Modifications to husbandry and housing conditions of laboratory rodents for improved well-being

For a change to be considered enriching, the change must enhance animal welfare and improve biological functioning of the animals. A review of the literature shows that a consensus on the definition of changes constituting "environmental enrichment" has yet to be reached. For this reason, the results of studies on the effects of rodent enrichment are inconsistent. In many cases, changes have not been shown to be real improvements. However, enrichment is increasingly appreciated as a way to improve the well-being of rodents, providing them with opportunities for species-specific behaviors that might be available to them in the wild. Frequently defined as "change to the environment," enrichment can be as complex as devices (frequently termed "toys") or as simple as the provision of tissues from which mice readily construct nests. Nest making is a learned behavior in rats, and laboratory rats do show preferences for chewable objects in their environment. Rather than attempting a comprehensive review of the entire literature on environmental enrichment and its effects on rodent physiology and behavior, this paper focuses on husbandry and housing alterations that may improve the welfare of laboratory rodents. The effects of beneficial changes in housing and husbandry on rodent well-being and on experimental variability--and thus cost--are discussed. Areas that require more research are suggested. Also suggested are possible inexpensive and effective enrichment schemes for laboratory mice that might include reducing the cage floor space per mouse combined with providing nesting material.     

Communal nesting and communal nursing in house mice,Mus musculus domesticus

The functional significance of communal nesting and nursing is poorly understood. Female house mice often communally nest, and within these communal nests females appear to indiscriminately nurse all pups, a rare trait for any mammal. In this study, the hypothesis that communal nesting provides protection from conspecific infanticide was tested and supported in semi-natural populations of house mice. Conspecific infanticide in single-mother nests (69%, N = 412) was twice that in communal nests (33%, N = 508). Because this major benefit of communal nesting does not require communal nursing, direct benefits to communal nursing itself were tested. Most proposed benefits should result in heavier weaning weights, but no differences were found between communal and single nests in the semi-natural populations. If communal nursing is to be avoided in communal nests, dams must recognize their own pups. Retrieval tests conducted in the laboratory produced equivocal results. Dams discriminated between pups that differed in age, but not between their own and other age-matched pups. The major survival advantage of communal nesting, coupled with the failure to find nutritional advantages for communally nursed pups, supports a recent suggestion that communal nursing is an unavoidable consequence of communal nesting. This hypothesis is further strengthened by data indicating that communal nesting partners tend to be kin, thereby providing inclusive fitness benefits to communal nursing. Although costs of communal nursing were proposed and tested, no such costs were found. We also show from 15 observations of infanticide that all classes of adults (territorial and non-territorial males, pregnant and non-pregnant females) are infanticidal. These observations are in conflict with previous laboratory studies.     

Nesting, courtship and kidney hypertrophy in Schistocephalus-infected male three-spined stickleback from an upland lake

The effect of Schistocephalus solidus infection on reproductive development and behaviour of male three-spined stickleback Gasterosteus aculeatus from an upland lake (Llyn Frongoch, Wales, U.K.) was investigated. Uninfected and infected males were collected on two separate occasions during the 2005 breeding season and encouraged to build nests under favourable laboratory conditions. Male nuptial colouration, courtship levels and nesting activity were recorded daily over two separate 21 day studies. Completed nests were removed, encouraging males to build multiple nests. On termination of each study, males were dissected and the kidney and any S. solidus plerocercoids were removed and weighed. In contrast to uninfected males, which readily built multiple nests, no infected males completed a nest during either study. Infected males also exhibited significantly reduced courtship levels, nesting activity, nuptial colouration, kidney development and body condition, compared with uninfected fish. Among infected three-spined stickleback, courtship and nesting behaviours tended to be more severely impacted in fish harbouring heavier plerocercoid burdens. In contrast to other populations, male three-spined stickleback from Llyn Frongoch appear to be unable to overcome the negative impacts of infections on reproductive development and behaviour when provided with short-term favourable conditions. Discrepancies between populations might have been related to the trophic status at the site of capture, emphasizing the importance of studying the biological effects of infections under different environmental conditions.     

Evaluation of social and physical enrichment in modulation of behavioural phenotype in C57BL/6J female mice

Housing conditions represent an important environmental variable playing a critical role in the assessment of mouse behaviour. In the present study the effects of isolation and nesting material on the behaviour of female C57BL/6J mice were evaluated. The mice were subjected to different rearing conditions from weaning (at the age of 3 weeks). The study groups were group- and single-housed mice, divided further into groups with or without nesting material (species-specific enrichment). After 8 weeks spent in respective conditions the behavioural testing began. Both factors (social conditions and nesting material) appeared to have a significant impact on the behavioural phenotype. However, it is important to stress that the interaction between the factors was virtually absent. We established that isolation increased locomotor activity and reduced anxiety-like behaviour in several tests of exploration. In contrast, absence of nesting material increased anxiety-like behaviour. Neither factor affected rota-rod performance, nociception and prepulse inhibition. Contextual fear memory was significantly reduced in single-housed mice, and interestingly, in mice with nesting material. Cued fear memory was reduced by single-housing, but not affected by enrichment. Mice from enriched cages displayed faster and better learning and spatial search strategy in the water maze. In contrast, isolation caused significant impairment in the water maze. In conclusion, both isolation and species-specific enrichment have profound effects on mouse behaviour and should be considered in design of the experiments and in assessment of animal welfare issues.     

Ecological and Behavioral Aspects of Nest Building in Wild Bonobos (Pan paniscus)

Data on nest building behavior were collected from free ranging bonobos (Pan paniscus). Considering the physical features of both nests and trees used for nest construction, bonobos appeared to show various preferences: (1) Trees with small leaves were chosen more frequently than expected; (2) The majority of nests were constructed in trees of the middle forest layer (15 to 30 m) and at a mean height of 16 m; (3) nests built during the day were located higher than night nests and differed in the general mode of construction (number of trees involved). Comparing nests made by mature males and females it became apparent that females (4) built nests higher, (5) constructed day nests more frequently and (6) used them for a longer time than males. Comparison of data collected at different study sites indicates site-specific differences concerning selection of trees and location of nesting sites. Behavioral observations made clear that nests were not only used for rest but also for other activities like feeding, social grooming and play. Moreover, the study provided the first evidence that bonobos build nests also in order to avoid potential or imminent conflicts with other party members. Field observations indicate that these nests symbolize a taboo zone and show another facet of the social abilities of bonobos for the solution of intra group conflicts. It is suggested that this behavior derives from the spatial intolerance shown by females during the process of weaning.     

筑巢材料在实验大、小鼠环境丰荣中的应用

相对于标准鼠盒,实验大、小鼠更喜好有筑巢材料的“环境丰荣”鼠盒.向标准鼠盒中放入筑巢材料,可以为大、小鼠提供筑巢、躲避、攀爬、休息的环境.因此,在实验大、小鼠饲养过程中,提供筑巢材料,是一个简单有效的丰富实验动物环境,提高实验动物福利的方法.本文就筑巢材料应用背景、近年来在啮齿类实验动物环境丰荣中的应用、对实验动物福利的影响三方面做以综述,并对今后筑巢材料在改善实验动物福利的应用前景作以展望.     

Experimental Study of Nest-site Selection in the Biscutate Swift (Streptoprocne biscutata, Aves: Apodidae) in Southern Brazil

Biscutate swift Streptoprocne biscutata nests are usually built on protected rocky cliff walls. Birds often renest at the same location. Remains of previous nests may offer information about potential nests and quality of nest-sites. Here, we experimentally study nest-site selection to test the hypothesis that information from previous nests is used in current nest-site selection. We placed old nest material at artificial nest-sites to test whether new nest-sites are chosen based on the presence of nesting material. We also tested whether the use of natural nest-sites is influenced by nesting material by creating two types of natural sites: previously used natural nest-sites with vestiges of old nests removed and never used natural nest-sites to which vestiges of old nests were added. In the first experiment, in 139 nest-use opportunities, 16 artificial nest-sites were used, all of which included vestiges. In the second experiment, in 91 nest-use opportunities, four nests were in previously unused but natural locations to which vestiges had been added, 22 nests were in previously used sites without vestiges, and the remaining 65 nests remained unused. Two processes are apparently in action: first, prior experience and memory; second, vestiges indicate where nesting has occurred, possibly useful for first breeding, or for imperfect memory. Previous nesting information may explain why swifts use nesting locations for many years and why new nesting colonies seldom form. This transmission of information suggests that swifts tend to be conservative and nest where previous nesting has occurred.     

Assessment of the use of two commercially available environmental enrichments by laboratory mice by preference testing

In the field of biomedical research, the demand for standardization of environmental enrichment for laboratory animals is growing. For laboratory mice, a wide variety of environmental enrichment items are commercially available. Most of these comply with the demands for standardization, hygiene and ergonomics. Whether these items also comply with their actual purpose, to enhance the well-being of the mice, is often not assessed scientifically. In this study, we tested the preference of mice for two commercially available nest boxes differing in shape and material: the Shepherd Shack/DesRes (SS/DR) and the Tecniplast Mouse House (TMH), in a simple preference test. To indicate strength of preference, both nest boxes were also tested against a highly preferred nesting material. Preference for the most preferred nest box was investigated further. Our results indicated a strong preference by mice for the SS/DR, but not for the TMH. Furthermore, nesting material was almost always combined with the SS/DR, but not with the TMH. More elaborate testing of the SS/DR in an automated preference test system confirmed that mice spent significantly more time in a cage in which an SS/DR is provided. Differences between both nest boxes are discussed with regard to their attractiveness to mice. It is also argued that enrichment should primarily be developed in concordance with the animals' needs prior to the marketing of enrichment tools.     

Sleeping tree choice by Bwindi chimpanzees

Unlike nearly all other nonhuman primates, great apes build sleeping nests. In Bwindi Impenetrable National Park, Uganda, chimpanzees build nests nightly and also build day nests. We investigated patterns of nest tree use by Bwindi chimpanzees to understand ecological influences on nest tree selection. We analyzed data on 3,414 chimpanzee nests located from 2000 to 2004. Chimpanzees at Bwindi were selective in their use of nest trees. Of at least 163 tree species known to occur in Bwindi [Butynski, Ecological survey of the Impenetrable (Bwindi) Forest, Uganda, and recommendations for its conservation and management. Report to the Government of Uganda, 1984], chimpanzees utilized only 38 species for nesting. Of these, four tree species (Cassipourea sp., Chrysophyllum gorungosanum, Drypetes gerrardii, and Teclea nobilis) accounted for 72.1% of all nest trees. There was considerable variation in nesting frequencies among the top four species between and within years. However, these species were used significantly more often for nesting than other species in 70.9% (39 of 55) of the months of this study. A Spearman rank correlation found no significant relationship between tree abundance and tree species preference. Ninety-three percent of all nests were constructed in food tree species, although not necessarily at the same time the trees bore food items used by chimpanzees. The results indicate that nesting tree species preferences exist. Bwindi chimpanzees' choice of nesting tree species does not appear to be dependent on tree species density or use of the tree for food. We discuss possible reasons for the selectivity in nest trees by the Bwindi population.     

Environmental modification and agonistic behavior in NIH/S male mice: nesting material enhances fighting but shelters prevent it

Outbred NIH/S male mice were housed from weaning in groups of 4 without enrichment (control) or with nesting material (nest), nesting material and a box (nest-and-box), or nesting material and a tube (nest-and-tube) as environmental modification. The aim of the study was to investigate effects of widely recommended nesting material and additional shelters on male mice. The aggressiveness of the mice in their home cages clearly increased in the nest group, as assessed by the number of wounds. In the nest group, fighting was a stressful situation for the mice, leading to changes in weight gain and in the weights of the thymus, adrenals, spleen, and epididymal adipose tissue. Moreover, the agonistic behavior of these mice toward an intruder was increased both in individual tests (an intruder with the individual mouse) and group tests (an intruder with a group of mice). The provision of a box or tube as a shelter, in addition to nesting material, prevented intracage fighting and did not lead to alterations in the weight gain or organ weights of the mice. However, the agonistic behavior of mice with shelters was slightly increased in behavioral tests. Anxiety in the elevated plus-maze was not affected by any of the housing systems. In conclusion, the agonistic behavior of NIH/S mice, an aggressive strain, seemed to be easily enhanced by these environmental modifications. The suitability of any enrichment should be carefully evaluated, especially when highly aggressive mice are used.     

Aspen wood-wool is preferred as a resting place, but does not affect intracage fighting of male BALB/c and C57BL/6J mice

Aspen wood-wool, provided as nesting material, was evaluated as a possible improvement of cage environment for 10-14-week-old inbred male mice maintained in groups of six (BALB/c n = 72 and C57BL/6J n = 36). The daily behaviour of mice was video recorded and their body weight, food consumption, weights of some organs and serum corticosterone concentrations were measured. Aggressive interactions between cage mates and against a strange intruder as well as the number of wounds on the back of the animals was monitored in order to evaluate the effect of nesting material on intermale aggression. Nesting material did not affect the daily active/passive behaviour patterns of mice, although animals clearly preferred it as a resting place. BALB/c mice given nesting material showed less weight gain and smaller brown adipose tissue weights than animals without nesting material. The other characteristics measured were not affected by the presence of nesting material in either strain. The presence of nesting material had no effect on fighting in cages. C57BL/6J mice were more aggressive than BALB/c mice according to the number of wounded animals in a cage. Wounded BALB/c mice had enlarged spleens and decreased epididymal adipose tissue weights. In conclusion, the nesting material used in this study did not adversely affect the animals. On the other hand, the material was clearly preferred to conventional bedding as a resting place. These findings suggest that nesting material may improve the cage environment of laboratory mice. Furthermore, there was an indication of strain differences in aggressive behaviour. It could be suggested that C57BL/6J mice are less tolerant towards intruders and housing six mice per cage is not suitable for this strain.     

Nest preference and ecology of cavity-nesting bees (Hymenoptera: Apoidea) in Punjab,Pakistan

Among bees, 85 % are solitary species, most of them are ground-nester and some are cavity-nesting and construct their nests in pre-existing cavities. This work was conducted to evaluate the substrate preference and nest architecture (acceptance, occupation percentage, seasonality and parasitism) of cavity-nesting bees in different substrates. Trap nests offered comprised five different materials (drilled cavities in wood, bamboo, cardboard tubes, plastic soda straws, and mud blocks). These were installed in four districts for two years. The nesting cavities of five different diameters (6, 8, 10, 12 and 14 mm) were provided in each nesting material with an average length of 180 ± 9.92 mm. In all the materials, 5400 nesting cavities were offered, out of which 628 were colonized by bees from two families (Megachilidae and Apidae) and six species (Megachile cephalotes, M. lanata, M. bicolor, Xylocopa basalis, X. fenestrata and Ceratina smaragdula), including one parasitic bee (Euaspis carbonaria). The bee species differed significantly in occupying five nesting materials. The most preferred diameters were 8 mm and 10 mm, with 52.20% and 29.45% of colonization, respectively. Nesting was done throughout the year except in winter. This study will serve as a baseline for future studies and conservation programs of cavity-nesting bees in Pakistan.     

Factors Affecting Nesting Site Choice in Chimpanzees at Tshibati,Kahuzi-Biega National Park: Influence of Sympatric Gorillas

We recorded 310 fresh chimpanzee night nests at 72 nest sites to determine their choice of tree and site for nesting vis-à-vis the effects of sympatric gorillas. Chimpanzees did not use trees for nesting according to their abundance, but instead tended to nest in fruit trees that they used as food sources. Nesting patterns of chimpanzees may vary with nesting group size, the type of vegetation, and fruit species eaten or not eaten by gorillas. When chimpanzees lodged as a small group in the secondary forest, they nested more frequently in trees bearing ripe fruits eaten only by themselves than in those with fruit eaten also by gorillas. When they lodged as a large group in the primary forest, they nested more frequently in trees bearing ripe fruits eaten by both apes. Nest group size is positively correlated with the availability of preferred ripe fruits in secondary forest. These findings not only reflect the larger foraging groups at the larger fruiting trees but also suggest that chimpanzees may have tended to occupy fruiting trees effectively by nesting in them and by forming large nest groups when the fruits attracted gorillas. Competition over fruits between gorillas and chimpanzees, due to their low productivity in the montane forest of Kahuzi, may have promoted the chimpanzee tactics.     

Sea Turtle Nesting as a Process Influencing a Sandy Beach Ecosystem

Sea turtle egg mortality, egg predation, and small organisms associated with turtle nests were studied at Playa Ostional, Costa Rica. Sites with concentrated sea turtle nesting were compared with solitary nesting sites as a function of place and time based on ANOVA, Akaike's Information Criterion, and Bayesian analyses. Results indicate that sea turtle egg mortality was significantly associated (P < 0.005) with flowing water that erodes or saturates nesting sites, and with overlapped nesting in which sea turtles disturb each other's nests. Sarcophagid and calliphorid fly larvae (Bayesian prior = 1.19; posterior = 2.27), fungi (prior = 1.14; posterior = 1.92), mites (prior = 0.51; posterior = 1.15), and several other types of small organisms increased in number after turtle egg laying (N= 303 nests; 34,451 turtle eggs). During peak sea turtle nesting periods, visitation to nesting sites by poachers and vertebrate predators was high, and relative number of nests disturbed by these predators was low (P < 0.02). In multimodel analysis, the three most parsimonious models were: (1) turtle egg mortality and distance from mean high tide; (2) turtle egg predation and distance from mean high tide; and (3) turtle egg mortality and nesting density, with Akaike weights of 0.224, 0.203, and 0.153 respectively. Intensive sea turtle nesting might result in upwelling and turnover of nesting debris and nest organisms, and may influence biotic community structure of sandy beach ecosystems.     

The cage preferences of laboratory rats

Preference tests were used to assess a range of enrichment options for rats kept under standard New Zealand (and similar) caging conditions. The rats did not show significant preferences for most of the options, over an empty cage. The exceptions were shredded paper, a nesting box and a semi-enriched condition incorporating a range of modifications. These cage modifications are recommended for the enrichment of laboratory rats.     

Discovery of a regular nesting area of loggerhead turtle <Emphasis Type="Italic">Caretta caretta</Emphasis> in southern Italy: a new perspective for national conservation

Loggerhead turtle Caretta caretta nesting in Italy had been reported to be limited to the Pelagian Islands and only sporadically elsewhere. As presence of loggerhead turtle nests had occasionally been reported (1988–1999) along about 200 km of the Ionian coast of Calabria, we carried out a project to assess the actual state of the nesting population between 2000 and 2004. We divided the coastline in two sectors (A: 52 km, and B: 146 km) that were monitored from mid-June to end of July for a total of n = 174 monitoring days and 1,813.6 km patrolled on foot with different intensities (extensive versus intensive). In sector B, through extensive monitoring we did not find any emergence tracks, but in sector A by intensive survey (2002–2004: one survey/3.64 days) we detected 3–8 nests/year. In total, 25 nests (both observed and reported), were recorded in our study area, and an assessment of a total of 15–16 nests/year was suggested. These figures, within the national scenario depicted from the review of known nesting events in the last 40 years (88 records concerning more than 143–144 nests), show that loggerhead turtle nesting has been underestimated in Italy, due to inadequate monitoring protocols, and that nesting is more frequent than expected (at least 30–40 nests/year). Conservation strategies in Italy should then focus not only on the reduction of mortality at sea, but also include large-scale actions to preserve scattered (but regular) nesting events.     

Does nesting habitat predict hatch synchrony between brood parasitic brown‐headed cowbirds Molothrus ater and two host species?

Nestling brown-headed cowbirds Molothrus ater typically hatch earlier and grow faster than young of the many host species of this generalist obligate brood parasite. However, a cowbird chick also benefits from the presence of some host nest mates as the parasite is provisioned disproportionately more with increasing brood size. Since asynchronous hatching affects both cowbird and host nestlings' growth and survival, mechanisms that optimize the timing of egg-laying by female parasites should be prevalent. Several habitat features might facilitate optimal timing of parasitic egg-laying and we examined whether aspects of host nesting habitat predicted cowbird hatching synchrony. We tested whether synchronous nests were less concealed, closer to perches, and located in areas of higher host density than asynchronous nests using a broad-scale information theoretic approach. There was no support for these predictions regarding song sparrow ( Melospiza melodia ; n=55) or yellow warbler ( Dendroica petechia ; n=67) nests parasitized by brown-headed cowbirds at Mono Lake, USA. For example, the best statistical models for predicting hatching synchrony in yellow warbler nests included nesting-patch width and nest-substrate shrub species. However, these relationships were relatively weak: both synchronous and asynchronous nests were in patches with statistically indistinguishable widths and the two dominant shrub species at our site contained similar proportions of synchronous and asynchronous nests. We conclude that the variability of host nesting habitats does not contribute to a biologically consistent effect on hatching synchrony by this generalist brood parasite.     

Nest site fidelity and clutch frequency of loggerhead turtles are better elucidated by satellite telemetry than by nocturnal tagging efforts: Implications for stock estimation

Satellite telemetry and ground-based tagging studies are complementary methods to define the spatial and temporal patterns of nesting behavior by migratory sea turtles. Estimates of site fidelity and clutch frequencies are compared for satellite telemetry versus ground truth patrols over a 6 km stretch at a southwest Florida loggerhead (Caretta caretta) rookery. Site fidelity ranged from 1.9 km to 109.1 km for all nests deposited by a female within a season. The mean site fidelity was 28.1 km for all nests, but declined to 16.9 km if omitting the first nest. Nest frequency ranged from 2 to 8 nests per season, with a modal value of 5 nests. Satellite telemetry documented a mean nest frequency of 5.4 nests per female in comparison to 2.2 nests detected by monitoring patrols. The remigrant females had higher clutch frequency, were larger in size, and had higher site fidelity compared to newly tagged females. Satellite telemetry provided improved measurements of site fidelity and reveals a need for revised fecundity estimates. If measures of clutch frequency are representative of loggerhead assemblages nesting elsewhere within the South Florida grouping, the confidence bounds on Western Atlantic loggerhead stocks are approximately 32% lower than currently accounted for annual nesting individuals.     

An investigation into the effectiveness of compressed straw blocks in reducing abnormal behaviour in growing pigs

The provision of manipulable material to pigs is a legal requirement to ensure their good welfare. Loose straw is edible, chewable, investigable and manipulable, and has been proven to be effective in reducing abnormal behaviour. However, it is incompatible with slatted systems and therefore not a viable option for many intensive units. Thus, there is a need to identify enrichment materials that are as effective as loose straw and compatible with slatted floors. This study investigated the viability of using compressed straw blocks on a commercial farm in terms of cost and effectiveness in reducing biting relative to plastic hanging toys. A total of 880 slaughter pigs were used, among which half (n = 440, 8 groups) were provided with commercial pig toys (TOY) and the other half (n = 440, 8 groups) were provided compressed straw blocks (STRAW BLOCK). Animals were separated according to sex to investigate whether there was a higher propensity to tail and ear bite depending on sex. Pigs were monitored from weaning to slaughter, with tails being examined post scalding and any condemnations recorded at the abattoir. Throughout their life, the tails and ears of all pigs were examined individually, and behavioural observations of each pig group were conducted fortnightly. Salivary cortisol was also obtained from a subsample of pigs from each group every fortnight to monitor stress levels. The highest straw usage was recorded in the second weaner stage (39 to 67 days after weaning). Enrichment type or sex had no effect on tail lesion scores or cold carcass weight recorded at the abattoir. There was also no effect of enrichment type or sex on body, tail, or ear lesion scores during either the weaner (0 to 39 days after weaning) or finisher stage (67 to145 days after weaning). Female pigs showed more biting behaviour than males, and female pigs that were provided STRAW BLOCK performed better than those provided TOY. In both the weaner and finisher accommodation, more instances of interaction were recorded with TOY pens as enrichment than with STRAW BLOCK, but the interaction duration was not recorded. There was no effect of sex on either stage. More instances of aggression were observed with the TOY than with STRAW BLOCK in the weaner stage (P < 0.05). Enrichment type or sex had no effect on cortisol levels. Thus, relative to plastic hanging toys, compressed straw blocks provided in this manner did not benefit pig welfare.     

Elaborate Nests in a Weaverbird: A Role for Female Choice?

Studies on sexual selection have focused on behaviour and morphology, but several groups of animals build elaborate structures associated with acquiring a mate. I investigated female choice for nests built by male baya weavers (Ploceus philippinus). Nest choice by females should be strong, as nests are obvious direct benefits provided by males. I used a field experiment supplemented with correlational information to ask whether females appear to base mate choice decisions on male behaviour, nest architecture, and nest location. When the nests of highly visited males were exchanged with those of poorly visited males, female visits remained highest at the original male and location. I found no relationship between female choice and male display or other behaviour. Correlational analyses show that nest location was a better predictor of female choice than was nest architecture. These data suggest that current female choice is driven more by access to safe nesting sites rather than to well‐built nests, possibly because all males are able to build nests of adequate quality. However, nest architecture is unlikely to be irrelevant to females, and its role deserves further investigation.