Long-term data on basic reproductive parameters and evaluation of endocrine,morphological, and behavioral measures for monitoring reproductive status in a group of semifree-ranging Tonkean macaques (Macaca tonkeana)

Cycle and gestation lengths, menstruation patterns, female genital swelling characteristics, and male-female consortship durations are reported in a semifree-ranging group of Tonkean macaques (Macaca tonkeana) studied over a 12 year period. In addition, profiles of urinary estrone conjugates (E1C) and immunoreactive pregnanediol glucuronide (PdG) throughout four complete menstrual cycles in two females and three full-term pregnancies are presented. Based on intermenstrual intervals, a mean cycle length of 37–41 days (n = 55 cycles in 10 females) was found. Gestation length averaged 173 days (n = 27 pregnancies in eight females). Measurement of PdG immunoreactivity in urine revealed a cyclic pattern with a 5–15-fold increase between follicular and luteal phase concentrations, suggesting that PdG is a reliable indicator of ovarian cyclicity and luteal function. In contrast to PdG, E1C excretion showed no clear pattern throughout the cycle; however, highest values of E1C were usually found shortly before the onset of the luteal phase PdG rise at the presumed time of ovulation. Levels of both hormones were elevated during the first half of gestation and showed a marked increase throughout the second half, with maximum E1C concentrations being up to 100-fold higher than nonpregnant levels. Consortships by the male and occurrence of female genital swelling were long lasting (on average 5–10 days and 13 days, respectively) and were restricted to the follicular phase of the cycle. The day of maximal swelling and day of detumescence as well as the end of male consortship were closely associated with the periovulatory period. Swellings and consortships were longer following lactational ammenorhea than for subsequent cycles. The evolutionary significance of the cyclical changes undergone by females upon their relations with males is discussed. © 1996 Wiley-Liss, Inc.     

Monitoring luteal function in the lion-tailed macaque (Macaca silenus) through urinary progesterone metabolite measurements

A direct radioimmunoassay for measuring urinary 20-hydroxyprogesterone cross-reactivity to monitor and assess luteal function and detect pregnancy in the lion-tailed macaque (Macaca silenus) is described. Urine samples were collected daily during ten nonconceptive and five conceptive ovarian cycles of five dult female lion-tailed macaques. Urine was analyzed for concentrations of 20α-hydroxypro-gesterone cross-reactivity, estrone conjugates, and creatinine. The strength of the luteal phase in normal nonconceptive cycles (n = 8) is characterized by a maximum sevenfold increase (day 9) in mean 20α-hydroxyprogesterone cross-reactivity over follicular phase levels; the duration, by a 13-day sustained elevation of mean 20α-hydroxyprogesterone cross-reactivity levels. Pregnancy is detectable from 20α-hydroxyprogesterone cross-reactivity values approximately 20 days after the periovulatory estrone conjugate peak (n = 4). Apparent anovulation (n = 1), extended follicular phase (n = 1), and early abortion (n = 1) also are detectable using 20α-hydroxyprogesterone cross-reactivity measurements.     

Fatal pulmonary acariasis in an aged indoor rhesus macaque (Macaca mulatta)

Pulmonary acariasis is a sporadic, incidental finding in colony‐raised rhesus macaques (Macaca mulatta). Prophylactic treatment in indoor‐raised and indoor‐housed macaques is not routine due to low prevalence, lack of clinical significance, and potential risk of toxicosis. This case is an unusually severe infestation of Pneumonyssus simicola in an indoor‐housed rhesus macaque, which ultimately resulted in this animal's death.     

Male-Induced Sociosexual Behavior by Vaginal Secretions in Macaca arctoides

Odor communication in Old World monkeys and apes is controversial, because most females have evolved visual and behavioral cues to signal fertility, e.g., sexual swellings. Female stump-tailed macaques (Macaca arctoides) do not have swellings, and mediation of chemical communication likely occurs because males engage in sexual behavior mostly throughout the periovulatory phase of the menstrual cycle. We tested whether vaginal secretions from different cycle phases, with saline solution as a control, promote changes in the frequency of male genital exploration, copulation, and coercive behavior toward females different from the donors, while female donors were apart from the group. Males explored more female genitals when exposed to follicular, periovulatory, and early luteal secretions in comparison to saline or menstrual or late luteal secretions. The increase in coercive behavior after exposure to follicular and periovulatory secretions most likely was a male response to the lack of cooperation of target females in engaging in copulation, as the latter were not receptive during the tests. The strength of male response to vaginal secretions varied significantly as a result of individual variability between donor females, yet the variability does not correlate either to dominance rank or to female age. Exploratory behavior of males correlates significantly with their social rank. Our results suggest that vaginal secretions are among the cues that male Macaca arctoides use to acknowledge the reproductive status of females in the absence of visual signals.     

Relationship between ovarian cycle phase and sexual behavior in female Japanese macaques (Macaca fuscata)

We conducted behavioral observations simultaneously with fecal sample collection on eight nonlactating females 2-3 times per week, October 1997-March 1998, to examine the relationship between ovarian hormones and the sexual behavior of female Japanese macaques (Macaca fuscata) during the mating season. We analyzed samples by enzyme immunoassay for fecal hormone levels. Hormone profiles of estrone-glucuronide (E1) and pregnanediol-glucuronide (PdG) were used to separate ovarian cycles into three phases (follicular, periovulatory, and luteal). Hormonal profiles indicate average cycle lengths of 27.6 +/- 4.2 days (+/- SD; n = 26). Average lengths of the luteal and follicular phases were 12.3 +/- 3.8 days (+/- SD) and 8.3 +/- 3.4 days (+/- SD), respectively. We observed female Japanese macaques engaging in sexual activity throughout the ovarian cycle, with the highest rates occurring during the follicular and periovulatory phases as compared to the luteal phase. The attractivity of female Japanese macaques increased significantly during the follicular and periovulatory phases of the ovarian cycle, when E1 levels are peaking and PdG levels drop to baseline. In addition, females displayed a significant increase in proceptive behavior during the follicular and periovulatory phases. Grooming bouts, as well as proximity between female and male macaques, also increased significantly during the follicular and periovulatory phases. We conclude that fluctuating levels of ovarian hormones in different phases of the cycle are significantly associated with variable rates of copulatory and pericopulatory behaviors in these Japanese macaque females.     

Conflict,displacement activities,and menstrual cycle in long-tailed macaques

This study was aimed at testing whether the female menstrual cycle is associated with changes in social tension experienced by eight heterosexual pairs of long-tailed macaques (Macaca fascicularis). Displacement activities were considered as a behavioral measure of social tension and recorded during the periovulatory and the menstrual flow phases of the cycle. While male yawning and body shake underwent nonsignificant increases, male autogrooming and scratching increased significantly during the periovulatory phase. Female displacement activities did not show any significant difference between the two phases. Male and female socio-sexual behavior was not affected by the menstrual cycle. These results suggest that the male conflictual state, but not the female one, is affected by female menstrual cycle.     

Estrogen and progesterone metabolites and follicle-stimulating hormone in the aged macaque female.

The study presented characterizes the ovarian and pituitary function of the aged female macaque through a complete annual reproductive cycle to compare hormone dynamics during the human and nonhuman primate menopausal transition. Data collected over an entire year from aged macaque females indicated that urinary FSHbeta subunit baseline levels statistically significantly increased in females after age-related abnormal menstrual cycles occurred. These abnormal cycles were followed by anovulation and complete cessation of follicular activity. No statistically significant difference in urinary FSHbeta subunit levels was seen between females that exhibited year-round normal ovarian cycles and those that exhibited seasonal ovarian cycles followed by an interval of anovulation during the nonbreeding season. Basal urinary estrogen metabolite levels were not observed to decrease until ovarian cycles became abnormal and FSHbeta subunit levels began to rise. Early follicular phase circulating inhibin beta levels were statistically significantly reduced only when ovariectomized females were compared to the year-round normally cycling females. A statistically nonsignificant trend toward decreased inhibin secretion, however, was apparent in aged females with normal cycles, aged females with abnormal cycles, anovulatory aged females, and finally, ovariectomized females. Whereas decreased circulating levels of dehydroepiandrosterone sulfate showed a general decline over the 1-yr study period in all groups, they were lowest in the year-round normally cycling group, progressively higher in the normal-to-anovulatory group and abnormal-to-anovulatory group, and highest in the anovulatory group. Finally, the nonbreeding season was associated with the highest number of abnormal cycles, suggesting that onset of complete ovarian senescence in these study macaques was more likely to occur during that time (i.e., females were less likely to return to normal ovarian cycles the following breeding season and more likely to exhibit permanent ovarian quiescence).     

Detecting ovulation in Macaca nemestrina by correlation of vaginal cytology, body temperature and perineal tumescence with laparoscopy

Vaginal cytology, basal body temperature, and perineal tumescence were correlated with laparoscopic observations during the menstrual cycles of five pigtail monkeys (Macaca nemestrina) of known fertility. Percentages of cells obtained in vaginal smears revealed systematic variation in the presence of cell types in relation to the menstrual cycle. Measuring the percentage of exfoliate vaginal epithelial cells containing pyknotic nuclei proved to be of little value for separating the menstrual cycle into its follicular and luteal phases, nor did body temperature provide an accurate index for the occurrence of ovulation. Perineal tumescence, however, measured from the first day of menses to onset of detumescence, was a reliable indicator of the lengths of the follicular and luteal phases as correlated with laparoscopic confirmation of ovulation. Maximal perineal tumescence usually occurred within 12 hours of ovulation, although on one occasion the two events were separated by 48 hours.     

Dynamics of circulating concentrations of gonadotropins and ovarian hormones throughout the menstrual cycle in the bonnet monkey: role of inhibin A in the regulation of follicle‐stimulating hormone secretion

In higher primates, increased circulating follicle‐stimulating hormone (FSH) levels seen during late menstrual cycle and during menstruation has been suggested to be necessary for initiation of follicular growth, recruitment of follicles and eventually culminating in ovulation of a single follicle. With a view to establish the dynamics of circulating FSH secretion with that of inhibin A (INH A) and progesterone (P₄) secretions during the menstrual cycle, blood was collected daily from bonnet monkeys beginning day 1 of the menstrual cycle up to 35 days. Serum INH A levels were low during early follicular phase, increased significantly coinciding with the mid cycle luteinizing hormone (LH) surge to reach maximal levels during the mid luteal phase before declining at the late luteal phase, essentially paralleling the pattern of P₄ secretion seen throughout the luteal phase. Circulating FSH levels were low during early and mid luteal phases, but progressively increased during the late luteal phase and remained high for few days after the onset of menses. In another experiment, lutectomy performed during the mid luteal phase resulted in significant decrease in INH A concentration within 2 hr (58.3±2 vs. 27.3±3 pg/mL), and a 2‐ to 3‐fold rise in circulating FSH levels by 24 hr (0.20±0.02 vs. 0.53±0.14 ng/mL) that remained high until 48 hr postlutectomy. Systemic administration of Cetrorelix (150 µg/kg body weight), a gonadotropin releasing hormone receptor antagonist, at mid luteal phase in monkeys led to suppression of serum INH A and P₄ concentrations 24 hr post treatment, but circulating FSH levels did not change. Administration of exogenous LH, but not FSH, significantly increased INH A concentration. The results taken together suggest a tight coupling between LH and INH A secretion and that INH A is largely responsible for maintenance of low FSH concentration seen during the luteal phase. Am. J. Primatol. 71:817–824, 2009. © 2009 Wiley‐Liss, Inc.     

Effects of ovarian tissue reduction on the menstrual cycle: persistent normalcy after near-total oophorectomy

These experiments were designed to evaluate whether removal of approximately 95% visible ovarian tissue would interrupt the short- or long-term regulation of cyclic ovarian function. On cycle Days 2 4 (onset of menses = Day 1), the entire left ovary and approximately 90% of the right ovary were removed from three cycling cynomolgus monkeys. After approximately 95% ovariectomy, there was an acute elevation of follicle-stimulating hormone (FSH) and luteinizing hormone (LH), which lasted 11 +/- 2 days. A midcycle-like gonadotropin surge occurred 20 +/- 3 days following approximately 95% ovariectomy; the next menses occurred 19 +/- 1 days later. Follicular phase patterns of estradiol preceded the midcycle gonadotropin surge, and luteal phase progesterone levels indicated subsequent ovulation. Two of three monkeys resumed normal menstrual cyclicity in the following cycle with follicular phase, luteal phase, and menstrual cycle lengths similar to pretreatment levels. Histological examination of the ovarian remnant removed on Day 21 of the next cycle revealed a morphologically normal corpus luteum and many small follicles. A second group of 6 rhesus monkeys also underwent approximately 95% ovariectomy for long-term evaluation of menstrual cyclicity; typical 28-day menstrual cycle patterns were observed in 4 of the 6 monkeys for 5 mo, with 2 of these 3 animals maintaining regular menstrual cycles for 1 yr. In summary, our data suggest that normal ovarian function, i.e. recruitment, selection, and dominance of the ovulatory follicle, ovulation, and subsequent corpus luteum function, is maintained with only approximately 5% of functional ovarian tissue remaining.(ABSTRACT TRUNCATED AT 250 WORDS)     

Laparoscopic observations of ovaries before and after ovulation in the Japanese monkey (Macaca fuscata)

Laparoscopic observations of ovaries before and after ovulation were made in 74 cycles in 39 Japanese monkeys (Macaca fuscata). Lengths of menstrual cycles, the follicular phase and the luteal phase in these animals were 25.3±2.8 days, 13.2±1.4 days, and 12.8±2.3 days, respectively. Ovarian morphology during the menstrual cycle in the Japanese monkey agreed in some features and disagreed in others, with findings inMacaca mulatta andMacaca fascicularis reported in the previous papers using laparotomy or laparoscopy. Follicular appearance before and after ovulation varied so considerably from ovary to ovary that it was hard to predict the accurate ovulation time or to estimate the precise age of corpus luteum by means of observations on the morphological appearance only. However, diagnosis of whether ovulation had already occurred or not, was possible when careful observations were made, although some ovaries without haemorrhagic and luteinized appearance after ovulation were very similar to pre-ovulatory follicle. Clear luteinization was detected without fail in many cases three days after ovulation. This study was supported by the Scientific Research Fund of the Ministry of Education of Japan (No. 856165).     

Estradiol-induced follicular atresia in rhesus monkeys is not prevented by exogenous gonadotropins

Estradiol-17β (E₂) induces atresia of the dominant preovulatory follicle (DF) when administered on day 6 of the menstrual cycle. The present study was designed to determine whether the atretogenic effect of E₂ could be averted by the administration of exogenous gonadotropins, in an attempt to determine whether E₂-induced atresia in primates is due to a direct action at the ovarian level or is mediated via pituitary secretion. After identification of the DF during laparoscopy, cyclic monkeys received Silastic capsules containing E₂ placed s.c. for 24 hours, plus one of the following treatments: phosphate-buffered saline, or 25 I.U. of either human urinary menopausal gonadotropin (hMG), FSH-rich hMG, human urinary FSH (uFSH), or human pituitary FSH (pFSH) injected i.m. twice daily for 2 days. The control treatment resulted in atresia of the DF and extended follicular phases (26.3 ± 5.9 days, x? ± S.D.), but in normal luteal phases following ovulation of a substitute DF. Similar results occurred in all animals receiving FSH-rich hMG or pFSH, and in 11 of 16 animals receiving hMG or uFSH (P > 0.05). Since all possible routes and regimens of gonadotropin administration were not attempted, a central action of E₂ cannot be ruled out. However, we believe that the experimental observations support our contention that the atretogenic action of E₂ is exerted in part at the ovary.     

Ovarian cycle phase and same-sex mating behavior in Japanese macaque females

The relationship of the ovarian cycle phase to same-sex mounting activity in adult female Japanese macaques (Macaca fuscata) was studied during the 1997/1998 mating season. Fecal samples were collected from eight female subjects two to three times per week and analyzed by enzyme immunoassay for fecal hormone levels. Hormone profiles of estrone (E1) and pregnanediol (PdG) were used to separate ovarian cycles into three phases: follicular, periovulatory, and luteal. Patterns of same-sex and heterosexual mounting behavior in the females were analyzed for phase variation during conceptive cycles. Same-sex mounting among female Japanese macaques occurred most frequently during the follicular and periovulatory phases of the cycle, and not at all during the luteal phase, paralleling the pattern found in heterosexual mounting behavior. These findings suggest a link between hormonal fluctuations and patterns of sexual mounting, regardless of whether the partner is of the same or opposite sex.     

Effect of enucleation of the corpus luteum at different stages of the luteal phase of the human menstrual cycle on subsequent follicular development

To investigate the mechanism of suppression of follicular development during the luteal phase of the human menstrual cycle, the corpus luteum was enucleated surgically from 10 women at various times after ovulation. In the 24 h after CL enucleation there was an immediate and rapid fall in the concentration of oestradiol and progesterone and a temporary decline in the concentration of FSH and LH. Within 3 days, however, all 10 women showed evidence of renewed follicular activity as indicated by a progressive rise in the concentration of oestradiol. This rise was preceded by a rise in the concentration of FSH and LH, and ovulation, as indicated by a mid-cycle surge in LH and rise in the concentration of plasma progesterone, occurred 16-19 days after enucleation. There was no significant difference in the time to ovulation following enucleation at different times of the luteal phase. The post-operative follicular phase, measured from the time of enucleation, was 3 days longer than that observed pre-operatively from the first day of menstrual bleeding. In the follicular phase of post-operative cycles the concentration of FSH was higher and that of oestradiol lower than the corresponding values before surgery. These results indicate that the absence of healthy antral follicles in the luteal phase of the cycle is due to the inhibitory effects of the corpus luteum. The fact that, after CL enucleation, emergence of the dominant follicle was always preceded by a rise in the concentration of FSH and LH suggests that suppression of gonadotrophins by ovarian steroids secreted by the corpus luteum is responsible for the inhibition of follicular development during the luteal phase of the cycle.     

Heterosexual interactions in laboratory-housed stumptail macaques (Macaca arctoides): observations during the menstrual cycle and after ovariectomy.

Heterosexual interactions of pairs of stumptail macaques (Macaca arctoides) were studied in relation to the female menstrual cycle and after ovariectomy. Five intact male and 10 tubal-ligated female macaques were observed in laboratory pair tests of 20-mins duration, and data were obtained on various male and female behaviors. Each male was tested with the same two females during four 40-day observation periods. Males were tested daily and females were tested every other day. After two 40-day testing periods, one female partner of each male was ovariectomized and the other was sham-operated. Blood was collected regularly from the females during the course of observation and serum levels of estradiol and progesterone were determined by radioimmunoassay. The midcycle peak of estradiol was observed to occur approximately 18 days prior to menses. A distinct secondary peak in estradiol was observed to occur during the luteal phase of all cycles examined. Of 28 different male and female behaviors studied only female presentation to male sexual contact showed a significant midcycle peak related to the endogenous estradiol surge. After ovariectomy a significant decrease in the frequency of several male copulatory behaviors was observed, but most males continued to copulate regularly with their spayed partners throughout the period of this study. Thus, the pattern of copulatory behavior observed in stumptail macaques over the cycle of the female partner and subsequent to ovariectomy differs from that observed in other macaque species studied in the laboratory. It is concluded that cyclical fluctuations in the level of ovarian hormones are not significantly related to measures of sexual interactions in laboratory tests of this species, although the maintenance of copulation and associated behaviors at high levels depends to some degree upon the ovary.     

Ovarian cycle of the captive formosan gem‐faced civets (Paguma larvata taivana)

Formosan gem‐faced civets are classified to be endemic sub‐species of Paguma larvata in Taiwan. Little about their reproductive physiology has been reported. This study was designed to characterize the ovarian activity throughout the year and define ovarian cycle length and the lengths of its component phases. Serum samples were collected for enzyme immunoassay (progesterone and estradiol) from seven captive civets twice weekly for 1 year. Meanwhile, periodic changes in external genitalia (vulva swelling) and vaginal cytology were examined and recorded. Results showed estrous cycles exhibited two types: 18‐day (18.5±1.1, n=64) and 28‐day (27.6±1.0, n=28) as shown by progesterone and estradiol fluctuations and corresponding changes in vulva morphology and vaginal cytology. Both types showed a similar 7‐day follicular phase, peaking progesterone at Day 7. The 18‐day cycle type prevails in the spring and summer whereas the 28‐day cycle type is significant in the autumn. In summary, female gem‐faced civets are polyestrous (approximately 13 cycles/year), and non‐typical seasonal breeders, with follicular phase and two distinct durations of luteal phases (diestrus) cycling throughout the year, but the frequency of ovarian cycles was remarkably gradually decreased from September to February of next year. Zoo Biol 0:1–11, 2007. © 2007 Wiley‐Liss, Inc.     

Cortisol response and ovarian hormones in juvenile and cycling female Cebus monkeys: effect of stress and dexamethasone

We examined cortisol profiles in relation to ovarian hormones and their response to a repeated composite stressor with and without dexamethasone suppression. To evaluate the day-to-day changes in circulating cortisol relative to ovarian hormones, we subjected five adult female Cebus apella monkeys daily to restraint, sedation, transport to a neighboring room for femoral venipuncture, and return to the cage throughout the menstrual cycle. The cortisol response to the repeated stressor for blood collection, its relationship with the ovarian function, and the effects of dexamethasone were evaluated in six juveniles (18-24 months old) and five adult females in the luteal phase. Blood was sampled at time 0; then the monkeys received the vehicle and their blood was sampled again at 1, 2, 4, and 24 hr. This experiment was repeated 3 weeks later, with dexamethasone (i.m. 2 mg/Kg) injected instead of vehicle. Plasma aliquots were assayed for cortisol, progesterone, and estradiol. The results revealed that from middle infancy and throughout adulthood, hypercortisolism is the norm in female Cebus monkeys. The high cortisol values remained unchanged across the cycle despite the cyclic changes in estradiol and progesterone levels. Juvenile monkeys exhibited a higher cortisol response to stress than adults, and both juvenile and adult monkeys exhibited the typical suppression by dexamethasone. A rapid suppression of progesterone co-occurred in parallel with cortisol after dexamethasone injection in juvenile monkeys, suggesting that most circulating progesterone originates in the adrenals. In contrast, adult females exhibited an overincrement of progesterone levels, in parallel with a rise in cortisol, in response to the stressor, and this effect was exacerbated by dexamethasone. The findings suggest that hypercortisolism is insufficient to disrupt ovarian development toward a normal cyclical function, and that ovarian steroids have no influence on day-to-day circulating cortisol levels. On the other hand, the overincrement of progesterone levels induced by stress and/or glucocorticoids during the early luteal phase is unlikely to interfere with the development of this phase and implantation in this monkey species.     

Development of an enzyme-immunoassay (EIA) for the measurement of follicle-stimulating-hormone (FSH) in callitrichid primates using a monoclonal antibody against the human-FSH-β-subunit

Despite the importance of Callitrichid primates in both biomedical and conservation research, practical and reliable immunoassays for the measurement of follicle-stimulating hormone (FSH) have not yet been described. A panel of monoclonal antibodies against specific peptide fragments within either the alpha or beta subunit of human FSH was evaluated for their ability to recognize FSH from Callitrichid and other New World primates. One of these, monoclonal antibody 46.3h6.b7 raised against human FSH, was selected due to its ability to recognize marmoset monkey FSH and its low crossreactivity with other gonadotrophins. The antibody formed the basis of an enzymeimmunoassay using a highly purified human urinary FSH (Metrodin®, Serono) preparation coupled to biotin as label and unmodified as standard. After 24 h incubation, antibody bound label was visualized by addition of streptavidin-peroxidase followed by the appropriate substrate. Parallelism was obtained between the standard and dilutions of pituitary extracts, urine and plasma from the common marmoset (Callithrix jacchus) as well as from two tamarin species (Saguinus fuscicollis and S. oedipus) and one squirrel monkey (Saimiri sciureus). Profiles of plasma and urinary FSH during the follicular phase are shown for two individual marmosets. The ability to measure FSH in Callitrichidae provides new opportunities for studies of the reproductive biology of these New World primate species. Am. J. Primatol. 41:179–193, 1997. © 1997 Wiley-Liss, Inc.     

Validation of an ELISA for urinary dopamine: applications in monitoring treatment of dopamine‐related disorders

Dopamine is a catecholamine that serves as a neurotransmitter in the central and peripheral nervous system. Non‐invasive, reliable, and high‐throughput techniques for its quantification are needed to assess dysfunctions of the dopaminergic system and monitor therapies. We developed and validated a competitive ELISA for direct determination of dopamine in urine samples. The method provides high specificity, good accuracy, and precision (average inter‐assay variation < 12%). The analysis is not affected by general urinary components and structurally related drugs and metabolites. The correlation between ELISA and LC‐MS/MS analyses was very good (r = 0.986, n = 28). The reference range was 64–261 μg/g Cr (n = 64). Week‐to‐week biological variations of second morning urinary dopamine under free‐living conditions were 23.9% for within‐ and 35.5% for between‐subject variation (n = 10). The assay is applied in monitoring Parkinson's disease patients under different treatments. Urinary dopamine levels significantly increase in a dose‐dependent manner for Parkinson's disease patients under l ‐DOPA treatment. The present ELISA provides a cost‐effective alternative to chromatographic methods to monitor patients receiving dopamine restoring treatment to ensure appropriate dosing and clinical efficacy. The method can be used in pathological research for the assessment of possible peripheral biological markers for disorders related to the dopaminergic system.