Live black soldier fly (Hermetia illucens) larvae in feed for laying hens: effects on hen gut microbiota and behavior.

This study examined the effects of including live black soldier fly (BSF, Hermetia illucens) larvae in the diet of laying hens on gut microbiota, and the association between microbiota and fearfulness. A total of 40 Bovans White laying hens were individually housed and fed 1 of 4 dietary treatments that provided 0, 10, 20%, or ad libitum daily dietary portions of live BSF larvae for 12 wk. Cecum microbiota was collected at the end of the experiment and sequenced. Behavioral fear responses to novel objects and open field tests on the same hens were compared against results from gut microbiota analyses. The results showed that the bacteria genera Enterococcus, Parabacteroides, and Ruminococcus torques group were positively associated with increased dietary portion of live larvae, while Lactobacillus, Faecalibacterium, Bifidobacterium, Subdoligranulum, and Butyricicoccus were negatively associated with larvae in the diet. Inclusion of larvae did not affect fear behavior, but the relative abundance of Lachnospiraceae CHKCI001 and Erysipelatoclostridium was associated with fear-related behaviors. Further studies are needed to determine whether the change in gut microbiota affects fearfulness in the long-term.

Health effects of feed dilution and roughage in Ross 308 broiler breeder cockerels.

To ensure reproductivity and prevent obesity, broiler breeder's growth rate is controlled by quantitative feed restriction. However, feed restriction is associated with chronic hunger, frustration, and abnormal behaviors, thus representing a welfare problem. Feed diluted with insoluble fiber is an alternative, allowing larger amounts of feed and more gut filling, increasing satiety without increasing the caloric intake. Previous research on feed dilution has focused on pullets not cockerels. In addition, the health effects of diluted feed are less explored. The aim of this study was to investigate the effects of feed dilution and roughage on health indicators in broiler breeder cockerels, 5 to 10 wk of age (WOA), a very feed restricted period. In total, 200 Ross 308 broiler breeder cockerels were housed in 12 pens (6 pens/treatment), 16 to 17 birds per pen. The treatments were standard feed (Control) and feed diluted with (20%) insoluble oat hulls and 150 g of roughage (lucerne/alfalfa) daily per pen (D + R). The D + R birds received 20% more feed per day. Body weight, pecking injuries, footpad dermatitis (FPD), and litter quality were recorded weekly between 5 and 9 WOA. At 10 WOA all birds were euthanized and subjected to postmortem assessment. D + R birds tended to weigh more than controls (P = 0.055), but diet did not affect flock uniformity (P = 0.57). The weekly assessment showed no effect of treatment on foot pad lesion scores (P = 0.31). However, an effect on FPD was observed postmortem (P = 0.04), where Control had slightly better footpad condition compared to the D + R group. Litter had to be completely changed in 2 control pens during the trial, due to quality deterioration. There was no effect of treatment on the weight of specific organs, the length of the gut, the intestinal mucosa, nor on the incidence of wounds on the head (P = 0.15). Overall, these results indicate that feed dilution and roughage have neither adverse nor beneficial health effects on Ross 308 broiler breeder cockerels.

Environmental enrichment in commercial flocks of aviary housed laying hens: relationship with plumage condition and fearfulness.

Management strategies can have positive effects on laying hen welfare, including prevention of damaging behavior, aggression, and fear, particularly by using environmental enrichment (EE). However, few studies have investigated the effects of the provision of EE in commercial aviary flocks. This knowledge gap is particularly significant considering the increasing numbers of non-beak trimmed hens worldwide kept in aviaries. The aim of this study was to survey and investigate the relationship between commercially applied EE and plumage condition and fearfulness in Norwegian flocks of loose-housed laying hens. Forty-five indoor multi-tiered aviary-system flocks of laying hens from across Norway were visited at the end of lay (range: 70-76 wk of age). The flocks consisted of either Lohmann LSL (n = 30) or Dekalb White (n = 15) non-beak-trimmed hens. During the visit, the researchers collected data on the farmers' use of the following five types of enrichment: pecking stones, gravel, oyster shells, grains scattered in the litter, and "toys". Feather loss was assessed individually in approximately 50 hens per flock and scores were awarded using a 3-point scale (0-2) for each of the following body parts: head, back/wings, breast, and tail. Finally, a novel object test was performed in 4 different locations in each house. The results showed that damage to the tail feathers was correlated positively to the first age of provision of toys (Pearson correlation coefficient = 0.41; P = 0.051) and negatively to the amounts of gravel stones provided (Pearson correlation coefficient = -0.43, P = 0.02). No other associations between the welfare indicators and the provision of EE objects were found, likely because of the low variation of enrichment provision. The present study showed that the provision of EE objects such as toys and gravel stones can have significant benefits to the condition of laying hen plumage. This study also adds to the body of literature supporting the importance of early life experiences on the behavioral development of laying hens and on their welfare at older ages.

Feeding live Black Soldier Fly larvae (Hermetia illucens) to laying hens: effects on feed consumption, hen health, hen behavior, and egg quality.

The use of insects in animal feed has the potential to reduce the demand for soybean production and reduce the deforestation and loss of natural resources. In particular, the black soldier fly (BSF, Hermetia illucens) larvae have received attention due to their ability to convert organic waste into high-value biomass. Several studies have investigated the effects of providing BSF larvae to both broilers and laying hens. However, knowledge gaps regarding hens' voluntary intake of live larvae and the effects of larvae consumption on egg production still remain. Therefore, the aim of the present study was to determine the effects of the consumption of 4 different amounts of live BSF larvae on laying hen feed consumption, hen health and fearfulness, and egg production and quality. To this end, 40 Bovans White laying hens were housed individually and provided with 0, 10, 20% or ad libitum daily portions of live larvae from 18 to 30 wk of age. The larvae consumption and concentrate consumption, hen weight, egg production, and egg quality were monitored. Overall, differences were found between the hens given ad libitum access to larvae compared to the other treatments. Ad libitum hens, consumed 163 ± 41 g larvae/hen/day, consumed less concentrate (P = 0.03) and gained more weight (P = 0.0002) than all other treatments. They also had an overall higher consumption of protein, fat and energy (P < 0.03). There was no effect of larvae consumption on egg production, egg weight, shell thickness, shell breaking strength, or Haugh unit (P > 0.05). There was also no effect on hen behavior toward a novel object or in an open field test. This study is the first to provide different amounts of live BSF larvae, including an ad libitum portion to laying hens. The 20% diet could promote sustainability in the egg industry and be economically advantageous if BSF larvae can be bought in bulk for less than 40% of the cost of the concentrate.

Post-mortem examination of fast-growing broilers with different degrees of identifiable gait defects.

BACKGROUND: The walking ability of many broilers is characterised by slight or definite defects categorised as gait scores (GS) 1 and 2. The present study aimed to examine potential relationships between GSs and indicators of body morphology, leg pathology, tibia strength and wooden breast in Ross 308 broilers assessed as GS ≤ 2. METHODS: At 38 days of age, GS and live body weight of 179 birds was recorded. Each bird was examined post-mortem for signs of wooden breast, contact dermatitis and a range of leg pathologies. Weights of different body parts and tibia strength were quantified. RESULTS: Within sex, GS increased with increasing live body weight (p = 0.020). There was a tendency for an effect of GS on prevalence of footpad dermatitis (p = 0.086) and dislocated femoral joint cartilage (p = 0.059) where both pathologies increased in frequency with increasing GS. Greater load was required to fracture tibia from GS2 than GS0 birds (p = 0.040). CONCLUSIONS: Within this relatively small data set, no strong relationships between GS ≤ 2 and indicators of body morphology, leg pathology, tibia strength and wooden breast in Ross 308 broilers were found, except for the live terminal body weight. Further studies, involving larger data sets are required for full clarification.

Are changes in behavior of fast-growing broilers with slight gait impairment (GS0-2) related to pain?

Impaired walking ability in terms of slight or definite defects is more common in broiler production than lameness that obviously hinders movement, but it has received limited scientific attention. This study aimed to compare behavior of conventional broilers with impaired walking ability (assessed as gait score (GS) 2) with those walking normally (GS0) and those with only a slight gait defect (GS1). Behavior in the home environment was registered, and an analgesic intervention to quantify changes in time budgets indicating pain relief was applied. The study included 192 Ross 308 broilers. On day 27 of age, the birds were distributed as evenly as possible into birds of GS0 and GS2 of each sex based on obtained gait score. Following this, each experimental bird was housed with 3 companion birds. On days 30 and 32 of age, the behavior in the home pens was recorded. All experimental birds were injected with the NSAID carprofen on one of the 2 d and saline on the other. The statistical analyses used the GS scored on the day of recording as explanatory factor. Compared to GS0 birds, GS2 birds tended to be more inactive (mean (CI): 4,193 (3,971-4416) vs. 4,005 (3,753-4,257) s; P = 0.074), spent more time sitting while feeding (306 (266-353) vs. 213 (180-251) s; P = 0.026), were less likely to perch (probability: 0.78 (0.69-0.85) vs. 0.91 (0.85-0.95); P = 0.012), and spent less time performing comfort behavior (749 (689-814) s vs. 875 (792-967) s; P = 0.043). Compared to GS1 birds, GS2 birds spent more time inactive (GS1: 4,022 (3,818-4225) s; P = 0.027), less time foraging (289 (253-329) vs. 347 (309-388) s; P = 0.047), and were less likely to perch (GS1: 0.90 (0.86-0.93); P = 0.001). For some of these behavioral variables, administration of carprofen led to behavioral changes across the GSs, which may suggest that the behavioral expression of the broilers was limited by pain. These findings are of relevance to animal welfare, but the underlying causes are still not fully clarified.

Effect of Qualitative Feed Restriction in Broiler Breeder Pullets on Stress and Clinical Welfare Indicators.

The feed restriction applied during rearing of broiler breeders inflicts chronic hunger, and frustration due to unfulfilled behavioural needs for feeding. To alleviate the welfare problems associated with feed restriction, qualitative feed restriction allows a larger amount of feed to be provided without increasing the energy intake. In the present study, the aim was to investigate the effect of scatter-fed qualitative feed restriction on a range of welfare indicators in broiler breeders at the end of the rearing period. In total, 1,200 female breeder chicks of the genotype Ross 308 were housed in 24 pens: six pens of initially 50 birds per dietary treatment. The treatments were: (1) standard feed (Control), (2) standard feed diluted with oat hulls (Insoluble), (3) standard feed diluted with oat hulls and sugar beet pulp (Mixed), and (4) standard feed plus maize silage (Roughage). At 15 weeks of age, a blood sample was taken from 40 birds (10/treatment) five times within 24 h. The plasma was analysed for corticosterone concentration. At 19 weeks of age, a clinical welfare assessment was performed on all birds before they were sacrificed. From each bird, three feathers were plucked and macroscopically examined for the presence of fault bars. Feather length and weight were also recorded. Mortality was registered on occurrence throughout the rearing period. Treatment affected the plumage condition, footpad dermatitis, plumage dirtiness, vent pasting, and number of severe fault bars (P ≤ 0.05) but not plasma corticosterone concentration, hock burns, hyperkeratosis and mortality (P ≥ 0.17). There was an effect of the interactions between treatment and feather type on the total number of fault bars per feather, average position of the fault bars relative to the base of the feather, and growth rates of feather mass and length (P < 0.0001). Overall, the results showed improved welfare of Roughage birds and reduced welfare of Mixed birds, whereas the welfare of Insoluble birds did not seem to differ noticeably from that of Control birds. We recommend to further develop a feeding strategy that includes daily allocation of roughage to broiler breeders during the rearing period.

Effects of environmental complexity on welfare indicators of fast-growing broiler chickens.

Increasing environmental complexity, e.g., by providing environmental enrichment, has been suggested as a way to increase activity levels and improve leg health in broilers. The aim of this study was to investigate the effects of different types of environmental complexity on leg health and measures of welfare of fast-growing broilers housed according to conventional European legislation. A total of 58 pens with approximately 500 broilers each (Ross 308), corresponding to a stocking density of 40 kg/m2, were used. A total of 8 treatment groups, of which 5 were enrichment treatments (roughage, vertical panels, straw bales, and elevated platforms at 5 and 30 cm) and 3 were standard resources manipulations [increased distances between feed and water (7 m and 3.5 m), stocking density reduced to 34 kg/m2, and 1 control group (1.5 m distance between feed and water and no enrichment objects)] were randomly assigned to each pen. At 35 D of age, 60 birds from each pen were assessed for gait, footpad dermatitis, hock burns, plumage cleanliness, presence of scratches, and leg deformities. Birds housed with 30 cm elevated platforms had worse gait compared to those housed with straw bales and at the lower stocking density of 34 kg/m2 (P = 0.004 and P = 0.001). Broilers from the control group also had worse gait compared to those housed at 34 kg/m2 stocking density. In addition, birds housed with access to a 30 cm elevated platform had healthier footpads compared to birds housed with access to straw bales (P = 0.0001) and with increased distance between feed and water (P = 0.011). Furthermore, birds housed with straw bales had worse footpad condition compared to birds with access to a 5 cm elevated platform (P = 0.002). There were no observed treatment effects on scratches, plumage cleanliness, leg deformities and body weight (P > 0.05). Based on the welfare indicators used in the present study, decreased stocking density has the potential of improving animal welfare, whereas the effects of elevated platforms need to be further studied before a final conclusion can be drawn, as footpad health was positively affected, but walking ability was impaired.

Effects of environmental enrichment on health and bone characteristics of fast growing broiler chickens.

Providing environmental enrichment for broilers is a potential strategy to increase welfare, activity, and health. The aim of this study was to evaluate the effect of environmental enrichment on health and leg bone characteristics of broilers. One control and 8 types of enrichment were included: 2 distances between food and water (7 and 3.5 m), roughage, vertical panels, straw bales, 2 platforms (30 and 5 cm), and a lowered stocking density (34 kg/m2). Birds were kept according to conventional Danish guidelines. The study included 58 pens with approximately 500 birds each. On day 35 of age, 25 birds per pen were killed and included in a postmortem analysis of wooden breast, body condition scores, pathological conditions (femoral head necrosis, arthritis, tenosynovitis, fractures, tibial dyschondroplasia, and twisted tibiotarsus), muscle width of the lower leg, and tibiotarsus properties (bone strength, weight, length, and proximal diameter, middle diameter, and distal diameter). It was predicted that environmental enrichment would have a positive effect on pathology with the exceptions that environmental enrichment that increased activity would pose a risk factor for wooden breast development, and straw bales would be a risk factor for bacterial infections (arthritis, tenosynovitis, and femoral head necrosis). Furthermore, it was hypothesized that enriched groups would have increased muscle width, bone strength, and dimensions of the tibiotarsus. Broilers with 7 m between food and water had a longer distal diameter of the tibiotarsus than those with straw bales (P = 0.04). The birds provided with vertical panels had wider leg muscle than the treatments with roughage (P = 0.045), 3.5 m distance (P = 0.049), and straw bales (P = 0.044). No effects were found for the remaining outcomes. These results suggest that provision of vertical panels and increased distance between resources can result in larger muscle and bone dimension, possibly having a positive effect on leg health. Furthermore, the provision of environmental enrichment does not appear to be a risk factor for wooden breast or bacterial infection.

The Productivity and Financial Impacts of Eight Types of Environmental Enrichment for Broiler Chickens.

Reduced mobility in broilers can contribute to leg health problems. Environmental enrichment has been suggested as one approach to combat this through stimulating increased physical activity. Past studies have tested the effect of environmental enrichments on bird behaviour, health and welfare, but few have estimated their financial impacts. This study tested the impact of eight types of environmental enrichment on enterprise net margin, accounting for direct intervention costs plus indirect effects via changes to bird mortality, weight, feed intake, feed conversion ratio, and foot pad dermatitis. The trial used 58 pens each containing approximately 500 broilers (Ross 308) at a stocking density of 40 kg/m2. The environmental enrichments were: roughage, vertical panels, straw bales, elevated platforms (5 and 30 cm), increased distances between feed and water (7 and 3.5 m) and stocking density reduced to 34 kg/m2, plus a control group. Mortality was recorded daily and feed intake and weight weekly. Footpad dermatitis was assessed on day 35. Only one intervention improved financial performance (3.5 m between feed and water) above the control, suggesting that most environmental enrichment would have a negative financial impact due to the additional intervention costs, unless consumers were willing to pay a price premium.

Welfare assessment of conventional and organic broilers in Denmark, with emphasis on leg health.

The aim of this study was to survey and report the walking ability in broilers housed in both conventional and organic production systems in Denmark. To this end, the authors assessed the walking ability, by using the Bristol scale, in 31 conventional broiler flocks and in 29 organic broiler flocks distributed across Denmark. In addition, assessment of contact dermatitis, leg abnormalities, scratches and plumage cleanliness, as well as postmortem analysis of tibial dyschondroplasia, was performed in conventional broilers. The survey found a prevalence of 77.4 per cent of impaired walking ability (gait score (GS) >0) in conventional broilers and 38.1 per cent in organic broilers. The prevalence of severe lameness (GS >2) was 5.5 per cent and 2.5 per cent for conventional and organic broilers, respectively. The prevalence of tibial dyschondroplasia in conventional broilers was 4.7 per cent. The results on other welfare indicators are also presented for conventional broilers. The results from the present and previous surveys indicate that the prevalence of impaired walking ability in broiler chickens in the Danish conventional production system is high, but the severity has been steadily decreasing over the last 19 years. Furthermore, the results from the survey of organic broilers suggest that lameness is less prevalent and severe in this system relative to conventional production.

Rearing Laying Hens in Aviaries Reduces Fearfulness following Transfer to Furnished Cages.

Appropriate rearing is essential for ensuring the welfare and productivity of laying hens. Early experience has the potential to affect the development of fearfulness. This study tested whether rearing in aviaries, as opposed to cages, reduces the fearfulness of laying hens after transfer to furnished cages. Fear responses were recorded as avoidance of a novel object in the home cage. Lohmann Selected Leghorns were reared in an aviary system or conventional rearing cages and then transported to furnished cages at 16 weeks, before the onset of lay. Observations of a selection of birds were conducted at 19 (N = 50 independent cages) and 21 (N = 48 independent cages) weeks of age. At 19 and 21 weeks, cage-reared birds showed higher levels of fearfulness indicated by spending more time away from the novel object compared to aviary-reared birds. These results suggest that rearing in an enriched aviary environment reduces fearfulness up to the fifth week after transfer to a new housing system, compared to rearing in cages.

Exposure to Increased Environmental Complexity during Rearing Reduces Fearfulness and Increases Use of Three-Dimensional Space in Laying Hens (Gallus gallus domesticus).

The complexity of the rearing environment is important for behavioral development and fearfulness. The aim of this study was to test the hypothesis that laying hens reared in a complex aviary system with exposure to mild intermittent stressors would be less fearful, less sensitive to stress, and would use elevated areas of the pen more often as adults than hens reared in a barren cage environment. Laying hens (N = 160) were housed in the same rearing house; half of the birds (n = 80) in an aviary and the other half (n = 80) in cages. At 16 weeks of age, the birds were transported to the experimental facilities. Their behavior was recorded at 19 and 23 weeks of age and analyzed by analysis of variance on individual scores for a fearfulness-related principal component generated using principal component analysis. The results indicate that aviary-reared birds have lower levels of fearfulness compared with cage-reared birds both at 19 weeks and at 23 weeks of age. When comparing the response induced by initial exposure to a novel object at 19 and 23 weeks of age, more aviary-reared birds tended to fly up at 19 weeks compared to the cage-reared birds, indicating a tendency toward a more active behavioral response in the aviary-reared birds than in cage-reared birds. There was no difference between treatments in the flight response at 23 weeks. The groups did not differ in defecation frequency or the concentration of fecal corticosterone metabolites at either age. At 19 weeks, observation of the spatial distribution in the home pens indicated that more aviary-reared birds spent time on the low perch, the elevated platform, and the upper perch, compared to the cage-reared birds. However, at 23 weeks of age, these differences were no longer detected. The results of this study support the hypothesis that increased environmental complexity during rearing reduces fearfulness of adult laying hens.

Does Early Environmental Complexity Influence Tyrosine Hydroxylase in the Chicken Hippocampus and "Prefrontal" Caudolateral Nidopallium?

In adult chickens, the housing system influences hippocampal morphology and neurochemistry. However, no work has been done investigating the effects of the early life environment on chicken brain development. In the present study, we reared 67 commercial laying hens (Gallus gallus domesticus) in two environments that differed in the degree of complexity (aviary or cage system). These two groups were further divided into two age groups. At 20 weeks of age, 18 aviary-reared birds and 15 cage-reared birds were humanely euthanized and their brains dissected. At 24 weeks of age, a further 16 brains from aviary-reared birds and 18 brains from cage-reared birds were collected. These brains were prepared for immunohistochemical detection of tyrosine hydroxylase (TH), the rate-limiting enzyme in the biosynthesis of dopamine, in the hippocampus and the caudolateral nidopallium (NCL). There were no differences between the treatment groups in TH staining intensity in the hippocampus or the NCL. In the medial hippocampus, the right hemisphere had higher TH staining intensity compared to the left hemisphere. The opposite was true for the NCL, with the left hemisphere being more strongly stained compared to the right hemisphere. The present study supports the notion that the hippocampus is functionally lateralized, and our findings add to the body of knowledge on adult neural plasticity of the avian brain.

Early Life in a Barren Environment Adversely Affects Spatial Cognition in Laying Hens (Gallus gallus domesticus).

Spatial cognition in vertebrates is adversely affected by a lack of environmental complexity during early life. However, to our knowledge, no previous studies have tested the effect of early exposure to varying degrees of environmental complexity on specific components of spatial cognition in chickens. There are two main rearing systems for laying hens in the EU: aviaries and cages. These two systems differ from one another in environmental complexity. The aim of the present study was to test the hypothesis that rearing in a barren cage environment relative to a complex aviary environment causes long-lasting deficits in the ability to perform spatial tasks. For this purpose, 24 white Dekalb laying hens, half of which had been reared in an aviary system and the other half in a conventional cage system, were tested in a holeboard task. Birds from both treatment groups learnt the task; however, the cage-reared hens required more time to locate rewards and had poorer levels of working memory. The latter finding supports the hypothesis that rearing in a barren environment causes long-term impairment of short-term memory in chickens.

Does rearing laying hens in aviaries adversely affect long-term welfare following transfer to furnished cages?

This study tests the hypothesis that hens that are reared in aviaries but produce in furnished cages experience poorer welfare in production than hens reared in caged systems. This hypothesis is based on the suggestion that the spatial restriction associated with the transfer from aviaries to cages results in frustration or stress for the aviary reared birds. To assess the difference in welfare between aviary and cage reared hens in production, non-beak trimmed white leghorn birds from both rearing backgrounds were filmed at a commercial farm that used furnished cage housing. The videos were taken at 19 and 21 weeks of age, following the birds' transition to the production environment at 16 weeks. Videos were analysed in terms of the performance of aversion-related behaviour in undisturbed birds, comfort behaviour in undisturbed birds, and alert behaviour directed to a novel object in the home cage. A decrease in the performance of the former behaviour and increase in the performance of the latter two behaviours indicates improved welfare. The results showed that aviary reared birds performed more alert behaviour near to the object than did cage reared birds at 19 but not at 21 weeks of age (P = 0.03). Blood glucose concentrations did not differ between the treatments (P>0.10). There was a significant difference in mortality between treatments (P = 0.000), with more death in aviary reared birds (5.52%) compared to cage birds (2.48%). The higher mortality of aviary-reared birds indicates a negative effect of aviary rearing on bird welfare, whereas the higher duration of alert behavior suggests a positive effect of aviary rearing.

Behavioural effects of the commonly used fish anaesthetic tricaine methanesulfonate (MS-222) on zebrafish (Danio rerio) and its relevance for the acetic acid pain test.

The pros and cons of using anaesthesia when handling fish in connection with experiments are debated. A widely adopted practice is to wait thirty minutes after anaesthesia before behavioural observations are initiated, but information about immediate effects of a treatment is then lost. This is pertinent for responses to acute stressors, such as acid injection in the acetic acid pain test. However, omission of anaesthetics in order to obtain data on immediate responses will compromise the welfare of fish and contribute to experimental noise due to stress. We therefore tested the effect of tricaine methanesulfonate on the behaviour of zebrafish. We predicted that tricaine (MS 222) would decrease swimming velocity and that the control fish would show an increased level of anxiety- and stress-related behaviours compared to the tricaine group. Following acclimatization to the test tank, baseline behaviour was recorded before immersion in either tricaine (168 mg l(-1), treatment group, N = 8) or tank water (control group, N = 7). Latencies to lose equilibrium and to lose response to touch were registered. The fish was then returned to the test tank, and the latency to regain equilibrium was registered in anaesthetized fish. When equilibrium was regained, and at five, thirty and sixty minutes after the fish had been returned to the test tank, behaviour was recorded. The tricaine fish showed the following responses (mean ± sd): latency to lose equilibrium 22.6 s±3.9; latency to lose response to touch 101.9 s±26.8; latency to regain equilibrium 92.0 s±54.4. Contrary to our predictions, neither treatment caused a change in any of the behaviours registered. This indicates that tricaine has no effect on several commonly used behavioural parameters, and that it may be unnecessary to postpone behavioural observations to 30 min after anaesthesia.