CHOLELITHIASIS IN CAPTIVE MOUNTAIN CHICKEN FROGS (LEPTODACTYLUS FALLAX): DIAGNOSTIC IMAGING AND HISTOPATHOLOGICAL FEATURES.

The mountain chicken frog (Leptodactylus fallax) is a critically endangered species highly reliant on ex situ conservation programs. In captivity, cholelithiasis has been reported during clinical examinations using diagnostic imaging and observed during postmortem examinations. Some individuals have presented with nonspecific clinical signs potentially associated with this condition, but little is known about its clinical relevance. A retrospective evaluation of ultrasonographic (n = 139) and radiographic (n = 156) images and histopathology samples (n = 32) obtained between 2014 and 2020 from 133 individuals was performed. An ultrasound scoring system was developed to evaluate changes in the gallbladder, with 38% (53/139) of the images showing abnormal contents. Gallbladder distension appeared associated with these findings (generalized estimating equation model, P < 0.001). In contrast, radiography proved to have low overall sensitivity (48%) when compared to ultrasonography for identifying abnormal gallbladder contents. A weak level of agreement (Cohen's kappa = 0.535) was observed between the imaging modalities. Histologically, cholecystitis was present in 86% (12/14) of the individuals, thickening of the gallbladder wall in 71% (10/14), and intrahepatic cholangitis in 90% (18/20). There was no evidence of cholestasis in any of the affected individuals. A significant difference was observed in the hepatic melanomacrophage density between animals with and without cholelithiasis (Welch two-sample t test, P < 0.001). This study provides an overview on the use of diagnostic imaging modalities to evaluate cholelithiasis and proposes a convenient ultrasound score to standardize monitoring of the condition in L. fallax. It is also the first systematic review of histopathological changes associated with cholelithiasis in this species.

Diagnosis and Treatment of Heavy Metal Toxicosis in Six Waldrapp Ibis (Geronticus eremita).

Six adult Waldrapp ibis (Geronticus eremita) were presented with heavy metal toxicosis secondary to paint chip ingestion after being placed in an indoor confinement enclosure because of a regional highly pathogenic avian influenza epornitic and government restrictions. Four of the ibis developed clinical disease signs between 45 and 64 days of confinement, including weakness, lethargy, inability to fly, low body condition, and bilateral wing droop while 2 birds appeared to remain normal during that period. Common clinicopathological findings included elevated plasma aspartate aminotransferase, creatine kinase, glutamate dehydrogenase, and bile acids values. Erythrocyte morphological changes included prominent hypochromasia, decreased size of polychromatophils with occasional cytoplasmic stippling, abnormal shapes, and D cells. Whole-body radiographic imaging revealed particulate radio-opaque material in the ventriculus of all affected birds. One bird died before chelation therapy could be instituted and heavy metal testing of the liver revealed increased concentrations of lead and zinc. Lead toxicosis was diagnosed in the 5 remaining birds by plasma lead analysis. These 5 birds were hospitalized for an average of 40 days and treated with sodium calcium edetate and fluid therapy, which resulted in resolution of the toxicosis. Plasma biochemistry, complete blood counts, and blood lead values were performed repeatedly on all 5 birds and were used to guide therapy and monitor treatment response. This case series emphasizes the importance of early contingency planning and reviews the treatment of lead toxicosis in birds with a focus on long-term clinical monitoring and hematology.

FERLAVIRUS-RELATED DEATHS IN A COLLECTION OF VIPERID SNAKES.

Between June and October 2013, 26 snakes of six viperid species kept in two adjoining rooms died ( n = 16) or were euthanized on medical (1) or welfare grounds (9). Two were from the main zoo collection, but the other 24 had been imported and quarantined for a minimum of 6 mo. Four of those that died and the single snake euthanized on medical grounds showed minor signs of respiratory disease prior to death, and five were weak, lethargic, and/or poor feeders. Frequent postmortem findings among all snakes were poor body condition (18) and respiratory disease (13). Seventeen cases were examined histologically, and pneumonia, sometimes with air sacculitis and/or tracheitis, was present in 15 individuals. Lung samples from 24 snakes were ferlavirus polymerase chain reaction (PCR) positive, and one of the two snakes for which only liver was available was also positive. The negative liver sample was from a snake that died of sepsis following anesthesia for surgical removal of a spindle cell sarcoma. Correlation with antemortem PCR testing of glottal and cloacal swabs in five cases was poor (sensitivity = 40%). Immunohistochemistry (IHC) for ferlaviruses on the tissues of 13 PCR-positive cases showed positive labeling in 7 only. Tissues samples from 22 ferlavirus PCR-positive snakes were examined for Chlamydia species by PCR, and 9 were positive, although DNA sequencing only confirmed two of three tested as Chlamydia pneumoniae. Immunohistochemistry for Chlamydia pneumoniae of seven cases (two Chlamydiales PCR positive, one of which was sequenced as C. pneumoniae, plus five negative) confirmed the Chlamydia PCR results. These two Chlamydiales PCR and IHC positive snakes were ferlavirus PCR positive, but IHC negative suggesting that, even though a ferlavirus was the predominant cause of the outbreak, in a few cases death may have been due to chlamydiosis with ferlavirus present, but not acting as the primary pathogen.

Analysis of biochemical markers of bone metabolism in Asian elephants (Elephas maximus).

Two human enzyme immunoassays (EIA) and one radioimmunoassay (RIA) were validated and used to measure osteocalcin (OC), bone alkaline phosphatase (BAP), and the cross-linked telopeptide domain of type I collagen (ICTP), in serum from Asian elephants (Elephas maximus). Sera from four adult females sampled on 7 consecutive days were also analyzed to assess the existence and magnitude of intraindividual day-to-day variability of the serum concentration of these markers. Sample dilution curves were parallel with assay standard curves, which demonstrated that excellent cross reactivity existed between assay antibodies and elephants marker antigens. Statistically significant inverse correlations were found between age and concentrations of all three markers: BAP, r = -0.862 (P < 0.01); OC, r = -0.788 (P < 0.002); and ICTP, r = -0.848 (P < 0.01). Strong positive correlations were found between BAP and OC (r = 0.797, P < 0.01), OC and ICTP (r = 0.860, P < 0.01), and between BAP and ICTP (r = 0.958, P < 0.01). No statistically significant intraindividual variability was found over 7 days in the four adult females for any of the markers assessed (OC: P = 0.089; ICTP: P = 0.642; BAP: P = 0.146; n=4 in each case). The overall coefficient of variability observed in this group of animals was 10.3%, 7.4%, and 5.5% for OC, BAP, and ICTP, respectively. These results suggest a potential role for biochemical markers of bone turnover in monitoring skeletal health and bone disease in Asian elephants.