Non-invasive endocrine monitoring of ovarian and adrenal activity in chinchilla (Chinchilla lanigera) females during pregnancy, parturition and early post-partum period.

The chinchilla is a rodent that bears one of the finest and most valuable pelts in the world. The wild counterpart is, however, almost extinct because of a drastic past and ongoing population decline. The present work was developed to increase our knowledge of the reproductive physiology of pregnancy and post-partum estrus in the chinchilla, characterizing the endocrine patterns of urinary progesterone, estradiol, LH and cortisol metabolites throughout gestation and post-partum estrus and estimating the ovulation timing at post-partum estrus. Longitudinal urine samples were collected once per week throughout pregnancy and analyzed for creatinine, cortisol, LH, estrogen and progesterone metabolite concentrations. To indirectly determine the ovulation timing at post-partum estrus, a second experiment was performed using pregnant females subjected to a post-partum in vivo fertilization scheme. Urinary progestagen metabolites increased above baseline levels in early pregnancy between weeks-8 and -11 respectively to parturition, and slightly declined at parturition time. Urinary estrogens showed rising levels throughout mid- and late pregnancy (weeks-9 to -6 and a further increase at week-5 to parturition) and decreased in a stepwise manner after parturition, returning to baseline levels two weeks thereafter. Cortisol metabolite levels were relatively constant throughout pregnancy with a tendency for higher levels in the last third of gestation and after the pups' birth. Parturition was associated with dramatic reductions in urinary concentrations of sex steroids (especially progestagens). Observations in breeding farms indicated that the females that resulted in a second pregnancy after mating, did so on the second day after parturition. These data were in agreement with an LH peak detected 24h after parturition. Urinary steroid hormone patterns of estrogen and progestagen metabolites provided valuable information on endocrine events during pregnancy and after parturition in the chinchilla. Results presented in this study enhance our understanding of natural reproductive dynamics in the chinchilla and support empirical observations of breeders that post-partum ovulation occurs ∼ 48 h after parturition.

Validation of a shed skin corticosterone enzyme immunoassay in the African House Snake (Lamprophis fuliginosus) and its evaluation in the Eastern Massasauga Rattlesnake (Sistrurus catenatus catenatus).

This study investigates the use of an enzyme immunoassay to measure keratin glucocorticoid concentrations in reptilian shed skins. Keratin glucocorticoid concentrations were compared to fecal glucocorticoid concentrations during the period of keratin growth in the African House Snake (Lamprophis fuliginosus) and the Eastern Massasauga Rattlesnake (Sistrurus catenatus catenatus). Biochemical validation was performed for the shed skin and fecal corticosterone enzyme immunoassays in the African House Snake. Biological and physiological validations were attempted in the African House Snake. A statistically significant positive association was detected between shed skin corticosterone and the mean fecal corticosterone metabolites from 3 weeks before to 1 week after the previous ecdysis in the African House Snake. A statistically significant difference was not detected between the shed skin corticosterone concentrations of the minimally handled control and the weekly handled (or experimentally stressed) African House Snakes. Adrenocorticotropic hormone stimulation did not result in the physiological validation anticipated for shed skin corticosterone concentrations in the African House Snake.

Noninvasive analysis of fecal reproductive hormone metabolites in female veiled chameleons (Chamaeleo calyptratus) by enzyme immunoassay.

The noninvasive technique of gonadal steroid metabolite measurement in feces for evaluation of reproductive activity has proven an effective and important tool for population management in various captive species, but has not yet been validated and used in reptile species. In this study, enzyme immunoassays (EIAs) were validated for the analysis of fecal samples from female veiled chameleons (Chamaeleo calyptratus) for estrogen (E2), testosterone (T), and progesterone (P) and their metabolites. High performance liquid chromatography and physiological methods (GnRH stimulation) were used for the validation of the assays. Biological events, such as skin color changes indicative of ovarian activity and oviposition, correlated with the cyclical pattern of E2, T and P metabolites in feces over a period of two reproductive cycles. This is the first study to report frequent longitudinal measurements of fecal hormone levels by EIA in a reptile species.

Effects of a phytosterol mixture on male fish plasma lipoprotein fractions and testis P450scc activity.

Plant sterols (phytosterols) have been identified as one potential source of reproductive effects in fish living downstream of pulp mills. beta-Sitosterol, the predominant plant sterol in pulp mill effluent, has previously been shown to decrease plasma sex steroid and cholesterol levels and in vitro gonadal steroid production in fish. In this study, male brook trout (Salvelinus fontinalis) and goldfish (Carassius auratus) were exposed to a phytosterol mixture (72% beta-sitosterol) via Silastic intraperitoneal implants to help elucidate the mechanisms of action of phytosterols on steroid depression. As cholesterol is exogenously supplied for gonadal steroidogenesis, changes in plasma cholesterol fractions were examined. In male brook trout, low-density lipoprotein cholesterol and triglyceride levels decreased significantly, 43 and 50%, respectively, in phytosterol-treated fish. It is improbable, however, that these decreases are linked to depressed gonadal steroidogenesis in fish. The activity of P450scc, which converts cholesterol to pregnenolone (the first step in the steroidogenic pathway), was not affected in testis mitochondria isolated from brook trout or goldfish. Further investigation of the mechanisms of action of phytosterols is required.

Isolation of compounds from bleached kraft mill recovery condensates associated with reduced levels of testosterone in mummichog (Fundulus heteroclitus).

Previous studies have identified chemical recovery condensates as a primary source of hormonally active substances within a Canadian bleached kraft pulp mill. Although reverse osmosis treatment of condensates raises the exposure threshold that alters circulating levels of testosterone in mummichog (Fundulus heteroclitus), the responsible chemicals have not been characterized. In this study, a solid-phase extraction (SPE) method was developed to isolate chemical recovery condensate extractives for evaluation of hormonal activity. Condensates generated during softwood and hardwood pulp production were investigated for their relative potential to affect both circulating and gonadal production of sex steroids in mummichog. Mummichog were exposed to whole condensates, extracts from suspended particulates (> 1 microm), two fractions from SPE, and residual condensates after SPE. The distribution of bioactivity among condensate fractions was similar for both wood furnishes. In both sexes, significant depressions in circulating testosterone and in in vitro gonadal testosterone production were associated with exposure to whole condensates, particulate extracts, one SPE fraction, and residual material after SPE. An optimized SPE method subsequently demonstrated complete recovery of polar, bioavailable chemicals that reduced testosterone levels in both sexes. Characterizations of active fractions by gas chromatography-mass spectrometry (GC-MS) showed the presence of extractives with molecular masses > or = 240 amu possessing functionalities consistent with lignin degradation products. This study provides the first isolation of chemicals derived from pulp production associated with impaired reproductive performance in fish.