Adrenal activity and anxiety-like behavior in fur-chewing chinchillas (Chinchilla lanigera).

Due to its complexity, in combination with a lack of scientific reports, fur-chewing became one of the most challenging behavioral problems common to captive chinchillas. In the last years, the hypothesis that fur-chewing is an abnormal repetitive behavior and that stress plays a role in its development and performance has arisen. Here, we investigated whether a relationship existed between the expression and intensity of fur-chewing behavior, elevated urinary cortisol excretion and anxiety-related behaviors. Specifically, we evaluated the following parameters in behaviorally normal and fur-chewing animals of both sexes: (1) mean concentrations of urinary cortisol metabolites and (2) anxiety-like behavior in an elevated plus-maze test. Urinary cortisol metabolites were higher only in females that expressed the most severe form of the fur-chewing behavior (P≤0.05). Likewise, only fur-chewing females exhibited increased (P≤0.05) anxiety-like behaviors associated with the elevated plus-maze test. Overall, these data provided additional evidence to support the concept that fur-chewing is a manifestation of physiological stress in chinchilla, and that a female sex bias exists in the development of this abnormal behavior.

Video Validation of Tri-Axial Accelerometer for Monitoring Zoo-Housed Tamandua tetradactyla Activity Patterns in Response to Changes in Husbandry Conditions.

Accelerometers are a technology that is increasingly used in the evaluation of animal behaviour. A tri-axial accelerometer attached to a vest was used on Tamandua tetradactyla individuals (n = 10) at Biodiversity Park. First, the influence of using a vest on the animals' behaviour was evaluated (ABA-type: A1 and A2, without a vest; B, with a vest; each stage lasted 24 h), and no changes were detected. Second, their behaviour was monitored using videos and the accelerometer simultaneously (experimental room, 20 min). The observed behaviours were correlated with the accelerometer data, and summary measures (X, Y and Z axes) were obtained. Additionally, the overall dynamic body acceleration was calculated, determining a threshold to discriminate activity/inactivity events (variance = 0.0055). Then, based on a 24 h complementary test (video sampling every 5 min), the sensitivity (85.91%) and precision (100%) of the accelerometer were assessed. Animals were exposed to an ABA-type experimental design: A1 and A2: complex enclosure; B: decreased complexity (each stage lasted 24 h). An increase in total activity (%) was revealed using the accelerometer (26.15 ± 1.50, 29.29 ± 2.25, and 35.36 ± 3.15, respectively). Similar activity levels were detected using video analysis. The results demonstrate that the use of the accelerometer is reliable to determine the activity. Considering that the zoo-housed lesser anteaters exhibit a cathemeral activity pattern, this study contributes to easily monitoring their activities and responses to different management procedures supporting welfare programs, as well as ex situ conservation.

Multivariate Analyses of the Activity Pattern and Behavior of the Lesser Anteater on Open and Closed Days at Córdoba Zoo, Argentina.

The human factor is one of the major determinants of animal welfare in zoos. Assessing changes in activity patterns, behavior and space use due to human presence might help understanding visitors' influence on these animal welfare indicators. In Córdoba (Argentina), we assessed the activity pattern and behavior of Tamandua tetradactyla (lesser anteaters; n = 5) during natural light/dark phases and the animals' use of space. We analyzed responses of lesser anteaters to humans in days open to the public (Sundays and Tuesdays) and days closed to the public (Mondays), collecting data at 5-minute intervals during 12 days, for each animal. Data were analyzed at two temporal scales: a 24 h period and visiting hours. Multivariate analyses showed no differences in activity pattern, behavior and space use among days, exhibiting a consistent response of each individual over the studied days. Principal Component Analysis showed differences between female and male behaviors. Based on these results, the visitor effect on these lesser anteaters would appear to be neutral, indicating that the presence of visitors is at most, a minor concern in the housing and management of these individuals in Córdoba Zoo.

Time-related changes in functional activity and capacitation of chinchilla (Chinchilla lanigera) spermatozoa during in vitro incubation.

The application of assisted breeding programs for chinchilla, an endangered species, requires detailed knowledge about their gamete physiology. Main purposes of the present study were to examine the time-related changes during 8h in vitro incubation in parameters that reflect chinchilla sperm functional activity (including sperm motility, viability, membrane and acrosome integrity), and to determine the incubation time required for achieving in vitro sperm capacitation, evaluated through the quantification of the percentages of sperm that underwent the acrosome reaction in response to progesterone (P, 20 microM) or another acrosome reaction inducer the calcium ionophore, A23187 (20 nM). Semen was obtained by electroejaculation, subjected to swim-up and incubated for 0, 2, 4 and 8h. After these periods, sperm functional activity was assessed. In all treatments percentages of motile, viable and viable sperm with intact acrosomes decreased (p<0.001) after 8h of incubation. The percentages of swollen gametes decreased (p<0.001) after 2h of incubation. Capacitation of chinchilla sperm could be achieved within 4h, as indirectly demonstrated by the increase of acrosome reacted cells in response to P or A23187 (time x treatment interaction: p=0.02).

Distribution and concentration of maternal progesterone in the yolk of Greater Rhea eggs (Rhea americana).

Progesterone is the most concentrated maternal yolk steroid characterized to date in birds; however, no information about it is available in ratite eggs. We collected freshly laid eggs from zoo-housed Greater Rhea females (Rhea americana) bred under similar rearing conditions during two breeding seasons to characterize concentration and distribution of maternal yolk progesterone. After high-performance liquid chromatography analysis, yolk hormone was measured using a commercial electrochemiluminescence immunoassay. Progesterone concentrations were found to vary significantly among the yolk layers, supporting a follicular origin for this steroid in Greater Rhea eggs. Additionally, highly similar mean absolute yolk progesterone concentrations were detected between 2013 and 2015 breeding seasons (1,332.98 ± 82.59 and 1,313.59 ± 85.19 ng/g, respectively). These values are also comparable to those found in some domestic carinate species. Findings suggest that at population level, when rearing conditions are similar, mean absolute yolk maternal progesterone concentrations also appear bounded. Future research on the factors and mechanisms that regulate progesterone deposition in Greater Rhea eggs is needed to better understand whether its levels depend on different rearing conditions.

Year-round testicular volume and semen quality evaluations in captive Chinchilla lanigera.

In mammals, reproductive performance is usually associated with seasons. Chinchilla lanigera, an endemic South American rodent, reproduces throughout the year in captivity but its seasonal breeding pattern is not fully understood. The present study was designed to evaluate (bi-weekly) over 1 year: (1) testicular volume variations and (2) seminal volume, sperm concentration and functional activity changes. Five animals were studied; they were individually housed indoors (22.2 +/- 1.0 degrees C) under natural photoperiod in Argentina (Córdoba, 31 degrees S-64 degrees W). Semen was obtained by electroejaculation; a total of 116 ejaculates was evaluated. Monthly values for paired testicular volume were less in the middle of the summer than in other seasons (p < 0.006), while those for seminal volume and total spermatozoa/ejaculate were not significantly different; these variables ranged between 7.2-30.9 cm(3), 10-130 microL and 0.9-432.6 x 10(6), respectively. Spermatozoa concentration was (x 10(6)/mL) 2145.9 +/- 365.3 and the pH of semen was 7.3 +/- 0.0. Spermatozoa functional activity showed no significant differences between monthly evaluations; confidence intervals were calculated for the means of: motility, 92.2-95.8%; viability, 92.2-96.1%; swollen cells (hypo-osmotic swelling test), 81.2-87.7% and viable intact acrosome, 83.5-89.0%. The present study represents the first longitudinal reproductive assessment in the chinchilla male. In conclusion, males produce spermatozoa continuously that exhibit high quality functional activity.

Assessment of urine and fecal testosterone metabolite excretion in Chinchilla lanigera males.

Endemic chinchilla (Chinchilla spp.) populations are nearly extinct in the wild (South America). In captive animals (Chinchilla lanigera and C. brevicaudata), reproduction is characterized by poor fertility and limited by seasonal breeding patterns. Techniques applied for studying male reproductive physiology in these species are often invasive and stressful (i.e. repeated blood sampling for sexual steroids analysis). To evaluate endocrine testicular function, the present experiments were designed to (a) determine the main route of testosterone excretion (14C-testosterone infusion in four males); (b) validate urine and fecal testosterone metabolite measurements (HPLC was used to separate metabolites and immunoreactivity was assessed in all metabolites using a commercial testosterone radioimmunoassay, and parallelism, accuracy and precision tests were conducted to validate the immunoassay); and (c) investigate the biological relevance of the techniques applied (quantification of testosterone metabolite excretion into urine and feces from five males injected with hCG and comparison between 10 males and 10 females). Radiolabelled metabolites of 14C-testosterone were excreted, 84.7+/-4.2 % in urine and 15.2+/-3.9 % in feces. A total of 82.7+/-4.2% of urinary and 45.7+/-13.6% of fecal radioactivity was excreted over the first 24 h period post-infusion (metabolite concentration peaked at 8.2+/-2.5 h and 22.0+/-7.0 h, respectively). Several urinary and fecal androgen metabolites were separated by HPLC but only fecal metabolites were associated with native testosterone; however, there was immunoreactivity in more than one metabolite derived from 14C-testosterone. After hCG administration, an increase in androgen metabolite excretion was observed (p<0.05). Males excreted greater amounts daily of urinary androgen metabolites as compared with females (p<0.05); this difference was not evident in feces. Results of the present study indicate that the procedure used is a reliable and non-invasive method to repeatedly monitor variations in testicular endocrine activity in this species. It can be a useful tool that would help ensure the survival of the wild populations as well as to provide the basis for a more efficient use by the fur industry.

Validation of a radioimmunoassay for measuring testosterone concentrations in plasma samples of the subterranean rodent Ctenomys talarum: outstandingly elevated levels in the wild and the effect of captivity.

We validated the Coat-a-Count radioimmunoassay (RIA) kit for measuring testosterone in plasma samples of the subterranean rodent Ctenomys talarum and evaluated testosterone levels in free-living and captive individuals. The performance of the assay was evaluated by the assessment of parallelism, accuracy and precision. Moreover, the high specificity of the assay antibody was confirmed by high-pressure liquid chromatography with ultraviolet detector, followed by electrospray ionization tandem mass spectrometry. Results indicated that plasma samples have to be treated with heat and diluted before the RIA for the optimization of the assay. Plasma testosterone concentrations in free-living animals were outstandingly elevated (up to 329 ng/mL), which are among the highest ever reported for mammals. On the other hand, captivity produced a 14-fold decrease in plasma testosterone concentrations, emphasizing that very significant changes in the endocrine milieu may occur in wild animals kept under laboratory conditions. Our results place tuco-tucos as an interesting model for the study of androgen regulation in mammals, suggesting that target tissues may have low sensitivity to the testosterone signal and agree with a scenario of elevated levels of sex hormone-binding globulin in plasma.