The tumor suppressor p53 is a central regulatory molecule of apoptosis and is commonly mutated in tumors. Kaposi's sarcoma-associated herpesvirus (KSHV)-related malignancies express wild-type p53. Accordingly, KSHV encodes proteins that counteract the cell death-inducing effects of p53. Here, the effects of all KSHV genes on the p53 signaling pathway were systematically analyzed using the reversely transfected cell microarray technology. With this approach we detected eight KSHV-encoded genes with potent p53 inhibiting activity in addition to the previously described inhibitory effects of KSHV genes ORF50, K10 and K10.5. Interestingly, the three most potent newly identified inhibitors were KSHV structural proteins, namely ORF22 (glycoprotein H), ORF25 (major capsid protein) and ORF64 (tegument protein). Validation of these results with a classical transfection approach showed that these proteins inhibited p53 signaling in a dose-dependent manner and that this effect could be reversed by small interfering RNA-mediated knockdown of the respective viral gene. All three genes inhibited p53-mediated apoptosis in response to Nutlin-3 treatment in non-infected and KSHV-infected cells. Addressing putative mechanisms, we could show that these proteins could also inhibit the transactivation of the promoters of apoptotic mediators of p53 such as BAX and PIG3. Altogether, we demonstrate for the first time that structural proteins of KSHV can counteract p53-induced apoptosis. These proteins are expressed in the late lytic phase of the viral life cycle and are incorporated into the KSHV virion. Accordingly, these genes may inhibit cell death in the productive and in the early entrance phase of KSHV infection.

The semelparous dasyurids display a unique life history, in that all males die within a few weeks of the completion of the breeding season. Studies of several semelparous species have revealed that the male die-off is stress-related, and accompanied by increased plasma androgen and cortisol levels and decreased corticosteroid binding capacity, resulting in suppression of immune and inflammatory responses. This study examines the endocrine profile of male brush-tailed phascogales (Phascogale tapoatafa) that survive beyond the breeding season in captivity. Plasma cortisol, corticosteroid binding globulin and albumin levels were monitored in both males and females and steroid partitioning calculated. Captive males surviving beyond the breeding season did not show the elevation in plasma cortisol and decrease in corticosteroid binding capacity reported in wild males. Plasma albumin concentrations also remained constant during the sampling period. These data indicate that captive males do not undergo the same stress response described in wild populations.

Wild rabbits (Oryctolagus cuniculus) in Australia are the descendents of 24 animals from England released in 1859. We surveyed rabbits and rabbit fleas (Spilopsyllus cuniculi) in Australia for the presence of trypanosomes using parasitological and PCR-based methods. Trypanosomes were detected in blood from the European rabbits by microscopy, and PCR using trypanosome-specific small subunit ribosomal RNA (SSU rRNA) gene primers and those in rabbit fleas by PCR. This is the first record of trypanosomes from rabbits in Australia. We identified these Australian rabbit trypanosomes as Trypanosoma nabiasi, the trypanosome of the European rabbit, by comparison of morphology and SSU rRNA gene sequences of Australian and European rabbit trypanosomes. Phylogenetic analysis places T. nabiasi in a clade with rodent trypanosomes in the subgenus Herpetosoma and their common link appears to be transmission by fleas. Despite the strict host specificity of trypanosomes in this clade, phylogenies presented here suggest that they have not strictly cospeciated with their vertebrate hosts. We suggest that T. nabiasi was inadvertently introduced into Australia in the 1960s in its flea vector Spilopsyllus cuniculi, which was deliberately introduced as a potential vector of the myxoma virus. In view of the environmental and economic damage caused by rabbits in Australia and other islands, the development of a virulent or genetically modified T. nabiasi should be considered to control rabbits.

Little is known about the trypanosomes of indigenous Australian vertebrates and their vectors. We surveyed a range of vertebrates and blood-feeding invertebrates for trypanosomes by parasitological and PCR-based methods using primers specific to the small subunit ribosomal RNA (SSU rRNA) gene of genus Trypanosoma. Trypanosome isolates were obtained in culture from two common wombats, one swamp wallaby and an Australian bird (Strepera sp.). By PCR, blood samples from three wombats, one brush-tailed wallaby, three platypuses and a frog were positive for trypanosome DNA. All the blood-sucking invertebrates screened were negative for trypanosomes both by microscopy and PCR, except for specimens of terrestrial leeches (Haemadipsidae). Of the latter, two Micobdella sp. specimens from Victoria and 18 Philaemon sp. specimens from Queensland were positive by PCR. Four Haemadipsa zeylanica specimens from Sri Lanka and three Leiobdella jawarerensis specimens from Papua New Guinea were also PCR positive for trypanosome DNA. We sequenced the SSU rRNA and glycosomal glyceraldehyde phosphate dehydrogenase (gGAPDH) genes in order to determine the phylogenetic positions of the new vertebrate and terrestrial leech trypanosomes. In trees based on these genes, Australian vertebrate trypanosomes fell in several distinct clades, for the mostpart being more closely related to trypanosomes outside Australia than to each other. Two previously undescribed wallaby trypanosomes fell in a clade with Trypanosoma theileri, the cosmopolitan bovid trypanosome, and Trypanosoma cyclops from a Malaysian primate. The terrestrial leech trypanosomes were closely related to the wallaby trypanosomes, T. cyclops and a trypanosome from an Australian frog. We suggest that haemadipsid leeches may be significant and widespread vectors of trypanosomes in Australia and Asia.

Figure 2: Pontine Micturition Centre (A) and Periaqueductal Grey (B) Figure 1: Comparison of the 3D anatomical dataset (top) and the corresponding co-registered T2*-weighted functional raw dataset (bottom) of a single subject. T2*-weighted images reveal a high quality even in the brainstem including the pons, known to be particularly vulnerable to susceptibility and flow artifacts. Functional Imaging of Brainstem Structures Involved in Voiding Control

Blood was collected from 31 Parma wallabies from five different captive populations. The morphology of erythrocytes, leucocytes and platelets was similar to that previously described for macropodids. The effects of location (population), anaesthesia and analyser on the haematological characteristics of the wallabies were considered. Location had an affect on many of the values, anaesthesia affected some of the leucocyte values, and analyser had little effect.

Trypanosome infections in their natural hosts are frequently difficult to detect by microscopy, and culture methods are unreliable and not suitable for all species of Trypanosoma. A nested PCR strategy for detecting and identifying Trypanosoma species, suitable for detecting both known and unknown trypanosomes, is presented. Thirty-two blood samples from 23 species of Australian birds and mammals were screened by a nested PCR for the presence of Trypanosoma sp. ssrRNA. Three infections were detected, one in an eastern grey kangaroo (Macropus giganteus), one in a common wombat (Vombatus ursinus) and one in a platypus (Ornithorhynchus anatinus). The kangaroo and wombat are new host records for Trypanosoma sp.; the platypus parasite was Trypanosoma binneyi. The three parasites could be distinguished by restriction fragment length polymorphisms of the amplified fragment of the ssrRNA gene. The kangaroo and wombat parasites were also isolated in a semi-solid blood agar medium. The culture forms of the kangaroo trypanosome had an expanded flagellar sheath in which structures similar to hemidesmosomes were detected by EM. The nested PCR was at least as sensitive as culture, and analysis of the PCR products gave parasite-specific fingerprints. Therefore this method could be suitable for rapidly screening host animals for the presence of trypanosomes and identifying the infecting strain.

The premise that drugs not be injected into the caudal body of reptiles because they will be carried by the renal portal system to the kidneys, where they may be nephrotoxic or rapidly excreted, was tested by comparing the pharmacokinetics of gentamicin (excreted via glomerular filtration in mammals) and carbenicillin (excreted partly via renal tubular secretion in mammals) following injection into the forelimb or hindlimb of red-eared sliders (Trachemys scripta elegans). Ten sliders received intramuscular gentamicin (10 mg/kg) in a forelimb (n = 5) or a hindlimb (n = 5), and plasma levels of the drug were assayed over time. Following drug clearance, the experiment was repeated with the site of injection reversed so that each animal acted as its own control. Another 10 sliders were similarly treated, using intramuscular carbenicillin (200 mg/kg). Injection site of gentamicin had no effect on any pharmacokinetic parameter (time to maximum plasma concentration, maximum plasma concentration, half-life, area under the curve, clearance, and volume of distribution). However, the area under the curve of plasma carbenicillin concentration vs. time was significantly lower following hindlimb injection, in comparison with forelimb injection, at 1, 4, and 8 hr, which may reflect reduced bioavailability of the drug, as would be expected with renal portal perfusion and tubular excretion on first pass through the kidney. This effect on carbenicillin likely is not clinically important because plasma levels remained above recommended minimum inhibitory concentrations. Because blood draining the caudal body of reptiles passes through the kidneys or the liver before reaching the central circulation, the effect on the pharmacokinetics of a drug injected in that region will vary with its renal or hepatic extraction rate. Generally, this effect is unlikely to be significant.

The anatomy of the renal portal system of the red-eared slider (Trachemys scripta elegans) is described, based on dissection of six double latex-injected specimens (three males, three females). The anatomy of these vessels, which had not previously been described in this species, was found not to differ significantly from the fundamental chelonian pattern. Fluoroscopic radioangiography revealed that venous blood returning from the hindlimbs flowed predominantly to the Liver and bypassed the kidneys. Blood from the tail either flowed to the kidneys or bypassed them and flowed directly to the liver. A putative valve is described that governs venous blood flow from the caudal body to or around the kidneys.

The long-acting tranquilizers zuclopenthixol decanoate and pipothiazine palmitate were used to calm recently captured red-necked wallabies (Macropus rufogriseus). The two drugs were administered in combination at dose rates of 7.5, 10, and 15 mg/kg, and the response of the wallabies was monitored and compared with an untreated group. Giving both drugs at a dose rate of 10 mg/kg produced the most consistent levels of tranquilization. Tranquilization at all dose rates declined after 10 days. No side effects were observed, although elevated plasma fibrinogen and alkaline phosphatase levels occurred in all animals.

Electrocardiographic (ECG) values were measured in nine red-eared sliders (Trachemys scripta elegans), anaesthetised with either ketamine, ketamine/xylazine or ketamine/midazolam, for the purpose of cardiocentesis. Lead II traces were then recorded through three cutaneous electrodes. The ECG complexes consisted of small upright P waves, larger R waves and no Q or S wave deflections, very long QT intervals, small upright T waves, and a short TP interval. No SV waves were recorded. This pattern was similar to the ECGs reported in other reptilian species, and differs from mammalian ECGs in having lower amplitude waveforms, a much longer repolarisation phase (longer ST and QT intervals), and a considerably shorter TP interval. A significant correlation was found between the heart rate and the QT and ST intervals, a lower heart rate being associated with a longer period of repolarisation without significantly prolonging the depolarisation (P, PR and QRS) intervals.

SUMMARY Electrocardiographic (EGG) values were measured in nine red-eared sliders (Trachemys scripta elegans), anaesthetised with either ketamine, ketamine/xylazine ketamine/midazolam, for the purpose of cardioeentesis. Lead II traces were then recorded through three cutaneous electrodes. The EGG complexes consisted of small upright P waves, larger R waves and no Q or S wave deflections, very long QT intervals, small upright T waves, and a short TP interval. No SV waves were recorded. This pattern was similar to the ECGs reported in other reptilian species, and differs from mammalian EGGs in having lower amplitude waveforms, a much longer repolarisation phase (longer ST and QT intervals), and a considerably shorter TP interval. A significant correlation was found between the heart rate and the QT and ST intervals, a lower heart rate being associated with a longer period of repolarisation without significantly prolonging the depolarisation (P, PR and QRS) intervals.

Nineteen red-eared sliders (Trachemys scripta elegans) were anesthetized using one of three regimes: 60 mg/kg ketamine, 60 mg/kg ketamine with 2 mg/kg xylazine, or 60 mg/kg ketamine with 2 mg/kg midazolam, all injected i.m. No significant differences in induction times, anesthetic depth, or recovery rates were noted among the various regimes, which produced a variable level of anesthesia suitable for minor procedures only. Differences did appear when sliders were divided on the basis of weight. When the smaller sliders (< 1 kg) were administered the same mg/kg dosage of the drug combinations as were the larger sliders, they took significantly longer to recover and to become active. Hematologic and biochemical values, blood gas readings, and EKG tracings were all within normal limits, with the exception of blood gas readings and EKG tracings in one slider that died under ketamine/midazolam anesthesia.

Sixteen gray wolves (Canis lupus) were immobilized with a combination of medetomidine and ketamine. A mean (+/-SD) dose rate of 55.3 +/- 10.7 mug/kg medetomidine and 2.76 +/- 0.53 mg/kg ketamine induced complete immobilization in 9.3 +/- 3.4 min. Eight of the 16 wolves were given 49.4 +/- 8.4 mug/kg atropine in addition to the medetomidine/ketamine combination. All non-atropinized wolves exhibited bradycardia and cardiac arrhythmias. and most showed excessive salivation (62.5%). Atropinized animals exhibited tachycardia, but no arrhythmias or excessive salivation were recorded. No differences in respiratory rates or rectal temperatures were seen between the two groups. Blood samples for hematologic, plasma biochemical, and blood gas analysis during the course of the immobilizations revealed only a mild acidosis. Following administration of 260 +/- 30 mug/kg atipamezole (half intramuscularly, half intravenously), the wolves regained their feet in 10.7 +/- 12.7 min, losing any residual ataxia within 45.4 +/- 23.8 min. Vomiting occurred in 56.3% of wolves on recovery.