H.A. van Lith

The influence of animals on the development of children.

There is a widespread belief that interaction with an animal is beneficial for the development of children, and several studies (most with methodological shortcomings) have investigated the influence of (companion) animals on the social-emotional and cognitive development of children. In this article, the 1984 model of Professor Jay Belsky has been used to describe which variables influence the development of children and how the companion animal-child interaction influences these variables. The value of the AAA/AAT (Animal Assisted Activities/Animal Assisted Therapy) programmes in children with a wide variety of clinical and social problems, such as behaviour problems and autistic spectrum symptoms, is discussed. The findings suggest that (companion) animals positively influence childrens development and have a valuable role in therapy.

Blood sampling from the retro-orbital plexus, the saphenous vein and the tail vein in rats: comparative effects on selected behavioural and blood variables:

We compared the behaviours of rats, and measured various blood parameters, after three blood sampling techniques: orbital puncture while they were under diethyl-ether anaesthesia, blood collection by tail vein puncture under O2-N 2O-halothane anaesthesia and puncture of the saphenous vein without anaesthesia. Twelve rats were subjected to the three treatments according to a Latin square design. After each treatment, the behaviour of the rats was automatically monitored using the so-called LABORASTM method, which discriminates between grooming, locomotion and inactivity in rats. Based on excitation scores and urine production, it was found that induction of diethyl-ether anaesthesia combined with orbital puncture caused more distress than did the other two blood sampling techniques. The three techniques had no differential effects on the behaviours of grooming, locomotion and inactivity. Collecting 0.5 ml of blood by orbital puncture was ± 7 times faster than doing so by saphenous vein puncture and ± 15 times faster than collecting blood by tail vein puncture while the rats were under O2-N 2O-halothane anaesthesia. The levels of some haematological and plasma variables differed significantly between the three blood collection techniques. These observations may help to select the most appropriate technique of blood sampling with respect to anticipated discomfort in the animals.

High-resolution genetic mapping of mammalian motor activity levels in mice

The generation of motor activity levels is under tight neural control to execute essential behaviors, such as movement toward food or for social interaction. To identify novel neurobiological mechanisms underlying motor activity levels, we studied a panel of chromosome substitution (CS) strains derived from mice with high (C57BL/6J strain) or low motor activity levels (A/J strain) using automated home cage behavioral registration. In this study, we genetically mapped the expression of baseline motor activity levels (horizontal distance moved) to mouse chromosome 1. Further genetic mapping of this trait revealed an 8.3‐Mb quantitative trait locus (QTL) interval. This locus is distinct from the QTL interval for open‐field anxiety‐related motor behavior on this chromosome. By data mining, an existing phenotypic and genotypic data set of 2445 genetically heterogeneous mice (http://gscan.well.ox.ac.uk/), we confirmed linkage to the peak marker at 79 970 253 bp and refined the QTL to a 312‐kb interval containing a single gene (A830043J08Rik). Sequence analysis showed a nucleotide deletion in the 3′ untranslated region of the Riken gene. Genome‐wide microarray gene expression profiling in brains of discordant F2 individuals from CS strain 1 showed a significant upregulation of Epha4 in low‐active F2 individuals. Inclusion of a genetic marker for Epha4 confirmed that this gene is located outside of the QTL interval. Both Epha4 and A830043J08Rik are expressed in brain motor circuits, and similar to Epha4 mutants, we found linkage between reduced motor neurons number and A/J chromosome 1. Our findings provide a novel QTL and a potential downstream target underlying motor circuitry development and the expression of physical activity levels.

Differences in response to anaesthetics and analgesics between inbred rat strains.

Differences in response to analgesic and anaesthetic drugs can partly be attributed to variations in the genetic background of experimental animals. This study was carried out to determine differences in the response of inbred rat strains to a selection of analgesics and drugs used in anaesthetic protocols. A cross between the most contrasting strains can then be phenotyped in future studies in order to localize quantitative trait loci (QTLs) involved in analgesic/anaesthetic drug sensitivity. Eight inbred strains (n = 6 rats/strain) were selected for the study: the pigmented ACI, BN and COP strains and the albino F344, LEW, SHR, WAG and WKY strains. Each rat was injected intravenously with two analgesics (buprenorphine 0.05 mg/kg and nalbuphine 1 mg/kg) and three drugs used in anaesthetic protocols (propofol 25 mg/kg, medetomidine 50 μg/kg and ketamine 10 mg/kg), respectively, using a crossover design. Analgesic responses were assessed using an analgesiometric procedure. The sleep time of the rat and, where applicable, the interval between injection and loss of righting reflex were used to determine the anaesthetic response. Six out of eight strains responded significantly different from each other to the analgesic effect of buprenorphine with the ACI strain as hyper-responder. The tail withdrawal latency at 55°C of the F344 and WKY rats using buprenorphine was not significantly different from baseline tail withdrawal latencies. In this study, all strains were non-responsive to the analgesic effects of nalbuphine. The response to all three drugs used in anaesthetic protocols differed significantly among the strains. The F344 and BN strains were relatively resistant to the sedative effects of medetomidine. Use of ketamine was abandoned in the ACI and BN strains when the first two animals of both strains died soon after induction. With all three drugs the sleep time of albino rats was significantly longer compared with that of the pigmented ones. We conclude that the results from this study can be used in future studies where QTLs for the sensitivity to anaesthetic/analgesic drugs are localized.

Orbital sinus blood sampling in rats: effects upon selected behavioural variables

The question addressed was whether the behaviour of rats is changed after orbital sinus blood sampling while they are under diethyl-ether anaesthesia. Twelve rats were subjected to sham anaesthesia, diethyl-ether anaesthesia and anaesthesia plus orbital puncture according to a Latin square. After each treatment, the behaviour of the rats was automatically monitored using the so-called LABORAS method, which discriminates between grooming, locomotion and inactivity. Treatment ended, and behaviour monitoring began, when the light period changed over to the dark period. The various behaviours were quantified as relative duration and frequency. Anaesthesia versus sham anaesthesia reduced the relative duration of grooming during the first 5 h after treatment. Anaesthesia plus orbital puncture versus anaesthesia alone did not significantly influence grooming, but orbital puncture did reduce the relative duration and the frequency of locomotion during the entire 20 h period, which was mainly due to a decrease in the dark period. After orbital puncture, the animals were also less frequently inactive. It is concluded that orbital puncture has an effect on behaviour superimposed on that of diethyl-ether anaesthesia. This observation may contribute new arguments to the debate on the acceptability of the orbital puncture technique.

Butyrylcholinesterase activity in plasma of rats and rabbits fed high-fat diets

1. Comparative studies with rats and rabbits were carried out to address the question as to whether the amount of dietary fat affects butyrylcholinesterase (EC 3.1.1.8.) activity in plasma. 2. Plasma butyrylcholinesterase activities were about 5-fold higher in rabbits than rats. 3. Ad libitum feeding of diets enriched with corn oil caused increased body weights in rabbits but not in rats 4. Plasma butyrylcholinesterase activities of rats were increased with increasing intakes of corn oil. In rabbits, such an effect could not be demonstrated conclusively. 5. Evidence is presented that in rats the substitution of dietary corn oil for isocaloric amounts of either carbohydrates or protein produces similar increases in plasma butyrylcholinesterase activity. 6. This suggests that among macronutrients the amount of fat primarily determines butyrylcholinesterase activity in the plasma of rats.

Orbital bleeding in rats while under diethylether anaesthesia does not influence telemetrically determined heart rate, body temperature, locomotor and eating activity when compared with anaesthesia alone

The question addressed was whether orbital bleeding in rats, while under diethylether anaesthesia, affects their locomotor activity, body core temperature, heart rate rhythm and eating pattern. Roman High Avoidance (RHA) and Roman Low Avoidance (RLA) rats were used to enhance generalization of the results. Orbital bleeding when the rats were under diethylether anaesthesia was compared with diethylether anaesthesia alone. To take into account any effects of handling, the rats were also subjected to sham anaesthesia. The RHA rats urinated more during anaesthesia, needed more time to recover from the anaesthesia and showed a greater endocrine stress response to diethylether anaesthesia when compared with the RLA rats. During anaesthesia, the RHA rats showed a greater fall of body temperature and bradycardia than did the RLA rats. Diethylether anaesthesia reduced locomotor activity in the RHA rats, but had no effect in the RLA rats. In neither RHA nor RLA rats did anaesthesia plus orbital puncture, versus anaesthesia alone, influence body temperature, heart rate rhythm, locomotor and eating activity. The lack of effect of orbital puncture occurred both in the short term (within 2 h) and long term (within 48 hours) and thus this study indicates that orbital puncture had, at least with respect to variables measured in the present study, no effect superimposed on that of diethylether anaesthesia.

Influence of amount of dietary fat and protein on esterase-1 (ES-1) activities of plasma and small intestine in rats

The objective of the present study was to characterize nutritionally esterase-1 (ES-1). For this purpose, the effects of replacement of dietary carbohydrates by isoenergetic amounts of either fat or protein on ES-1 activities of plasma and small intestine were studied in male rats. Purified diets differing in the amounts of maize starch plus dextrose, casein and various types of fat were used. Plasma and jejunal ES-1 activities were found to be increased with increasing fat intakes. As to the type of fat, increasing plasma ES-1 activities were induced by coconut fat, olive oil, maize oil and medium-chain triacylglycerols, in this order. Maize oil induced higher jejunal ES-1 activities than coconut fat and olive oil, but had similar effects to medium-chain triacylglycerols. Maize oil was more powerful in increasing plasma ES-1 activity than isoenergetic amounts of casein, but with respect to jejunal ES-1 activity these dietary components were equally effective. It is concluded that the amounts of fat and protein in the diet are important determinants of ES-1 activities in plasma and jejunum.

Plasma esterase-1 (ES-1) activity is increased in rats fed high-fat diets

The question addressed is whether the amount and type of dietary fat affects esterases in plasma. Rats were fed semipurified diets containing 2.0 to 19.4% (w/w) of fat in the form of coconut fat or corn oil. Fat was added to the diets at the expense of isocaloric amounts of carbohydrates. Plasma total esterase activities measured with 4-nitrophenylacetate as substrate were slightly increased with increasing fat intakes. However, an increase in fat concentration of the diet was associated with a pronounced increase in the activity of the so-called ES-1 isozyme in plasma. ES-1, which represents very little plasma total esterase activity, was quantified densitometrically as the high-mobility, anodal esterase band on polyacrylamide gel electrophoresis. The positive association between amount of dietary fat and ES-1 activity was identical for coconut fat and corn oil.

Evidence for epigenetic interactions for loci on mouse chromosome 1 regulating open field activity

The expression of motor activity levels in response to novel situations is under complex genetic and environmental control. Several genetic loci have been implicated in the regulation of this behavioral phenotype, but their relationship to epigenetic and epistatic interactions is relatively unknown. Here, we report on a quantitative trait locus (QTL) on mouse chromosome 1 for novelty-induced motor activity in the open field, using chromosome substitution strains derived from a high active host strain (C57BL/6J) and a low active donor strain (A/J). The QTL for open field (horizontal distance moved) peaked at the location of Kcnj9, however, QTL detection was initially masked by an interplay of both grandparent genetic origin and genetic co-factors influencing behavior on chromosome 1. Our findings indicate that epigenetic interactions can play an important role in the identification of behavioral QTLs and must be taken into consideration when applying behavioral genetic strategies.