Effect of a Dummy Audience on Male–Male Interactions in Siamese Fighting Fish, Betta splendens

Recent research on Siamese fighting fish, Betta splendens, and other taxa has demonstrated that an audience can cause males to alter their behavior in aggressive interactions. One factor not taken into account in these studies is how exactly the audience influences these interactions. It is possible that a live audience may interact with the subjects, creating an active communication network rather than a signaler–receiver dyad with a passive audience. Here, we used a dummy audience to control for information exchange between the audience and the interactants that might cause them to modify their behavior. Audience treatments included dummies of male and female B. splendens, a dummy cichlid, and a control condition with no audience present. The presence of a dummy audience did not influence male–male interactions. However, males spent the most time near the audience tank when the audience was a dummy of a B. splendens. This suggests that some factor other than the physical presence of the audience is responsible for the modification of behavior found in previous audience effect studies in Siamese fighting fish. However, we cannot rule out definitively that our dummy audience is viewed as unimportant by the opponents and, thus, ignored. Further research is necessary to determine which component of the audience is important for producing audience effects.     

Male <Emphasis Type="Italic">Betta splendends</Emphasis> are equally aggressive toward neighbors and strangers

Male Siamese fighting fish, Betta splendens, are known for their ferociousness when defending their territories against male or female conspecific intruders. The aim of this study was to investigate whether this species exhibits the dear enemy phenomenon, where territorial males would be less aggressive toward neighbors with already established territories than toward complete stranger males. In experiment 1, a male Betta was placed in an aquarium. A second male was placed in a glass jar that was fitted in one corner of the aquarium. After 24 h, a cover was placed around the jar, and then removed after 1 h. Alternatively, the jar male was replaced with a different male of a different color while the jar was covered. The frequency and duration of opercular expansion by the aquarium male were recorded. In experiment 2, two male Bettas were placed, each in a half of a large aquarium that was equally divided by a clear, perforated Plexiglas divider. After 24 h, one male (intruder) was transferred into the other male’s (resident) half. In another variation of the experiment, the intruder was a stranger male Betta that the resident male had never encountered before. The opercular expansion and duration were scored for the resident male. The results indicated that male Siamese fish reacted similarly to familiar and stranger males. This lack of dear enemy effect in Siamese fish could be due to an inability to discriminate between neighboring males and non-neighboring males. Alternatively, territorial males could be equally aggressive to all intruders because all intruders represent equal danger.     

Social conditioning and dominance in male betta splendens

To determine whether social conditioning modifies relative dominance in the Siamese fighting fish ($\text{Betta splendens}$), individual fish were matched for gill cover erection frequency (GCEF) to a mirror. One member of each pair experienced a series of four winning encounters (dominants) while the other of the pair experienced four losing encounters (subordinates). After the fighting experience, matched pairs were placed together and aggressive responses of both fish were recorded. Fish experiencing wins displayed significantly more and were more aggressive than fish experiencing losses which showed significantly higher levels of avoidance. However, no change in mirror GCEF occurred as a result of differential experience.     

Alarm reaction and alert state inGambusia Affinis (Pisces,Poeciliidae) in response to chemical stimuli from injured conspecifics

We studied the behavior of the Poeciliid fishGambusia affinis after the introduction of 3 substances into their tank: a homogenization ofGambusia affinis, a homogenization of the Anabantid fishBetta splendens, and a blank made of distilled water. The response of the fish was measured as a change in their spatial distribution in the tank after the introduction of the substance. Two sizes of fish were used, and theGambusia homogenization produced clear alarm reactions in both, the fish fleeing to the bottom of the tank. This is one of a few examples available of recognition of alarm substances in non-ostariophysian fish. In addition, we found that the small fishes that had recently been exposed to the alarm substance stayed in an ‘alert state’, in which they had an increased sensitivity to mechanical and visual fright stimuli.     

The effect of body coloration and group size on social partner preferences in female fighting fish (Betta splendens)

Females of the fighting fish Betta splendens have been shown to associate with other B. splendens females in a manner reminiscent of shoaling behavior. Since body coloration varies dramatically in this species, and since body coloration has been shown to affect shoalmate choice in other species of fish, we examined the influence of body coloration on association preferences in female B. splendens. In dichotomous choice tests, B. splendens females spent more time swimming near groups of females (regardless of coloration) than swimming near an empty chamber, and chose to swim near fish of similar coloration to their own when choosing between two distinctly colored groups of females. When examining the interplay between body coloration and group size, focal fish spent more time swimming near larger groups (N = 5) of similarly colored fish than swimming near an individual female of similar coloration. However, focal fish showed no preference when presented with an individual female of similar coloration and a larger group of females of dissimilar coloration. These results suggest that association choices in B. splendens females are strongly affected by both body coloration and by group size.     

Male Siamese fighting fish use gill flaring as the first display towards territorial intruders

The Siamese fighting fish (Betta splendens) is well known as an aggressive fish with unique spawning and parental care behavior. During reproduction, male fish construct a bubble nest, court females, protect the brood, and defend the territory through aggressive displays. Aggression in male Siamese fighting fish has long been the subject of investigation; however, the kinematics of aggression during contests have been largely overlooked. Here we investigated how nest-holding, male Siamese fighting fish use two different types of displays, gill flaring and fin spreading, towards intruders during various reproductive phases; before (BB) and after bubble nest building, and after spawning (AS), and hatching (AH). Males were more aggressive towards male than female intruders and the level of aggression changed significantly between reproductive phases. Gill flaring, the more energetically costly display, was the dominant initial display towards male and female intruders in BB, AS, AH phases. However, defending males switched to fin spreading after prolonged exposure to intruders. The results suggest that Siamese fighting fish use gill flaring as an acute response in order to defend their territory; this response may be replaced by fin spreading as a chronic response, probably to reduce the energetic costs during the contest.     

Hypoxia-induced developmental plasticity of larval growth,gill and labyrinth organ morphometrics in two anabantoid fish: The facultative air-breather Siamese fighting fish (Betta splendens) and the obligate air-breather the blue gourami (Trichopodus trichopterus)

Larval and juvenile air breathing fish may experience nocturnal and/or seasonal aquatic hypoxia. Yet, whether hypoxia induces respiratory developmental plasticity in larval air breathing fish is uncertain. This study predicted that larvae of two closely related anabantid fish—the facultative air breather the Siamese fighting fish (Betta splendens) and the obligate air breathing blue gourami (Trichopodus trichopterus)—show distinct differences in developmental changes in body, gill, and labyrinth morphology because of their differences in levels of dependency upon air breathing and habitat. Larval populations of both species were reared in normoxia or chronic nocturnal hypoxia from hatching through 35–38 days postfertilization. Gill and labyrinth variables were measured at the onset of air breathing. Betta splendens reared in normoxia possessed larger, more developed gills (~3× greater area) than T. trichopterus at comparable stages. Surface area of the emerging labyrinth, the air breathing organ, was ~ 85% larger in normoxic B. splendens compared to T. trichopterus. Rearing in mild hypoxia stimulated body growth in B. splendens, but neither mild nor severe hypoxia affected growth in T. trichopterus. Condition factor, K (~ 1.3 in B. splendens, 0.7 in T. trichopterus) was unaffected by mild hypoxia in either species, but was reduced by severe hypoxia to <0.9 only in B. splendens. Severe, but not mild, hypoxia decreased branchial surface area in B. splendens by ~40%, but neither hypoxia level affected Trichopodus branchial surface. Mild, but not severe, hypoxia increased labyrinth surface area by 30% in B. splendens. However, as for branchial surface area, labyrinth surface area was not affected in Trichopodus. These differential larval responses to hypoxic rearing suggest that different larval habitats and activity levels are greater factors influencing developmental plasticity than genetic closeness of the two species.     

Red fish, blue fish: trade-offs between pigmentation and immunity in Betta splendens

Carotenoid pigments are responsible for many examples of sexuallyattractive red, orange, and yellow coloration in animals andplay an important role in antioxidant and immune defenses. Becausevertebrates cannot synthesize carotenoids, limited dietary availabilitymay impose a trade-off between maintaining ornamental colorationand health. We used an experimental approach to test the carotenoidtrade-off hypothesis in the fighting fish Betta splendens, byexamining whether carotenoid allocation strategies differ amongconspecifics that exhibit a gradient of body coloration fromblue to red. We found that male redness is underlain by carotenoidsand that females preferred to associate with red males overblue males, suggesting a sexually-selected advantage to beingred. Moreover, we found strong experimental support for thecarotenoid trade-off hypothesis, as individuals that variedin color did not appear to allocate carotenoids equally to bothimmune response and coloration. Redder fish given supplementalcarotenoids increased in both immune response (to a phytohemagglutinationchallenge) and redness compared with controls. In contrast,bluer fish given supplemental carotenoids did not become morered but instead benefited immunologically more so than eithercontrol or redder supplemented fish. These results enhance ourunderstanding of the evolution and plasticity of carotenoidmobilization and utilization pathways in animals.     

Characteristics of the Achnatherum splendens community along the Qinghai-Tibet Railway, China

The Achnatherum splendens community is an important type of warm steppe vegetation which plays a very important role in environmental protection in the area along the Qinghai-Tibet Railway. In order to understand its dynamic features, the characteristics of the A. splendens community, such as spatial distribution pattern, species richness and biomass, were examined near Haiyan County, Qinghai Province. Results showed that the A. splendens community consisted of two types of mosaic patches, the microaggregation dominated by A. splendens and the microaggregation dominated by Stipa sp. The number and size of the two patches showed an interesting distribution pattern. The A. splendens patch decreased with increasing distance away from the railway, while species diversity increased. The total coverage of A. splendens was significantly higher than that of Stipa sp. The number and size of patches may indicate the degradation of the A. splendens community, which could be accelerated by human activities. Based on the characteristics and status quo of the A. splendens community, we suggest that some measures be taken to control human activities and rodents to conserve the A. splendens community. __________ Translated from Acta Ecologica Sinica, 2007, 27(2): 497–503 [译自:生态学报]     

Benthic fish exhibit more plastic crypsis than non-benthic species in a freshwater spring

Cryptic coloration reduces the ability of predators to detect prey, but the plasticity of this defense varies. Some organisms possess static and permanent cryptic coloration, whereas in other species color changes may be induced. Depending upon the species, induced color changes may be reversible or irreversible. In this study, we examined a subtle, rapid, and reversible crypsis in which small fish exhibit muted changes in brightness to match varying substrates in clear spring water. In the laboratory, we visually measured the changes in brightness, using a ten-point brightness scale, of five abundant small species in our study spring. Two species, Lucania goodei and Heterandria formosa, exhibited no change, but the other three species exhibited changes in brightness to match background brightness. Two species, Gambusia holbrooki and Poecilia latipinna, exhibited only slight shifts, whereas Lucania parva exhibited relatively large shifts in brightness and color pattern—from virtually white to tan interspersed with dark-brown bands. In the field, L. parva also exhibited significant shifts in brightness and color pattern, both when swimming freely and when enclosed in an open-bottomed cage. These results suggest that rapid cryptic changes in brightness may augment other forms of defense in small vulnerable fish.     

Audience effect is context dependent in Siamese fighting fish, Betta splendens

Investigations of communication networks in animals have focusedprimarily on determining whether animals extract informationfrom peripheral contests (eavesdropping) or respond to the presenceof bystanders (audience effect). The possibility that an animal'sresponse to being watched might be context dependent, however,has been explored in far less detail. This study investigatedthe influence of two contexts, exposure to audiences of differentsexes and presence or absence of a nest, on the aggressive behaviorof interacting male Siamese fighting fish, Betta splendens.Males interacted in the presence (male, female) or absence ofan audience in three different nest conditions (0, 1, or 2 nests).Audience sex and territorial status influenced aggressive behaviorin the interacting males, but a strong audience x nest interactionalso was uncovered. Males were more aggressive when neithermale had a nest and a male audience was present than when afemale or no audience was present. Males also were more aggressivewhen only one male had a nest and a male audience was presentthan when a female or no audience was present. When both maleshad nests and a male audience was present, however, males wereless aggressive than when only one male or neither male hada nest. In sum, aggressive behavior was influenced by the interactionbetween audience and nest; neither nest nor audience alone wassufficient to explain the results. Male Siamese fighting fishalter their behavior based on both external cues, the sex ofthe audience, and internal cues, reproductive state and resourcepossession. Our results emphasize the importance of consideringaspects of an animal's environment when examining audience effectsand communication networks in general.     

Virtual plankton: A novel approach to the investigation of aquatic predator‐prey interactions

Small‐scale zooplankton swimming behaviors can affect aquatic predator‐prey interactions. Difficulties in controlling prey swimming behavior however, have restricted the ability to test hypotheses relating differences in small‐scale swimming behavior to frequency of predation by fish. We report here a Virtual Plankton (VP) system that circumvents this problem by allowing the observation of fish “preying"on computer‐generated prey images whose size, shape, color and swimming behavior can be precisely controlled. Two experiments were performed in which bluegill sunfish (Lepomis macrochirus) were given a choice of either two VP images, one of which moved twice as fast as the other, or six VP, one of which moved either faster (1.25 x, 1.5 x or 2 x ) or slower (0.5 x) than the other five. Current predator‐prey models based on encounter probabilities and prey visibility predict that moving faster increases predation risk and conversely, moving slower decreases predation risk. In agreement with existing predator‐prey models, in both experiments, fish chose faster moving VP significantly more often than their slower moving neighbors. Contrary to the predictions of existing models, in the second experiment with six VP, the rate at which fish chose a prey image moving half as fast as the five surrounding images did not differ significantly from the rate predicted by chance(l/6). These results suggest that current fish‐zooplankton predation models would benefit by the incorporation of small‐scale swimming behavior and assessments of its influence on overall prey visibility.     

Mate Choice and Spawning Success in the Fighting Fish Betta splendens: the Importance of Body Size, Display Behavior and Nest Size

Courtship displays should be exaggerated enough to attract mates and yet tempered so as not to deter them. We tested this hypothesis in the fighting fish Betta splendens by studying courtship displays and body size and their relationships with male parental quality and female fecundity, as well as the effects of display behavior and body size on mate choice decisions and spawning success. Because of their high degree of parental investment, males are expected to be discriminating in their choice of mates. Males who displayed more frequently built larger nests, a measure of parental quality, but larger males did not. When females were paired with males with high display rates, however, the pair had fewer eggs in their nest, even when accounting for female body mass. In a mate choice test using computer‐generated male stimuli that differed only in display behavior, females showed no preferences for displaying males vs. non‐displaying males, or for males with higher display rates vs. lower display rates. In similar tests in which the computer‐generated males differed only in size, females preferred larger males, but also preferred males that differed with respect to body size (negative assortative mating). Males preferred computer‐generated females that performed courtship displays over non‐displaying females, but showed no preferences for female body size. Neither a female's body size nor her display behavior was a significant predictor of her fecundity as estimated by the number of eggs released during spawning. Thus, our results suggest that female B. splendens must balance male parental quality (nest size) with the risk of potentially disruptive or dangerous behavior during spawning, and that females may minimize these risks through negative size‐assortative mating. Female display behavior, while unrelated to fecundity in our study, may attract males because it indicates reproductive readiness or serves a species‐recognition function.     

Visual Sensation of an Ethological Stimulus, the Agonistic Display of Betta splendens, Revealed using Multi-Unit Recordings from Optic Tectum

We made multi-unit recordings from the optic tectum of Betta splendens while they viewed the agonistic display of conspecifics. Movement of displaying animals evoked brisk, arrhythmic bursting in tectal units. Particularly high frequency spike bursts coincided with the onset of full display, a behavioral element known to be of ethological significance. These results show that it is possible to obtain informative electrophysiological recordings from optic tectum using behaving conspecifics as stimuli. More frequent use of natural stimuli can be expected to contribute to a greater understanding of the link between the neural substrates of vision and visually guided behavior.     

Differences in aggressive behavior between convict cichlid color morphs: amelanistic convicts lose even with a size advantage

Convict cichlids (Archocentrus nigrofasciatus) are a territorial, monogamous, and biparental Central American cichlid fish. Convicts exist in two common color morphs: the wild-type (WT) black-barred form and an amelanistic (AM) barless morph. Color morphs affect aggressive interactions in other species of fish. We staged fights between males of each color morph with varying size asymmetries and found that WT males were able to overcome a size disadvantage by increasing their rate of aggressive behavior. AM males lost more often when smaller than their opponent, apparently because they did not increase their rate of aggressive behavior when at a size disadvantage. We discuss two possible hypotheses to explain these findings: (1) that there are genetic differences in aggressive behavior between the morphs and (2) that AM fish are disadvantaged in staged contests because they are unable to signal via changes in bar coloration.     

High molecular mass glycoproteins associated with the siliceous scales and bristles of Mallomonas splendens (Synurophyceae) may be involved in cell surface development and maintenance

The elaborate scale case of Mallomonas splendens (Synurophyceae) consists of an overlapping arrangement of siliceous scales. In addition, siliceous bristles are attached to specialized base plate scales located at both the anterior and posterior ends of the cells. We have generated monoclonal antibodies against molecules associated with the scale case of M. splendens. One of these antibodies, designated MsS.H9, labelled a proteinaceous epitope of high-molecular-mass cell surface glycoproteins. Immunofluorescence and immunoelectron microscopy demonstrated that only two regions of M. splendens scale cases were labelled by MsS.H9, namely, the upper surface of the scales that contact neighboring scales and the bases of the bristles. Immunoelectron microscopy using thin sections of M. splendens cells showed these labelling sites corresponded to the amorphous material at the sites of scale-to-scale overlap and to a fibrillar complex located at scale-to-bristle attachment sites. Scales and bristles of M. splendens are formed within the cell, in silica deposition vesicles. Immunolabelling of cell sections containing developing scales and bristles showed that MsS.H9 labelling sites were present very early in the formation of these cell surface components. MsS.H9 labelling was also found associated with developing flagellar hairs whereas no labelling was detected on these structures after their deployment onto the flagellum. The location of MsS.H9 labelling sites strongly suggests that the molecule(s) recognized by the antibody plays a role in the adhesion of the individual components making up the scale case of M. splendens.Abbreviations CER chloroplast endoplasmic reticulum - ER endoplasmic reticulum - SDV silica deposition vesicle This work was supported by a grant from the Australian Research Council to R.W. We thank Dr. P. L. Beech for Fig. 13, Dr. L. Perasso for technical assistance and the Plant Cell Biology Group for the use of their monoclonal facilities.     

Development of the cranium and paired fins in Betta splendens (Teleostei: Percomorpha): Intraspecific variation and interspecific comparisons

The development of the chondrocranium and the relative timing of ossification of the osteocranium is described in the teleost fish Betta splendens from a large series of cleared and differentially stained specimens. General trends in ossification patterns are examined from developmental, phylogenetic, and functional contexts. As in many other vertebrates, dermal bones form before cartilage bones. Ossification sequence conforms to functional need in a very general way, but there are many inconsistencies in the details of order. For example, some bones that are directly involved in feeding ossify no earlier than bones more indirectly involved. Comparisons of ossification sequence within specific cranial regions are made among Betta splendens, Oryzias latipes (Atherinomorpha), and Barbus barbus (Ostariophysi) within a phylogenetic framework. Many evolutionary changes in relative sequence of ossification are evident within regions among these taxa, yet many other sequences are conserved. The logistic difficulty of comparing entire cranial ossification sequences (vs. regional sequences) makes evident the need for new methods for identifying and quantifying sequence changes. Intraspecific variation in order of ossification is described for the first time in teleost fishes. To the extent that ossification sequence varies intraspecifically, conclusions drawn from previous interspecific comparisons are compromised. Understanding the importance of changes in ossification order within and among taxa will require experimental, functional, and evolutionary work. © 1996 Wiley-Liss, Inc.     

Analysis of early growth of guppy strains,Poecilia reticulata,with different color patterns

Summary The guppy, Poecilia reticulata, is economically the most important species of freshwater ornamental fish cultured in Singapore. About 30 strains with different color patterns and fin shapes are reared in guppy farms practising monoculture in Singapore. To compare the growth rates of domesticated strains with different color patterns, newborn fry of 11 strains were obtained on the same day from a single farm in Singapore and were reared experimentally in the laboratory for about 100 days. Each strain was distributed randomly into 4 tanks with 30 fish/tank. Weekly weighings of 10 fish/tank were made from 17 to 100 days of age. Three strains were homozygous for the autosomal recessive blond gene which gives rise to a pale yellow background pigmentation (bb). These blond strains had significantly smaller body weights than corresponding ones with the same color pattern but with the wild-type grey-brown background coloration due to the dominant allele (BB). The strains with the red tail pattern due to a dominant X-linked gene (Rdt) had more rapid growth than those with other tail color patterns including the blue, black, green snakeskin and variegated. However, no significant differences were detected among the other color pattern strains. Thus among the strains studied, the blond strains were associated with slower growth while those with the red tail color were associated with faster growth.     

The influence of social status on the rate of growth, eye color pattern and insulin-like growth factor-I gene expression in Nile tilapia, Oreochromis niloticus

Many aspects of teleost physiology are subject to regulation by social interactions. To evaluate the relationship of social status with growth, eye color pattern and hepatic Insulin-like Growth Factor-I (IGF-I) mRNA expression, 30 Oreochromis niloticus were isolated for 10 days and were used in a social pair study. Results revealed that growth of both dominant (except 1 day after social interaction) and subordinate individuals was suppressed, but growth suppression was greater in the subordinates. The dominant fish completely inhibited the feeding of the subordinate individuals during and 1 day after they were introduced into the aquaria together. After that, a pattern of highly aggressive attacks by dominant fish only partially inhibited feeding by the subordinates. Differential alterations in growth rate between dominants and subordinates were attributed more to behavioral changes (i.e., feeding) as transduced by physiological regulators (i.e., IGF-I level and possibly serotonin and/or neuropeptide Y) but may also be due to changes in metabolism. The fish's relative position in the social hierarchy consistently influenced the levels of IGF-I mRNA in the liver and the eye color pattern. Lower social status depressed hepatic IGF-I levels while dominant status stimulated hepatic IGF-I production, possibly in response to inhibition of somatostatin release in the hypothalamus, leading to greater secretion of pituitary growth hormone (GH). A significant positive association was detected between the IGF-I mRNA expression of the dominant fish and the level of aggression (number of attacks) during the encounter. Social status also influenced the eye color pattern of the fish. During aggressive interactions, most of the fish (22 out of 24) displayed decreased eye darkening. At the later part of the encounter, all subsequent subordinates displayed eye-darkening patterns which acted as a social signal announcing social submission. After the encounter dominant fish had paler eye color pattern than subordinates.     

Parameters of rewards on choice behavior in Siamese fighting fish (Betta splendens)

Five experiments were conducted with Siamese fighting fish (Betta splendens) to investigate how choices in a T-maze were affected by parameters of a social reward (aggression display to another male): presence or absence, amount, delay and distance traveled. Bettas showed a preference for the side associated with the presence of another male rather than the side associated with nothing (Exp 1), a greater length of time of the reward (Exp 2) and shorter delay (Exp 3). The animals were indifferent when one side offered a longer delay to a longer reward time compared with a shorter delay to a shorter reward time (Exp 4). What was most surprising, however, was that fish preferred to choose the side that was associated with swimming a greater distance to reach an opponent male (Exp 5). These experiments demonstrate that, while some parameters of a visual reward affect behavior in predicable ways (greater amount, shorter delay), the complex motivations underlying inter-male aggression can produce what appear to be paradoxical results.