Vermamoeba vermiformis resides in water-based heater-cooler units and can enhance Mycobacterium chimaera survival after chlorine exposure.

BACKGROUND: Mycobacterium chimaera colonizes water-based heater-cooler units (HCUs), from which it can spread to patients during surgery. Vermamoeba vermiformis is a free-living waterborne amoeba, which was consistently present within HCUs. AIM: To determine whether these amoebae can be involved in the persistent presence of M. chimaera. METHODS: An in-vitro disinfection model. FINDINGS: Increased survival of M. chimaera was observed after chlorine exposure in the presence of V. vermiformis. Confocal microscopy demonstrated the intracellular presence of M. chimaera in V. vermiformis. CONCLUSION: In this way, V. vermiformis can contribute to the persistent presence of M. chimaera in HCUs. Cleaning and disinfection protocols should take this phenomenon into account.

Acanthamoeba castellanii interferes with adequate chlorine disinfection of multidrug-resistant Pseudomonas aeruginosa.

Verona-Integron-encoded-Metallo-ß-lactamase-positive Pseudomonas aeruginosa (VIM-PA) is a cause of hard-to-treat nosocomial infections, and can colonize hospital water networks alongside Acanthamoeba. We developed an in-vitro disinfection model to examine whether Acanthamoeba castellanii can harbour VIM-PA intracellularly, allowing VIM-PA to evade being killed by currently used hospital disinfectants. We observed that A. castellanii presence resulted in significantly increased survival of VIM-PA after exposure to chlorine for 30 s or for 2 min. This undesirable effect was not observed after disinfection by 70% alcohol or 24% acetic acid. Confocal microscopy confirmed the presence of VIM-PA within A. castellanii pseudocysts. Our data indicate that A. castellanii contributes to persistent VIM-PA colonization of water systems after chlorine treatment.

Automated spherical aberration correction in scanning confocal microscopy.

Mismatch between the refractive indexes of immersion media and glass coverslips introduces spherical aberrations in microscopes especially for high numerical aperture objectives. This contribution demonstrates an automated adjustment of the coverslip correction collar in scanning confocal microscopy to compensate for spherical aberrations due to coverslip thickness mismatch. With a motorized coverslip correction collar, the adjustment procedure consists of xz image scans, image processing, correction quality evaluation, the mismatch estimation, and eventually the optimal adjustment of the correction collar. For fast correction with less photodamage, coarse-fine Gaussian fitting algorithms are proposed and evaluated with various specimen for their estimation accuracy. The benefits of the proposed automated correction are demonstrated for various coverslips with biological specimens, showing the optimized resolution of the confocal microscope.

DNA damage induced Pol eta recruitment takes place independently of the cell cycle phase.

When DNA is damaged in cells progressing through S phase, replication blockage can be avoided by TLS (Translesion DNA synthesis). This is an auxiliary replication mechanism that relies on the function of specialized polymerases that accomplish DNA damage bypass. Intriguingly, recent evidence has linked TLS polymerases to processes that can also take place outside S phase such as nucleotide excision repair (NER). Here we show that Pol eta is recruited to UV-induced DNA lesions in cells outside S phase including cells permanently arrested in G(1). This observation was confirmed by different strategies including global UV irradiation, local UV irradiation and local multi-photon laser irradiation of single nuclei in living cells. The potential connection between Pol eta recruitment to DNA lesions outside S phase and NER was further evaluated. Interestingly, the recruitment of Pol eta to damage sites outside S phase did not depend on active NER, as UV-induced focus formation occurred normally in XPA, XPG and XPF deficient fibroblasts. Our data reveals that the re-localization of the TLS polymerase Pol eta to photo-lesions might be temporally and mechanistically uncoupled from replicative DNA synthesis and from DNA damage processing.

Distinct effects of SP-B and SP-C on the uptake of surfactant-like liposomes by alveolar cells in vivo and in vitro.

The effects of surfactant protein B (SP-B) and SP-C on the uptake of surfactant-like liposomes by alveolar type II cells and alveolar macrophages were studied both in vivo and in vitro. In vivo, mechanically ventilated rats were intratracheally instilled with fluorescently labeled liposomes that had SP-B and/or SP-C incorporated in different concentrations. Consequently, the alveolar cells were isolated, and cell-associated fluorescence was determined using flow cytometry. The results show that the incorporation of SP-B does not influence the uptake, and it also does not in the presence of essential cofactors. The inclusion of SP-C in the liposomes enhanced the alveolar type II cells at a SP-C to lipid ratio of 2:100. If divalent cations (calcium and magnesium) were present at physiological concentrations in the liposome suspension, uptake of liposomes by alveolar macrophages was also enhanced. In vitro, the incorporation of SP-B affected uptake only at a protein-to-lipid ratio of 8:100, whereas the inclusion of SP-C in the liposomes leads to an increased uptake at a protein-to-lipid ratio of 1:100. From these results, it can be concluded that SP-B is unlikely to affect uptake of surfactant, whereas SP-C in combination with divalent cations and other solutes are capable of increasing the uptake.

Induction of superovulation in cyclic rats by administration of decreasing doses of recombinant follicle stimulating hormone (Org32489).

The objective of this study was to set up a superovulation protocol in adult cyclic rats by using recombinant human follicle stimulating hormone (rhFSH; Org32489). Good results were obtained by treatment with decreasing doses of rhFSH (2.5 to 0.5 IU) during the dioestrus period. The number of corpora lutea (CL) found in rats treated with this protocol was 43.5 +/- 3.4; this is more than three times the number in saline-treated control rats (13.0 +/- 0.4). Fertilization of oocytes after superovulation was as good as after normal ovulation in terms of number of 2-cell stage embryos found 2 days after mating. The absolute number of implantations was twice the number observed in saline-treated control rats (23.3 +/- 1.8 versus 10.6 +/- 0.5); therefore the number of implantations per CL was lower in superovulated rats. The serum concentrations of luteinizing hormone (LH), endogenous FSH and oestradiol-17beta were decreased during rhFSH treatment, while the inhibin serum concentration was increased. The progesterone serum concentration was increased on the days of pro-oestrus and oestrus after treatment. No difference was observed in the testosterone serum concentration. Pretreatment with 10 IU rhFSH at oestrus before giving the decreasing doses of rhFSH during dioestrus reduced the ovulatory response. Finally, treatment with a constant low dose of rhFSH instead of a decreasing dose of rhFSH did not result in spontaneous ovulation. However, ovulation induction by means of a human chorionic gonadotrophin bolus resulted in superovulation in six out of eight rats. It is concluded that superovulation in cyclic rats can be achieved using rhFSH treatment. However, it was found that the type of rhFSH regimen was very important to achieve appropriate stimulation. The optimal protocol was treatment with decreasing doses of rhFSH during dioestrus. The oocytes retrieved could be fertilized as well as oocytes of saline-treated control rats. The results also indicate that treatment with higher doses of rhFSH might induce a desensitization for FSH and LH.

Inhibin and oestradiol-17 beta in antral follicles of various size classes of cyclic rats.

On the various days of the 5-day oestrous cycle of the rat, ovarian antral follicles were dissected out and grouped in five size classes. Four follicles of the same size class were homogenized jointly in medium, after which inhibin-like bioactivity, inhibin immunoreactivity and oestradiol-17 beta content were measured. In general, there was a significant correlation between immunologically and biologically active inhibin levels in the different size classes; overall correlation was 0.85 (n = 87, P < 0.00001). In the smallest antral follicles (classes 1 and 2) inhibin bioactivity was detected only during the first three days of the cycle. With increasing follicle size, inhibin bioactivity and immunoreactivity increased, with maximal activity present in the largest, i.e. preovulatory, follicles (class 5) during the last three days of the cycle (the day of oestrus denotes day 1 of the cycle). These results indicate that only follicles which reach the antral stage at oestrus, and are known to be recruited by the periovulatory FSH peak, acquire the potency to produce biologically active inhibin. This is the cohort of follicles from which selection of ovulatory follicles will normally take place. In contrast to inhibin, follicular oestradiol-17 beta concentrations were negligible until the last days of the cycle when oestradiol-17 beta was present in follicles larger than class 2; levels increased with increasing follicle size and a maximal level was found in preovulatory follicles at pro-oestrus.(ABSTRACT TRUNCATED AT 250 WORDS)

Recombinant FSH (Org32489) induces follicle growth and ovulation in the adult cyclic rat.

The effects of a single injection of recombinant human FSH (rhFSH; Org32489) on ovulation rate and timing and on antral follicle growth were studied in adult 5-day cyclic rats. Rats injected at 1700 h on dioestrus-2 with a dose of 10 IU rhFSH showed, on average, no increase in ovulation rate on the day of expected oestrus. However, an additional, precocious ovulation resulting in a normal number of corpora lutea (13.3 +/- 0.4, n = 6) was found to take place on the night after injection, i.e. dioestrus-3. No mating behaviour, as shown by the absence of vaginal plugs the next morning, was observed at this ovulation. Follicle counts showed a loss of large antral follicles due to ovulation and increased numbers of healthy small antral follicles at 17 and 41 h after injection, indicating a decrease of atresia of growing follicles as well as additional recruitment of new antral follicles. The endogenous serum FSH concentration on the subsequent day of oestrus (65 h after the rhFSH injection) as well as recruitment of small antral follicles were lower in the rhFSH-treated rats than in saline-treated controls. The ovulation at oestrus, 48 h after the precocious, rhFSH-induced ovulation showed large differences in the number of oocytes between the rats in one treatment group. Similar results in terms of immediate ovulation induction were obtained by using a highly purified human urinary FSH preparation (i.e. metrodin). Furthermore, the direct induction of ovulation by rhFSH or metrodin could not be prevented by the injection of an LHRH antagonist.(ABSTRACT TRUNCATED AT 250 WORDS)

Model of antral follicle dynamics during the 5-day cycle in rats based on measurement of antral follicle inflow.

Antral follicles were counted in ovaries from young adult Wistar rats, collected on the 5 days of the ovarian cycle. Follicles were classified as healthy, early atretic or late atretic and divided into five volume classes. From these data, a model was developed in which the inflow of healthy follicles into the various size classes was quantified. This model describes the follicle dynamics during a normal 5-day cycle. It was concluded that the stage of early atresia takes between 20 and 24 h. The inflow of follicles into the antral stage (volume > or = 100 x 10(5) microns2) was continuous but not constant. The highest inflow was found during pro-oestrus and oestrus, at about the time of the first and second FSH surge. The total inflow during each cycle was about 120 follicles of which only 10% ovulated. These ovulating follicles were recruited during the previous pro-oestrus and oestrus. Follicle selection took place in volume classes 1 and 2 (volume 100-350 x 10(5) microns3) during oestrus and dioestrus 1. At dioestrus 2, the follicles that will ovulate have been selected and can be recognized on the basis of their bigger size.

Prepubertal reduction of the ovarian follicle population by combined LH-releasing hormone antagonist treatment and unilateral ovariectomy influences follicle characteristics but not the ovulation rate at first oestrus.

The effect on first ovulation of the massive reduction of the total pool of ovarian follicles during the infantile and late juvenile period was studied in rats. Treatment with an LH-releasing hormone antagonist (LHRH-A) during infancy (5 mg/kg body weight on days 6, 9, 12 and 15 of life) was combined with unilateral ovariectomy performed on either day 15 (early ULO) or 2-5 days before the expected day of first ovulation (late ULO). Rats were killed on the day of first or second oestrus, when blood was collected and the (remaining) ovaries were prepared for differential counting of follicles and corpora lutea. In addition, blood was sampled 8 h after ULO and the ovaries studied histologically in the group of rats which were unilaterally ovariectomized 2-5 days before first ovulation. The time of first ovulation was not influenced by treatment with LHRH-A, early or late ULO, or a combination of LHRH-A treatment and ULO. Ovulation rate after LHRH-A treatment was decreased, but was still within the normal range in intact rats and in early ULO rats compared with saline-treated controls. Serum FSH concentrations 8 h after ULO performed 2-5 days before first ovulation were similar in saline- and LHRH-A-treated rats (845 +/- 59 and 801 +/- 99 (S.E.M.) micrograms/l respectively) and had increased compared with intact controls (216 +/- 15 micrograms/l). Treatment with LHRH-A resulted in a reduction of more than 50% in healthy and atretic follicles, and late ULO reduced the number of healthy follicles even further.(ABSTRACT TRUNCATED AT 250 WORDS)

Initial ovulation rate and follicle population after injection of inhibin-neutralizing antiserum in the late-prepubertal rat.

In late-prepubertal female rats passive immunoneutralization of endogenous inhibin was achieved by injection of inhibin antiserum. Effects on follicle population, timing of sexual maturation, ovulation rate at first and second oestrus and serum FSH levels were studied. Rats were injected with antiserum, (non-immune) control serum from castrated sheep (castrated serum) or their IgG fractions, or with saline on day 33 or 3 or 2 days (days -3/-2) before the expected day of first ovulation, day 38.5 +/- 0.2 (n = 70). Blood was collected from different subgroups at 8, 24 and 48 h, and at first and second oestrus after injection. At necropsy, ovaries were histologically prepared for differential counting of follicles (48 h and first oestrus) and counting of corpora lutea (CL; first and second oestrus) as an index of ovulation rate. Results from rats injected with either serum or its IgG fraction were not different, as was the case when rats were injected with either castrated serum or saline. Thus, results from groups treated with antiserum and antiserum IgG were combined and labelled 'antiserum', and the castrated serum, castrated serum IgG and saline-treated groups were combined and labelled 'control'. The activity of inhibin-neutralizing antibodies in the circulation of antiserum-treated rats was reduced by 43% between 8 h and second oestrus after injection, as determined by the binding of purified bioactive radioiodinated 31 kDa bovine inhibin.(ABSTRACT TRUNCATED AT 250 WORDS)

Ovulation rate, follicle population and FSH levels in cyclic rats after administration of an inhibin-neutralizing antiserum.

Ovulation rate, follicle growth, serum FSH and oestradiol concentrations were studied after a single intraperitoneal injection of inhibin antiserum in 5-day-cyclic rats. Control rats received (non-immune) serum from castrated sheep or saline. Rats were injected at 10.00 h on dioestrus-1 (D1), i.e. the day following the day of oestrus, or at 17.00 h on dioestrus-2 (D2). The ovaries were excised at necropsy 48 h after injection, or at first or second oestrus after injection. After routine histology fresh corpora lutea were counted and/or differential follicle counts were made. Results from rats injected with either (non-immune) serum from castrated sheep or with saline were not different and were therefore combined to form the control group. The activity of inhibin-neutralizing antibodies in the circulation of antiserum-treated rats was reduced by approximately 39% between 8 h and second oestrus after injection, as determined by the binding of purified bioactive radioiodinated 31 kDa bovine inhibin. Rats were injected on D1 and killed at first oestrus. The number of fresh corpora lutea was significantly higher in antiserum-treated rats than in controls (13.9 +/- 0.4 vs 11.8 +/- 0.4; P less than 0.05). Other rats injected on D1 were killed either 48 h or at the second oestrus after injection. Blood was collected 8, 16, 24 and 48 h and at first and second oestrus after injection.(ABSTRACT TRUNCATED AT 250 WORDS)

Effects of a luteinizing hormone-releasing hormone antagonist in late-juvenile female rats: blockade of follicle growth and delay of first ovulation following suppression of gonadotropin concentrations.

Subcutaneous injections of an antagonist against luteinizing hormone-releasing hormone (LHRH-A, Org, 30276) were administered to late-juvenile female rats. The effects on timing of vaginal opening and first ovulation on serum gonadotropin concentrations and on follicle growth were studied. The dose of 100 micrograms LHRH-A/100 g body wt, given on Days 28, 31, and 34, did not influence timing of first ovulation. After administration of 500 micrograms LHRH-A/100 g body wt, ovulation was retarded by 4.7 days if injections were given on Days 28 and 31; by 6.7 days if given on Days 28, 31, and 34; and by 11.5 days if given on Days 28, 31, 34, and 37. Serum LH and FSH concentrations 3 days after the first, second, and third injections of 500 micrograms LHRH-A were significantly (p less than 0.01) lower than in saline-treated controls. Ovarian follicle counts showed decreased numbers of (antral) Class 2, 3, and 4 follicles 3 days after injection of 500 micrograms LHRH-A/100 g body wt on Day 28; a significantly higher number of Class 1 follicles and a further decrease in Class 2, 3, and 4 follicles 3 days after the second LHRH-A injection; and total absence of Class 3, 4, and 5 follicles 3 days after the third LHRH-A injection. Six days after the third LHRH-A injection, Class 3 and 4 follicles reappeared in the ovaries.(ABSTRACT TRUNCATED AT 250 WORDS)

Short- and long-term effects of an LHRH antagonist given during the prepubertal period on follicle dynamics in the rat.

The effects of the suppression of the high gonadotrophin concentrations normally present by the end of the second week of life on ovarian follicle dynamics were studied in immature rats. Gonadotrophins were suppressed by treatment with an LHRH antagonist (LHRH-A; Org. 30276) on days 6, 9, 12 and 15, and the total population of ovarian follicles was studied at 15 and 28 days, on the day of first oestrus and on the day of oestrus at or following 90 and 300 days of age. Primordial follicles were counted and growing follicles were counted and measured. In rats treated with LHRH-A, follicle recruitment into the growing pool was clearly diminished; the number of growing follicles was significantly (P less than 0.01) lower up to the day of first oestrus and the pool of primordial follicles was significantly (P less than 0.05) larger at 15 and 28 days. Ovarian weights were significantly lower in rats treated with LHRH-A until at least 90 days of age. However, on the day of oestrus at or after 90 and 300 days of age, there were no differences in either the pool of primordial follicles or the pool of growing follicles between rats treated with LHRH-A and control rats. There was also no difference between groups in the number of fresh corpora lutea at these ages. It was concluded that the early peak in gonadotrophin concentrations in immature rats causes substantial recruitment of follicles into the growing pool.(ABSTRACT TRUNCATED AT 250 WORDS)

Ovarian follicle dynamics in immature rats treated with a luteinizing hormone-releasing hormone antagonist (Org. 30276).

The high concentrations of gonadotropins present in immature female rats by the end of the second week of life were suppressed by treatment with an antagonist against luteinizing hormone-releasing hormone (LHRH-A; Org. 30276) on Days 6, 9, 12, and 15 of life. Differential ovarian follicle counts were made on Days 15, 22, 28, and on the day of first estrus of all growing follicles and follicles greater than or equal to 100 x 10(5) microns 3 (mostly antral). In LHRH-A-treated rats, a retardation of follicle growth was noted on Day 15, followed by a gradual loss of growing follicles that amounted to 20% on Day 22 and 40% on Day 28; at first estrus, the total population of growing follicles was only 50% of that present in control rats. Antral follicles, first present at 22 days of age, were lower in number at 28 days of age and at first estrus in LHRH-A-treated rats; this was true for both healthy and atretic follicles. Ovarian weights were significantly reduced in LHRH-A-treated rats at 15 and 28 days of age and on the day of first estrus. However, the numbers of corpora lutea following the first, and normally timed, ovulation were the same in both groups. It was concluded that for early recruitment of follicles to reach a full-sized pool of growing follicles at the age of puberty, high concentrations of gonadotropins early in life have a significant role.

Inhibition of first ovulation: administration of an LHRH antagonist to immature female rats.

Subcutaneous injections of an LHRH antagonist (ALHRH; Org.30093) were administered to immature female rats. Neither a single high dose (50 micrograms) nor repeated daily doses of 5-30 micrograms ALHRH/day, administered between 28 and 38 days of age, influenced the age and body weight at the time of vaginal opening or first ovulation. If repeated daily doses of 2 X 10 micrograms ALHRH were given from 32 to 42 or from 37 to 47 days of age, first ovulation was delayed by 3.0 and 6.3 days respectively. Administration of 10 micrograms ALHRH at 09.00 h and again at 17.00 h on the day of first pro-oestrus was found to be sufficient to block the expected first ovulation in 36 out of 38 rats. This effect could be repeated by administering the same doses of ALHRH at pro-oestrus and again on the next day: ovulation was blocked in eight out of eight rats. A single dose of 10 micrograms ALHRH, administered on the morning of pro-oestrus, blocked ovulation in five out of twelve rats. Both the preovulatory LH and FSH surge, as measured at 16.00 h on pro-oestrus, were found to be inhibited by ALHRH treatment. On the day after pro-oestrus no recruitment of new small antral follicles had occurred in rats with ovulatory blockade. Delayed ovulation took place 2-5 days after ALHRH injection at pro-oestrus; until 3 days after injection rats were able to ovulate their original preovulatory follicles, thereafter newly developed follicles ovulated and large ovarian cysts were found in the ovaries, next to fresh corpora lutea.(ABSTRACT TRUNCATED AT 250 WORDS)

Periovulatory changes in serum concentration and ovarian content of 5 alpha-androstane-3 alpha, 17 beta-diol in the adult rat.

The high amounts of 5 alpha-androstane-3 alpha, 17 beta-diol (3 alpha-diol) present in immature female rats decline towards first ovulation, but on the day of first proestrus a peak is seen. This raises the possibility that during adulthood similar proestrous peaks may occur. Therefore, serum concentrations and ovarian content of 3 alpha-diol were estimated every two hours between 0900 and 2100 h in adult cyclic rats on the day of proestrus. In the same rats, serum concentrations of estradiol (E2), progesterone (P) and luteinizing hormone (LH) were measured, as were ovarian contents of E2 and P. A significant elevation in ovarian 3 alpha-diol was found between 0900 and 1700 h proestrus, whereas serum concentrations of 3 alpha-diol were elevated from 1300 to 2100 h. The high morning values of ovarian 3 alpha-diol correlated with those for ovarian E2 (p less than 0.005); the elevated serum concentrations of 3 alpha-diol during the afternoon correlated with serum P (p less than 0.005) and with serum LH concentrations (p less than 0.005). Serum and ovarian values were positively correlated for P and E2, but not for 3 alpha-diol. The rise in serum 3 alpha-diol could be prevented by blocking the LH surge with sodium pentobarbitone (Nembutal; 35 mg/kg b.w.) administered at 1300 h. In Nembutal-treated rats, the concentration of 3 alpha-diol at 1700 h (886 pg/ml) was significantly lower than in saline-treated control rats (1135 pg/ml; p less than 0.005).(ABSTRACT TRUNCATED AT 250 WORDS)

Inhibin increases in the ovaries of female rats approaching first ovulation: relationships with follicle growth and serum FSH concentrations.

In order to relate various prepubertal events in a group of 95 late prepubertal female rats, the following data were obtained during the last 10 days before the day of first ovulation: amounts of ovarian inhibin-like activity (ILA) in some animals (n = 47); size and numbers of healthy (antral) follicles with a volume greater than or equal to 100 X 10(5) microns3 (or diameter greater than or equal to 260 microns) present per ovary in their litter-mates (n = 48); serum FSH concentrations in both groups. Rats were unilaterally ovariectomized to obtain an ovary for either estimation of ILA content or for histological procedures and counting of follicles. At the time of unilateral ovariectomy they were bled to obtain serum for estimation of FSH concentrations. Rats were kept until the day after the day of first ovulation to determine the time-interval between the day of unilateral ovariectomy and first ovulation. They were studied between 10 and 1 days (days -10 to -1, maturational age) before first ovulation. In addition, adult cyclic rats were bilaterally ovariectomized on different days of the oestrous cycle for estimation of ovarian ILA content. The amount of ovarian ILA was estimated in steroid-free ovarian cytosols using an in-vitro bioassay system with dispersed anterior pituitary cells and subsequent measurement of FSH and LH in the spent medium. The amount of ovarian ILA was about 83 units/ovary from days -10 to -5, and subsequently increased (P less than 0.005) to reach a maximum on day -1, the day of pro-oestrus (213 units/ovary).(ABSTRACT TRUNCATED AT 250 WORDS)

Changes in ovarian steroidogenesis in prepuberal rats induced to ovulate by low doses of human chorionic gonadotrophin.

To determine whether the decrease in ovarian 5 alpha-reduced androgen production before first ovulation might be caused by an increase in serum LH, prepuberal female rats were injected at 28-31 days of age with low doses of human chorionic gonadotrophin (hCG) (0.05-0.075 i.u., four times daily). This treatment resulted in ovulation of six to ten ova per rat on day 32 in all animals. Treatment with hCG resulted in a gradual decrease in ovarian content and production (i.e. content in ovary and medium after 4h of incubation) of 5 alpha-dihydrotestosterone (DHT) and 5 alpha-androstane-3 alpha, 17 beta-diol. The ovarian content of DHT and the production of 5 alpha-androstane-3 alpha, 17 beta-diol decreased within 24 h after the first injection of hCG. Oestradiol content and production increased between 24 and 48 h after the start of treatment and was maximal on day 31 (day of pro-oestrus). Activities of 5 alpha-reductase and aromatase were measured in ovarian homogenates obtained on days 29-31. Activity of 5 alpha-reductase in hCG-treated rats was lower than that in control rats on all days studied. Aromatase activity in hCG-treated rats increased between days 29 and 31. It was concluded that multiple injections of low doses of hCG, which may induce ovulation, cause a decrease in 5 alpha-reduced androgen production, which is probably due to a decrease in 5 alpha-reductase activity. The subsequent increase in oestradiol production corresponds with an increase in aromatase activity.(ABSTRACT TRUNCATED AT 250 WORDS)