Morphological evidence for the physiological nature of follicular atresia in veiled chameleons (Chamaeleo calyptratus).

Follicular atresia (FA) has been assumed to serve different functions in reptiles, e.g. helping to develop hierarchies, limiting clutch size, and regression of ovarian structures. Reproductive output is dependent on a balance between ovulations and FA. Excessive rates of FA may not only be detrimental for the survival of a population, but have also been associated with pathological conditions. In order to gain insights into the physiological and potentially pathological processes of FA, we performed a decriptive study on the morphological features of the ovaries in sexually mature female veiled chameleons (Chamaeleo calyptratus, VC). Of 60 clinically healthy female VC with continuous ovarian cycling and at least one confirmed cycle with FA over at least 1.5 years, 30 were selected for macroscopic evaluation of ovarian appearance and 7 were subjected to histology and immunohistology. While FA of previtellogenic follicles happened at a low rate, expected for a species with two germinal beds per ovary and polyautochronic reproductive pattern, atresia in the late vitellogenic stage affected entire generations of follicles, consequential to ovulatory failure. Histologically, no pathological processes were identified in any of the animals. Rather, three stages of FA (early, middle, late) were defined and vitellogenic follicles showed two distinct morphological types of FA: yolky and cystic. Yolky FA was found in 21/30 (70%) animals, while cystic FA co-occurred in 9/30 (30%) of the animals.

ULTRASONOGRAPHIC AND COMPUTED TOMOGRAPHIC CHARACTERISTICS OF THE REPRODUCTIVE CYCLE IN FEMALE VEILED CHAMELEONS (CHAMAELEO CALYPTRATUS).

Female veiled chameleons, Chamaeleo calyptratus, have a high fecundity and fast maturation, which makes them a suitable model species for squamate reproduction. The authors investigated the morphological follicular development of a group of 20 healthy adult animals over a 12-mon period using ultrasonography (US) and CT. Four stages of follicular development could be distinguished by imaging diagnostics and were confirmed by histology: previtellogenesis, vitellogenesis, gravidity, and atresia. Using a linear ultrasound transducer (18 MHz), previtellogenic follicles could be visualized as small, round, hypoechoic structures. Identification of this stage was unreliable on CT. On US, vitellogenic follicles remained round and showed increasing echogenicity from the hypoechoic center outwards, displaying vinyl-like hyperechoic banding in later stages. On CT, early vitellogenic follicles were round, hyperdense structures, which reduced in density as they grew. A hyperdense inner ring with a hypodense central point characterized late vitellogenesis. Following ovulation, eggs became distinctly oval on both CT and US, with formation of a hyperdense or hyperechoic outer ring, respectively. Atresia followed in cases where no ovulation occurred, and was divided into yolky and cystic atresia. Sonographically, early yolky atretic follicles became unevenly shaped, packed against one another, and developed heterogenous content. Late atretic follicles were homogenous and reduced in size. Reduction of density and uneven shape were also observed on CT. Cystic atretic follicles developed an anechoic cavity with a dense peripheral accumulation of content. In many animals 2-3 generations of atretic follicles were observed without indication of impairment to the development of the newest batch of follicles. Thus, follicular atresia need not necessarily lead to a pathological condition in veiled chameleons, at least not within a few consecutive cycles.

Fecal adrenal hormone patterns during ovulatory and non-ovulatory reproductive cycles in female veiled chameleons (Chamaeleo calyptratus).

The relationship between the reproductive (hypothalamic-pituitary-gonadal; HPG) and adrenal (hypothalamic-pituitary-adrenal; HPA) hormone axes is complex and can vary depending on the species and environmental factors affecting an individual. In an effort to understand this relationship in female veiled chameleons (Chamaeleo calyptratus), the patterns of fecal metabolites of corticosterone (C), estradiol (E), testosterone (T), and progesterone (P) were analyzed by enzyme immunoassay (EIA) during ovulatory (OC; eggs laid) and non-ovulatory cycles (NOC; no eggs laid). Glucocorticoid (GC) metabolites in the fecal extracts were characterized by HPLC and corticosterone EIA performance was assessed by parallelism, accuracy and precision tests. The results indicated that the assay chosen reliably measured the hormone metabolites present in the fecal extracts. Regular, cyclical hormone metabolite patterns consisting of an E peak followed by peaks of T, P and C in close succession were observed during both ovulatory and non-ovulatory cycles; relative levels of P and C, however, were higher during ovulatory cycles. Corticosterone metabolite levels, in particular, increased throughout vitellogenesis and peaked in late vitellogenesis (in non-ovulatory cycles) or around the time of ovulation, and remained elevated throughout the gravid period, falling just prior to oviposition. The results provide evidence of variation in glucocorticoid production throughout different stages of the reproductive cycle, including a role in the ovulatory process; the physiology, however, remains unclear.

Orchiectomy in Testudo species: technical aspects and effect on courtship behaviour.

Courtship behaviour of tortoises includes biting, ramming, chasing, same-sex sexual behaviour, and it commonly is excessive in European tortoise species kept as pets. As a result, males are kept individually. To assess the effect of orchiectomy on excessive courtship behaviour in European tortoises, 50 tortoises (Testudo graeca , T hermanni and T marginata ) underwent endoscopically assisted orchiectomy from April through September 2013 to 2017. Statistical analyses were performed in relation to species, body mass, testicular size and season and owners were invited to fill out a follow-up questionnaire. Body mass of the tortoises ranged from 334 to 2645 g (mean 1056 g) and the age from 5 to estimated 60 years. Testicular length ranged from 1.4 to 7.0 cm (mean 2.7 cm) and testicular mass from 0.6 to 12.6 g (mean 3.9 g). A complete or partial reduction (allowing group-housing) of the excessive courtship behaviour was noted by 95 per cent of owners and 59 per cent of the owners reported a noticeable change of behaviour within a month of surgery. Given the seasonal variation in testicular size, the best period for castrations in male European tortoises is shortly after brumation (April, May), when testes are proportionally smaller.

Neoplasia of captive yellow sea horses (Hippocampus kuda) and weedy sea dragons (Phyllopteryx taeniolatus).

Syngnathidae is the family of fish that includes sea horses, pipefish, and sea dragons. To date, only a single publication has described neoplasia in syngnathids, a fibrosarcoma of the brood pouch in an aquarium-reared lined sea horse (Hippocampus erectus). From 1998 until 2010, the Toronto Zoo submitted 172 syngnathids for postmortem; species included the spotted or yellow sea horse (Hippocampus kuda), the pot-bellied sea horse (Hippocampus abdominalis) and the weedy sea dragon (Phyllopteryx taeniolatus). Seven neoplasms and two neoplastic-like lesions were identified from these cases. Under light microscopy, the neoplasms had morphological characteristics of a cardiac rhabdomyosarcoma, renal adenocarcinoma, renal adenoma, renal round cell tumors, which were likely lymphomas, exocrine pancreatic carcinoma, and intestinal carcinoma. Of these neoplasms, four had clear evidence of metastasis: the pancreatic and intestinal carcinomas and both round cell tumors. As syngnathids are highly fastidious animals, they can be difficult to maintain in captivity. In order to improve their husbandry, preventative and palliative care, as well as treatment, it is important to investigate and document the types of diseases affecting syngnathids.

Serum protein electrophoresis by using high-resolution agarose gel in clinically healthy and Aspergillus species-infected falcons.

Serum protein electrophoresis has gained importance in avian medicine during the past decade. Interpretation of electrophoretic patterns should be based on species-specific reference intervals and the electrophoresis gel system. In this study, serum protein electrophoresis by using high-resolution agarose gels was performed on blood samples collected from 105 falcons, including peregrine falcons (Falco peregrinus), gyrfalcons (Falco rusticolus), saker falcons (Falco cherrug), red-naped shaheens (Falco pelegrinoides babylonicus), and hybrid falcons, that were submitted to the Dubai Falcon Hospital (Dubai, United Arab Emirates) between 2003 and 2006. Reference values were established in clinically healthy birds and compared with values from falcons infected with Aspergillus species (n = 32). Falcons with confirmed aspergillosis showed significantly lower prealbumin values, which is a novel finding. Prealbumin has been documented in many avian species, but further investigation is required to illuminate the diagnostic significance of this negative acute-phase protein.

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.

Cerebral xanthomatosis in three green water dragons (Physignathus cocincinus).

Cerebral xanthomatosis was diagnosed in three female green water dragons (Physignathus cocincinus), all of which presented with progressive neurologic signs. No antemortem evidence for xanthomatosis was identified, but on postmortem examination cholesterol granulomas, composed of cholesterol clefts surrounded by macrophages and multinucleated giant cells, were found in the forebrain of each animal and were associated with significant displacement and pressure on the adjacent brain. Although the cause of xanthomatosis in these animals is unknown, nutrition and trauma may be involved in the pathogenesis of this condition. Cerebrum, cholesterol, green water dragon, Physignathus cocincinus, xanthoma.

Fecal hormone patterns during non-ovulatory reproductive cycles in female veiled chameleons (Chamaeleo calyptratus).

Reproductive failure, in particular preovulatory stasis, is a common problem in captive female reptiles. For a better understanding of its pathogenesis, the patterns of fecal estradiol (E2), testosterone (T), progesterone (P) and their metabolites of 21 anovulatory female veiled chameleons, Chamaeleo calyptratus, were compared with those of 25 animals which ovulated during at least one cycle over the study period. Regular, cyclical hormone patterns, consisting of E2 peaks followed by simultaneous T and P peaks, were observed in all animals, regardless of whether oviposition occurred, indicating continuous ovarian activity. P concentrations were; however, significantly lower in animals undergoing anovulatory cycles. The average time period between the E2 and the T peak was 24.8 days, between the E2 and the P peak 26.5 days, and between the T and P peak 2.6 days. The mean length of reproductive cycles between two sequential ovulatory complexes (oviposition to oviposition) was 132.5 days (range 112-156 days). The results provide evidence that reproductive "failure" occurs more frequently than suspected with some females alternating between ovulatory and anovulatory cycles without any outward evidence of the variation in ovarian cycles. It is proposed that this may be related to physiological adaptation to adverse environmental conditions for breeding and that husbandry factors in captivity are primarily responsible for the progression from a physiological to pathological process.

Anesthetic management of a white rhinoceros (Ceratotherium simum) undergoing an emergency exploratory celiotomy for colic.

OBSERVATIONS: A 26-year-old male white rhinoceros (Ceratotherium simum), weighing approximately 2000 kg was anesthetized for an exploratory celiotomy. Sedation was achieved with intramuscular butorphanol (0.04 mg kg(-1)) and detomidine (0.025 mg kg(-1)) and induction of anesthesia with intravenous glyceryl guaiacolate (50 g) and three intravenous boluses of ketamine (200 mg, each); the trachea was then intubated and anesthesia maintained with isoflurane in oxygen using a circle breathing system. Positioning in dorsal recumbency for the surgery and later in sternal recumbency for the recovery represented challenges that added to the prolonged anesthesia time and surgical approach to partially correct an impaction. The rhinoceros recovered uneventfully after 10.4 hours of recumbency. CONCLUSIONS: Anesthetic management for an exploratory celiotomy with a midline approach is possible in rhinoceroses, although planning and extensive staff support is necessary to adequately position the patient.

Characterization of fecal hormone patterns associated with the reproductive cycle in female veiled chameleons (Chamaeleo calyptratus).

Reptiles have gained popularity in the North American and European pet trade. Large numbers of captive-born veiled chameleons, Chamaeleo calyptratus, are produced annually but knowledge of their reproductive cycle has been limited to anecdotal observations. This study describes the hormonal changes associated with reproductive cycling in female veiled chameleons using non-invasive fecal evaluation of metabolites of the three principal ovarian steroids, estradiol (E2), testosterone (T), progesterone (P), and their metabolites, by enzyme immunoassays. The hormone patterns were compared with follicular development and ovulation as determined by magnetic resonance imaging (MRI). Three main cycle stages were identified on MRI: the previtellogenic stage (PV) with the absence of visible follicular structures, vitellogenic stage (V) with the presence of round follicular structures >2mm diameter, and the gravid stage with the presence of oval egg structures. Although the absolute values of the baselines and peaks for each hormone varied among animals, approximately 24-fold increases over mean P baseline values and 7.5-fold increases over mean E2 and T baseline values were associated with biological events. E2 rose during vitellogenesis, peaked in late vitellogenesis and fell shortly thereafter. P rose during the late vitellogenic stage, peaked in mid-gravidity and fell to baseline values at oviposition. Ovulation occurred with the decreasing E2:P ratio. T levels varied during the pre- and vitellogenic stages then mirrored P with a distinct peak during the time of ovulation and gravidity. These data provides us with the necessary background for future studies on the reproductive biology of this species.