The effects of prenatal PCBs on adult social behavior in rats

Endocrine disrupting chemical (EDC) exposures during critical periods of development may influence neuronal development and the manifestation of sexually dimorphic sociability and social novelty behaviors in adulthood. In this study, we assessed the effects of gestational exposure to PCBs on the social behavior of males and females later in adulthood. A weakly estrogenic PCB mixture, Aroclor 1221 (A1221, 0.5 or 1 mg/kg) was administered to pregnant Sprague–Dawley rat dams. Both a positive control (estradiol benzoate; EB, 50 μg/kg) and negative control (dimethylsulfoxide; DMSO in sesame oil vehicle) were similarly administered to separate sets of dams. The sexes responded differently in two tasks essential to sociality. Using a three-chamber apparatus that contained a caged, same-sex, gonadectomized stimulus animal and an empty stimulus cage, we found that both sexes showed a strong preference for affiliating with a stimulus animal (vs. an empty cage), an effect that was much more pronounced in the males. In the second task, a novel and a familiar stimulus animal were caged at opposite ends of the same apparatus. Females displayed a higher degree of novelty preference than the males. During both tests, females had significantly higher social approach behaviors while male engaged in significantly more interactive behaviors with the conspecific. Of particular interest, males born of dams that received prenatal A1221 (0.5 mg/kg) exhibited an overall decrease in nose-to-nose investigations. These behavioral data suggest that the males are more sensitive to A1221 treatment than are females. In addition to behavioral analysis, serum corticosterone was measured. Females born of dams treated with A1221 (0.5 mg/kg) had significantly higher concentrations of corticosterone than the DMSO female group; males were unaffected. Females also had significantly higher corticosterone concentrations than did males. Overall, our results suggest that the effects of gestational exposure to PCBs on adult social behavior are relatively limited within this particular paradigm.     

Behavioral defeminization by prenatal androgen treatment in rats can be overcome by sexual experience in adulthood

Exposure to testosterone during a critical period of prenatal development disrupts the normal display of sexual behaviors in adult ovariectomized (OVX) rats treated with estradiol benzoate (EB) followed by progesterone (P). The organizational hypothesis posits that prenatally androgenized females (PNAFs) are desensitized to EB. We tested this hypothesis by first treating PNAFs with varying doses of EB (2.5, 5, 10, 20 μg) followed by P (500 μg), and second by subjecting females to an established EB behavioral sensitization paradigm where females are first given sexual experience with EB (10 μg) and P prior to repeated sexual behavior testing with EB alone. Long-Evans females were androgenized in utero by a s.c. injection of 500 μg testosterone propionate or the oil control to pregnant dams on gestational day 18. Female offspring were OVX on postnatal day 80 and tested one week later in the unilevel 4-hole pacing chamber. Genital tissue was defeminized in PNAFs, and the lordosis quotient (LQ) and partial (i.e., hops/darts) and full solicitations were significantly lower, while defensive behaviors were higher, in PNAF females, relative to non-PNAF females regardless of the acute EB priming dose. However, repeated testing with EB alone (10 μg), or EB and P eliminated the differences between groups on LQ and hops/darts, indicating that the behavioral deficit can be overcome by sexual experience. These results suggest that PNAFs are not desensitized to EB, and despite disruptions in sexual differentiation of anatomical structures, the deficiency in sexual behavior in response to acute EB and P can be experientially overcome. PNAFs appear, however, to have a chronic deficit in the expression of full solicitations.     

RU486 facilitates or disrupts the sensitization of sexual behaviors by estradiol in the ovariectomized Long–Evans rat: Effect of timecourse

An acute injection of estradiol benzoate (EB) to the ovariectomized (OVX) rat activates low levels of lordosis, and subsequent progesterone (P) administration augments lordosis and recruits a complete pattern of sexual behavior including appetitive behaviors (e.g., hops/darts and solicitations). However, repeated injections of 5 μg or 10 μg EB (but not 2 μg EB), administered every 4 days to sexually-experienced OVX rats results in a behavioral sensitization, such that lordosis quotients (LQs) and appetitive behaviors progressively increase. We have shown that adrenal P does not play a critical role because behavioral sensitization to EB is not prevented by adrenalectomy. Here we tested whether P receptors play a role by examining the effect of chronic administration of the P receptor antagonist RU486 at a dose that reliably inhibits sexual behavior in fully primed OVX rats. Females were treated with EB (5 or 10 μg), and 5 mg RU486 dissolved in 0.4 mL vehicle (VEH; 80% sesame oil, 15% benzyl benzoate, 5% benzyl alcohol) 48 h and 5 h prior to each of 7 tests, respectively, occurring at 4-day intervals in unilevel 4-hole pacing chambers. Control animals were treated with 2, 5, or 10 μg EB + VEH. As expected, sensitization did not occur in females treated with 2 μg EB + VEH, and those females received fewer intromissions and ejaculations than all other groups. RU486 did not prevent the sensitization of LQ, moderate and high lordosis magnitudes (LM2 and LM3) or appetitive sexual behaviors on early tests, and in fact potentiated appetitive behaviors, LQ, LM2 and LM3, consistent with its facilitative actions in females treated with EB-alone, as we and others have reported previously. However, despite the initial facilitation, blocking P receptors by chronic administration of RU486 inhibited the maintenance of behavioral sensitization to EB.     

The inhibitory effects of corncob bedding on sexual behavior in the ovariectomized Long–Evans rat treated with estradiol benzoate are overcome by male cues

The mechanisms underlying the sensitization of sexual behaviors by repeated administration of estradiol benzoate (EB) to ovariectomized (OVX) rats are not well understood. Here we tested whether two housing conditions play a role. Sexual behavior in the female rat is dependent on the activation of ERα (estrogen receptor alpha) by estradiol. Corncob (CC) bedding has been reported to have adverse effects on the reproductive behavior and physiology of rats, and to disrupt ERα signaling in mice. In addition, some rodent behaviors are stimulated by olfactory stimuli and enhanced in the presence of estradiol. Upon arrival to the facilities OVX Long–Evans rats were housed on either Sani-Chips (SC) or CC in a room that housed only females (F) or males and females (M). Females were first given four sexual training sessions with 10 μg EB + 500 μg progesterone (P; administered 48 h and 4 h prior to training, respectively), followed by a 2-week hormone washout period. Next, 10 μg EB was administered s.c. every 4 days, 48 h prior to each of 8 test sessions in a unilevel 4-hole pacing chamber. On the final training day (i.e., when primed with EB + P), no inhibitory effects of corncob bedding were found, however a facilitation of the lordosis quality occurred in SC/F. Although all groups appear to have sensitized to the repeated administration of EB, CC/F animals displayed fewer high quality lordosis magnitudes and hop/darts, and received fewer mounts and intromissions overall. They also had a lower lordosis quotient (LQ) on tests 2–4 although this effect disappeared by test 5. These results suggest that although CC may inhibit some components of female sexual behavior when primed with EB alone, cues from sexually vigorous males can overcome that inhibition. Moreover, they suggest that male cues can facilitate mechanisms of estradiol sensitization. We recommend that quality control studies be conducted at individual institutions to assess any impact of corncob bedding on animal physiology and behavior.     

Effects of uterine and lactational exposure to di-(2-ethylhexyl) phthalate on spatial memory and NMDA receptor of hippocampus in mice

Di-(2-ethylhexyl) phthalate (DEHP) is an environmental endocrine disrupter. Currently, little is known about neurodevelopmental toxicity of DEHP in wildlife and humans. The present study investigated the effects of DEHP, focusing on the changes in the behavior of offspring mice at the ages of 6 and 12 w, respectively, following utero and lactational exposure to DEHP (10, 50, and 200 mg/kg/d) from gestation day 7 through postnatal day 21. The results of open field tasks showed that DEHP increased the grooming of males at age 6 w and females at age 12 w but decreased the frequency of rearing of 6-w-old females and the number of grid crossings of 12-w-old females. In the Morris water maze task, 50 and 200 mg/kg/d DEHP significantly prolonged the time of searching the hidden platform in water maze and reduced the time staying in the target quadrant during a probe trial of 6-w-old male mice, but not of 6-w-old females nor 12-w-old mice of both sexes, suggesting an impaired spatial learning and memory among younger males after perinatal exposure to DEHP. Western blot analyses further showed that DEHP at 50 and 200 mg/kg/d decreased the levels of the N-methyl-d-aspartic acid (NMDA) receptor subunits NR1 and NR2B in the hippocampus of 6-w-old males. These results suggest that uterine and lactational exposure to low doses of DEHP sex-specifically impacted behaviors, including locomotion activity and spatial memory, via the concomitant inhibition of the NMDA receptor of the hippocampus in offspring mice.     

Genetic differences in the elevated plus-maze persist after first exposure of inbred rats to the test apparatus

The elevated plus-maze (EPM) is an anxiety model thought to assess different types of emotional states depending on whether or not the animals have been previously exposed to the test apparatus. Accordingly, benzodiazepine-treated rodents generally differ from controls in the first but not in the second EPM trial. Inbred Lewis and SHR rats of both sexes (N = 10) were submitted twice (test and retest) to the EPM with a 24 h interval between trials. Overall strain differences (Lewis < SHR) were observed in both males and females concerning anxiety-related measures (time spent and percent of entries in the open arms) regardless of previous maze experience. Moreover, prior exposure to the test apparatus produced an overall decrease in the approach towards the open arms in both strains and sexes. The fact that genetic differences did not diminish or disappear in the second trial, suggests that test and retest in the EPM are likely to share some common emotional components and that differences between naïve LEW and SHR rats are not similar to those observed between control and benzodiazepine-treated animals.     

Olfactory conditioned same-sex partner preference in female rats: Role of ovarian hormones

The dopamine D2-type receptor agonist quinpirole (QNP) facilitates the development of conditioned same-sex partner preference in males during cohabitation, but not in ovariectomized (OVX) females, primed with estradiol benzoate (EB) and progesterone (P). Herein we tested the effects of QNP on OVX, EB-only primed females. Females received a systemic injection (every four days) of either saline (Saline-conditioned) or QNP (QNP-conditioned) and then cohabited for 24 h with lemon-scented stimulus females (CS +), during three trials. In test 1 (female-female) preference was QNP-free, and females chose between the CS + female and a novel female. In test 2 (male-female) they chose between the CS + female and a sexually experienced male. In test 1 Saline-conditioned females displayed more hops & darts towards the novel female, but QNP-conditioned females displayed more sexual solicitations towards the CS + female. In test 2 Saline-conditioned females displayed a clear preference for the male, whereas QNP-conditioned females displayed what we considered a bisexual preference. We discuss the effect of dopamine and ovarian hormones on the development of olfactory conditioned same-sex preference in females.     

Tobacco and alcohol as risk factors for oesophageal cancer in a high incidence area in South Africa

BackgroundThe Eastern Cape Province of South Africa, which includes the former Transkei has high rates of squamous cell oesophageal cancer (OC), thought to be caused mainly by nutritional deficiencies and fungal contamination of staple maize. A hospital-based case-control study was conducted at three of the major referral hospitals in this region to measure, among other suspected risk factors, the relative importance of tobacco smoking and alcohol consumption for the disease in this population.MethodsIncident cases (n = 670) of OC and controls (n = 1188) were interviewed using a structured questionnaire which included questions on tobacco and alcohol-related consumption. Odds ratios (ORs) with 95% confidence intervals for each of the risk factors were calculated using unconditional multiple logistic regression models.ResultsA monotonic dose-response was observed across the categories of each tobacco-related variable in both sexes. Males and females currently smoking a total of >14 g of tobacco per day were observed to have over 4-times the odds of developing OC (males OR = 4.36, 95% CI 2.24–8.48; females OR = 4.56, 95% CI 1.46–14.30), with pipe smoking showing the strongest effect. Similar trends were observed for the alcohol-related variables. The quantity of ethanol consumed was the most important factor in OC development rather than any individual type of alcoholic beverage, especially in smokers. Males and females consuming >53 g of ethanol per day had approximately 5-times greater odds in comparison to non-drinkers (males OR = 4.72, 95% CI 2.64–8.41; females OR = 5.24, 95% CI 3.34–8.23) and 8.5 greater odds in those who smoked >14 g tobacco daily. The attributable fractions for smoking and alcohol consumption were 58% and 48% respectively, 64% for both factors combined.ConclusionTobacco and alcohol use are major risk factors for OC development in this region.ImpactThis study provides evidence for further reinforcement of cessation of smoking and alcohol consumption to curb OC development.     

Perinatal effects of exposure to PCBs on social preferences in young adult and middle-aged offspring mice

In social species, social interactions between conspecifics constitute a fundamental component to establish relations, provide best chances to reproduce, and even improve survival rates. In this study, a three-chambered social approach test was used to estimate the level of sociability and level of preference for social novelty in both male and female young adult (postnatal day (PND) 50) and middle-aged (PND 330) offspring mice (n = 10 per group) that were perinatally exposed to a mixture of six polychlorinated biphenyls (PCBs), 28, 52, 101, 138, 153, and 180, at environmentally low doses (10 and 1000 ng/kg b.w. for dams during gestation and lactation), a profile that closely mimics human exposure to contaminated fish. Our results showed that PCBs bidirectionally modulated social preferences in offspring mice, and the effects were sex and age dependent. However, increased levels of social interactions were rather frequently detected in both assays of the three-chambered test. Reduced social interaction was only induced in 1000 ng/kg PCB-exposed middle-aged males, which exhibited similar preferences to social and non-social stimuli when compared to middle-aged controls. Furthermore, results showed that plasma levels of both corticosterone and acetylcholinesterase activity were higher in all PCB-exposed middle-aged males and females than in their control counterparts. In summary, although the effects of PCBs were only of moderate magnitude, our results suggest that a PCB mixture can act as an endocrine disruptor in offspring mice, disturbing the formation of normal social habits.     

Effects of short- and long-term changes in spatial density on the social behavior of captive chimpanzees (Pan troglodytes)

The constraints of captivity may often require non-human primates to experience restrictions in space for both long- and short-term periods of time. The tension-reduction model predicts that great apes should increase affiliative behaviors and decrease aggressive behaviors as a coping strategy. The conflict avoidance model, however, predicts that great apes should decrease all social interactions (affiliative and aggressive) as a coping strategy. The purpose of this study was to test the conflict-avoidance and tension-reduction models by examining the effects of both short-term (1–2 days) and long-term (6 months) changes in spatial density on social behavior in 23 adult captive chimpanzees (6 males, 17 females) housed at the Primate Foundation of Arizona. Affiliative (i.e., social groom, social play) and agonistic (i.e., charging display, attack) were assessed using scan-sampling techniques while subjects were rotated from high-density to low-density conditions for varying lengths of time. Results of short-term increases in spatial density (crowding) supported the conflict-avoidance strategy for females, through reduced levels of aggressive (F_1,16 = 17.11, p = 0.001) and affiliative (F_1,16 = 21.13, p < 0.001) behaviors. Males, however, supported the tension-reduction model during short-term high-density by decreasing aggression (F_1,5 = 10.53, p = 0.02) while increasing affiliation (F_1,5 = 9.10, p = 0.03). Females partially supported the tension-reduction model during long-term high-density by increasing affiliative behaviors (F_1,16 = 14.19, p = 0.002) compared to short-term high-density levels, while rates of aggression remained low. Finally, males supported the tension-reduction by reducing aggression (F_1,5 = 7.668, p = 0.04) and increasing affiliation (F_1,5 = 6.08, p = 0.05) during long-term high-density. Results confirm that chimpanzees use different strategies during short-term versus long-term increases in spatial density and that these strategies may be influenced by individual sex. However, sample sizes are small and additional research on male chimpanzees is needed.     

Estrogenic plant consumption predicts red colobus monkey (Procolobus rufomitratus) hormonal state and behavior

Numerous studies have examined the effects of anthropogenic endocrine disrupting compounds; however, very little is known about the effects of naturally occurring plant-produced estrogenic compounds (i.e., phytoestrogens) on vertebrates. To examine the seasonal pattern of phytoestrogen consumption and its relationship to hormone levels (407 fecal samples analyzed for estradiol and cortisol) and social behavior (aggression, mating, and grooming) in a primate, we conducted an 11-month field study of red colobus (Procolobus rufomitratus) in Kibale National Park, Uganda. The percent of diet from estrogenic plants averaged 10.7% (n = 45 weeks; range: 0.7–32.4%). Red colobus fed more heavily on estrogenic Millettia dura young leaves during weeks of higher rainfall, and the consumption of this estrogenic item was positively correlated to both their fecal estradiol and cortisol levels. Social behaviors were related to estradiol and cortisol levels, as well as the consumption of estrogenic plants and rainfall. The more the red colobus consumed estrogenic plants the higher their rates of aggression and copulation and the lower their time spent grooming. Our results suggest that the consumption of estrogenic plants has important implications for primate health and fitness through interactions with the endocrine system and changes in hormone levels and social behaviors.     

Differential patterns of home-range,net displacement and resting sites use of Conepatus chinga in southern Brazil

Studies on the spatial ecology of Molina's hog-nosed skunk were conducted in southern Brazil. Between April 2008 and April 2009, 18 individuals were tagged with radio collars and monitored by radio-telemetry (VHF). The average home-range estimated for 12 skunks was 1.63 km² ± 1.17 km². Males had home-ranges 2.5 times larger than females, and also showed significantly larger patterns of displacement. These differences in space probably were related not only to sexual size dimorphism, but also to differential behavior between sexes. We identified six basic types of resting sites, among which holes in the ground were the most common and comprised more than 50% of the total. Reuse of resting sites was high (32%), especially for females, which had a significantly higher rate than males. Although they showed a large proportion of home-range overlap even in the core areas, skunks exhibited a solitary life style, rarely sharing resting sites and maintaining relatively long distances from each other, equivalent to twice the distance of daily movements. The almost strictly nocturnal activity pattern observed in this study is a common characteristic of the family. Among practically all of the ecological features analyzed, males and females showed significant differences, denoting the importance of a sex-specific approach in ecological studies regarding this species.     

Effects of developmental exposure to bisphenol A on spatial navigational learning and memory in rats: A CLARITY-BPA study

Bisphenol A (BPA) is a ubiquitous industrial chemical used in the production of a wide variety of items. Previous studies suggest BPA exposure may result in neuro-disruptive effects; however, data are inconsistent across animal and human studies. As part of the Consortium Linking Academic and Regulatory Insights on BPA Toxicity (CLARITY-BPA), we sought to determine whether female and male rats developmentally exposed to BPA demonstrated later spatial navigational learning and memory deficits. Pregnant NCTR Sprague–Dawley rats were orally dosed from gestational day 6 to parturition, and offspring were directly orally dosed until weaning (postnatal day 21). Treatment groups included a vehicle control, three BPA doses (2.5 μg/kg body weight (bw)/day—[2.5], 25 μg/kg bw/day—[25], and 2500 μg/kg bw/day—[2500]) and a 0.5 μg/kg/day ethinyl estradiol (EE)-reference estrogen dose. At adulthood, 1/sex/litter was tested for seven days in the Barnes maze. The 2500 BPA group sniffed more incorrect holes on day 7 than those in the control, 2.5 BPA, and EE groups. The 2500 BPA females were less likely than control females to locate the escape box in the allotted time (p value = 0.04). Although 2.5 BPA females exhibited a prolonged latency, the effect did not reach significance (p value = 0.06), whereas 2.5 BPA males showed improved latency compared to control males (p value = 0.04), although the significance of this result is uncertain. No differences in serum testosterone concentration were detected in any male or female treatment groups. Current findings suggest developmental exposure of rats to BPA may disrupt aspects of spatial navigational learning and memory.     

Use of space by Necromys lasiurus (Rodentia,Sigmodontinae) in a grassland among Atlantic Forest fragments

The spatial patterns presented by the rodent Necromys lasiurus were studied through capture-mark-recapture in a grassland among Atlantic Coastal Forest fragments in Southeastern Brazil. Males moved farther than females. Males moved similar distances in the dry and wet seasons, whereas females moved significantly longer distances in the dry season. Home ranges sizes were estimated for 32 individuals, and varied from 0.02 to 0.52 ha. Males had larger home ranges than females when all data are pooled and in the wet season; in the dry season home ranges of both sexes were of similar sizes. Home range sizes did not vary significantly between seasons for either sex. Home ranges of males overlapped extensively among themselves and with females, whereas females showed mutually exclusive home ranges. The spatial patterns described here are consistent with the results found for this species in different habitats and they suggest that N. lasiurus has a promiscuous mating system.     

Embryo production in superstimulated llamas pre-treated to inhibit follicular growth

Llamas are monotocous and the length of their gestation period varies between 342 and 350 days. Thus the average number of offspring any female can produce throughout her reproductive life is very limited to spread a desired genome. The multiple ovulation and embryo transfer (MOET) technique allows an alternative to this limitation and reduces the generation interval. The objective of this study was to evaluate embryo recovery in superstimulated llamas which had previously been hormone-treated to inhibit follicular growth. A total of 50 female llamas were monitored daily via rectal palpation and ultrasound and divided according to their ovarian follicular growth into four phases. The females in each phase were then randomly divided into two groups: A (n = 20) received a single dose of 1 mg of estradiol benzoate (EB) on the first day of the treatment + 100 mg of progesterone (P4) i.m. for 5 days with 5 animals per phase and B (n = 20) received 1 mg EB at onset + 150 mg P4 i.m. for a period of 5 days with 5 animals per phase. Group C (n = 10) or control did not receive any prior hormonal treatment and the females were in follicular phase I. All groups were monitored daily and, in the presence of ovarian follicles smaller than the dominant size at the end of treatment, all were superstimulated with 1000 IU eCG. For plasma progesterone concentration recording, daily blood samples were collected from days ?1 to 5 in the treated females in Group A and B. No significant differences were observed regarding the inhibition of follicle growth and in the plasma progesterone concentrations between Group A and B. The ovarian response to superstimulation was 56.2%, 71.4% and 90%, with the average number of dominant follicles produced per female being 4.4 ± 0.9; 4.8 ± 0.7 and 4.6 ± 0.6 in Groups A, B and C, respectively. The embryo recovery rate was 77.7%; 90% and 66.7% and the average number of embryos recovered per female was 2.9 ± 0.9; 2.6 ± 0.9 and 2.4 ± 0.8 for Groups A, B and C, respectively. In Groups A and B, the static follicular phase (III) seemed to be ideal for initiating the assisted reproductive technique of MOET. Although prior administration of P4 + EB seems to have no effect on the number of females that responded to the superstimulation treatments, the number of embryos recovered showed a tendency to be higher when ovarian follicle growth inhibition was performed beforehand.     

Nutritional supplementation improves the sexual response of bucks exposed to long days in semi-extensive management and their ability to stimulate reproduction in goats

In confined management systems, well-nourished bucks rendered sexually active by exposure to long days are efficient in fertilizing out-of-season goats. However, underfeeding is common in semi-extensive management systems and may reduce the reproductive efficiency of bucks. The objective of the present study was to determine whether nutritional supplementation improved the sexual activity of bucks submitted to long days in semi-extensive management systems and their ability to stimulate the reproduction of goats in semi-extensive or confined conditions. In experiment 1, three groups of bucks were placed in different flocks and grazed daily with females for 7 h. Each day after grazing, males were separated from females and moved into open pens. One group did not receive any treatment (control group; n = 6). Two other groups were submitted to artificially long days from 15 November to 15 January. From 16 January, one group did not receive nutritional supplementation (long-day group; n = 5), whereas bucks from the other group each received 600 g of a commercial concentrate (long-day + supplementation group; n = 5). The fourth group was kept in confined conditions, exposed to long days and fed alfalfa hay (long-day confined group; n = 6). On 26 March, anovulatory goats from other flocks were assigned to four groups (n = 27 each) and confined separately in open pens. Three bucks of each group were housed with the females. Pregnancy rates were greater in the goats housed with the long-day group than those housed with the control group (P < 0.01). However, pregnancy rates did not differ between the long-day confined group (89%) and long-day + supplementation group (70%; P = 0.09), but these rates were greater than those from the long-day (37%) and control groups (0%; P < 0.05). In experiment 2, two groups of males (n = 3 each) were incorporated into two flocks under semi-extensive management and grazed daily with females for 7 h. One group of males did not receive any treatment (control group). The other group was submitted to long days and nutritional supplementation as in experiment 1 (long-day + supplementation group). Males remained with females during the whole study. The pregnancy rate was greater in the goats joined by males of the long-day + supplementation group (78%) than in those from the control group (0%; P < 0.001). We conclude that long days and nutritional supplementation improve the ability of bucks kept in semi-extensive management to stimulate reproduction of out-of-season goats in confined or semi-extensive management systems.     

Sexual performance of Awassi ram lambs reared in different sex composition groups

The objective of this study was to evaluate the effect of sex composition of the group of lambs before puberty on later sexual performance in Awassi ram lambs. Thirteen Awassi ram lambs of the same age were raised in either all-male group (n = 7) or in a group mixed with females (n = 6) from weaning to puberty. Blood samples, body weights (BW) and scrotal circumferences (SC) of ram lambs were recorded between eight and 10 months of age. Sexual performance testing was performed at 9 months of age by individually exposing ram lambs to oestrous females on five occasions. Body weight and SC were greater (P < 0.01) in the all-male compared with the mixed group. Plasma testosterone concentrations were not influenced (P > 0.05) by treatment. Plasma testosterone concentrations were significantly correlated (P ≤ 0.05) with BW (r = 0.3) and SC (r = 0.4). No treatment effects were detected regarding bouts of leg-kicking. Bouts of anogenital sniffing, the frequency of mount attempts and mounting frequency were greater (P < 0.05) in the all-male group. Even though the frequency of raising the fat tail of females was similar between the two treatments, the mixed group tended to be (P = 0.08) more efficient in doing so than the all-male group (higher tail raising/mount). Results of the current study indicate that mixing groups of ram and ewe lambs before puberty may be insufficient to improve later sexual performance of ram lambs.     

Genetic variation in fertility of heat-stressed male mice

The damaging effects of heat stress on male fertility are evident in developing spermatozoa expressed in ejaculates 18–28 days post-stress in mice. Our objectives were to: (1) assess genetic variation in fertility of heat-stressed male mice and (2) determine response to selection for fertility after heat stress in male mice. Mature male mice were exposed to heat stress (35 ± 1 °C; n = 50) or control (21 ± 1 °C; n = 10) conditions for 24 h (day 0) and then hemicastrated for tissue collection. Two periods of mating tests followed, period 1 (from days 3 to 11) when no reductions in fertility were anticipated, and period 2 (days 18–26) when variation in fertility was expected. Period 2 pregnant females were sacrificed in late gestation. Males were indexed by multiplying overall mean ovulation rate by pre-implantation survival and number of pregnant period 2 mates. The five highest and five lowest ranking males were identified as heat stress resistant and susceptible, respectively. Resistant males were 61.2 units superior in the index, 57.5% greater in pregnancy rate, and 57.6 total fetuses greater than susceptible males. Progeny of resistant sires were superior to progeny of susceptible sires in estimated breeding value by 4.5 units for the index, 4.1% for pregnancy rate, and 5.2 fetuses (P < 0.0001). Heritability estimates for the index, pregnancy rate, and number of fetuses ranged from 0.09 to 0.13, suggesting male fertility following heat stress is heritable and responds to selection.     

Changes in the swimming behavior of Pseudodiaptomus annandalei (Copepoda,Calanoida) adults exposed to the diatom toxin 2-trans, 4-trans decadienal

Diatoms are broadly present in marine habitats and often dominate seasonal phytoplankton blooms in polar and temperate latitudes. Certain species produce polyunsaturated aldehydes upon mechanical wounding caused by mesozooplankton grazing. Ample evidence is available on toxin-induced reproductive failure in copepods, yet their behavioral effects remain unclear. Here we present results of laboratory experiments in which we investigated the immediate effects of the diatom-derived aldehyde 2-trans, 4-trans decadienal on the three-dimensional swimming behavior of the calanoid copepod Pseudodiaptomus annandalei. Short-term direct exposure to the toxin at 3 μM, 6 μM and 12 μM induced hyperactivity in the three adult states, as evidenced by a marked and dose-dependent increase in the number of trajectories. It also caused a higher proportion of vertical movements. In males and ovigerous females exposed to decadienal at 3 μM and 6 μM, hyperactivity came with an equally specific dose-dependent decrease in swimming speed. Males and ovigerous females swam faster at 12 μM than at 6 μM, suggesting a complex mode of action of the toxin. In non-ovigerous females, decadienal had little effects on swimming speed, supporting the assumption that female copepods are less affected by certain environmental stressors. Multifractal analysis revealed differences in the statistical properties of the swimming behavior between experimental conditions. The moment structure function of the displacement appeared to be moderately multifractal in the three adult states swimming in control water. Ethanol as carrier solvent at 200 ppm caused an increase in swimming speed and a switch toward a more ballistic motion in males and ovigerous females. On the opposite, exposure to the toxin reduced or cancelled the effects of ethanol and resulted in a more Brownian motion for high moment values. Decadienal had little effects on the behavior of non-ovigerous females except at the highest concentration. Our results demonstrate that decadienal, a model diatom aldehyde, impairs the behavior of adult copepods. They provide further information on the interaction between diatoms and their main predator.     

Cognition in female rats after blocking conversion of androgens to estrogens

Women and non-human females have surprisingly high levels of circulating testosterone, yet the effects of androgens on non-reproductive behaviors, including cognition, of females are not well characterized. The current project used an aromatase inhibitor, letrozole, to block conversion of androgens to estrogens. Adult female rats were ovariectomized and administered either vehicle only, testosterone propionate only (400 μg/kg, TP only), letrozole only (1 mg/kg, Letro only), or the combination of letrozole and testosterone (TP + Letro) over 4 weeks. A gonadally intact group was used for comparisons. During the last 3 weeks, the animals were tested for working memory in both a spatial task (radial arm maze) and a non-spatial task (object recognition). At sacrifice, uterine weights and serum testosterone and estradiol were determined. Behavioral results were the intact animals showed better working memories on the object recognition task, but that there were no differences among the ovariectomized groups. In the radial arm maze task, groups with best to worst performance were TP only > Intact = TP + Letro > vehicle = Letro only. Highest to lowest serum titers, for testosterone, were TP + Letro > TP only > Intact = Letro only > vehicle and, for estradiol, Intact > TP only > Vehicle > Letro only = TP + Letro. Our interpretation is that testosterone enhanced spatial performance when bioavailability of both TP and E2 are high, and high testosterone can rescue spatial memory when E2 bioavailability is low.