min. In rats with doses 0.25/60 and 0.5/75 other reflexes disappeared, reappearing after 20-25 min and 60-70 min in males, the respective values being 120-140 min and 150-180 in females. The righting reflex reappeared after 130-135 and 160-190 min in males and 240 and greater than 300 min in females in the order above. In guinea pigs irrespective of route of ketamine administration all reflexes remained. In rabbits, a dose 0.5/25 caused disappearance of ear pinch reflex. We conclude that the medetomidine-ketamine combination can be used in rats and rabbits as an anaesthetic, and in guinea pigs only for immobilization.

Experimental compound OR-K-242-HCl an anilide derivative slows conduction velocity of the heart in the conscious beagle dog.

OR-K-242-HCl is an antitussive compound. Because it resembles lidocaine structurally, one of its target organs may be the heart. In anaesthetized guinea pigs OR-K-242-HCl depressed function of the heart. The aim of the present study was to test effects of OR-K-242-HCl on the conduction velocity of the heart in the conscious beagle dog. OR-K-242-HCl was administered 6 mg/kg and 12 mg/kg i.v. and 6 mg/kg, 12 mg/kg, 24 mg/kg, and 36 mg/kg orally. The doses for lidocaine (reference substance) were 6 mg/kg, and 12 mg/kg i.v. and 6 mg/kg, 12 mg/kg, and 24 mg/kg orally. Five consecutive noisefree cycles of ECG were used before and every 15 min up to three hours after the drug administrations. For each cycle PQ-, QTC-, RR-interval, as well as, QRS-complex and heart rate were calculated. Both OR-K-242-HCl and lidocaine prolonged the conduction velocity in the atria and ventricles. After i.v. and oral administration OR-K-242-HCl induced stronger and more uniform responses than lidocaine. The oral drug administrations produced more sustained responses than the intravenous injections for both compounds.