THE DIAGNOSIS AND EPIDEMIOLOGY OF AN INFERTILITY DISEASE IN THE FEMALE KOALA

A radiographic technique, incorporating pneumoperitoneum, was developed to aid in identification of cyst-like structures in the reproductive tract of female koalas. These lesions, including pyometra and fluid-filled cysts associated with the upper reproductive tract, were viewed as radiopacities with clearly demarcated margins lateral to the caudal lumbar vertebrae. This technique provided a means of assessing with a high degree of reliability the incidence and distribution of this condition in various populations of koalas throughout eastern and southern Australia. A radiographic survey of 237 adult female koalas revealed a 43% (101/237) incidence of this condition, which is closely correlated with the lack of reproductive success observed in some populations of koalas in the wild. Although the etiology of this condition is little understood at present, the isolation of Chlamydia psittaci from the reproductive tracts of affected koalas, both male and female, is recorded.     

INTRAVENOUS UROGRAPHY IN THE KOALA (Phascolarctos cinereus)

Urograms were performed on 16 adult koalas to evaluate this procedure for use in the koala. Because there is only small amount of intraabdominal fat, radiographs of the abdomen of koalas lack detail. Contrast medium was excreted more slowly in the koala than in the dog and there was considerable variation in the rate of excretion between koalas. The results of this study indicated that urography was valuable technique for assessing renal anatomy, but there was considerable variation in the time for excretion of the contrast medium.     

Plasma concentrations of chloramphenicol after subcutaneous administration to koalas (Phascolarctos cinereus) with chlamydiosis

Nine mature koalas with chlamydiosis, typically keratoconjunctivitis and/or urogenital tract infection, were treated with daily subcutaneous injections of chloramphenicol at 60 mg/kg for 45 days (five koalas), or for a shorter duration (four koalas). All koalas were initially positive for Chlamydia pecorum as determined by real-time polymerase chain reaction (qPCR). Plasma chloramphenicol concentrations were determined at t = 0, 1, 2, 4, 8, and 24 h after the day 1 injection (nine koalas) and after the day 15 injection (seven koalas). Chloramphenicol reached a median (and range) maximum plasma concentration of 3.03 (1.32-5.03 μg/mL) at 4 (1-8 h) after the day 1 injection and 4.82 (1.97-27.55 μg/mL) at 1 (1-2 h) after day 15. The median (and range) of AUC(0-24) on day 1 and day 15 were 48.14 (22.37-81.14 μg·h/mL) and 50.83 (28.43-123.99 μg·h/mL), respectively. The area under the moment curve (AUMC)(0-24) median (and range) for day 1 and day 15 were 530.03 (233.05-798.97 h) and 458.15 (291.72-1093.58 h), respectively. Swabs were positive for chlamydial DNA pretreatment, and all koalas except one, produced swabs negative for chlamydial DNA during treatment and which remained so, for 2-63 days after treatment, however whether chloramphenicol treatment prevented long-term recrudescence of infection was not established. At this dose and dosing frequency, chloramphenicol appeared to control mild chlamydial infection and prevent shedding, but severe urogenital disease did not appear to respond to chloramphenicol at this dosage regime. For koalas affected by severe chlamydiosis, antibiotics alone are not sufficient to effect a cure, possibly because of structural or metabolic changes associated with chronic disease and inflammation.     

Schirmer tear tests and intraocular pressures in conscious and anesthetized koalas (Phascolarctus cinereus)

Objective To estimate mean Schirmer tear test (STT) and intraocular pressure (IOP) values in healthy koalas both conscious and anesthetized. Methods Data were gathered from koalas in Victoria, Australia. Conscious examinations were performed on captive koalas. Free‐ranging (wild) koalas were examined under anesthesia. Anesthesia was induced using alfaxalone, and animals were maintained on oxygen and isoflurane if required. All animals were healthy and had no surface ocular pathology detectable during slit lamp biomicroscopy. STT I tests were performed using commercial STT test strips placed in the lower fornix for 1 min. IOP was measured using an applanation tonometer after topical anesthesia. The higher value of the two eyes for both STT and IOP was analyzed. STT was measured in 53 koalas (34 conscious, 19 anesthetized) and IOP was measured in 43 koalas (30 conscious, 13 anesthetized). A two‐sample t‐test was used to compare means. A P‐value <0.05 was regarded as significant. Mean ± SD is presented. Results The mean higher STT in conscious koalas was 10.3 ± 3.6 mm wetting/min and in anesthetized koalas it decreased to 3.8 ± 4.0 mm wetting/min (P < 0.0001). The mean higher IOP in conscious koalas was 15.3 ± 5.1 mmHg, and in anesthetized koalas it was 13.8 ± 3.4 mmHg (P = 0.32). There was no effect of sex on either STT or IOP. Conclusions The mean and SD of STT and IOP values for koalas both conscious and anesthetized were reported. The mean STT was significantly reduced by alfaxalone anesthesia.     

Histological survey for oxalate nephrosis in Victorian koalas (Phascolarctos cinereus)

The Mount Lofty Ranges koala (Phascolarctos cinereus) population in South Australia has a high prevalence of the renal disease oxalate nephrosis, for which an underlying genetic cause is suspected. South Australian koalas primarily originate from those in French Island, Victoria; however, oxalate nephrosis has not previously been reported in Victorian koalas. Examination of kidney tissue sections from 63 koalas across Victoria found that nine koalas were affected by oxalate nephrosis (14.3%). These included 2/5 koalas from French Island (40%), 4/14 koalas from the western regions (29%), 2/11 Raymond Island koalas (18%), and 1/13 Cape Otway koalas (8%). There were no cases of oxalate nephrosis identified in the Strzelecki koalas (n = 12). These findings suggest that oxalate nephrosis occurs in koalas from French Island and populations that have received significant influx of koalas from French Island, but not in the Strzelecki region, which has little to no French Island input. This lends support to the theory that an inherited abnormality of oxalate metabolism could underlie the high prevalence of oxalate nephrosis in the Mount Lofty Ranges koala population, and molecular investigations are currently underway to investigate a genetic cause.     

Haematological and biochemical investigations of diseased koalas (Phascolarctos cinereus)

Haematological and biochemical investigations were performed on 14 koalas with uncomplicated cystitis, 8 with complicated cystitis, 8 with conjunctivitis, 8 with lymphosarcoma, and 14 with miscellaneous diseases. Changes were limited and inconsistent in individual koalas with uncomplicated cystitis and conjunctivitis. In contrast, individual koalas with complicated cystitis were more likely to have anaemia, leukocytosis due to neutrophilia, hypoproteinaemia due to hypoalbuminaemia, and azotaemia due to elevated urea concentration. Although these changes were non-specific they did allow assessment of prognosis for survival and response to treatment. Koalas with lymphosarcoma were invariably anaemic, leukaemic, azotaemic and hypoalbuminaemic. Elevated enzymes (aspartate transaminase [AST]. lactate dehydrogenase [LD] and gamma glutamyl transferase [GGT]) were more common in koalas with lymphosarcoma. Koalas affected by miscellaneous conditions showed variable changes but once again anaemia, leukocytosis, azotaemia, elevated AST and LD, and hypoalbuminaemia were not uncommon. On the basis of these findings a minimal profile is suggested for the investigation of sick koalas and would include haematocrit, total and differential leukocyte counts, urea, total protein and albumin concentrations and AST, GGT and LD activities.     

PNEUMONIA ASSOCIATED WITH BORDETELLA BRONCHISEPTICA IN CAPTIVE KOALAS

Bordetella bronchiseptica was isolated from 12 cases of purulent bronchopneumonia from a captive koala colony near Brisbane, Queensland. An initial epizootic in March 1967 causing 13% mortality was followed by annual outbreaks causing 2% mortality usually in newly weaned and aged koalas in late winter to early spring. High population density and a low plane of nutrition in winter were thought to predispose to the occurrence of the disease.     

Symmetric dimethylarginine values in koalas (Phascolarctos cinereus) based on oxalate nephrosis status

Oxalate nephrosis is a prevalent renal disease in koalas (Phascolarctos cinereus) of the Mount Lofty Ranges population in South Australia. The symmetric dimethylarginine (SDMA) assay is widely used in companion animals to diagnose renal disease, particularly in the early stages. This study aimed to determine: (1) reference intervals for SDMA in koalas and (2) SDMA values of koalas with oxalate nephrosis. Blood samples were collected from 41 Mount Lofty Ranges koalas euthanased on welfare grounds. Koalas were necropsied and, based on renal histopathology, were classified as unaffected (n = 22) or affected (n = 19) by oxalate nephrosis. Serum or plasma samples were analysed for creatinine, urea and SDMA and urine samples for urine specific gravity (USG). The reference interval for SDMA in unaffected koalas was 2.4–22.9 μg/dL. In koalas with oxalate nephrosis, SDMA was elevated in 74% of cases above the upper limit of the confidence interval. SDMA was elevated in three affected koalas with normal creatinine values. A positive correlation was found between SDMA and creatinine (R = 0.775, P < 0.001) and SDMA and urea (R = 0.580, P < 0.001) and a negative correlation between SDMA and USG (R = −0.495, P = 0.027). In conclusion, SDMA correlates well with other commonly used tests of renal function in koalas and should be included as part of the standard diagnostic process to increase the accuracy of oxalate nephrosis diagnosis in koalas.     

Epizootiology of Chlamydia infections in two free-range koala populations

The prevalence of Chlamydia pecorum and Chlamydia pneumoniae infections in two free-range koala populations was assessed using genus-specific PCR combined with species-specific DNA probe hybridisation. Population A had a very high overall level of chlamydial infection (85%) with significantly more of these infections being due to C. pecorum (73%) compared to C. pneumoniae (24%). The second population had a much lower prevalence of infection (10%) with equal levels of both species. An important finding of this study was that. while five of 24 C. pecorum-infected koalas had clinical signs of the disease (both ocular and urogenital sites), none out of seven C. pneumoniae-infected koalas had signs of clinical disease. This suggests that C. pecorum may be the more pathogenic of the two chlamydial species infecting this host. The level of infection (assessed by intensity of the specific hybridisation signal) also differed between chlamydial species, with C. pecorum infections ranging from low to high grade whereas C. pneumoniae infections were always low grade. When the age of infected koalas was examined, 58% of young, sexually immature koalas were found to have C. pecorum infections, increasing to 100% of koalas in the older age groups. This suggests that, in this population at least, young koalas are readily infected with C. pecorum from their mothers. While the infection levels with C. pneumoniae were too low to be statistically significant, again, sexually immature koalas were found to be infected. The recent separation of chlamydial infections in koalas into two species is beginning to indicate different epizootiologies for koala C. pecorum compared to koala C. pneumoniae.     

Chlamydial infection in a colony of captive koalas

Forty three koalas in a captive colony were investigated for the presence of Chlamydia psittaci infection and associated disease. Swabs were taken from conjunctivae and urogenital sites for cell culture isolation of C psittaci and for cytological examination (direct smears) for chlamydial inclusions and evidence of inflammation. On the basis of cell culture isolation, 28 samples from 25 koalas were positive for C psittaci (that is, infected). Three koalas were positive from both sites, 5 from conjunctivae alone and 17 from urogenital sites alone. Seven of the 8 koalas with positive conjunctival swabs had overt signs of conjunctivitis, but only 3 of the 20 koalas with positive urogenital swabs had overt signs of 'wet bottom' (continual urine soiling due to cystitis) or purulent discharge. However, 5 of the 20 with positive urogenital swabs had past episodes of 'wet bottom'. Moreover, examination of direct cytological smears showed evidence of inflammation (neutrophils) in 7 of 8 koalas with positive conjunctival swabs and 17 of 20 with positive urogenital swabs. Chlamydial inclusions were rarely identified with surety on direct cytological smears. In the 18 koalas without chlamydia, one had overt conjunctivitis while 2 had past episodes of conjunctivitis. The koala with conjunctivitis at the time of sampling had a prior history of 'wet bottom'. Examination of direct cytological smears revealed 2 of the chlamydial negative koalas had high numbers of neutrophils in urogenital smears. It was concluded that C psittaci infection may cause overt or sub-clinical disease, with the former developing when the koalas were stressed through management procedures or concomitant disease.     

Use of quantitative real-time PCR to monitor the shedding and treatment of chlamydiae in the koala (Phascolarctos cinereus)

The aim of this study was to monitor chlamydial shedding patterns in clinically affected koalas before, during and following treatment using quantitative real-time PCR. Swab samples were obtained from 14 koalas presented for treatment at the Australian Wildlife Hospital. Four of these animals were followed over a period of 8–9 weeks. Primers were designed based on the consensus signature sequence of the 16S rRNA chlamydial gene. Additional primers were designed based on the sequence of the koala beta-actin gene and used to normalize chlamydial values when comparing results from different swab samples. Chlamydial 16S rRNA gene copy number was highest in swab samples from clinically affected sites. Daily injections of chloramphenicol resulted in a marked and rapid reduction in the numbers of chlamydiae being shed from all sites. In general, chlamydial copy number was no longer detectable by the end of the 2nd week of treatment. No evidence of relapse of infection was detected at 2 weeks after the cessation of treatment. In contrast, topical chloramphenicol treatment of the eyes required a longer treatment period and had little effect on the shedding of chlamydiae from other sites of the body. Further studies are required to confirm the efficacy of a shorter treatment period.     

Review of some pharmacokinetic and pharmacodynamic properties of anti‐infective medicines administered to the koala (Phascolarctos cinereus)

Although koalas are iconic Australian animals, no pharmacokinetic studies of any first‐line medicines used to treat diseased or injured koalas had been published prior to 2010. Traditionally, medicine dosages suggested for this species underwent linear extrapolation from those recommended for domesticated species. The koala, a specialist folivore whose natural diet consists of almost exclusively Eucalyptus spp. foliage has anatomical and physiological adaptations for detoxifying their diet which also affect medicine pharmacokinetic profiles. This review addresses aspects of medicine absorption, clearance, and other indices (such as medicine binding to plasma proteins) of enrofloxacin/marbofloxacin and chloramphenicol used for the systemic treatment of chlamydiosis, and fluconazole ± amphotericin, and posaconazole for the treatment of cryptococcosis. Based on observations from published studies, this review includes suggestions to improve therapeutic outcomes when administering medicines to diseased koalas.     

Hip dysplasia in koalas (Phascolarctos cinereus) at the San Diego Zoo.

A retrospective/prospective radiographic study documented 55 cases of moderate to severe hip dysplasia, with varying degrees of shallowing of the acetabulum, flattening or loss of the femoral head, widening or loss of the femoral neck, and femoral diaphyseal abnormalities in northern koalas (Phascolarctos cinereus) in the San Diego Zoo (San Diego, California, USA) collection. For the retrospective study, historic radiographs were examined when availble. For the prospective study, three standard views (ventrodorsal extended leg, ventrodorsal frog leg, and lateral extended leg) were used. A scoring system was developed using four areas (acetabulum, femoral head, femoral neck, and femur) and ranges of 0 to 5 (0 = not affected to 5 = severely affected) for each area, creating a total score out of 40. Scores were graded as follows: 0-2 = normal dysplasia; 3-6 = mild dysplasia; 10-19 = moderate dysplasia; and 20-40 = severe dysplasia. Thirty koalas were graded as severe, 25 koalas as moderate, and 38 koalas as excellent or mild. Affected koalas may or may not demonstrate gait abnormalities. Mild to severe degenerative joint disease may develop and symptoms may be alleviated with glucosamine/chondroitin sulfate and nonsteroidal anti-inflammatory drugs. The etiology of hip dysplasia in koalas is not currently understood.     

INTERNAL FIXATION OF FRACTURES OF THE ULNA IN THE HORSE

SUMMARY Surgical repair of ulnar fracture was attempted in 7 horses. Of 6 animals in which tension band plates were used, 4 recovered, one was killed after implant failure and one died of respiratory failure during post-operative recovery. The treatment of one horse with a Steinman Pin and figure-8 tension band wire was unsuccessful. One horse with bilateral ulnar fractures was given only stall rest and it recovered.     

Chronic vascular catheterisation of the koala and the rabbit

SUMMARY: Vascular access ports were surgically placed, first in rabbits maintained under laboratory conditions, and then in koalas maintained in a wildlife park. The ports remained patent for 9 months in rabbits and for up to 13 months in the koalas and were removed successfully. They allowed collection of blood samples without assistance or disturbance in koalas, and without stress as reflected by plasma cortisol concentration. The use of retention rings on the cannula tubing is recommended.     

Within-population diversity of koala Chlamydophila pecorum at ompA VD1-VD3 and the ORF663 hypothetical gene

Infection of koalas by Chlamydophila pecorum is very common and causes significant morbidity, infertility and mortality. Fundamental to management of the disease is an understanding of the importance of multi-serotype infection or pathogen virulence in pathogenesis; these may need consideration in plans involving koala movement, vaccination, or disease risk assessment. Here we describe diversity of ompA VD1-3, and ORF663 hypothetical gene tandem repeat regions, in a single population of koalas with diverse disease outcomes. We PCR amplified and sequenced 72 partial ompA segments and amplified 25 tandem repeat segments (ORF663 hypothetical gene) from C. pecorum obtained from 62 koalas. Although several ompA genotypes were identified nationally, only one ompA genotype existed within the population studied, indicating that severe chlamydial disease occurs commonly in free-ranging koalas in the absence of infection by multiple MOMP serotypes of C. pecorum. In contrast, variation in tandem repeats within the ORF663 hypothetical gene was very high, approaching the entire range reported for pathogenic and non-pathogenic C. pecorum of European ruminants; providing an impetus for further investigation of this as a potential virulence trait.     

Pyometra and pyovagina in koalas

SUMMARY Four female koalas were found to have pyometra and pyovagina at autopsy. All had soiling of fur around the cloaca and were in poor condition. One koala had cystic ovaries in addition to the pyometra while another had extensive peritonitis due to rupture of one of the uterine horns. Chronic cystitis was present in all koalas while one also had pyelonephritis. Microbiological examination revealed a mixed flora of both aerobes and anaerobes.     

THE INFLUENCE OF THE TIME OF TREATMENT IN RELATION TO FEEDING ON THE EFFICACY OF AMOXYCILLIN THERAPY IN DOGS AND CATS

SUMMARY A field trial comparing the clinically assessed response to amoxycillin therapy in 148 fasted and 129 non-fasted dogs and cats suffering clinically diagnosed bacterial infections is reported. A satisfactory response to treatment was obtained in 88.8% of these cases. The results indicate that the clinical response to amoxycillin therapy was not affected by the concurrent administration of the antibiotic with food.     

Season- and Age-related Reproductive Changes Based on Fecal Androgen Concentrations in Male Koalas,Phascolarctos cinereus

The purposes of the present study were to clarify age- and season- related androgen patterns, and to compare the reproductive physiology between Japanese captive koala populations and Australian populations. To measure fecal androgens, feces were collected from male koalas (4.2 to 13.8 years of age) kept in Japanese zoos. Fecal androgens were extracted with methanol from the lyophilized samples and determined by enzyme immunoassay using 4-androstene-3,17-dione antibody. Fecal androgen concentration in male koalas increased after sexual maturation and remained relatively high until old age. In the survey with the Japanese zoo studbook of koalas, copulation (conception) month showed a pyramid shape with a peak in March to June (60.7%) in koalas born and reared in Japanese zoos and from July to April with the highest concentration in September to January (69.7%) in Australian institutes. Japanese zoo koala populations have a characteristic physiological cycle adapted to Japan''s seasonal changes. The suitable month of year for copulation or conception in Japan is diametrically opposed to that in Australia. Mean fecal androgen concentrations by month in the males born and reared in Japan indicated annual changes with the highest concentration in May and the lowest value in November. Fecal androgen analysis may be a noninvasive alternative tool to monitor circulating testosterone and may be helpful in understanding reproductive activity and physiology in male koalas.     

宿主细胞磷酸激酶在新城疫病毒感染中的作用

为探讨磷酸化通路在新城疫病毒（Newcastle disease virus,NDV）感染中的作用,使用多种蛋白磷酸激酶抑制剂和激活剂预处理细胞,比较处理后NDV在细胞上生长状况的差异。结果显示NDV在PKC蛋白激酶抑制剂处理的细胞上的生长受到抑制,这种抑制作用能够被PKC蛋白激酶激活剂中和,而PKA蛋白激酶抑制剂和p38MAPK蛋白激酶抑制剂对其影响不大。通过PKC蛋白激酶抑制剂的细胞毒性试验证实,这种抑制作用不是由于药物对细胞的毒性或者因药物加入而改变培养基pH值而造成的。病毒在staurosporine处理的细胞上的生长曲线试验显示,浓度在20~200 nmol/L能对病毒感染产生抑制作用,并且药物浓度越高,上清中病毒的滴度相对越低。本研究结果表明PKC磷酸化通路在新城疫病毒的感染过程中发挥了重要作用。