Eye darkening as a reliable, easy and inexpensive indicator of stress in fish.

We expand the use of eye darkening (ED) to indicate non-social stress in the fish Nile tilapia Oreochromis niloticus (L.). ED is easily estimated, not requiring any sophisticated equipment, and is non-invasive, facilitating the collection of several measures of stress over time. In the current study, we showed the following: (i) high- and low-ED occur spontaneously, indicating different fish reactions to adjustments to a novel environment; (ii) fish confinement or air exposure clearly increases ED (air exposure is a stronger stressor than confinement), and the time to restore basal values indicates the severity of the impact of the stressor on the fish (this response is not affected by period of the day, e.g., morning or afternoon); and (iii) in adults, females were more responsive (slower recovery) to 2-min air exposure than to 30-min confinement.

Chemical communication of predation risk in zebrafish does not depend on cortisol increase.

We investigated chemical cues among groups of zebrafish (Danio rerio) when communicating information about the risk of predation. We found that visual cues of the predator (tiger Oscar, Astronotus ocellatus) did not increase whole-body cortisol levels in groups of zebrafish but that water conditioned by these (donor) zebrafish stressed (target) conspecifics, thereby increasing whole-body cortisol. This finding was confirmed when these zebrafish groups were in different aquaria and communicated exclusively via water transfer. This result indicates that the stress induced in the target zebrafish does not depend on an increase in whole-body cortisol levels in the donor zebrafish. Because cortisol participation is rejected in this predation-risk communication, other chemicals from the stress systems should be investigated.

Red light stimulates feeding motivation in fish but does not improve growth.

Nile tilapia fish were individually reared under similar light levels for 8 weeks under five colored light spectra (maximum wavelength absorbance): white (full light spectrum), blue (∼452 nm), green (∼516 nm), yellow (∼520 nm) or red (∼628 nm). The effects of light on feeding, latency to begin feeding, growth and feed conversion were measured during the last 4 weeks of the study (i.e., after acclimation). We found that red light stimulates feeding, as in humans, most likely by affecting central control centers, but the extra feeding is not converted into growth.

A case study: the effect of visitors on two captive pumas with respect to the time of the day.

This study analyzed the effects of noise levels and number of visitors on the behaviors of a mother puma and her daughter in a zoo environment with respect to the time of day. The study monitored visitation (noise and number) over two 1-week periods (4 weeks between periods) and frequency of various puma behaviors (videorecorded). The study analyzed videotaped behavior of the pumas based on the time of day and visitors' number and noise levels. There was a direct association of puma behaviors with visitors' number and noise levels. The daughter puma was the most affected and behavioral changes of both pumas correlated more strongly with the noise level than the number of visitors. The noise level and number of visitors affected the behavior of the mother and her daughter only in the morning. The results of this study indicate that both noise level and number of visitors affected these pumas but revealed the noise as predominant. Moreover, the response of these pumas to visitors depended on the time of the day.

Ventilation rates indicate stress-coping styles in Nile tilapia.

Behavioural responses to stress can form distinct profiles in a wide range of animals: proactive and reactive profiles or coping styles. Stress responsiveness can also differentiate between the behavioural profiles. The tendency to regain feed intake following transfer to a novel social-isolation tank (the speed of acclimation) can discriminate between proactive or reactive profiles. Consequently, differential stress responsiveness can be linked to this feeding behaviour trait. This study shows that ventilation rates of Nile tilapia, Oreochromis niloticus (L.), correlate with the rate of feeding resumption, following transfer to a novel social-isolation aquarium. Therefore, ventilation rate (VR) indicates coping styles; consequently, VR is a proxy for the way fish will deal with environmental challenges.

The aggressive behavior of Nile tilapia introduced into novel environments with variation in enrichment.

Many studies show environmental enrichment is correlated with benefits to captive animals; however, one should not always assume this positive relationship given that enrichment increases the amount of resources that a territorial animal must defend and possibly affects its aggressive dynamics. In this study, we tested if environmental enrichment affects aggressive interactions in the aggressive fish Nile tilapia (Oreochromis niloticus). We compared fights staged between pairs of male tilapia of similar size (= matched in resource holding potential) in a novel arena that was either barren or enriched, to examine whether enrichment enhances territory value in line with theoretical predictions, with the potential for compromised welfare. We evaluated time elapsed until the first attack (latency), frequency of aggressive interactions and fight duration. We detected fight dynamic differences at the pair level. Higher resource value generated increased aggression but had no effect on fight duration or latency. This conclusion is in line with game theory predictions concerning resource value and contradicts the theory that enrichment of the environment will serve welfare purposes.

The correlation between subordinate fish eye colour and received attacks: a negative social feedback mechanism for the reduction of aggression during the formation of dominance hierarchies.

Eye darkening has been linked to social status in fish. The subordinate's eyes darken, while the eyes of the dominant fish become pale. Although this phenomenon has been described in salmonid fishes and in the African cichlid Nile tilapia Oreochromis niloticus, it is unclear whether eye darkening correlates with a reduction in aggressive behaviour. Thus, we evaluated the link between social status and eye darkening. We evaluated whether the eye colours of subordinate fish correlate with the frequency of received attacks in a neotropical fish, the pearl cichlid Geophagus brasiliensis. We paired pearl cichlids and quantified both the aggressive behaviour and the eye darkening of each fish. As has been described for Nile tilapia and Atlantic salmon, a clear-cut hierarchical relationship formed, where dominance and subordination were associated with pale and dark eye colours, respectively. Initially, eye colour darkening was positively correlated with the frequency of received attacks; however, a negative association occurred following eye darkening, in which the intensity of aggressive interactions decreased. Thus, fish that initially received a high number of attacks signalled subordination more rapidly and intensely (rapid and dramatic eye darkening), thereby inducing a negative social feedback mechanism that led to reduced aggression.

Challenges in assessing fish welfare.

Descriptions of feeling states in nonhuman animals have relied on indirect evidence from empirical data. Assumptions that fish do not experience suffering lack evidence and in fact contradict a large body of indirect scientific evidence and ethical concern. Why should the burden of proof rest on those defending the hypothesis that fish feel pain and other discomfort? In this article I address this controversy and describe typical methodsand the problems associated with themto identify animal welfare (feeling-based, physiological, and behavioral approaches intended to demonstrate feelings and welfare states). Then I urge a shift in scientific focus from efforts to either identify an internal state of well-being or determine whether an organism suffers, to efforts to identify conditions that promote a "good state" for an animal (i.e., a state it would choose). For this approach, I discuss preference tests and their implications for scientific research, teaching, aquarism, and fishing.