Brain and spinal cord lesions in encephalitozoonosis in mink

Central nervous system lesions were studied by light microscopy in 43 farmed mink, aged 5 months to 2 1/2 years, with spontaneous encephalitozoonosis and showing cataractous eye changes. Lesions were found in the brain and spinal cord of all animals examined but were generally mild and chronic. The lesions were consistent with those previously described in spontaneous encephalitozoonosis in other carnivores. Parasites in parasitophorous vacuoles and free or phagocytosed in necrotic and granulomatous lesions were demonstrated in animals aged 5 months to 1 year. The occurrence of arterial lesions of the polyarteritis nodosa type found in the youngest animals probably indicates fetal infection. In animals aged 1 1/2 and 2 1/2 years active lesions were usually lacking and the changes were characterized by arterial sclerosis, sometimes with aneurysmal formations, small perivascular lympho-plasmacytic cuffings and focal gliosis.     

Brain and spinal cord lesions in encephalitozoonosis in the blue fox

The central nervous system lesions in encephalitozoonosis were studied in 11 naturally infected blue foxes. Immunohistochemical staining was employed in the demonstration of the parasites and identification of host cells. Consistent findings in acute to subacute stages included granulomatous meningoencephalomyelitis and vasculitis, with arterial lesions similar to polyarteritis nodosa. Chronic stages were dominated by arteriosclerosis, perivascular mononuclear infiltrations and gliosis, with less prominent granulomatous reaction. Parasites were almost constantly observed in association with active lesions both in vessels and nervous tissue. Parasitophorous vacuoles (PV) occurred in endothelial cells and in the medial layer of arteries. Neurons, including even their processes, were widely parasitized, a fact that may account for the brain being an important target organ. PV also occurred in ependymal cells of the choroid plexus and occasionally in macrophages. A great number of parasites was demonstrated in phagolysosomes of macrophages, a fact that probably reflects a relative resistance to digestion, and may consequently contribute to the development of the prominent granulomatous reaction. It was concluded that the brain lesions, except in chronic stages of mild infections, seem to be pathognomonic for encephalitozoonosis in the blue fox.     

Quantitation of meloxicam in the plasma of koalas (Phascolarctos cinereus) by improved high performance liquid chromatography

An improved method to determine meloxicam (MEL) concentrations in koala plasma using reversed phase high performance liquid chromatography equipped with a photo diode array detector was developed and validated. A plasma sample clean-up step was carried out with hydrophilic-lipophilic copolymer solid phase extraction cartridges. MEL was separated from an endogenous interference using an isocratic mobile phase [acetonitrile and 50 mM potassium phosphate buffer (pH 2.15), 45:55 (v:v)] on a Nova-Pak C₁₈ 4-µm (300 × 3.9 mm) column. Retention times for MEL and piroxicam were 8.03 and 5.56 min, respectively. Peak area ratios of MEL to the internal standard (IS) were used for regression analysis of the calibration curve, which was linear from 10 to 1,000 ng/mL (r² > 0.9998). Average absolute recovery rates were 91% and 96% for MEL and the IS, respectively. This method had sufficient sensitivity (lower quantitation limit of 10 ng/mL), precision, accuracy, and selectivity for routine analysis of MEL in koala plasma using 250-µL sample volumes. Our technique clearly resolved the MEL peak from the complex koala plasma matrix and accurately measured MEL concentrations in small plasma volumes.     

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.     

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.     

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.     

Pharmacokinetic profile of amoxicillin and its glucuronide-like metabolite when administered subcutaneously to koalas (Phascolarctos cinereus)

Amoxicillin was administered as a single subcutaneous injection at 12.5 mg/kg to four koalas and changes in amoxicillin plasma concentrations over 24 hr were quantified. Amoxicillin had a relatively low average ± SD maximum plasma concentration (C_max) of 1.72 ± 0.47 µg/ml; at an average ± SD time to reach C_max (T_max) of 2.25 ± 1.26 hr, and an elimination half-life of 4.38 ± 2.40 hr. The pharmacokinetic profile indicated relatively poor subcutaneous absorption. A metabolite was also identified, likely associated with glucuronic acid conjugation. Bacterial growth inhibition assays demonstrated that all plasma samples other than t = 0 hr, inhibited the growth of Escherichia coli ATCC 25922 and Staphylococcus aureus ATCC 29213 to some extent. Calculated pharmacokinetic indices were used to predict whether this dose could attain a plasma concentration to inhibit some susceptible Gram-negative and Gram-positive pathogens. It was predicted that a twice daily dose of 12.5 mg/kg would be efficacious to inhibit susceptible bacteria with an amoxicillin minimum inhibitory concentration (MIC) ≤ 0.75 µg/ml such as susceptible Bordetella bronchiseptica, E. coli, Staphylococcus spp. and Streptococcus spp. pathogens.     

Fatal disseminated encephalitozoonosis in a captive, adult Goeldi's monkey (Callimico goeldii) and subsequent serosurvey of the exposed conspecifics

A captive, adult male Goeldi's monkey (Callimico goeldii) (GM) presented in acute respiratory distress 4 yr after importation into the United States from Europe. Radiographs and echocardiogram were consistent with heart failure. The monkey died within 24 hr of presentation. Necropsy findings included multicentric arteritis and aortitis with aneurysm associated with microsporidian organisms morphologically consistent with Encephalitozoon species. Polymerase chain reaction confirmed organisms were Encephalitozoon cuniculi. Sequence analysis of amplicons generated by using primers specific for the polar tube protein of E. cuniculi determined the organism to be genotype II. An E. cuniculi serosurvey of potentially exposed conspecifics that represented approximately 40% of the captive GM population in the United States was conducted. Multiple individuals that had been imported from Europe with the individual of this report were seropositive via an immunofluorescent antibody assay for E. cuniculi. Multiple samples were available from 3 individuals that demonstrated a decrease in titer or reversion to seronegative status within 3 yr of initial positive status. All other GM were negative on serology. This case is unique in that the genotype identified (genotype II) was different than the genotype (genotype III) reported in other New World primate (NWP) species, the patterns of arteritis were different from the typical pattern of microsporidial vasculitis described in other species, and clinical disease was observed in an adult. Most reported cases of clinical disease secondary to E. cuniculi in NWP have been in neonates and juveniles.     

综述蜜蜂孢子虫病的防治方法

蜜蜂微孢子虫可感染蜜蜂,引起蜜蜂孢子虫病,严重影响蜜蜂的生长繁殖,对养蜂业造成巨大损失,是感染蜜蜂的主要病原微生物之一。结合实验室经验,主要从感染前预防和感染后治疗两方面出发,介绍了蜜蜂孢子虫病的防治方法,为防治蜜蜂孢子虫病提供一个系统而全面的参考。     

獭兔脑原虫病的诊断

对发生脑原虫病的獭兔进行尸体剖检和组织病理学检查，尸体剖检发现肾脏有2-4mm灰白色凹陷区，组织病理学检查可见典型的肉芽肿性脑炎及间质性肾炎的变化，这是该病特征性的病理形态学的变化，并在大脑胶质结节内和肾脏髓质见到虫体的剖面。     

Squamous cell carcinoma in a digit of the hind limb with systemic metastasis in a 17-year-old female koala

We encountered a case of cutaneous squamous cell carcinoma (SCC) in a 17-year-old female koala at a zoo. A fragile, papillary, elevated mass was found on the third digit of the right hind limb. SCC was identified histopathologically: squamous cell-like polygonal tumor cells showed a nest-like growth pattern with epidermal down growth, central keratinization and necrotic foci, and invaded dermal connective tissues. Metastatic lesions were observed in various organs, including the lung and axillary lymph node: in the lung, multiple metastatic foci similar to the primary lesion, and in the axillary lymph node, individual polygonal tumor cells infiltrated the sinusoids. Immunohistochemistry revealed that the tumor cells were positive for proliferating cell nuclear antigen, which exhibited 32–33% of labeling indices in the tumor cells. To our knowledge, this is the first report of a case of SCC in a digit of a koala.     

Fatal suppurative meningoencephalitis caused by Klebsiella pneumoniae in two calves

One calf died (No. 1) and another was euthanized following astasia (No. 2). Histopathological examination revealed suppurative meningoencephalitis in these calves. Klebsiella pneumoniae antigens were detected in lesions. Thymocytes were decreased in the thymus cortex in both cases. 16S rRNA gene sequencing of the No. 1 isolate and bacterial extracts from formalin fixed paraffin embedded sections of No. 2 revealed that both samples were K. pneumoniae. The No. 1 isolate showed multidrug resistance against penicillin antibiotics, fosfomycin, streptomycin, macrolide antibiotics, tetracycline antibiotics, and clindamycin. Immunosuppression is a significant septicemic K. pneumoniae infection risk factor. Our study provides new aspects regarding K. pneumoniae infections in cattle, bacterial meningoencephalitis differentiation, and K. pneumoniae and bacterial meningoencephalitis treatments.     

Extraglomerular lesions in kidneys of mink with encephalitozoonosis.

Extraglomerular renal lesions were studied by light and electron microscopy in 13 farmed mink which showed cataractous eyes associated with spontaneous encephalitozoonosis. The extraglomerular renal lesions consisted of multiple renal cysts, multifocal-to-coalescing interstitial nephritis and vasculitis. Tubular cysts of varying size were present in the corticomedullary junction and medulla. The inflammatory infiltrates were composed mostly of lymphocytes and plasma cells and usually accompanied an interstitial fibrosis. Vasculitis, perivasculitis and sclerotic arteries were frequently seen.     

兔脑炎微孢子虫病的免疫学研究进展

兔脑炎微孢子虫病是由兔脑炎微孢子虫引起的一种隐性、慢性感染或临床经过的人畜共患的原虫病。近十余年来 ,出现大量有关兔脑炎微孢子虫引起艾滋病患者致死性临床感染的报道 ,此病日益引起医学界的广泛关注和重视。就兔脑炎微孢子虫病的免疫学检测方法、细胞免疫应答、感染兔脑炎微孢子虫后人及动物的免疫学比较的研究状况进行了综述。     

Immunohistochemical identification of Encephalitozoon cuniculi in phacoclastic uveitis in four rabbits

PURPOSE: Encephalitozoon cuniculi is a microsporidium with a wide range of mammalian hosts. In rabbits it can be responsible for cataract and lens-induced uveitis (LIU). The aim of this study was to provide specific immunohistochemical demonstration and localization of E. cuniculi within the eye, in rabbits with LIU. MATERIAL AND METHODS: Four rabbits were presented with a white mass in the eye and iris discoloration. Complete ophthalmic examinations were performed and a presumptive diagnosis of LIU was made in all cases. Initial therapy with a topical steroid, atropine and systemic enrofloxacin was instituted while serologic (IFA or ICA tests) and cytologic lab results were pending. The final outcome in all cases was enucleation. Routine histology and immunohistochemistry (ABC method) with an antiserum anti-Encephalitozoon cuniculi were performed. RESULTS: Indirect immunofluorescence performed on one rabbit serum expressed a titer of 1 : 32; carbon immunoassay on the serum of the other three rabbits expressed a titer of 1 : 5120 in one, and a titer of 1 : 2560 in the other two cases. Histologically, an intraocular, locally extensive pyogranulomatous infiltration that partially filled the posterior chamber, encasing a wide anterior lens capsule break, was detected in all cases. Immunohistochemically, spores reacting with anti-Encephalitozoon cuniculi antiserum were present in all specimens, occasionally within macrophages and lens epithelial cells. CONCLUSION: Detection of E. cuniculi in rabbits with phacoclastic uveitis has been investigated in the past with different methods. Based on our results, we suggest that immunohistochemistry should be regarded as a useful tool both for specific demonstration of E. cuniculi and for its localization within tissues.     

Serological survey for antibodies to Encephalitozoon cuniculi in pet rabbits in Italy

Pet rabbits (n = 125) from Southern Italy were submitted to a serological screening for Encephalitozoon cuniculi, using an enzyme-linked immunosorbent assay (ELISA) and a carbon immunoassay (CIA). Seventy-eight examined rabbits showed clinical signs suggestive of encephalitozoonosis (head tilt, ataxia, paralysis, cataracts, uveitis, polyuria and polydipsia), whereas 47 were healthy rabbits. Antibodies anti-E. cuniculi were found in 84/125 (67.2%) sera analysed. The results of the chi-squared test showed that sex and health status had no significant effect (P > 0.05) on E. cuniculi seropositivity; however, rabbits older than 4 months had a seropositivity for E. cuniculi significantly (P < 0.05) higher than that of rabbits aged up to 4 months. The results of the present survey reinforce the assumption that rabbit may be indicated as the main reservoir of E. cuniculi; therefore, routine screening examinations in pet rabbits are strongly advised considering the zoonotic potential of this parasite.