Applied Animal Behaviour Science 78 (2002) 329335

Preliminary study of the development of the Flehmen response in Equus caballus

J.W. Weeksa,*, S.L. Crowell-Davisb, G. Heusnerc
b a 651 Fox Farm Road, Asbury, NJ 08802, USA Department of Anatomy and Radiology, College of Veterinary Medicine, University of Georgia, Athens, GA 30602, USA c Department of Animal Science, Franklin College of Arts and Sciences, University of Georgia, Athens, GA 30602, USA

Abstract The ehmen response is commonly seen in most ungulates as well as in several other species (e.g. felids). The behavior is most often thought to be part of the sexual behavioral repertoire of males. One reigning hypothesis suggests that this behavior allows the male to determine the estrous state of a female through the chemosensory functions of the vomeronasal organ. However, females and young of both sexes also exhibit this behavior. Horse foals most frequently show the ehmen response during their rst month of life with colts showing the behavior more often than llies. This study tested the ehmen response on male and female foals throughout their pre-pubertal period. Foals were separately presented estrous and non-estrous urine weekly during the rst month of life and then monthly until they were approximately 7 months of age. No signicant differences were found between male and female foals for the following variables: latency to ehmen, duration of ehmen, frequency of ehmen and sniffs. # 2002 Elsevier Science B.V. All rights reserved.
Keywords: Horse behavior; Flehmen; Foal development

1. Introduction The ehmen response is a behavior commonly seen in a diverse array of species including most ungulates (Ladewig and Hart, 1980; Ladewig et al., 1980; Hradecky et al., 1983; Lindsey and Burton, 1983; Houpt and Guida, 1984; Bland and Jubilan, 1987; Hart et al., 1988; Mariner et al., 1988; Houpt et al., 1989; Stahlbaum and Houpt, 1989; Thompson, 1991; Gastal et al., 1996) and felids (DeBouer and Verberne, 1981; Hart and Leedy, 1987).
* Corresponding author. Tel.: 1-908-730-9451; fax: 1-908-231-9692. E-mail address: jenwweeks@aol.com (J.W. Weeks).

0168-1591/02/$ see front matter # 2002 Elsevier Science B.V. All rights reserved. PII: S 0 1 6 8 - 1 5 9 1 ( 0 2 ) 0 0 1 1 0 - 7

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The behavior pattern involves the retraction of the upper lip, wrinkling of the nose and baring of the gums. This is always accompanied by deep breathing and may also include the elevation and extension of the head. The ehmen response is exhibited by both males and females as well as by juveniles of both genders (Crowell-Davis and Houpt, 1985). The main function of ehmen has been hypothesized to be the discrimination of odors that signal estrous states in females (Estes, 1972). This has been much studied with conicting results. In horses it has been reported that stallions did not sniff or show ehmen more to estrous urine or feces than to non-estrous urine or feces (Mariner et al., 1988; Houpt et al., 1989). Stallions did, however, show ehmen more to estrous mares than diestrous mares (Anderson et al., 1996). Bulls, in contrast, showed a higher rate of ehmen to estrous urine than to non-estrous urine (Hradecky et al., 1983; Houpt et al., 1989), showed ehmen for a longer duration in response to estrous urine (Houpt et al., 1989), and showed repeated ehmen responses to estrous urine as opposed to non-estrous urine (Hradecky et al., 1983). These conicting results indicate that the function of the ehmen response is still unclear. The ontogeny of the ehmen response has been studied very little. The main focus of ehmen research has been the function of the behavior. This has been tested mostly through discrimination of estrous and non-estrous bodily uids. Reinhardt (1983) studied the ontogeny of ehmen in a semi-wild cattle herd. This study found that ehmen occurred in calves of both genders older than 1 week and was oriented toward both male and female herd mates. Flehmen only became frequent after 4 months of age. At that time, male calves showed the behavior more frequently and predominantly toward female counterparts. A similar study in equids (Crowell-Davis and Houpt, 1985) reported that Welsh ponies began to show the ehmen response as early as one day of age. Fillies showed no peaks in ehmen while colts expressed the behavior most often during the rst 4 weeks of life. Fillies showed a signicantly lower rate of ehmen than colts during the rst 12 weeks. The study was conducted in the eld. This study was designed to assess any potential differences in the ehmen behavior exhibited by male and female horse foals in response to presentations of both estrous and non-estrous urine. It was hypothesized that, similar to the eld study (Crowell-Davis and Houpt, 1985), colts would express the behavior more often than llies and the behavior would peak during the rst 4 weeks of life. 2. Materials and methods The subjects of this study were the foals of a herd of research mares housed at The University of Georgia Smithonia Breeding Center in Colbert, Georgia. There were 13 foals of varying breeds: 10 Quarter Horse, 1 Arabian, 1 Hanoverian and 1 Saddlebred. Of the 13 foals, 6 were male and 7 were female. The mares and foals were housed in a 10 ha pasture. Water was available ad libitum and supplemental grain was provided twice daily. 2.1. Urine collection Urine was collected prior to each testing period from one estrous and one non-estrous mare from a different research herd, using a free catch method. Estrus was determined by a

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combination of palpation and overt behavior. Testing began approximately 30 min after collection. In the interim time period, urine was stored in airtight glass containers. 2.2. Testing procedure For each test period (Fig. 1), the foal was placed in an open-air pipe stall with its dam in an open-air pipe stall immediately adjacent to it. Mares and foals were in visual and vocal contact at all times. It was also possible for mare and foal to be in physical contact, though nursing was not possible. Foals were allowed unlimited access to all areas of the pipe stall while dams were tied to the middle partition common to each stall. A plastic test bucket (standard commercial water bucket) was tied to the middle partition in the foals' stall at a height of approximately 3 ft. Prior to each testing session the foal was allowed a 5-min habituation period before the urine presentation. This was done to allow the foal to acclimatize to being separated from its dam. After the 5-min habituation period, estrous and non-estrous urine were presented for 5 min each for each individual subject. After the 15-min testing period, the mare and foal were returned to the herd. Presentation of each type of urine was counterbalanced for order of presentation during each test day for each individual. Foals were never forced into proximity with the bucket containing the urine. Each foal was tested once a week for the rst month of life, as this is when the behavior has been reported to occur most frequently (Crowell-Davis and Houpt, 1985). Foals were tested once a month for months 24. They were tested again at month 7. The nal test session was post-weaning therefore the mares were not present. 2.3. Statistical analysis All testing sessions were videotaped for later analysis. Analyses were conducted using the Observer (Noldus) program. Behavioral measures assessed were: frequency of sniffs, frequency of ehmen, latency to rst ehmen, and ehmen duration. A general linear

Fig. 1. Schematic drawing of experimental set-up.

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Table 1 Effect of urine type on behavioral measures (mean S:E:) Estrous Frequency of Sniff Latency to Flehmen (s) Frequency of Flehmen Flehmen duration (s) 1.37 278.95 0.17 0.42 1.64 66.20 0.55 1.43 Non-estrous 1.14 168.11 0.27 0.54 1.20 117.70 0.81 1.54

model (SPSS) procedure was run in order to assess any effects of age, urine type or subject gender on the ehmen response. 3. Results The foals' interest in the test bucket varied. During each test session most foals would approach the bucket to sniff, chew or manipulate it. Foals showed ehmen to estrous urine 0:17 0:55 times per test session. The foals showed ehmen 0:27 0:82 times to nonestrous urine. Males and females displayed ehmen nearly equally in response to estrous versus non-estrous urine (Tables 1 and 2). No signicant effects of gender were found for any of the behavioral measures associated with the ehmen response. Male and female foals did not differ in the frequency of sniffs or showing of ehmen. When ehmen was shown the duration of and latency to ehmen did not differ between the genders. Additionally, there were no signicant effects of age on any of the measures recorded. Male and female foals did not show any signicant discrimination of urine type. The ehmen response was exhibited nearly equally to estrous and non-estrous urine. 4. Discussion This study failed to nd any behavioral differences based on age, gender or urine type. This lack of effect of age runs counter to the ndings of Crowell-Davis and Houpt (1985). The present study also found no identiable peak in ehmen exhibited by colts, whereas the eld study found colts to show ehmen maximally in the rst 4 weeks of life. Similar to previous studies focusing on the discrimination of urine type in the horse (Mariner et al., 1988; Stahlbaum and Houpt, 1989) this study found no difference in sniffs and ehmen exhibited to estrous versus non-estrous urine. There are many possible explanations for why this study failed to replicate the eld ndings in Welsh ponies (Crowell-Davis and Houpt, 1985). Flehmen in the eld is commonly seen after urination by a conspecic. This urination is accompanied by several other sensory cues such as body elongation, tail raising and tail deviation. Providing only the olfactory cue may not have provided enough sensory input. Previous studies have shown that both olfactory and visual cues are involved in the initiation of the ehmen response. Anderson et al. (1996) found that the events associated with ehmen were lower

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Table 2 Effect of age on behavioral measures for the first month of life in response to estrous urine (mean S:E:) Week 1 Male Female Week 2 Male Female Week 3 Male Female 2.14 247.93 0.29 1.4 Week 4 Male Female

Frequency of Sniff 3.2 2.05 2 2.16 Latency to Flehmen 249.78 112.30 300 Frequency of Flehmen 0.2 0.45 0 Flehmen duration 0.6 1.34 0

2.17 2.99 1.5 1.87 2 2.10 300 276.42 57.77 287.43 30.78 0 0.17 0.41 0.33 0.82 0 0.33 0.82 1.63 4.00

0.22 1.67 1.03 2.5 1.87 108.86 238.13 109.46 257.14 113.39 0.49 0.33 0.52 0.29 0.76 2.40 1.35 2.43 1.29 3.4

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when the horse's vision was blocked and the frequency of ehmen was lower when olfaction was blocked. Houpt and Guida (1984) have suggested that stallions do not need to depend on olfactory cues as mares provide many visual and auditory cues that indicate they are in estrous. Another factor that may have contributed to the low expression of the ehmen response is stress. All testing periods, except the nal one, were conducted prior to the foals being weaned. Therefore, the foals had little experience with being separated from their dams. During the testing, foals were allowed considerable access (olfactory, auditory, visual and limited tactile) to their mothers. They were not, however, able to nurse, which is a comfort behavior in times of stress (Crowell-Davis, 1985). As the foals were not forced into contact with the bucket containing the urine, the stress of separation may have distracted the foals from the urine, thus limiting the response. Many foals spent numerous test sessions without even approaching the test bucket. As this study did not conrm our hypothesis nor replicate the ndings of Crowell-Davis and Houpt (1985) in ponies, further studies are needed that combine eldwork and experimental testing. Additional studies should also address other, non-sexual functions of this behavior such as motheroffspring olfactory recognition. References
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