Correlation of seasonal changes in sperm output with endocrinological changes in the adult male bonnet monkey,Macaca radiata

We have examined the monthly variations in sperm output and attempted to correlate the profiles of endocrine hormones secreted with the sperm counts throughout the year in the adult male bonnet monkey. As previously reported, there was a distinct spurt in sperm output beginning September through December months. A concomitant increase in serum testosterone and prolactin concentrations were also noted during September through November (mid and post-monsoon season). Although there was a marked increase in gonadotropin releasing hormone stimulated testosterone secretion, the peak testosterone concentrations post gonadotropin releasing hormone injection did not vary significantly (P > 0.05) throughout the year. Basal serum follicle stimulating hormone concentrations did not vary significantly (P > 0.05) during April to June months compared to September-November months. Serum inhibin concentration remained unaltered throughout the year, except in the month of March. The results of this study provide evidence for annual rhythms in prolactin and testosterone secretion and a distinct seasonality in the sperm output of the adult male bonnet monkey, but the pituitary responsiveness to exogenous gonadotropin releasing hormone remains unaltered throughout the year. Because of the existence of seasonality as noted in the present study, future studies which utilize the adult male bonnet monkey as an experimental model need to take into consideration the seasonal effects on reproductive function in this species.     

Seasonal thyroidal activity and reproductive characteristics of Iranian fat-tailed rams

An experiment was conducted to study seasonal variations of thyroidal activity, serum testosterone concentration and seminal characteristics of two breeds of Iranian fat-tailed sheep. Eight 3 to 4-year-old rams of Ghezel and Mehraban breeds (4 rams/breed) were randomly selected from a flock of fertile rams. Semen was collected by using an artificial vagina, and blood samples were obtained via jugular vein. Seminal characteristics, scrotal length, width, and circumference, and blood parameters were measured at 3-week intervals. Time of the year significantly (P < 0.05) affected the volume of semen, sperm concentration, percent live sperm, percent normal sperm, total sperm number, seminal pH, seminal lactate dehydrogenase (LDH) concentration, scrotal circumference, scrotal width, and serum levels of testosterone, cholesterol, triiodothyronine (T(3)), tetraiodothyronine (T(4)), free T(4) index (FT(4)I), and TSH. Scrotal length was not affected by the time of sampling (P > 0.05). The effect of breed on the serum concentrations of hormones and cholesterol was not significant. Volume of semen, percent normal sperm, and total number of normal sperm were significantly (P < 0.05) higher in Mehraban but scrotal circumference, scrotal width and scrotal length were greater in Ghezel rams. Breedxtime interaction effect was significant for T(4), FT(4)I, sperm concentration, and total sperm number. The largest values for TSH, T(4), FT(4)I, testosterone, total sperm number, percent normal sperm, percent live sperm, sperm concentration, volume of semen and scrotal circumference were found from early summer to winter and the lowest values were recorded at the end of spring and in early summer. In both breeds, high and negative correlations (r = -0.7 to -0.8) were found between LDH levels in semen and percent live sperm, percent normal sperm and total sperm number (P < 0.01). It is unlikely that such variations in seminal quality would affect the fertility under natural mating.     

The effect of season on serum testosterone concentrations in dogs

Photoperiod and environmental temperature are two important factors that may influence the reproductive cycle of various species. The objective of this study was to investigate seasonal influences on serum testosterone concentrations in dogs in a tropical zone, where the variation in day length between winter and summer solstice was approximately 2.5 h. Blood samples were collected every 15 days from seven adult dogs over a 14-month interval and serum testosterone concentrations were determined by radioimmunoassay. The year was divided into four seasons and mean testosterone concentrations for each season were related to the mean environmental temperature and rainfall during that season. Mean testosterone concentrations were 1.81 ng/mL (winter 2002), 1.93 ng/mL (spring 2002), 1.31 ng/mL (summer 2003), 2.02 ng/mL (autumn 2003) and 1.93 ng/mL (winter 2003). The temperature ranged from 10.2 to 32.8 degrees C and the rainfall from 33 to 476 mm. Serum testosterone concentrations were lower in summer 2003 than in both spring 2002 (P = 0.05) and autumn 2003 (P = 0.016). In a tropical zone, a combination of high temperature and substantial rainfall may have reduced serum testosterone concentrations in dogs.     

Comparison of semen characteristics,sperm freezability and testosterone concentration between Duroc and Yorkshire boars during seasons

This study was carried out to investigate the effects of Duroc and Yorkshire boars and seasons influencing semen characteristics, frozen-thawed sperm viability and serum testosterone concentration. Yorkshire boars produced higher semen volume compared to Duroc boars among seasons. However, sperm concentration did not differ significantly between Duroc and Yorkshire boars among seasons. Sperm motility of frozen-thawed sperm in Yorkshire boars was higher than in Duroc boars in spring and summer. Normal acrosome of frozen-thawed sperm in Yorkshire boars was higher than in Duroc boars in winter. Serum testosterone concentrations in Duroc and Yorkshire boars were higher in spring than in summer, autumn and winter. Serum testosterone concentrations in spring and summer were higher in Yorkshire boars than in Duroc boars. In conclusion, when serum testosterone concentrations were higher in Duroc and Yorkshire boars among seasons, semen volume, sperm concentration and frozen-thawed sperm viability were higher.     

Characterization of male killer whale (Orcinus orca) sexual maturation and reproductive seasonality

Longitudinal serum testosterone concentrations (n=10 males) and semen production (n=2 males) in killer whales were evaluated to: (1) characterize fluctuations in serum testosterone concentrations with respect to reproductive maturity and season; (2) compare morphologic changes to estimated age of sexual maturity, based on changes in serum testosterone concentrations; and (3) evaluate seasonal changes in sperm production. Classification of reproductive status and age class was based on differences (P < 0.05) in serum testosterone concentrations according to age; juvenile males ranged from 1 to 7 years (mean+/-S.D. testosterone, 0.13+/-0.20 ng/mL), pubertal males from 8 to 12 years (2.88+/-3.20 ng/mL), and sexually mature animals were 13 years and older (5.57+/-2.90 ng/mL). For captive-born males, serum testosterone concentrations, total body length and height to width ratio of the dorsal fin were 0.7+/-0.7 ng/mL, 495.6+/-17.5 cm and 1.14+/-0.13c m, respectively, at puberty; at sexual maturity, these end points were 6.0+/-3.3 ng/mL, 548+/-20 cm and 1.36+/-0.1cm. Serum testosterone concentrations were higher (P<0.05) from March to June than from December to February in pubertal animals (4.2+/-3.4 ng/mL versus 1.4+/-2.6 ng/mL) and than from September to December in sexually mature animals (7.2+/-3.3 ng/mL versus 4.0+/-2.0 ng/mL). Ejaculates (n = 90) collected from two males had similar (P > 0.05) sperm concentrations across all months. These data represent the first comprehensive study on male testosterone concentrations during and after sexual maturation, and on reproductive seasonality in the killer whale.     

Reproductive patterns and reproductive pathologies of stray bitches in the tropics

The objectives of this study were to determine the annual reproductive pattern and to estimate the frequency of reproductive pathologies in female mongrel stray dogs under tropical conditions. The genital tracts of 300 mongrel bitches from a municipal dog pound were examined post-mortem from January to December 2003. Season of the year, age, size, and body condition score (BCS) were recorded for each dog. The year was divided into three seasons: warm-dry (March-June), warm-humid (July-October), and fresh-humid (November-February). Distribution of estrus periods was not influenced by any of the factors studied (i.e. season, age, size, BCS). A significantly lower number of pregnancies were recorded during the warm-dry season, probably as a consequence of embryo resorption. Underweight animals had a significantly lower percentage of pregnancies than bitches of ideal BCS. More ovulations per bitch occurred during the warm-humid season than during the other seasons of the year, probably due to climatic factors. Bitches of medium and large size had more ovulations than those of small size. Of the 300 bitches examined, 43.5% had one or more genital pathologies. The most frequent pathologies found in the ovary, uterus, and vagina were epo?phoron cysts (6.7%), serosal inclusion cysts (5.0%), and transmissible venereal tumors (15.3%), but the capacity of the females to come into estrus or to become pregnant was not affected by these conditions. We concluded that stray domestic bitches in the tropics were not seasonal breeders, but their reproductive pattern was apparently modified by environmental factors such as temperature and probably photoperiod. Although several bitches in the present study had reproductive pathology, the most prevalent pathologies did not adversely reproductive capacity.     

Reproductive exocrine and endocrine profiles and their seasonality in male langur monkeys (Presbytis entellus entellus)

The reproductive exocrine and endocrine profiles in male langurs are reported with an emphasis on seasonality. The animals showed positive response to electroejaculation throughout the year. The sperm concentration varied between 10–383 × 10⁶/ejaculation with wide fluctuations all through the year. No appreciable changes in the motility and percent live sperm were observed throughout the year. The levels of seminal fructose and magnesium remained unchanged throughout the year, while acid phosphatase showed wide fluctuations. Citric acid showed elevation during February and March and LDH showed elevated levels during April and May. The annual range of serum testosterone was 6–34 nMol/l with a peak during July. Cortisol ranged between 575–1587 nMol/l and prolactin ranged between 107–900 mU/l. Wide fluctuations were observed in hormonal levels. No seasonality was exhibited in the seminiferous tubule diameter, nuclear diameter of Sertoli cells and Leydig cells, and the cholesterol, glycogen, and sialic acid contents of testis. None of the parameters studied have shown any correlation with season. The results suggest that the male langurs lack seasonality in their reproductive exocrine and endocrine profiles and thus could be used as model for research in human reproduction.     

Characterization of reproductive activity in captive male Eurasian lynx (Lynx lynx)

This study characterizes the reproductive activity of male European lynx kept in Central Russia. Four captive adult males were subjected to an electroejaculation prior to (November), during (March) and after (June) the breeding season. Concentration, motility and morphological integrity of spermatozoa as well as testis diameter and testosterone levels in serum were evaluated. Additionally, fecal samples were collected for 2 years to determine the fecal testosterone secretion. Testis sizes and serum testosterone concentrations were characterized by little changes with highest levels in March (2.98 mm³; 1.96 ng/ml testosterone) and lowest in June (2.34 mm³; 0.75 ng/ml testosterone). In faeces, the highest testosterone concentrations were measured in February followed by a second increase in May. The volume of ejaculates and percentages of motile and intact spermatozoa reached the maxima in March. By performing two-male mating experiments, we could prove multiple paternity within three litters. Paternity analysis of litter also revealed that 26 of 31 cubs (84%) were sired from the same male, independently from being the first or second mating partner of the respective female. This particular male showed the most developed and activated reproductive tract and also had the best semen quality, which seems to be important for sperm competition.     

Effects of altrenogest on total scrotal width, seminal characteristics, concentrations of LH and testosterone and sexual behavior of stallions

Twenty stallions (3 to 18 yr old) were used in a study between June 1993 and March 1994. The stallions were divided into 5 groups of 4 each, and, within groups, were randomly assigned to 1 of 4 treatments: 1) untreated controls; 2) once-a-day oral altrenogest (0.088 mg/kg BW) treatment for 150 d; 3) daily altrenogest treatment at the same dose for 240 d; and 4) daily oral altrenogest treatment for 240 d plus subcutaneous GnRH (80 microg) every 4 h from Days 151 to 240. Total scrotal width (TSW) was recorded and semen was collected and evaluated for gel free volume, concentration, sperm motility and sperm morphology. Sexual behavior (libido) was measured as times to first erection and ejaculation. Serum LH and testosterone (T) were measured at various periods throughout the study. Altrenogest decreased serum concentrations of LH and T, TSW, daily spermatozoa output (DSO), the percentage of normal spermatozoa and libido. There was a significant decrease in sperm motility in the Alt-240 and Alt-240+GnRH group, but not the ALT-150 group. The suppression appeared to be partially reversible because DSO, TSW and serum concentrations of LH increased after cessation of progestin treatment. Administration of GnRH during altrenogest treatment resulted in increased (P < 0.05) TSW, DSO and serum concentrations of LH but did not alter sperm morphology or behavior. In summary, the suppressive effects of altrenogest were apparently mediated primarily through a negative feedback inhibition of LH secretion.     

Effect of tamoxifen citrate on reproductive parameters of male dogs

Tamoxifen is a synthetic, nonsteroidal Type I antiestrogenic compound that competitively blocks estrogen receptors with a mixed antagonist-agonist effect. The manifestation of these different actions depends on each species, organ, tissue and cell type considered. Very little is known about the effect of antiestrogens in dogs. The objectives of this study were to determine the effects of tamoxifen citrate on some testis, prostate, hormone, and semen parameters in seven Beagle dogs with uncomplicated spontaneous benign prostatic hyperplasia. Two dogs were normospermic, four were oligozoospermic, and one was azoospermic. The dogs were allocated to a control pre-treatment period, followed by a treatment period, and five post-treatment periods (the duration of each period was 4 weeks). During the treatment period, 2.5mg tamoxifen citrate was given p.o. daily for 28 days to all the dogs. Maximum scrotal width, testicular consistency, libido semen parameters, prostatic volume, serum testosterone concentrations, and side effects were assessed. Tamoxifen negatively affected testis size and libido (P<0.01), and decreased prostatic volume (P<0.01) and testosterone concentrations during treatment. Semen quality deteriorated to nadir values (P<0.01) approximately one spermatic cycle after treatment and returned to pre-treatment values on the second cycle after treatment in all the dogs, except one young oligoazoospermic dog, in which the sperm count was higher ( P<0.01 ) at that time. No side effects were observed and fertility was conserved at the end of the study. Tamoxifen acted more like an agonist than antagonist on the gonadal axis and, therefore, upon both the prostate and testis. Therefore, tamoxifen may have therapeutic applications in dogs.     

Comparison of testosterone and insulin-like peptide 3 secretions in response to human chorionic gonadotropin in cultured interstitial cells from scrotal and retained testes in dogs

Levels of testosterone and insulin-like peptide 3 (INSL3) secretions in response to different doses of human chorionic gonadotropin (hCG) in cultured interstitial cells were compared between retained and scrotal testes in dogs. Retained (n=10) and scrotal (n=9) testes were obtained from small-breed dogs. The testicular tissues were dispersed in Dulbecco's Modified Eagle Medium with Ham's nutrient mixture containing 2000 PU/ml dispase II and 10% fetal bovine serum. The cells were plated with differing concentrations (0-10 IU/ml) of hCG for 18 h in multiwell-plates. Testosterone and INSL3 in the same spent medium were measured by enzyme-immunoassays (EIA). A new EIA with a reliable detection range of 0.025-5 ng/ml was developed in order to measure canine INSL3 in culture medium. Dose-dependent stimulation of testosterone by hCG was observed in the cells of both retained and scrotal testes. The incremental rate of testosterone secretion was significantly lower at 0.1, 1 and 10 IU/ml hCG in the cells of retained testes than in scrotal testes, however. INSL3 secretion was significantly stimulated at 10 IU/ml hCG relative to unstimulated controls comprising cells of scrotal testes; no such stimulation was observed in the cells of retained testes. At 10 IU/ml hCG, the incremental rate of INSL3 was significantly lower in the cells of retained testes than scrotal testes. These results suggest that LH-induced secretory testosterone and INSL3 responses are lower in the interstitial cells of retained testes than of scrotal testes. Furthermore, the high concentrations of LH may acutely stimulate INSL3 release in scrotal testes of dogs, but not in retained testes.     

Photoperiod entrainment of testosterone, luteinizing hormone, follicle-stimulating hormone, and prolactin cycles in rams in relation to testis size and semen quality

Adult rams were exposed to photoperiod treatments over 2 years to study the influence of light regimes on pituitary-testicular activity and semen quality. Initially, all rams (12 per group) were exposed to 3 months of long days (16L:8D). Group 1 was then exposed to a regime of continuous short days (8L:16D) and Groups 2, 3, and 4 were exposed to 4 months of short days alternated with 1, 2, or 4 months, respectively, of long days. Every 2 weeks, serum hormone levels and scrotal circumference were determined and semen quality was evaluated. Regular cycles in pituitary and testicular activities corresponding to the period of the lighting regime resulted in Groups 2, 3, and 4, but not in Group 1. In general, the change from long days to short days induced increases in follicle-stimulating hormone (FSH), luteinizing hormone (LH), and testosterone levels, scrotal size and sperm numbers and a decrease in prolactin. The reverse occurred after subsequent exposure to long days. After 4 months of long days, testicular regression was complete, but when long-day exposure was reduced, less regression occurred. With continuous exposure to short days, FSH and testosterone remained above basal levels, prolactin levels were depressed, scrotal size remained near the maximum, and elevated numbers of motile sperm were sustained.     

Seasonal variation in circulating testosterone and oestrogens of wild-caught California ground squirrels (Spermophilus beecheyi).

Blood samples were obtained each month for 18 months in 1984-85 from wild-caught California ground squirrels. Circulating testosterone and total oestrogens were extracted from the plasma, measured by radioimmunoassay and compared with concurrent changes in plasma progesterone and prolactin in the same individuals. Male plasma testosterone concentrations peaked in January, shortly before mating, whereas female testosterone concentrations were low throughout the year. Female plasma oestrogen concentrations peaked in February, during the mating period. Juvenile males exhibited a transient increase in circulating testosterone in September, followed by testis growth, preputial separation, and the appearance of epididymal spermatozoa. Juvenile females exhibited a transient increase in circulating oestrogens in November. By the start of the first mating season after their births, neither juvenile males nor females differed significantly from same-sex adults with respect to plasma concentrations of oestrogen or testosterone. Plasma concentrations of progesterone and prolactin in killed individuals were similar to those obtained in previous studies of free-living S. beecheyi. Mean plasma concentrations of all measured hormones varied significantly with reproductive condition in adults of both sexes.     

Year round plasma leptin and androgen concentrations in a tropical bat

The detailed reproductive patterns and their associated endocrine characteristics have been documented only for a few species of bats. The objective of this study was to examine seasonal changes in plasma concentrations of leptin and compare it with the changes in body mass, circulating concentrations of testosterone, androstenedione and its correlation with prolonged survival of sperm during winter dormancy in the male sheath-tailed batTaphozous longimanus Hardwicke, 1825. Six bats were captured every month for three consecutive years during 2002 to 2005 from Varanasi, a subtropical part of India. The changes in the body mass were positively correlated with circulating concentration of leptin. Leptin concentration reached a peak (14 ng/ml) in November coinciding with peak body mass. Leptin levels declined during other months of the year except for a rise in March and August. Plasma leptin was positively correlated with androstenedione concentration, but did not show significant correlation with testosterone level. We noticed a significant increase in testosterone secretionin vitro in response to leutinizing hormone (LH) stimulation. However, we did not notice any increase in testosterone or androstenedione secretionin vitro in response to leptin stimulation. Plasma leptin concentration did not show any correlation with testis mass in this study. The higher concentration of testosterone and androstenedione may be responsible for the prolonged survival of sperm in the epididymidies and higher levels of leptin in November may be responsible for maintaining reproductive function during winter dormancy. We suggest that inT. longimanus, higher leptin concentrations in November may be responsible for the gonadal recrudescence and reproductive response during winter dormancy is modified by energy availability and by changing leptin concentrations during this period.     

Effectiveness of exposure to longday followed by melatonin treatment on semen characteristics of Damascus male goats during breeding and non-breeding seasons

Eight mature Damascus male goats were randomly divided into two equal groups. The first group served as control and the second group was used to study the effect of exposure to longday (16L/8D) followed by melatonin treatment (2mg/(animal/day)) on semen characteristics and blood hormonal levels during breeding and non-breeding seasons. During breeding season, longday exposure followed by melatonin treatments resulted in significant increases (P<0.01) in means of sperm motility, ejaculate volume, sperm concentration, total sperm output, total functional sperm fraction and blood testosterone concentration, while means of reaction time, dead sperm, abnormal sperm and blood triiodothyronine (T(3)) concentration were significantly decreased (P<0.01). During non-breeding season, longday exposure followed by melatonin treatment exhibited improvements in some reproductive parameters by reducing (P<0.05) reaction time and percentage of dead sperm (P<0.01) and increasing (P<0.05) total functional sperm fraction. Furthermore, longday exposure followed by melatonin treatment resulted in significant increases (P<0.01) in both of blood testosterone and T(3) concentrations. It is concluded that longday exposure followed by melatonin treatment successfully improve semen characteristics of Damascus male goats during breeding and non-breeding seasons. However, the beneficial effects were more pronounced during the breeding season.     

Seasonal changes in the reproductive organs and plasma and pituitary hormone content of the male Bennett's wallaby (Macropus rufogriseus vufogriseus)

In a number of species of seasonally breeding marsupial, the male is fertile throughout the year but there is a marked seasonal change in weight of the accessory sexual glands. In this study, body weight, prostate, epididymis and testis weights and plasma concentrations of testosterone, LH and prolactin and pituitary content of LH and prolactin were determined in male Bennett's wallabies shot at 1–2 month intervals over a period of 17 months. There was a highly significant increase in prostate weight which was coincident with the breeding season for this species. A small but significant increase in testis weight was also observed but epididymis weight remained unchanged. Plasma testosterone concentrations were significantly increased at a time coincident with the increase in prostate weight. Plasma prolactin and LH concentrations were low in most animals and remained unchanged during the study. In contrast, pituitary prolactin and LH contents showed highly significant changes, with increasing and peak hormone content preceding maximum prostate weight and plasma testosterone concentrations by several months. While these latter results suggest a role for prolactin and LH in the seasonal control of the reproductive organs in the male wallaby, a more intensive study of the pattern of secretion of these hormones and possibly more sensitive hormone assays are required to understand their relative roles in regulating the annual cycle of prostate growth.     

Investigations on reproductive physiology in the male Eurasian lynx (Lynx lynx)

This study characterized (in vivo) morphological and functional parameters of reproductive organs of adult male lynx (n = 3) prior to, during, and after the breeding season (n = 3). Size and morphology of the reproductive tract were monitored by transcutaneous (testes) and transrectal (accessory sex glands) ultrasonography. Semen was collected by electroejaculation. Ejaculate volume, sperm number, motility, and morphology of spermatozoa as well as testosterone concentrations in blood serum and feces were evaluated. The testes and prostate had seasonal changes in size and echotexture. The mean (+/- S.D.) maximum and minimum testicular volume were 2.8 +/- 0.8 cm3 and 1.5 +/- 0.3 cm3, respectively. Fecal testosterone concentrations were highest in February (1240 +/- 393 ng/g feces), with a second increase in May (971 +/- 202 ng/g feces), but concentrations were lowest in January (481 +/- 52.9 ng/g feces). Ejaculate volume, total sperm number and percentage of motile, and intact spermatozoa were maximal in March (the middle of the breeding season). In one of the eight litters, multiple paternity was proven; however, in the remaining seven litters, all 16 cubs were sired by the same male. This particular male had the most developed and active testes and best semen quality, which may be important for sperm competition.     

Seasonal variation in reproductive physiological status in the Iberian ibex (Capra pyrenaica) and its relationship with sperm freezability

The present work examines the relationship between seasonal changes in testicular function, accessory gland size, and horn growth in Iberian ibexes, as well as the relationship between these changes and the resistance of ibex spermatozoa to freezing-thawing. The size of the bulbourethral glands and seminal vesicles showed pronounced monthly variation (P < 0.001), which was correlated positively with the plasma testosterone concentration (P < 0.001) and scrotal circumference (P < 0.001). The size of the accessory sex glands peaked during the autumn. Overall, semen quality was markedly improved during autumn and winter. When horn growth was at a minimum during autumn and winter, semen quality and accessory gland size were all increased compared to in spring and summer. However, increased plasma testosterone levels in the autumn were strongly associated with reduced sperm freezability; thus, the cryosurvival of spermatozoa collected during the autumn was poorer than at other times of the year. In winter, however, when the plasma testosterone concentration fell to baseline, the negative effects of cryopreservation on the percentage of motile spermatozoa and on the integrity of the plasma membrane of frozen-thawed sperm cells were significantly less intense (P < 0.05). These findings show a clear relationship between the functional and morphological status of the different parts of the reproductive tract that optimises reproductive function during the breeding season in the ibex male. They also show that winter is the most suitable season for the collection and cryopreservation of ibex spermatozoa.     

赤腹松鼠血清性激素水平的季节差异

正季节性繁殖是动物适应环境的具体表现,也是动物维持其种群发展的重要策略。生殖激素和动物的繁殖活动关系密切,协调实现繁殖后代的机能(赖平等,2012),如配子的发生、成熟与排出及受精、妊娠、分娩与泌乳等性行为活动。例如雄性甘肃鼢鼠(Myospalax cansus)血清中睾酮(程志兴,2009)和黄山短尾猴(Macaca thibetana)血清中孕酮含量(夏东坡,2007)在交配期间显著升高,     

Role of steroid hormones and prolactin in canine mammary cancer

In several animal studies, prolactin has been found to be essential for mammary epithelial development, and its administration has been consistently shown to increase the rate of mammary tumours. High levels of steroid hormones have also been suggested to enhance mammary cancer development. The present study investigates the levels of the following hormones in serum and in tissue homogenates in dogs bearing canine mammary tumours: prolactin (PRL), progesterone (P4), dehydroepiandrosterone (DHEA), androstenedione (A4), testosterone (T), 17beta-estradiol (17beta-E2) and estrone sulfate (S04E1). Eighty mammary tumours (40 dysplasias and benign and 40 malignant tumours) from 32 female dogs, and 10 normal mammary glands from eight female dogs without history of mammary tumours, were analysed. Prolactin and steroid hormones in serum and tissue homogenates, were analysed by enzyme immunoassays (EIA) techniques, previously validated for this animal species. Levels of prolactin in tissue homogenates were significantly different between malignant and benign mammary tumours (p<0.01). Serum prolactin concentrations were lower in the control group as compared with the group of dogs with benign tumours and in dogs with malignant tumours (p=0.01). Serum prolactin levels in dogs with benign lesions were not significantly different than those obtained from dogs with malignant tumours. Levels of steroid hormones were significantly higher in malignant tumours compared with the benign tumours and normal mammary glands (p<0.01) both in serum and homogenate determinations. Our results suggest that the canine neoplastic mammary gland could be a source of prolactin. Our hypothesis is that both prolactin and steroid hormones are involved in the growth of canine mammary cancer, and that they might have an autocrine/paracrine role in the maintenance of this disease.