The electrocardiogram of the Beagle dog: reference values and effect of sex, genetic strain, body position and heart rate.

The aim of the study was to establish a database for electrocardiographic parameters of Beagle dogs used for toxicological studies and to evaluate the influence of supplier, sex, heart rate (HR) and body position for electrocardiogram (ECG) recording on ECG parameters. Peripheral ECG leads were recorded from 934 female and 946 male dogs from Marshall Farms and 27 females and 30 males from Harlan, either standing on a table or restrained in a hammock. HR, RR, PQ and QT intervals, P and QRS duration and P-wave amplitude were measured. There were no major differences between sexes for ECG parameters. The axis of the heart was shifted to the left when the animals were restrained in a hammock compared to when they were standing on a table. The PQ interval was higher (about 9%) in Harlan than in Marshall dogs. HR was negatively correlated with QT (coefficient of linear correlation: r=-0.61 to -0.74), which emphasizes the need for a formula correcting QT interval for HR when interpreting changes in QT interval. HR was also negatively correlated with PQ intervals (r=-0.26 to -0.11), whereas a positive correlation was found between HR and the amplitude of the P wave (r=0.21-0.34). The level of the respiratory sinus arrhythmia (SA) was quantified by calculating the ratio of maximum to minimum RR interval measured over a 10 s period. This ratio was negatively correlated with HR (r =-0.49 to -0.33). Therefore, at high HRs, SA was less marked than at low HRs, but it did not completely disappear. Analysis of beat-to-beat variation indicated that QT and PQ intervals and the amplitude of P wave fluctuated over time and the degree of this variability was positively correlated with the level of SA. In conclusion, we have established reference values for the duration and/or amplitude of some ECG parameters both in terms of means and variability over the recording period, and we have evaluated the influence of body position, genetic strain and HR on the ECG parameters. These data can be used as baseline for the interpretation of the ECG of Beagle dogs.

A good practice guide to the administration of substances and removal of blood, including routes and volumes.

This article is the result of an initiative between the European Federation of Pharmaceutical Industries Associations (EFPIA) and the European Centre for the Validation of Alternative Methods (ECVAM). Its objectives are to provide the researcher in the safety evaluation laboratory with an up-to-date, easy-to-use set of data sheets to aid in the study design process whilst at the same time affording maximum welfare considerations to the experimental animals. Although this article is targeted at researchers in the European Pharmaceutical Industry, it is considered that the principles underpinning the data sets and refinement proposals are equally applicable to all those who use these techniques on animals in their research, whether in research institutes, universities or other sectors of industry. The implications of this article may lead to discussion with regulators, such as those responsible for pharmacopoeial testing. There are numerous publications dealing with the administration of test substances and the removal of blood samples, and many laboratories also have their own "in-house" guidelines that have been developed by custom and practice over many years. Within European Union Directive 86/609EEC1 we have an obligation to refine experiments to cause the minimum amount of stress. We hope that this article will provide background data useful to those responsible for protocol design and review. This guide is based on peer-reviewed publications whenever possible, but where this is not possible we have used "in-house" data and the experience of those on the working party (as well as helpful comments submitted by the industry) for a final opinion. The guide also addresses the continuing need to refine the techniques associated with the administration of substances and the withdrawal of blood, and suggests ways of doing so. Data-sharing between laboratories should be encouraged to avoid duplication of animal work, as well as sharing practical skills concerning animal welfare and scientific problems caused by "overdosing" in some way or another. The recommendations in this guide refer to the "normal" animal, and special consideration is needed, for instance, during pregnancy and lactation. Interpretation of studies may be confounded when large volumes are administered or excessive sampling employed, particularly if anaesthetics are used.

Effects of acute blood removal via the sublingual vein on haematological and clinical parameters in Sprague-Dawley rats.

Haematological and clinical effects were evaluated after multiple blood samplings over 24 h in the laboratory rat and for up to 22 days after collection to determine the time required for a return to baseline values. Male and female Sprague-Dawley rats (n = 7/sex/group) were bled 4 times over 24 h and blood samples of 1.2, 1.6, 2.4 and 3.2 ml were taken. There were no adverse clinical signs and no statistically significant differences in body weight, body weight gain and food consumption. The acute effect on main haematological parameters of blood removal over 24 h consisted of reductions of red blood cell count, haemoglobin and haematocrit which were similar for blood removal exceeding 7.5% and less than 15% of circulating blood volume, and the reductions were positively related to the amount of blood loss beyond 15%. Time to recover to baseline values was also proportional to the initial blood collection volume and was estimated to range from 48 h for amounts from 5% to below 7.5%, 12 days for amounts from 7.5% to up to 20% and 19 days for amounts above 20%. Recommendations are made concerning suitable blood collection regimes.

Malignant fibrous histiocytoma in rats at sites of implanted millipore filters.

Soft-tissue tumors developing in rats at the site of implanted Millipore filters showed the structural, cytochemical, and biologic characteristics of the malignant fibrous histiocytomas occurring in man. Evidence from this tumor model suggests that malignant fibrous histiocytomas can arise from pluripotential mesenchymal stem cells which are not derived from the mononuclear/phagocytic system and that chronic inflammation and scarring may predispose to their development.

Dazoxiben, a prototype inhibitor of thromboxane synthesis, has little toxicity in laboratory animals.

Dazoxiben, an orally active specific inhibitor of thromboxane synthetase, was administered by mouth daily to dogs and rats for 6 months. Dogs showed no evidence of toxicity up to 300 mg day-1 kg-1, the highest dose level used. Rats showed no evidence of toxicity after 100 mg day-1 kg-1, but at 300 mg day-1 kg-1 there were slight increases in plasma calcium and urea concentrations and a moderate incidence of focal nephrosis; males showed a slightly increased platelet count. Studies in rats and rabbits at dose levels up to 400 mg day-1 kg-1, by mouth, revealed no adverse effects on male or female fertility, embryogenesis, parturition or postnatal development. As dazoxiben is well absorbed after oral administration, the generally negative outcome to these toxicity studies suggests that selective inhibitors of thromboxane synthesis may be largely free of adverse effects which might impede their therapeutic or prophylactic use in clinical medicine.

Choice of rat strain: a comparison of the general pathology and the tumour incidence in 2-year old Sprague-Dawley and Long-Evans rats.

As the choice of species and strain of laboratory animal is of considerable importance in carcinogenicity studies, a two-year, spontaneous carcinogenicity study was performed to compare Sprague-Dawley (Crl: Cobs CD(SD)BR) and Long-Evans rats (CRL: Cobs (LE)BR) under the laboratory conditions at Amboise. Of the 108 animals per strain (54 per sex), no overall differences were noted in survival. Nearly half of the premature deaths in both strains appeared to be due to large pituitary adenomas compressing brain parenchyma. Although the incidence of benign mammary fibroadenomas was similar for both female groups, the incidence of invasive mammary carcinoma was higher in Sprague-Dawley females. Other differences were related to the higher incidence of pancreatic atrophy and nephrosis among Long-Evans rats. Studies of this type can help in the understanding of the pathology of laboratory rodents which may aid in the choice of strain in carcinogenicity studies.