The short-term effects of increasing meal frequency on stereotypic behaviour of stabled horses

In this study, we investigated the effect of increasing the number of meals of concentrate (whilst maintaining the same daily intake) on the behaviour of stabled horses with particular reference to stereotypic activities. The study was carried out on a working equestrian yard with stables for up to 50 horses. A pilot study was used to record incidence of stereotypic behaviour and to select subjects for the main study. In this, the behaviour of 30 warm-blooded horses was recorded during their morning (08:30 h) and afternoon (16:30 h) concentrate feeds. Whilst there was a low incidence of stereotypic behaviour (5.6% of scans) in the population, they were more commonly observed in the afternoon (7.1%) than the morning observations (4.2%; P < 0.05). The higher incidence in the afternoon observation appeared to be related to the lower availability of high fibre forage during the afternoon meal. In the main study nine horses were fed their normal ration of concentrate divided between two, four or six equally sized meals. Their behaviour was compared with seven control horses, which received two meals per day throughout the trial. As the number of meals increased, the treatment horses showed a decrease in oral stereotypies (P < 0.01), but an increase in weaving (P < 0.05) and nodding (P < 0.01) prior to feeding. The control group increased weaving, nodding and oral stereotypies (all P < 0.05) as their yard-mates received more meals. Consequently there was an overall increase in incidence of stereotypy in both treatment and control horses with the increase in meal frequency. The study, therefore, suggests that dividing the stabled horses’ concentrate ration into a number of smaller meals may be an effective means of reducing oral stereotypies, but that pre-feeding stereotypies may persist and that the practise may increase the frequency of stereotypic behaviour on unfed horses in visual contact.     

The effect of hunger on mating behaviour and sexual selection for male body size in Gerris buenoi

The set of mating behaviours expressed by an individual may depend upon the state of that individual and local environmental conditions. Understanding how these factors affect mating behaviours may elucidate how a mating system operates, and its consequences for the form and strength of sexual selection. We conducted two experiments on the water striderGerris buenoi to (1) determine the effect of hunger on the mating behaviour of both sexes and (2) examine female choice for large males. In our first experiment, we manipulated hunger (20 h starvation) in both sexes and recorded mating, male harassment, copulation duration and guarding duration. We predicted that hunger would increase female reluctance to mate because mating conflicts with foraging. Female hunger (20 h starvation) decreased mating rate by two-thirds but had no significant effect on male mating behaviour. In a second experiment, we examined the effect of female hunger, and resulting reluctance, on sexual selection for large male size. Hungry females (5 h starvation) were placed with two fed males (one large, one small) and we recorded male premating and mating behaviours. We observed significant large-male mating advantage when females were hungry, but not when satiated. Mating efforts (harassment, premating struggles) were similar for both male phenotypes in both female hunger treatments, suggesting that the mating advantage of large males resulted from increased reluctance of hungry females to mate. Neither male body size nor female hunger explained a significant amount of variation in copulation duration or guarding duration. We discuss our results in light of two competing hypotheses for female choice (active and passive) on male body size and suggest that passive choice for large males acts in this system. Copyright 2002 Published by Elsevier Science Ltd on behalf of The Association for the Study of Animal Behaviour     

Age‐related activity patterns are moderated by diet in Queensland fruit flies Bactrocera tryoni

Life‐history parameters and the fitness of tephritid flies are closely linked to diet. Studies of locomotor behaviour can provide insights to these links, although little is known about how locomotor behaviour is influenced by diet. In the present study, video recordings of Queensland fruit flies Bactrocera tryoni Froggatt (Diptera: Tephritidae) (‘Q‐flies’) that are maintained individually in cages are used to determine how diet affects the activity patterns (flight, walking, grooming, inactivity) of males and females at ages ranging from 4 to 30 days. The frequency and total duration of activities over 10‐min trials are affected by diet, age and sex. Supplementation of diet with hydrolysed yeast results in a higher frequency and duration of flight in flies of all ages and both sexes. The effect of diet on other activities varies with age. Q‐flies fed sugar only increase walking frequency steadily from 4 to 30 days post‐eclosion, whereas flies fed sugar + yeast have higher walking frequencies at 4 and 10 days than flies fed sugar only, although they then exhibit a sharp decline at 30 days post‐eclosion. The frequency and duration of inactivity remain consistent in flies fed sugar + yeast, whereas flies fed sugar only exhibit a marked increase in inactivity from 4 to 30 days post‐eclosion. Compared with older flies, 4 day‐old Q‐flies fed sugar only spend considerably more time grooming. The potential of activity monitoring as a quality control test for flies that are mass‐reared for use in sterile insect technique programmes is discussed.     

Sexual selection in a tropical fruit fly: role of a plant derived chemical in mate choice

The specific mechanisms by which selective pressures affect individuals are often difficult to resolve. In tephritid fruit flies, males respond strongly and positively to certain plant derived chemicals. Sexual selection by female choice has been hypothesized as the mechanism driving this behaviour in certain species, as females preferentially mate with males that have fed on these chemicals. This hypothesis is, to date, based on studies of only very few species and its generality is largely untested. We tested the hypothesis on different spatial scales (small cage and seminatural field‐cage) using the monophagous fruit fly, Bactrocera cacuminata. This species is known to respond to methyl eugenol (ME), a chemical found in many plant species and one upon which previous studies have focused. Contrary to expectation, no obvious female choice was apparent in selecting ME‐fed males over unfed males as measured by the number of matings achieved over time, copulation duration, or time of copulation initiation. However, the number of matings achieved by ME‐fed males was significantly greater than unfed males 16 and 32 days after exposure to ME in small cages (but not in a field‐cage). This delayed advantage suggests that ME may not influence the pheromone system of B. cacuminata but may have other consequences, acting on some other fitness consequence (e.g., enhancement of physiology or survival) of male exposure to these chemicals. We discuss the ecological and evolutionary implications of our findings to explore alternate hypotheses to explain the patterns of response of dacine fruit flies to specific plant‐derived chemicals.     

On the toxicokinetics and the metabolism of deoxynivalenol (don) in the pig

Eleven castrated male pigs weighing 88.1?±?3.9?kg on average were adapted to a diet containing DON (4.2?mg DON/kg) over a period of 7 days. Feed was given restrictively with 1.1?kg per meal (two meals per day). On the day of measurement, all pigs were slaughtered at different time intervals following the morning meal containing DON (1, 2, 3, 4, 5, 6, 8, 15, 18 and 24?h after feeding), with the exception of one pig which was slaughtered unfed. DON and de-epoxy-DON were analysed in serum and digesta from consecutive segments of the digestive tract (stomach, small intestine divided into three parts of a similar length, caecum, colon, rectum). DON was rapidly and nearly completely absorbed while passing through the stomach and the proximal small intestine. Maximum serum concentration appeared 4.1?h after the DON-containing meal and half of the systemically absorbed DON was eliminated after 5.8?h. De-epoxy-DON appeared in increasing proportions from the distal small intestine and reached approximately 80% of the sum of DON plus de-epoxy-DON in faeces collected from the rectum. It was concluded that de-epoxydation of DON, which primarily occurs in the hindgut, probably does not contribute much to a detoxification in the pig.     

Time-of-day effect on cardiac responses to progressive exercise

This study was designed to examine time-of-day effects on markers of cardiac functional capacity during a standard progressive cycle exercise test. Fourteen healthy, untrained young males (mean?±?SD: 17.9?±?0.7 yrs of age) performed identical maximal cycle tests in the morning (08:00-11:00?h) and late afternoon (16:00-19:00?h) in random order. Cardiac variables were measured at rest, submaximal exercise, and maximal exercise by standard echocardiographic techniques. No differences in morning and afternoon testing values at rest or during exercise were observed for oxygen uptake, heart rate, cardiac output, or markers of systolic and diastolic myocardial function. Values at peak exercise for Vo(2) at morning and afternoon testing were 3.20?±?0.49 and 3.24?±?0.55?L min(-1), respectively, for heart rate 190?±?11 and 188?±?15?bpm, and for cardiac output 19.5?±?2.8 and 19.8?±?3.5?L min(-1). Coefficients of variation for morning and afternoon values for these variables were similar to those previously published for test-retest reproducibility. This study failed to demonstrate evidence for significant time-of-day variation in Vo(2)max or cardiac function during standard progressive exercise testing in adolescent males.     

Alpha-COPI coatomer protein is required for rough endoplasmic reticulum whorl formation in mosquito midgut epithelial cells

Background

One of the early events in midgut epithelial cells of Aedes aegypti mosquitoes is the dynamic reorganization of rough endoplasmic reticulum (RER) whorl structures coincident with the onset of blood meal digestion. Based on our previous studies showing that feeding on an amino acid meal induces TOR signaling in Ae. aegypti, we used proteomics and RNAi to functionally identify midgut epithelial cell proteins that contribute to RER whorl formation.

Methodology/Principal Findings

Adult female Ae. aegypti mosquitoes were maintained on sugar alone (unfed), or fed an amino acid meal, and then midgut epithelial cells were analyzed by electron microscopy and protein biochemistry. The size and number of RER whorls in midgut epithelial cells were found to decrease significantly after feeding, and several KDEL-containing proteins were shown to have altered expression levels. LC-MS/MS mass spectrometry was used to analyze midgut microsomal proteins isolated from unfed and amino acid fed mosquitoes, and of the 127 proteins identified, 8 were chosen as candidate whorl forming proteins. Three candidate proteins were COPI coatomer subunits (alpha, beta, beta''), all of which appeared to be present at higher levels in microsomal fractions from unfed mosquitoes. Using RNAi to knockdown alpha-COPI expression, electron microscopy revealed that both the size and number of RER whorls were dramatically reduced in unfed mosquitoes, and moreover, that extended regions of swollen RER were prevalent in fed mosquitoes. Lastly, while a deficiency in alpha-COPI had no effect on early trypsin protein synthesis or secretion 3 hr post blood meal (PBM), expression of late phase proteases at 24 hr PBM was completely blocked.

Conclusions

alpha-COPI was found to be required for the formation of RER whorls in midgut epithelial cells of unfed Aa. aegypti mosquitoes, as well as for the expression of late phase midgut proteases.     

Female sexual receptivity and male copula guarding during prolonged copulations in the damselflyIschnura senegalensis (Odonata:Coenagrionidae)

Laboratory experiments were conducted to clarify the relationship between female sexual receptivity and male copula guarding inI. senegalensis, a species that copulates for several hours. In insectaries, most copulations were initiated early in the morning, and terminated relatively synchronously between 11 00 and 13 00. Females refused males with wing-flutter display and oviposited alone in the afternoon regardless of copulation events of that morning. Females could sexually receive males only in the morning. Males copulated for several hours until 12 00 after which females could oviposit. To determine whether copulations that last for hours function as male copula guarding or only of sperm displacement, emerged males were kept at various densities and permitted to copulate with virgin and mated females in insectaries. Both with virgin and mated females, “social” (not solitary; 2–4 males / insectary) males initiated copulations early in the morning and always terminated at around 12 00. However, both with virgin and mated females, solitary (one male / insectary) males terminated copulations in the morning. In both cases, duration of copulations did not significantly differ for virgin females and mated females. Therefore, long (several hour) copulation is more likely to function as male copula guarding than as sperm displacement, and duration of copulations is predicted to be shortened when male density is very low.     

Energetic constraints on mating performance in the sand goby

I tested the effect of food addition on reproductive successin male sand gobies, Pomatoschistus minutus, by comparing food-supplementedmales with unfed, control males. The sand goby is a small marinefish with paternal egg care. The males were breeding in artificialnest sites in otherwise natural conditions in the field. I quantifiedenergy reserves by extracting nonpolar lipids. The food supplementimproved the fat reserves of the fed males as compared to unfedmales. Fed males spent more time at the nest, whereas unfedmales spent a much smaller proportion of their time at the nest.As a consequence, fed males mated sooner than unfed males andtended to get more eggs. In the unfed group, mating speed wascorrelated to body length so that bigger males mated sooner.The results suggest that the reproductive success of breedingsand goby males is constrained by the availability of energybut that this constraint is most severe for small males andless severe for bigger males. Energy availability through itseffect on condition will affect the investment in reproductiveeffort.     

Grooming as a secondary behavior in the shrimp Macrobrachium rosenbergii (Crustacea,Decapoda, Caridea)

The giant freshwater prawn, Macrobrachium rosenbergii, is a large shrimp extensively used in aquaculture whose grooming behaviors were analyzed in this study. Macrobrachium rosenbergii exhibits three unique male morphotypes that differ in their behavior, morphology and physiology: small-clawed males (SM), orange-clawed males (OC) and blue-clawed males (BC). The largest and most dominant males, BC males, are predicted to have significantly different grooming behaviors compared to females and the other two male morphotypes. These BC males may be too large and bulky to efficiently groom and may dedicate more time to mating and agonistic interactions than grooming behaviors. Observations were conducted to look at the prevalence of grooming behaviors in the absence and presence of conspecifics and to determine if any differences in grooming behavior exist among the sexes and male morphotypes. Significant differences in the grooming behaviors of all individuals (females and male morphotypes) were found. BC males tended to have the highest grooming time budget (percent of time spent grooming) while SM males had a relatively low grooming time budget. The grooming behaviors of the male morphotypes differed, indicating while these males play distinct, separate roles in the social hierarchy, they also have different grooming priorities. The conditions in which Macrobrachium rosenbergii are cultured may result in increased body fouling, which may vary, depending on the grooming efficiencies and priorities of these male morphotypes. Overall, grooming behaviors were found to be a secondary behavior which only occurred when primary behaviors such as mating, feeding or fighting were not present.     

Pen size and parity effects on maternal behaviour of Small-Tail Han sheep

The aim of this experiment was to study the effects of pen size and parity on maternal behaviour of twin-bearing Small-Tail Han ewes. A total of 24 ewes were allocated to a 2×2 design (six per pen), with parity (primiparous or multiparous) and pen size (large: 6.0×3.0 m; small: 6.0×1.5 m) as main effects at Linyi University, Shandong Province, China. Behaviour was observed from after parturition until weaning. All ewes were observed for 6 h every 5 days from 0700 to1000 h and from 1400 to 1700 h. Continuous focal animal sampling was used to quantify the duration of maternal behaviours: sucking, grooming and following as well as the frequency of udder accepting, udder refusing and low-pitched bleating. Oestradiol and cortisol concentrations in the faeces (collected in the morning every 5 days) were detected using EIA kits. All lambs were weighed 24 h after parturition and again at weaning at 35 days of age. The small pen size significantly reduced following (P<0.005), grooming (P<0.001) and suckling durations (P<0.05), as well as the frequency of udder refusals (P<0.001). However, there was a significant interaction with ewe parity, with decreased grooming and suckling in the small pen largely seen in the multiparous ewes (P<0.001). Independent of pen size, multiparous ewes accepted more sucking attempts by their lambs (P<0.05) and made more low-pitched bleats than primiparous ewes (P<0.001). Multiparous ewes had higher faecal oestradiol concentrations than primiparous ewes (P<0.001), and ewes in small pens had higher faecal cortisol levels compared with ewes in larger pens (P<0.001). As lambs increased in age, the duration of maternal grooming, following and suckling as well as frequency of udder acceptance and low-pitched bleating all declined, and the frequency of udder refusing increased (P<0.001 for all). Ewe parity, but not pen size, affected lamb weight gain during the period of observation (P<0.001). This is the first study to show that pen size, interacting with parity, can affect the expression of maternal behaviour in sheep during lactation. The study is also the first to report on the maternal behaviour of Chinese native sheep breeds (Small-Tail Han sheep), with implications for the production of sheep in China.     

Inhibition of mating behaviour before feeding in the tick Aponomma hydrosauri

When on their host, both male and female adults of the reptile tick Aponomma hydrosauri require at least five days of feeding before normal mating behaviour will start. In off-host trials, fed females attract significantly more male contact than do unfed females, but unfed males show normal mating behaviour. All aspects of female mating behaviour are inhibited before feeding. Male mating behaviour has two stages: one comprises detachment and searching, the other, courtship behaviour. Only the former is inhibited before male feeding. This suggests the female signal has two components: one initiating searching, and the other initiating courtship. Males will always respond to the second component, but only after feeding will they respond to the first.     

Patterns of parental care in two cricetid rodents, Onychomys torridus and Peromyscus leucopus

Daily observations were made of the maternal and paternal behaviour of Onychomys torridus and Peromyscus leucopus in laboratory cages. In both species, the females spent more time in the nest area with the young and groomed the young more frequently than their male partners, but the males played a surprisingly active role in nest attendance. Paternal nest attendance was more frequent in O. torridus males than in P. leucopus males. Onychomys males were also more active in grooming the young than Peromyscus males. These species-specific patterns of parental care are discussed in relation to the behavioural and ecological characteristics of Onychomys and Peromyscus.     

Time-of-Day Effect on Cardiac Responses to Progressive Exercise

This study was designed to examine time-of-day effects on markers of cardiac functional capacity during a standard progressive cycle exercise test. Fourteen healthy, untrained young males (mean?±?SD: 17.9?±?0.7 yrs of age) performed identical maximal cycle tests in the morning (08:00–11:00?h) and late afternoon (16:00–19:00?h) in random order. Cardiac variables were measured at rest, submaximal exercise, and maximal exercise by standard echocardiographic techniques. No differences in morning and afternoon testing values at rest or during exercise were observed for oxygen uptake, heart rate, cardiac output, or markers of systolic and diastolic myocardial function. Values at peak exercise for Vo₂ at morning and afternoon testing were 3.20?±?0.49 and 3.24?±?0.55?L min^?1, respectively, for heart rate 190?±?11 and 188?±?15?bpm, and for cardiac output 19.5?±?2.8 and 19.8?±?3.5?L min^?1. Coefficients of variation for morning and afternoon values for these variables were similar to those previously published for test-retest reproducibility. This study failed to demonstrate evidence for significant time-of-day variation in Vo₂max or cardiac function during standard progressive exercise testing in adolescent males. (Author correspondence: thomas.rowland@bhs.org)     

Daily variation in body core temperature using radio-telemetry in aluminium industry shift-workers

The aim of this study was to investigate the diurnal variation in core temperature in aluminium shift-workers exposed to hot ambient conditions. Core temperature was continuously recorded via an ingestible radio-telemetry thermistor in 29 shift-workers. Data from the morning, afternoon and night shifts were aggregated for each participant to obtain 24-h recordings during work duties. Complete data were obtained from 10 participants. Results showed that body core temperatures recorded in the afternoon (from 12:00 h to 20:00 h) were significantly higher (P<0.05) than in the late evening, night and early morning (from 21:00 h to 08:00 h). In addition, core temperature displayed a circadian variation with a mesor of 37.45 (±0.19) °C, an amplitude of 0.23 (±0.12) °C and an acrophase at 16:36 h (±3:37 h). The peak values of core temperature recorded at each hour of the day on the work site followed the same pattern with an acrophase in the early afternoon. In summary, our data showed that shift-workers present higher core temperatures in the afternoon than in the morning or during the night. In addition, it was not the work duration but the hour-of-day that triggered the variation in core temperature. This result partly explains previous observations that workers under heat stress have a higher probability of heat illness during daytime shifts than during the night shift, and suggests that special care should be given to the afternoon shift and to the end of the morning shift.     

Influence of male competition on male mating behaviour in the cave molly, Poecilia mexicana

In many species, male mating behaviour is correlated with male body size, with large males often being preferred by females. Small surface-dwelling Poecilia mexicana males compensate for this disadvantage by being more sexually active and using sneaky copulations. In a cave-dwelling population, however, small males do not show this behaviour. Do small males alter their behaviour in the presence of a large rival? Here, we investigated the influence of male competition on male mating behaviour in the cave form. Two males of different sizes were mated with a female either alone or together with the other male. No aggressive interactions were observed between either fish. There was no statistically significant difference in the frequency of sexual behaviours between the two treatments. In both treatments, large males were more sexually active than small males. Thus, small cave molly males do not switch to an alternative mating behaviour in the presence of a larger rival. Possibly, the extreme environmental conditions in the cave (e.g. low oxygen content and high levels of hydrogen sulphide) favour saving energetic costs, resulting in the absence of alternative mating behaviour in small males.     

Mate Choice in Female Red-necked Pademelons,Thylogale thetis (Marsupialia: Macropodoidea)

Changes in the behaviour of captive female red-necked pademelons, Thylogale thetis, before, during and after oestrus were observed relative to the location of potential male mates. A penned female was presented with four adjacent, individually caged males representing the size classes large, medium, small and subadult. The female's behaviour was videotaped for 8–9 d spanning oestrous (3 d), post-oestrous (3 d) and normal (3 d) sessions of time. Changes in the activity, orientation and location of six penned females, relative to the caged males, were transcribed from videotape. A model of female pademelon behaviour pattern was developed. Results suggest that the females spent a significantly greater amount of time directly in front of the largest available male, relative to the other three smaller males during the oestrous period only, and that a significant proportion of this time was spent involved in active behaviours (grooming and locomotion, compared with inactive behaviours of sitting and scan/sniffing). Although there was a significant preference for females to orientate their bodies away from the location of the males, they faced the males significantly more during oestrus than any other session. Furthermore, the females faced the large male significantly more frequently than any other male. This study provides evidence for mate choice by female red-necked pademelons.     

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.     

Coping with social tension: Sex differences in the effect of food provision to small rhesus monkey groups

In previous research, male chimpanzees (Pan troglodytes) were found to be more ‘conciliatory’ than females, in that after aggressive incidents males more often engaged in socially positive contact with former opponents. The first part of the present paper presents similar results for a large breeding troop of rhesus monkeys (Macaca mulatta). To investigate further the possible sex difference in postconflict behaviour, a series of experiments was carried out on small isosexual groups of young rhesus monkeys, three male and three female groups. Food competition was created by providing either a single apple piece (Monopoly test) or a handful of small pieces (Equality test). Observations lasted for 30 min, and behaviour in tests without food provisioning was also recorded. The predicted response to food was an increase in aggression followed by restorative behaviour, such as grooming. In Equality tests both sexes showed similar responses, i.e. increased aggression and decreased grooming and cohesiveness. In Monopoly tests, however, their responses differed. After the aggression increase, which occurred in all groups, males showed a significant increase in grooming and cohesiveness, whereas females showed the (non-significant) opposite trend. Another sex difference was that changes in grooming and aggression frequencies were related to the rank order in female groups, but not in male groups. Since there was no evidence for a direct causal connection between grooming and aggression, a new model is introduced which links grooming behaviour to the type of food provision rather than to the social disturbance by aggression. According to this model inequality in food distribution causes social tensions. Males actively try to reduce these tensions, whereas females do not. Some alternative explanations are also discussed.     

Mediterranean Fruit Fly leks: factors affecting male location

1. The mating system of Mediterranean Fruit Flies ( Ceratitis capitata , Diptera: Tephritidae) is based on male leks which form within the canopies of certain trees. In this study the following hypotheses are tested: (a) fly location depends on microclimate and illumination and (b) larger and heavier males occupy preferred locations in leks. Accordingly, systematic quantitative observations of diel three-dimensional (3-D) locations of lekking C. capitata males were performed in field and field cage studies.
2. Fly locations were found to vary significantly during the activity hours.
3. Medflies were generally found calling from highest and most exterior locations during early morning and late afternoon hours. During the hottest hours (1200–1500) flies occupied lower locations within the tree canopy, and interior locations from 1000 to 1500.
4. Fly location (from 1000 to 1700) was correlated with the azimuth of the sun. However, the mean azimuth range of fly location was limited to 85° (59·45°–143·94°).
5. Both in the field and in the field cage the temperature, relative humidity and light intensity beneath the leaves on which the males perched were closer to the microclimate beneath fully shaded leaves than to microclimates beneath leaves exposed to direct sunlight.
6. It is concluded that male medflies occupy locations which confer suitable microclimates for calling and copulating, and suitable 3-D locations within the canopy which provide protection from predators, wind, direct sunlight and water loss.