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.

Effect of the Prolactin Inhibitor Cabergoline and the Gonadotropin Releasing-Hormone Agonist Deslorelin in the Suppression of Plasma Prolactin Concentrations and Egg Laying in Quail (Coturnix japonica).

Egg binding and excessive laying frequently affect avian patients, and in many cases the treatment includes suppression of egg production. Currently, for the suppression of egg production in avian patients, a gonadotropin-releasing hormone agonist, in the form of a deslorelin implant, is often used. However, the commercially available deslorelin implants have an undesired delayed onset, as well as a potential brief increase in gonadotropin secretion after administration ("flare-up" effect) that can lead to oviposition before the actual suppression of gonadotropins. The objective of this study was to investigate whether the prolactin inhibitor cabergoline suppresses ovulation and whether it could be used to bridge the time until the onset of effect by the deslorelin implant. We measured the effect of cabergoline (30 µg/kg PO q24h × 14 days), deslorelin implants (4.7 mg SC), and a combination of both on egg laying and plasma prolactin concentrations in 37 quail (Coturnix japonica) over 6 weeks. Quail were divided into 4 groups: group DesCab (deslorelin implant and cabergoline oral; n = 9); group DesPlac (deslorelin implant and placebo oral; n = 9); group PlacCab (placebo implant and cabergoline oral; n = 9); and group PlacPlac (placebo implant and placebo oral; n = 10). Regular egg laying stopped in 100% (9/9) of birds in group DesCab and 78% (7/ 9) of birds in group DesPlac within 5 days of placing the deslorelin implant. No bird ceased egg production in group PlacCab (0/9), and 10% of birds ceased egg production intermittently in group PlacPlac (1/10). Treatment with the deslorelin implant (P < .001) and with cabergoline (P = .04) had a significant (negative) influence on plasma prolactin concentrations compared with the baseline. The interaction of deslorelin and cabergoline treatment, as well as time after initiation of treatment, did not have a significant effect on plasma prolactin concentrations. These results show that daily oral cabergoline has no significant influence on egg laying and only a minor biologically nonsignificant effect on lowering the relative plasma prolactin concentrations in quail.

Metagenomic analysis of fecal and tissue samples from 18 endemic bat species in Switzerland revealed a diverse virus composition including potentially zoonotic viruses.

Many recent disease outbreaks in humans had a zoonotic virus etiology. Bats in particular have been recognized as reservoirs to a large variety of viruses with the potential to cross-species transmission. In order to assess the risk of bats in Switzerland for such transmissions, we determined the virome of tissue and fecal samples of 14 native and 4 migrating bat species. In total, sequences belonging to 39 different virus families, 16 of which are known to infect vertebrates, were detected. Contigs of coronaviruses, adenoviruses, hepeviruses, rotaviruses A and H, and parvoviruses with potential zoonotic risk were characterized in more detail. Most interestingly, in a ground stool sample of a Vespertilio murinus colony an almost complete genome of a Middle East respiratory syndrome-related coronavirus (MERS-CoV) was detected by Next generation sequencing and confirmed by PCR. In conclusion, bats in Switzerland naturally harbour many different viruses. Metagenomic analyses of non-invasive samples like ground stool may support effective surveillance and early detection of viral zoonoses.

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.

Evaluation of Two Miniplate Systems and Figure-of-eight Bandages for Stabilization of Experimentally Induced Ulnar and Radial Fractures in Pigeons ( Columba livia ).

Although plate fixation has advantages over other fixation methods for certain indications, it is rarely used in avian surgery, especially in birds that weigh less than 1000 g. Exceptionally small plating systems for these birds are required, which are relatively expensive and difficult to insert. To study avian fracture healing after repair using miniplates, we evaluated 2 steel miniplate systems in 27 pigeons ( Columba livia ) divided into 4 groups. In each pigeon, the left ulna and radius were transected and the ulna was repaired with a bone plate. In groups A and B, a 1.3-mm adaption plate was applied without and with a figure-of-eight bandage; in groups C and D, a 1.0-mm compression plate was applied without and with a bandage, respectively. Healing was evaluated with radiographs after 3, 14, and 28 days; flight tests were conducted after 14, 21, and 28 days; and the wing was macroscopically examined after euthanasia of birds on day 28. Fractures healed without bending or distortion of the plate in all 27 birds, and no significant differences in healing were found between treatment groups. At the end of the study, 23 pigeons (85.2%) showed good or very good flight ability. Results show the 1.3-mm adaption plate and the 1.0-mm compression plate meet the requirements for avian osteosynthesis and can be recommended for fracture repair of the ulna or other long bones in birds weighing less than 500 g. The application of a figure-of-eight bandage might be beneficial in fracture healing.

Transurethral Cystoscopy and Endoscopic Urolith Removal in Female Guinea Pigs (Cavia porcellus).

Urinary diseases are commonly found in guinea pigs. Diagnostic workup includes clinical examination, blood testing, imaging studies, urine culture, and urinalysis. This article describes the use of transurethral cystoscopy in female guinea pigs as an ancillary tool to diagnose abnormalities within the bladder and urethra. In addition, the transurethral cystoscopic removal of uroliths measuring up to 5 mm and situated within the urinary bladder is described.

Peripheral nerve sheath tumor in a subadult golden eagle (Aquila chrysaetos).

A 5-year-old, female golden eagle (Aquila chrysaetos) was admitted with tetraplegia that progressed to a nonambulatory, spastic tetraparesis after a few days of treatment. Clinical and radiologic examinations, including radiography, computed tomography scan, and myelography, were indicative of neoplasia involving a spinal nerve root. Postmortem magnetic resonance imaging and necropsy findings confirmed the diagnosis of a peripheral nerve sheath neoplasia, not, to our knowledge, previously reported in a raptor.

Blood values of captive beira antelope (Dorcatragus megalotis) prior to and during an outbreak of fibrinous pleuropneumonia syndrome (FPPS).

Currently the only captive population of beira antelope (Dorcatragus megalotis) is held at the Al Wabra Wildlife Preservation, Qatar. An outbreak of a severe respiratory disease--fibrinous pleuropneumonia syndrome, most likely caused by Mycoplasma ovipneumoniae--led to a marked population decline. Reactive systemic inflammatory (AA) amyloidosis was noted as a chronic manifestation of the disease. Blood samples had been collected for biochemistry and hematology baseline values prior to the outbreak. Population-level changes were analyzed before and during the course of the outbreak in selected blood parameters (white blood cells [WBC], blood urea nitrogen [BUN], and creatinine). The annual population WBC increased and decreased concurrently with the population size, with a significant correlation between the two measures (R = 0.92; P = 0.001). Both BUN and creatinine values were higher during the outbreak. These values peaked at the same time as mortality, which was 1 yr after the WBC peak. These changes were interpreted as the transition from an acute disease with a primary respiratory manifestation into a chronic condition where renal amyloidosis led to chronic renal failure and death. Also, elevated liver values in diseased animals were attributed to amyloidosis. Parallels to a literature report on a lung disease complex caused by M. ovipneumoniae in bighorn sheep (Ovis canadensis) were found. Trends in population-level blood values of the beira antelopes implicate amyloidosis as a significant, long-term consequence of the putative Mycoplasma infection.

Comparison of fluid types for resuscitation in acute hemorrhagic shock and evaluation of gastric luminal and transcutaneous Pco2 in Leghorn chickens.

The objective of this study was to compare the effects of 3 different fluid types for resuscitation after experimentally induced hemorrhagic shock in anesthetized chickens and to evaluate partial pressures of carbon dioxide measured in arterial blood (Paco2), with a transcutaneous monitor (TcPco2), with a gastric intraluminal monitor (GiPco2), and by end tidal measurements (Etco2) under stable conditions and after induced hemorrhagic shock. Hemorrhagic shock was induced in 40 white leghorn chickens by removing 50% of blood volume by phlebotomy under general anesthesia. Birds were divided into 4 groups: untreated (control group) and treated with intravenous hetastarch (haes group), with a hemoglobin-based oxygen carrier (hemospan group), or by autotransfusion (blood group). Respiratory rates, heart rates, and systolic arterial blood pressure (SAP) were compared at 8 time points (baseline [T0]; at the loss of 10% [T10%], 20% [T20%], 30% [T30%], 40% [T40%], and 50% [T50%] of blood volume; at the end of resuscitation [RES]; and at the end of anesthesia [END]). Packed cell volume (PCV) and blood hemoglobin content were compared at 6 time points (T0, T50%, RES, and 1, 3, and 7 days after induced hemorrhagic shock). Measurements of Paco2, TcPco2, GiPco2, and Etco2 were evaluated at 2 time points (T0 and T50%), and venous lactic acid concentrations were evaluated at 3 time points (T0, T50%, and END). No significant differences were found in mortality, respiratory rate, heart rate, PCV, or hemoglobin values among the 4 groups. Birds given fluid resuscitation had significantly higher SAPs after fluid administration than did birds in the control group. In all groups, PCV and hemoglobin concentrations began to rise by day 3 after phlebotomy, and baseline values were reached 7 days after blood removal. At T0, TcPco2 did not differ significantly from Paco2, but GiPco2 and Etco2 differed significantly from Paco2. After hemorrhagic shock, GiPco2 and TcPco2 differed significantly from Paco2. The TcPco2 or GiPco2 values did not differ significantly at any time point in birds that survived or died in any of the groups and across all groups. These results showed no difference in mortality in leghorn chickens treated with fluid resuscitation after hemorrhagic shock and that the PCV and hemoglobin concentrations increased by 3 days after acute hemorrhage with or without treatment. The different CO2 measurements document changes in CO2-values consistent with poor perfusion and may prove useful for serial evaluation of responses to shock and shock treatment.

Comparison of ventriculotomy closure with and without a coelomic fat patch in Japanese quail (Coturnix coturnix japonica).

Removal of foreign bodies from the ventriculus in birds may necessitate ventriculotomy. Complications with this intervention include leakage and adhesion formation. To investigate if the use of a coelomic fat patch and a tension-relieving suture in addition to a simple interrupted pattern would improve the healing process after ventriculotomy, 2 groups of 9 Japanese quail (Coturnix coturnix japonica) underwent ventriculotomy. In group 1, only simple-interrupted and tension-relieving sutures were used for closure of the ventriculotomy. In group 2, a coelomic fat patch from the surrounding adipose tissue was applied to the incision site in addition to the sutures. All quail recovered normally and were considered clinically healthy after surgery. Three birds from each group were euthanatized at days 7, 14, and 21 after surgery. On histologic examination, the suture techniques used for closure of the ventriculotomy led to minimal inflammation of the surrounding tissues in both groups. Serosal inflammation was significantly greater in group 2 birds that had the adipose patch at closure compared with group 1 birds. Therefore, the use of a coelomic fat patch to cover the site of ventriculotomy did not result in an improved healing process and its use is not recommended in quail.

Use of deferiprone for the treatment of hepatic iron storage disease in three hornbills.

CASE DESCRIPTION: 3 hornbills (2 Papua hornbills [Aceros plicatus] and 1 longtailed hornbill [Tockus albocristatus]) were evaluated because of general listlessness and loss of feather glossiness. CLINICAL FINDINGS: Because hepatic iron storage disease was suspected, liver biopsy was performed and formalin-fixed liver samples were submitted for histologic examination and quantitative image analysis (QIA). Additional frozen liver samples were submitted for chemical analysis. Birds also underwent magnetic resonance imaging (MRI) under general anesthesia for noninvasive measurement of liver iron content. Serum biochemical analysis and analysis of feed were also performed. Results of diagnostic testing indicated that all 3 hornbills were affected with hepatic iron storage disease. TREATMENT AND OUTCOME: The iron chelator deferiprone was administered (75 mg/kg [34.1 mg/lb], PO, once daily for 90 days). During the treatment period, liver biopsy samples were obtained at regular intervals for QIA and chemical analysis of the liver iron content and follow-up MRI was performed. In all 3 hornbills, a rapid and large decrease in liver iron content was observed. All 3 methods for quantifying the liver iron content were able to verify the decrease in liver iron content. CLINICAL RELEVANCE: Orally administered deferiprone was found to effectively reduce the liver iron content in these 3 hornbills with iron storage disease. All 3 methods used to monitor the liver iron content (QIA, chemical analysis of liver biopsy samples, and MRI) had similar results, indicating that all of these methods should be considered for the diagnosis of iron storage disease and monitoring of liver iron content during treatment.

Multiple papillomas in a diamond python, Morelia spilota spilota.

A 4-yr-old male diamond python (Morelia spilota spilota) was evaluated for multiple black papillated exophytic skin proliferations and signs of pneumonia. The histopathologic structure of the skin biopsy specimens led to the diagnosis of a benign papilloma-like neoplasia. In this case, papillomavirus DNA could be amplified from a biopsy sample with a broad range polymerase chain reaction. Nested pan-herpes polymerase chain reaction was negative, and herpesvirus inclusion bodies were not found. Because of the histologically benign nature of the papilloma, the skin proliferations were left untreated. Ten mo after the first presentation, the skin lesions had regressed almost completely; 34 mo later, only scars from the biopsies were left.

Evaluation of three miniplate systems for fracture stabilization in pigeons (Columba livia).

Bone plates are rarely used in avian fracture management for several reasons, and until recently, there was no plating system considered appropriate for use in birds with a body mass less than 500 g. To evaluate 3 different miniplate systems in avian fracture repair, 3 groups (A, B, and C) of 6 pigeons (Columba livia) each were used. The left ulna and radius of the pigeons were transected, and the ulna was stabilized. In group A, a 1.3-mm adaption plate was used. In group B, a limited contact system was created with washers that were placed between a 1.3-mm adaption plate and the bone. The intention was to reduce the compression of the periosteum and vascular damage to the bone. In group C, a 1.0-mm maxillofacial miniplate was used. Healing was evaluated with radiographs after 14 and 28 days. A flight test was conducted on day 28; the birds were then euthanatized, and the wing was dissected. Birds in group A with the adaptation plate achieved the best flight results (100%). In group B birds, no effect of the limited contact concept was visible at necropsy, and a high percentage of the screws had loosened, leading to failure (33%). The maxillofacial miniplates of group C birds were too weak and bent (100%). These results indicate that the adaption plate 1.3 met the desired requirements. To improve the system, further trials, with smaller drill bits and with screws having a smaller thread pitch, are recommended.

Bispectral index reveals death-feigning behavior in a red kite (Milvus milvus).

Red kites (Milvus milvus) are birds of prey known to feign death in the presence of humans. An adult wild red kite was anesthetized with isoflurane for coelioscopy. During surgery, heart rate and respiratory rate ranged from 240 to 260 beats per minute and from 16 to 28 breaths rates per minute, respectively. Pupil and corneal reflexes remained present, and body temperature was maintained at 40.4 degrees C (104.7 degrees F). Suppression ratio was 0 during the anesthetic episode. The bispectral index was 44 immediately after intubation, ranged from 44 to 57 during maintenance of anesthesia, and was 59 at the moment of extubation. The index increased to 85 while the kite remained immobile, which was suggestive of feigning death in sternal recumbency. Once the bird was perched upright, it immediately kept the upright position, which confirmed the correspondence of the bispectral index value (85) with a fully conscious patient. Although behavioral or cardiorespiratory variables remained unchanged, the degree of hypnosis was indicated by the bispectral index, which anticipated a possible sudden awakening episode of this bird.

Radiographic measurement of internal organs in Spix's macaws (Cyanopsitta spixii).

Radiology is an important diagnostic instrument in avian medicine, but standard measurement ranges for the objective evaluation of radiographs of birds are rare. To establish radiographic reference ranges for the critically endangered Spix's macaw (Cyanopsitta spixii), we measured radiographic silhouettes of the heart, liver, kidneys, spleen, proventriculus, and keel of the sternum on 29 radiographs taken under standardized conditions in adult and juvenile, clinically healthy birds. Ratios were determined for the proventricular diameter-to-keel height, the width of the heart to the width of the thorax, and for the "hourglass shape" (ratio of the width of the heart to the width of the liver). No significant differences were found between the sexes among the adult birds. Compared with adult birds, juvenile females had a significantly larger heart width (19.8 +/- 1.4 mm versus 21.2 +/- 0.7 mm), ratio of the heart width to the thorax width (0.86 +/- 0.08 versus 0.94 +/- 0.09), and horizontal width of the spleen (7.7 +/- 0.6 mm versus 8.5 +/- 0.4 mm). Results of radiographic measurements in the Spix's macaws were comparable to those published from other psittacine species. These reference ranges will facilitate a more objective radiographic evaluation of captive Spix's macaws.

Clinical course of a malignant peripheral nerve sheath tumor in a Siberian tiger (Panthera tigris altaica).

A 14-year-old male Siberian tiger (Panthera tigris altaica) was admitted with an ulcerating mass on the right thoracic wall. Radiographic and computed tomographic evaluation indicated 2 isolated cutaneous masses without any signs of metastasis. Histology of a Tru-Cut biopsy revealed an anaplastic sarcoma with giant cells. Both tumors were resected with appropriate normal tissue margins. The size of the defect did not allow primary closure of the wound; therefore, a mesh expansion technique was attempted. Three months later, the tiger had to be euthanized due to extensive metastasis to the lungs. Histomorphological features and immunohistochemical results confirmed the diagnosis of malignant peripheral nerve sheath tumor. In contrast to domestic animal experience, the tumor had spread extensively to the lungs without local reccurrence in a short period of time. Correct diagnosis requires various immunohistochemical evaluations of the tumor tissue.

Analysis of exhaled breath condensate in a mixed population of psittacine birds.

Collection of exhaled breath condensate (EBC) and the measurement of inflammatory markers contained therein (eg, hydrogen peroxide [H2O2], leukotriene B4 [LTB4], and pH) have been reported to be noninvasive tools for the investigation of respiratory disease in various species. In this study, the EBC of clinically healthy psittacine birds (n = 15) and psittacine birds with respiratory tract disease (n = 19) was examined, and inflammatory markers contained in the EBC were analyzed and compared. Awake birds were placed in an acrylic container from which the outflow passed through a condensation system that collected the EBC. All samples were analyzed for pH, H2O2, and LTB4. The mean values for each of these components, as well as the mean volume of the total EBC, measured from the apparently healthy birds did not differ significantly from those measured in birds with signs of respiratory tract disease. However, LTB4 in the EBC of diseased birds was higher than that of the apparently healthy birds and showed a trend toward significance. The study demonstrated the establishment of a standardized method for collecting and analyzing EBC in psittacine birds and a measurement protocol for pH, H2O2, and LTB4.

Bilateral vision loss in a captive cheetah (Acinonyx jubatus).

The following case report describes a 1-year-old female cheetah (Acinonyx jubatus) with bilateral blindness and unresponsive pupils. For comparison, a second healthy 2.5-year-old male cheetah without visual deficits was also examined. Clinical examination of both animals included biomicroscopy, indirect ophthalmoscopy, tonometry, and electroretinography. The young female cheetah showed no menace response, no direct or indirect pupillary light reflex, and no dazzle reflex in either eye. Fundus lesions, as detected by indirect ophthalmoscopy, are described for the female animal. In both eyes, the fundus color was green/turquoise/yellow with multiple hyperpigmented linear lesions in the tapetal area around the optic nerve. The optic nerve head was dark gray and about half the normal size suggesting bilateral optic nerve hypoplasia and retinal dysplasia or differentially optic nerve atrophy and chorioretinal scarring. The ERG had low amplitudes in the right eye but appeared normal in the left eye compared with the male cheetah. Blood levels did not suggest current taurine deficiency. This is addressed to some degree in the discussion. Bilateral optic nerve hypoplasia or optic nerve atrophy is a rare anomaly in cats and has not yet been described in a cheetah.