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Granulomatous amoebic encephalitis caused by the free-living amoeba Balamuthia mandrillaris is a highly fatal disease that was first isolated from a mandrill (Mandrillus sphinx), and has since been diagnosed in several nonhuman primates including orangutans. Indirect immunofluorescence antibody (IFA) techniques for Balamuthia have been used in the fields of human medicine and epidemiology both for exposure assessment and screening of clinical patients for antemortem diagnosis. Stored serum samples from five captive Northwest Bornean orangutans (Pongo pygmaeus pygmaeus), including one who had died from B. mandrillaris infection, housed at a single facility were screened with a human IFA assay for B. mandrillaris. Only the single, clinically affected individual was seropositive, and the results suggest that the use of the available human B. mandrillaris IFA assay is a novel diagnostic option for detection of Balamuthia antibodies in this species. A validated screening serological test could be used in individuals exhibiting signs consistent with granulomatous amoebic encephalitis to facilitate earlier antemortem diagnosis of Balamuthia infection, which is critical if treatment is to be pursued. This pilot study presents the use of serological detection methods for B. mandrillaris screening in a nonhuman primate. Subsequent use of the B. mandrillaris IFA assay in the larger captive population should be pursued for validation of the test and to provide further information on seroprevalence and evaluation of risk factors for exposure to Balamuthia and subsequent development of disease.

Granulomatous amoebic encephalitis caused by the free-living amoeba

A female Bornean orangutan (Pongo pygmaeus) aged 11 years and 6 months was examined by veterinarians after caretakers observed lethargy and facial grimacing. Within 72 h the primate had left-sided hemiparesis that worsened over the next week. An MRI revealed a focal right-sided cerebral mass suspected to be a neoplasm. Ten days after onset of clinical signs, the orangutan died. On postmortem exam, the medial right parietal lobe was replaced by a 7 × 4 × 3.5 cm focus of neuromalacia and hemorrhage that displaced the lateral ventricle and abutted the corpus callosum. Histopathology of the cerebral lesion revealed pyogranulomatous meningoencephalitis with intralesional amoeba trophozoites and rare cysts. Fresh parietal lobe was submitted to the Centers for Disease Control and Prevention lab for multiplex free-living amoebae real-time PCR and detected Balamuthia mandrillaris DNA at a high burden. Mitochondrial DNA was sequenced, and a 760-bp locus 19443F/20251R was compared to several human infections of B. mandrillaris and shown to be identical to the isolates from four human cases of encephalitis: 1998 in Australia, 1999 in California, 2000 in New York, and 2010 in Arizona. Indirect immunofluorescent antibody testing of stored serum samples indicated exposure to B. mandrillaris for at least 2 years prior to death. Within 1 week of the orangutan’s death, water from the exhibit was analyzed and identified the presence of B. mandrillaris DNA, elucidating a possible source of exposure. B. mandrillaris, first reported in a mandrill in 1986, has since occurred in humans and animals and is now considered an important emerging pathogen.

The objective of this study was to obtain an estimate of the pharmacokinetic parameters of moxidectin administered at a dosage of 1 mg/kg orally and topically to healthy adult giraffe (Giraffa camelopardalis). The maximum plasma concentration (CMAX) of moxidectin after oral and topical administration was 69.2 ± 4.6 and 18.6 ± 16.1 ng/ml (P = 0.045), respectively. The areas under the plasma curve (AUC), a measure of total drug exposure, was 532.0 ± 232.3 and 209.1 ± 180.0 day*ng/ml (P = 0.308) for the oral and topical administrations, respectively. These data suggest moxidectin achieves higher peak plasma concentrations following oral administration compared with topical (transdermal) administration using the cattle pour-on formulation. Additionally, the percent coefficient of variation, a measure of variability, was smaller for the oral formulation (CMAX %CV = 7%; AUC %CV = 44%) compared with the topical formulation (CMAX %CV = 86%; AUC %CV = 86%). The smaller variability suggests that oral admin...

New biochemically-derived products for the removal of paraffin wax from oil wells do not require additional capex nor heat and do not utilize bacteria. They contain inactivated microbial cells, biosurfactants and biosolvents, and other components harvested as microbial byproducts that emulsify and dissolve paraffin from rock pores and from the well surfaces over wide temperature, salinity, depth, and pH ranges. Additionally, they increase oil recovery by remediating near-wellbore formation damage, reducing interfacial tension, altering rock surfaces and changing their wettability, and reducing oil viscosity. The product application is environmentally superior to well treatments using hot oil/water and aromatic solvents and is economical due to low capital and operating costs required for product synthesis. Specifically, product preparation is achieved using a modular fermentation system that is installed near the points of application. This insures highly efficient and low-cost production and logistics, as well as reducing time from generation to application which maximizes potency. With sufficient space, water, and electricity, the initial manufacture of the dispersal products can occur within a few weeks. The treatment products utilized were initially developed and tested in laboratory studies, which showed that dispersion rates of the relevant paraffin samples were comparable to those achieved with toluene. The paraffin dispersal products exhibit a very high level of efficacy and safety when deployed in the Appalachian and Permian Basins. The potency of these products has led to outstanding paraffin removal results as indicated by reduced well failures in both vertical and horizontal wells and by visual observation of sucker rods removed from the wells. In addition, tank sludge and wax deposits in pipelines can be removed through either residual product flowing from the well or through direct application. Growth of detrimental bacteria and formation of biofilms are inhibited by the product application thereby reducing corrosion risk. Specifically, details of an almost two-year 70-well study in the Appalachian Basin are reported in which no well failures were observed due to paraffin buildup and 95% of the wells exhibited an enhanced oil recovery effect during the paraffin remediation treatments. This resulted in an approximate 50% average increase in sustained production rate over baseline. Analysis of the results forecasts a substantial increase in future production, thereby significantly enhancing the value of the producing wells. Importantly, longer times between required treatments and the increased recovery rates have transformed the paraffin maintenance program into a documented revenue generator for the operator.

Baseline information about the reproductive physiology of an endangered species is vital to captive breeding programs. This study analyzed reproductive parameters from eleven captive Chacoan peccaries (Catagonus wagneri) (mean age: 10.5 ± 1.2 years old) in Paraguay. After immobilization, testis length, width and firmness (hard to flaccid, 1–3) were assessed, followed by electroejaculation and analysis of semen. Samples were examined for percentage and progressive motility, total cell count, percentage live spermatozoa and morphology. Mean (±SEM) testis volume and firmness were 24.7 ± 1.8 cm3 and 2.1 ± 0.1, respectively. Mean ejaculate volume was 2.9 ± 0.7 ml with a pH of 7.7 ± 0.3. Few male peccaries had motile spermatozoa (n = 5/9) with a mean percentage and progressive motility of 18.3 ± 8.5% and 0.6 ± 0.3, respectively. The mean percentage of live spermatozoa was 25.1 ± 5.6%. Male peccaries had a low percentage of normal spermatozoa (12.4 ± 2.5%). The mean total count of spermatozoa per ejaculate was also quite low at 1.58 ± 1.01 million total spermatozoa per ejaculate. Spermatozoa defects were predominantly primary (77.7%) with the most common spermatozoa defects being tapered head (19.0 ± 7.4%), diadem/crater (17.7 ± 2.8%), and excess residual cytoplasm (9.6 ± 2.5%). Male age was not correlated with semen parameters (percent live: r=-0.19; motility percentage: r = 0.01; percent normal spermatozoa: r = 0.38; total count: r = 0.29; p>0.05). Evaluation of additional males from this population as well as other captive populations at various time points during the year is warranted.

Sinusitis is a common finding in avian species; however, it presents unique treatment challenges in ratites. A 6-yr-old male greater rhea (Rhea americana) presented acutely with right infraorbital sinus swelling and chemosis. Oral antibiotics were prescribed based on bacterial culture results with incomplete resolution of clinical signs. A computed tomography scan of the skull demonstrated a mixed-density soft tissue mass with extensive destruction of the right nasal bone, hard palate, maxilla, and frontal bone. The affected tissue was surgically resected and histopathology was consistent with inspissated necrotic tissue. Aerobic culture of the mass grew a multidrug-resistant Escherichia coli and was treated with amikacin. Ten months postsurgery, no further clinical signs were observed. Advanced diagnostic imaging with computed tomography and surgical therapy facilitated complete resolution of this resistant bacterial sinusitis in a species that is not typically amenable to frequent handling and ...

Infection with the fungal organism Batrachochytrium dendrobatidis was documented in two captive populations of aquatic caecilians (Typhlonectes natans), including 24 confiscated animals at the Bronx Zoo's Wildlife Health Center and two captive-born animals at the Phoenix Zoo. The animals at the Bronx Zoo were asymptomatic, and infection was discovered during quarantine polymerase chain reaction screening. Both animals at the Phoenix Zoo were clinically ill, and Batrachochytrium dendrobatidis infection was confirmed in both animals, but it is unclear what role, if any, the infection played in the animals' clinical signs. All of the Bronx Zoo's caecilians were successfully cleared of infection by elevating to and then holding water temperatures at 32.2 degrees C (90 degrees F) for 72 hr. One animal at the Phoenix Zoo died before treatment could be instituted, and the second died despite treatment with a 0.01% itraconazole bath. Chytridiomycosis has only been very recently first reported in caecilians, and much remains unknown about its behavior in this amphibian order. This is the first published report of Batrachochytrium dendrobatidis infection in captive-born caecilians and also the first describing details of treatment of chytridiomycosis in caecilians. It appears that raising tank temperature to 32.2 degrees C for 72 hr is a safe and effective treatment for aquatic caecilians with chytridiomycosis.

An 8-yr-old male buff-cheeked gibbon (Nomascus gabriellae) acutely developed abnormal behavior, decreased appetite, and dull mentation. Mild generalized muscle wasting and weight loss were the only other abnormalities noted on examination. Routine immunodiffusion serology for Coccidioides spp. were IgG and IgM positive. Magnetic resonance imaging of the brain was suggestive of an infectious meningoencephalitis with secondary obstructive hydrocephalus. A ventriculoperitoneal shunt was placed in standard fashion to reduce the imminent risk of mortality from increased intracranial pressure. Postoperative treatment included oral fluconazole, a tapered course of prednisolone, and physical therapy. Clinical signs improved steadily and the gibbon was fit to return to exhibit 8 wk post-shunt placement. This case of coccidioidomycosis demonstrates the complications that can occur with dissemination to the central nervous system and its management. It is the first published report describing the use of ventriculoperitoneal shunt placement in this species.

Parasitism is common in wild and captive amphibians; however, pharmacologic data are lacking for anthelmintic drugs. This study was developed to determine the plasma pharmacokinetics of selamectin after topical administration in bullfrogs. Thirty-two adult American bullfrogs (Rana catesbeiana) were randomly assigned into eight groups of four with each group representing a different collection time point. Seven groups received selamectin (6 mg/ kg) topically and the remaining group served as the untreated control group. One group of frogs was euthanized and blood samples immediately collected on days 0 (control), 1, 5, 10, 15, 20, 25, and 30. Plasma was analyzed for selamectin using high performance liquid chromatography with fluorescence detection. Individual samples were analyzed, then data were reported as the mean of the four frogs at each time point. A histologic evaluation of the lung, liver, kidney, and skin tissues was performed and none of the frogs showed histologic evidence of toxicity due to selamectin administration. The mean peak plasma concentration was 162.5 +/- 42.3 ng/ml, area under the curve was 2,856 ng day/ml, mean residence time was 12.2 days, and disappearance half-life was 1.87 days. Based on the plasma pharmacokinetics, bullfrogs appear to absorb selamectin very efficiently, concentrations reach high levels in the plasma, and there were no apparent histologic effects from single dose administration.

Many species of birds kept in captivity must be rendered flightless to prevent escape from open enclosures. In this study, we evaluated the use of diode laser and cryosurgery as methods of ablating primary feather follicles in domestic pigeons (Columba livia). In group 1 birds (n = 6), primary feather 10 of both wings was treated with the diode laser (10 W, 2 seconds) and primary feather 6 was treated with the cryoprobe (5 seconds, 3 cycles). In group 2 birds (n = 6), primary feathers 10 and 6 were treated with the cryoprobe for 20 and 30 seconds, respectively, for 3 cycles. In all birds, primary feather 8 on both wings was manually pulled as the control follicle. Results showed that in group 1 birds, diode laser ablation prevented feather regrowth in 83% of follicles, however, freezing with the cryoprobe for 5 seconds did not prevent feather regrowth. In group 2 birds, treatment with the cryoprobe prevented feather regrowth at 100% and 42% of the treated sites (treatment times of 20 and 30 s...

A 16-yr-old, female, captive-bred bontebok (Damaliscus pygargus) estimated to be 75 kg, was presented with a 1-mo history of chronic diarrhea and weight loss. A cause for the deteriorating condition was not found antemortem, and the bontebok was humanely euthanatized when it failed to respond to empirical therapy. On necropsy, numerous nodules were present on the pleural surfaces and within the lung parenchyma, the pancreas appeared inflamed and fibrotic. Histologic examination revealed lymphoma in the mesenteric lymph nodes, alimentary tract, reproductive tract, and liver. In the small and large intestine, rumen, esophagus, bile duct, and vagina, the neoplastic cells had a tropism for epithelium. The neoplastic cells within the epithelium and subepithelium of the alimentary tract stained positively for CD3, consistent with T cell differentiation.

An eight year old, wild-caught, male rough-necked monitor, Varanas rudicollis, was examined for hind limb partial paresis. Clinical pathology revealed a marked heterophilic leukocytosis. Spondylosis of several thoracolumbar vertebral bodies was present on spinal radiographs and computed tomography (CT) scan. A blood culture yielded a pure growth of Clostridium perfringens while an aspirate from the affected vertebrae yielded a pure growth of Clostridium sordelii. Long-term antibiotic therapy successfully reduced the leukocyte count within normal range however the vertebral lesions remained unchanged and the leukocytosis returned with discontinuation of antibiotics. Radiographic and CT appearance of the vertebral lesions suggest chronic inflammation from a persistent Clostridium sp. infection.

Captive refugia for four species of plethodontid salamanders were established. Following this successful maintenance, two of the species presented with kyphosis. In one of these species (San Marcos salamander, Eurycea nana), microsporidial organisms within the epaxial musculature were identified in post-mortem histopathology and confirmed by polymerase chain reaction rDNA sequence as most closely similar to Encephalitozoon helium. However, ultrastructural analysis and further rDNA sequencing suggested a new species of Pleistophora. In these affected individuals, the microsporidial infection produced kyphosis resulting from inflammation and fibrosis of the epaxial musculature to the parasite.

The results of long-term administration of leuprolide acetate (LA) depot in a 52-yr-old Asian elephant bull (Elephas maximus) for control of musth are presented. Twelve injections were administered for 6 yr during our interpretation of early musth or "premusth." Intervals between musth periods during the study varied from 2 to 34 mo. Blood samples, drawn weekly, were assayed for serum testosterone concentrations; mean levels were 11.78 +/- 1.97 nmol/L throughout the first 26 mo of the study, 7.28 +/- 1.28 nmol/L during the following 21 mo, and 0.45 +/- 0.035 nmol/L in the last 34 mo of this study. Early musth signs ceased within 3 days of drug administration after 10 of 12 injections. The mean serum testosterone concentrations were significantly decreased by the last 34 mo of the study. The results suggest leuprolide is a suitable alternative for controlling or preventing (or both) musth in captive Asian elephants, although permanent reproductive effects may occur. Zoos and wildlife conservation institutions could benefit from the use of LA in Asian elephants to increase the male availability in captivity, consequently ensuring genetic diversity and the perpetuation of the species.

Necropsy records for 102 Attwater's prairie chickens (Tympanuchus cupido attwateri) that had been captive-reared and had died at the San Antonio Zoo over a 4-yr period were evaluated to determine the causes of mortality. Gross necropsy, histopathology, and bacterial culture findings are summarized. Mortality was highest in young birds (

An adult female, wild-caught red coachwhip snake (Masticophis flagellum piceus) was euthanized at the Phoenix Zoo due to severe neurologic signs. Necropsy and histopathology revealed an invasive liposarcoma of the vertebral column, which likely caused the neurologic signs. Histology of the small intestine revealed a granuloma with intralesional yeasts morphologically compatible with the genus Coccidioides. The diagnosis of coccidioidomycosis was confirmed with immunohistochemistry staining. Coccidioides posadasii is endemic to Arizona and is an important cause of disseminated fungal infections in mammals in this region. This is the first known report of intestinal coccidioidomycosis in a veterinary species and the second report of coccidioidomycosis in a reptile.

An adult male Boelen's python, Morelia boeleni, presented with acute neurologic disease and was euthanatized. Histologic examination revealed nonsuppurative meningoencephalitis. Occasional eosinophilic intracytoplasmic inclusions were noted in glial cells. On the basis of clinical signs and histopathology, inclusion body disease of boid snakes was suspected, but inclusions were not seen in other organs commonly affected with the disease. Moreover, electron microscopy revealed that the inclusions contained stacks of filaments 13–14 nm wide. With the use of a generic paramyxovirus cDNA probe, sections of brain and esophageal ganglion demonstrated hybridization. The findings indicate that paramyxovirus was the likely cause of the encephalomyelitis in this python, and this virus should be included in the differential diagnosis of pythons exhibiting central nervous system disease.

The deaths of two Asian elephants (Elephas maximus) in August 1996 led the United States Department of Agriculture to require the testing and treatment of elephants for tuberculosis. From August 1996 to September 1999. Mycobacterium tuberculosis infection was confirmed by culture in 12 of 118 elephants in six herds. Eight diagnoses were made antemortem on the basis of isolation of M. tuberculosis by culture of trunk wash samples; the remainder (including the initial two) were diagnosed postmortem. We present the case histories, epidemiologic characteristics, diagnostic test results, and therapeutic plans from these six herds. The intradermal tuberculin test, enzyme-linked immunosorbent assay serology, the blood tuberculosis test, and nucleic acid amplification and culture are compared as methods to diagnose M. tuberculosis infection in elephants.