Beyond Morbidity and Mortality in Reintroduction Programmes: Changing Health Parameters in Reintroduced Eastern Bettongs Bettongia Gaimardi

Beyond morbidity and mortality in reintroduction programmes: changing health parameters in reintroduced eastern bettongs Bettongia gaimardi

T IMOTHY J. PORTAS,ROSS B. CUNNINGHAM,DAVID S PRATT,JOANNE D EVLIN P ETER H OLZ,WILLIAM B ATSON,JANE O WENS and A DRIAN D. MANNING

Abstract The eastern bettong Bettongia gaimardi, a potor- metrics for assessing the response of macropodoids to oid marsupial, has been extinct on the Australian mainland reintroduction. since the s. Sixty adult bettongs were reintroduced from Keywords Bettongia gaimardi, biochemistry, disease, east- the island of Tasmania to two predator-free fenced reserves ern bettong, haematology, health evaluation, parasite, on mainland Australia. We examined baseline health para- reintroduction meters (body weight, haematology and biochemistry, parasites and infectious disease exposure) in a subset of  ( male,  female) individuals at translocation and again at – months post-reintroduction. The mean body Introduction weight increased significantly post-reintroduction but there were no significant differences in body weight between he underlying health status of individual animals can the two reintroduction sites or between the sexes in response Tinfluence survival during translocation and establish- to reintroduction. Differences were evident in multiple ment in reintroduction programmes, and disease may influ- haematological and biochemical variables post-reintroduc- ence the persistence of populations in the longer term (Kock tion but there were few differences between the two reintro- et al., ; Cabezas et al., ; Clarke et al., ). Disease duced populations or between the sexes in response to may also have an impact on sympatric species at reintroduc- reintroduction. Ectoparasite assemblages differed, with tion sites, and detailed recommendations for assessing and  five of species failing to persist, and an additional four managing disease risk in wildlife translocations have been species were identified post-reintroduction. None of the bet- published (Leighton, ; Travis et al., ; Jakob-Hoff tongs had detectable antibodies to the alphaherpesviruses et al., ). Despite this, comprehensive health evaluations   Macropodid herpesvirus and post-reintroduction, in- are not routinely undertaken, and the potential for disease cluding one individual that was seropositive at transloca- to influence outcomes is often not considered nor adequate- tion. Similarly, the novel gammaherpesvirus potoroid ly managed in reintroduction programmes (Mathews et al.,  herpesvirus was not detected by polymerase chain reaction ; Deem et al., ). Various health parameters in free- (PCR) in any of the bettongs post-reintroduction, including ranging wildlife species, including haematology and bio- one individual that was PCR-positive at translocation. None chemistry, and parasite assemblages, are influenced by a of the bettongs had detectable antibodies to Toxoplasma range of environmental and host factors and could be ex- gondii either at translocation or post-reintroduction. Our pected to change when species are reintroduced to environ- data demonstrate changing baseline health parameters in ments from which they have been extirpated (Schultz et al., eastern bettongs following reintroduction to the ; Robert & Schwanz, ; Webster et al., ). Australian mainland are suggestive of improved health in Evaluation of changes to health parameters pre- and post- the reintroduced populations, and provide additional reintroduction could potentially be used to assess the longer term physiological response of species to reintroduction and may be a useful adjunct to traditional metrics (survival, dis- persal and reproductive success) used to assess reintroduc- TIMOTHY J. PORTAS (Corresponding author) and PETER HOLZ Veterinary and  Research Centre, Tidbinbilla Nature Reserve, RMB 141, Via Tharwa, tion outcomes (Ewen et al., ; Nichols & Armstrong, Australian Capital Territory 2620, Australia. E-mail [email protected] ; Maceda-Veiga et al., ).

ROSS B. CUNNINGHAM,WILLIAM BATSON and ADRIAN D. MANNING Fenner School of The eastern bettong Bettongia gaimardi is a small, noc- Environment and Society, Australian National University College of Medicine, turnal, predominantly mycophagous, potoroid marsupial Biology and Environment, Canberra, Australia that has been extinct on the Australian mainland since the DAVID SPRATT Australian National Wildlife Collection, CSIRO National Research   −  Collections Australia, Canberra, Australia s (Claridge et al., ). In adults were reintroduced from the island of Tasmania to two predator- JOANNE DEVLIN and JANE OWENS Faculty of Veterinary Science, the University of Melbourne, Parkville, Victoria, Australia free fenced reserves, Tidbinbilla Nature Reserve and Received  July . Revision requested  October . Mulligans Flat Woodland Sanctuary, in the Australian Accepted  November . First published online  March . Capital Territory (Batson et al., in press). Comprehensive

Oryx, 2016, 50(4), 674–683 © 2016 Fauna & Flora International doi:10.1017/S0030605315001283 Downloaded from https://www.cambridge.org/core. IP address: 170.106.33.19, on 28 Sep 2021 at 11:19:01, subject to the Cambridge Core terms of use, available at https://www.cambridge.org/core/terms. https://doi.org/10.1017/S0030605315001283 Bettong health following reintroduction 675

health assessments were undertaken during translocation, and baseline health and disease parameters were established for the species (Portas et al., ). We hypothesized that health parameters of eastern bettongs would change post- reintroduction and could therefore be useful measures for assessing the effect of reintroduction on bettong health beyond direct observations of morbidity and mortality. Here we compare body weight, haematology and biochemistry, parasite assemblages, and disease exposure (Toxoplasma gondii, macropodoid herpesviruses) of a subset ( males,  females) of the population pre- and post-reintroduction. We also compare the haematology and biochemistry of the populations at the two reintroduction sites and between the sexes in response to reintroduction.

Study area

Collection sites for free-ranging bettongs in Tasmania have been described previously and included eight locations re- presenting a mix of remnant native forest, forestry planta- tions and agricultural land (Portas et al., ; Batson et al., in press). Mulligans Flat Woodland Sanctuary, in the Australian Capital Territory, is a  ha remnant of FIG. 1 Location of the two reintroduction sites, Mulligans Flat highly threatened box–gum grassy woodland that is the sub- Woodland Sanctuary and Tidbinbilla Nature Reserve within the ject of intensive experimental restoration efforts (Manning Australian Capital Territory. et al., ; Shorthouse et al., ). Tidbinbilla Nature Reserve, also in the Australian Capital Territory, incorpo- At – months post-reintroduction (May–November rates a fenced sanctuary in an area of wet sclerophyll forest ) bettongs were captured at both reintroduction sites and open grassland for the conservation management of at night, using padded cage traps baited with a mixture of threatened Australian species (Fig. ). oats and peanut butter. Traps were set at dusk and checked – hours later. Bettongs were removed from traps and Methods placed in cloth bags before being processed on site. Anaesthesia was induced and maintained using isoflurane Previously we established baseline health and disease in oxygen delivered via a mask. Post anaesthesia, bettongs parameters in  adult (.  year, on the basis of dentition) recovered in cloth bags for up to  hour before being released eastern bettongs ( male,  female) live-trapped during at the capture site. July–October  and April–September  in Tasmania The physical examination process, sample and data col- and reintroduced to Tidbinbilla Nature Reserve and lection, and sample processing are described in detail in Mulligans Flat Woodland Sanctuary (Portas et al., ). Portas et al. (). Briefly, the following were performed: Bettongs released into Mulligans Flat ( males,  females) physical examination; body weight was recorded; pes length were monitored via global positioning system/VHF collars was measured; ticks and fleas were collected using forceps and live-trapping approximately every  months. They re- and fixed in % ethanol; mites and lice were collected ceived no supplemental food and the population was un- from skin and hair scrapings in glycerine; blood was col- managed. Bettongs at Tidbinbilla ( males,  females) were lected from the lateral coccygeal vein for assessment of housed in small groups in natural bushland enclosures (.– haematological and biochemical parameters and serology . ha) and managed to maximize genetic diversity of off- (Toxoplasma gondii, Macropodid herpesviruses  and  spring; they received supplemental food, including fresh [MaHV-, MaHV-]); pooled swabs from the conjunctival, locally available produce and a commercially available nasal and urogenital mucosa were collected for the detection pellet, at least weekly. Monitoring was limited to remote of herpesvirus DNA using polymerase chain reaction observation of feed stations via camera, and live-trapping (PCR); and faecal samples were collected and assessed approximately every  months. using the sodium nitrate floatation technique for endopara- Trapping, sedation and anaesthetic procedures at trans- site ova. Bettongs were weighed to the nearest gram using location have been described previously (Portas et al., ). electronic scales. For females with pouch young present,

weights were adjusted by subtracting the estimated weight of the pouch young (Batson et al., ). We calculated body condition index using the residuals of a linear regression of body weight against pes length (Johnson, ). Individuals were identified by the subcutaneous placement of a microchip. Haematological and biochemical analyses were performed by Vetnostics (North Ryde, Australia) within  hours of collection. The following were measured: haematocrit, haemoglobin, red blood cell count, mean corpuscular volume, mean corpuscular haemoglobin, mean corpuscular haemoglobin concentration, platelet count, neutrophils, lymphocytes, monocytes, eosinophils, basophils, sodium, potassium, chloride, bicarbonate, anion gap, urea, creatinine, glucose, bilirubin, aspartate amino transferase, alanine transaminase, gamma glutamyl transferase, alkaline phosphatase, total protein, globulin, albumin, albumin/globulin ratio, calcium, phosphate, creatine kinase, cholesterol and triglyceride. Any haematological and biochemical variables that were significantly different post reintroduction, and potentially influenced by nutrition, were reassessed after stat- FIG. 2 Fitted and observed relationship from regression analysis istically adjusting for body weight. of body weight vs pes length in eastern bettongs Bettongia gaimardi reintroduced from Tasmania to the Australian Capital Toxoplasma gondii serology was performed at the Territory, Australia (Fig. ). Department of Primary Industries, Water and Environment, Mount Pleasant Laboratories, Launceston, Australia, using the direct and modified agglutination tests for antibodies to translocation to . ± SE . kg post-reintroduction. T. gondii (Johnson et al., ). Both tests were performed There were no significant differences between mean body on all sera at translocation, after which time the laboratory weights at the two reintroduction sites or between males ceased to offer the direct test, and sera collected post- and females in response to reintroduction. reintroduction were assayed using the modified test only. Haematocrit, haemoglobin, red blood cell count, mean Herpesvirus serology and detection of herpesvirus DNA by corpuscular haemoglobin and platelets increased whereas PCR were performed at the Faculty of Veterinary Science, white blood cell count, neutrophils, lymphocytes and UniversityofMelbourne,Parkville,Australia,usingpreviously monocytes decreased post-reintroduction (Table ). There described techniques (Vaz et al., ). were fewer differences between haematological parameters Haematological and biochemical values (response vari- at the two reintroduction sites, with higher platelets, white ables) varied at two levels: between individuals and within in- blood cell count, neutrophils and monocytes in the dividuals. Candidate explanatory variables such as weight Tidbinbilla population than the Mulligans Flat population varied at both the individual and within-individual levels, (Table ). There were limited differences in response to re- whereas the factors sex and site (Tidbinbilla vs Mulligans introduction between the sexes other than for platelets, Flat) varied only at the individual level. The design variable which increased more (P = .), and white blood cell reintroduction (pre- vs post-) varied only at the within- count, which decreased more (P = .) in males. individual level. Given the multilevel sampling design and Potassium, anion gap, urea, creatinine, glucose, globulin data structure, these data were analysed within the framework and triglyceride were all higher post-reintroduction. of general linear mixed models using restricted maximum Sodium, chloride, bicarbonate, aspartate amino transferase, likelihood, with significance assumed at P , .. For over- alanine transaminase, gamma glutamyl transferase, albu- all inference, information was combined across the two le- min/globulin ratio, phosphate, creatine kinase and choles- vels. Statistical computation was performed using GenStat terol were all lower post-reintroduction (Table ). There  (VSN International Ltd., Hemel Hempstead, UK). were few differences between biochemical parameters at the two reintroduction sites, with lower sodium and bicar- Results bonate, and higher anion gap, creatinine, glucose and alkaline phosphatase in the Tidbinbilla population (Table ). We calculated the body condition index of the reintroduced The only difference between the sexes in response to re- eastern bettongs but found no significant relationship be- introduction was their phosphate level (P = .), which tween body weight and pes length (Fig. ). Body weight in- decreased more in females. Haematocrit (P , .), creased (F pr. , .) from a mean of . ± SE . kg at haemoglobin (P = .), red cell count (P = .),

TABLE 1 Comparison of haematological variables (mean ± SE) in  ( male,  female) eastern bettongs Bettongia gaimardi reintroduced from Tasmania to Mulligans Flat Woodland Sanctuary (MFWS) and Tidbinbilla Nature Reserve (TNR) in the Australian Capital Territory (ACT), Australia, at translocation, post-reintroduction and between populations at the two reintroduction sites.

At translocation Post-reintroduction Reintroduction site Variable1 Tasmania ACT P MFWS TNR P Haematocrit (%) 0.399 ± 0.006 0.456 ± 0.006 , 0.001* 0.445 ± 0.009 0.475 ± 0.011 0.106 − Hb (gL 1) 133.4 ± 2.24 154.9 ± 2.24 , 0.001* 152.5 ± 2.8 159.0 ± 3.6 0.337 − RBC (× 1012 L 1) 9.52 ± 0.156 10.80 ± 0.156 , 0.001* 10.56 ± 0.24 11.22 ± 0.3 0.161 MCV (fL) 41.50 ± 0.53 42.50 ± 0.53 0.195 41.86 ± 0.96 43.61 ± 1.17 0.275 MCH (pg) 14.13 ± 0.10 14.46 ± 0.10 0.029* 14.65 ± 0.21 14.14 ± 0.25 0.104 − MCHC (gL 1) 335.7 ± 1.99 339.6 ± 1.99 0.177 342.1 ± 3.6 335.4 ± 4.4 0.206 − Platelets (× 109 L 1) 387.9 ± 13.13 436.9 ± 13.13 0.016* 405.9 ± 28.9 490.4 ± 3.7 0.014* − WBC (× 109L 1) 3.58 ± 0.15 2.64 ± 0.15 , 0.001* 2.09 ± 0.24 3.59 ± 0.03 0.002* − Neutrophils (× 109 L 1) 2.20 ± 0.13 1.34 ± 0.13 , 0.001* 0.96 ± 0.19 2.00 ± 0.24 0.010* − Lymphocytes (× 109 L 1) 1.38 ± 0.09 1.15 ± 0.09 0.026* 1.00 ± 0.14 1.34 ± 0.17 0.216 − Monocytes (× 109 L 1) 0.15 ± 0.08 0.09 ± 0.08 0.004* 0.06 ± 0.017 0.14 ± 0.021 0.040* − Eosinophils (× 109 L 1) 0.03 ± 0.01 0.05 ± 0.01 0.446 0.035 ± 0.014 0.075 ± 0.018 0.278

 Hb, haemoglobin; RBC, red blood cell count; MCV, mean corpuscular volume; MCH, mean corpuscular haemoglobin; MCHC, mean corpuscular haemoglobin concentration; WBC, white blood cell count *Denotes statistically significant difference between values

TABLE 2 Comparison of biochemical variables (mean ± SE) in  ( male,  female) eastern bettongs reintroduced from Tasmania to Mulligans Flat Woodland Sanctuary (MFWS) and Tidbinbilla Nature Reserve (TNR) in the Australian Capital Territory (ACT), Australia, at translocation, post-reintroduction and between populations at the two reintroduction sites.

At translocation Post-reintroduction Reintroduction site Variable1 Tasmania ACT P MFWS TNR P − Sodium (mmol L 1) 147.20 ± 0.56 143.30 ± 0.56 , 0.001* 144.92 ± 0.8 140.50 ± 1.0 0.012* − Potassium (mmol L 1) 3.67 ± 0.32 4.74 ± 0.32 0.028* 4.78 ± 0.47 4.67 ± 0.56 0.914 − Chloride (mmol L 1) 106.87 ± 0.63 101.30 ± 0.63 , 0.001* 102.77 ± 0.9 98.77 ± 1.1 0.041 − Bicarbonate (mmol L 1) 29.10 ± 0.62 23.60 ± 0.62 , 0.001* 25.48 ± 0.77 20.36 ± 0.99 0.009* − Anion gap (mmol L 1) 15.03 ± 0.80 22.38 ± 0.80 , 0.001* 20.26 ± 1.04 26.05 ± 1.26 0.021* − Urea (mmol L 1) 6.05 ± 0.40 8.26 ± 0.40 , 0.001* 7.52 ± 0.54 9.52 ± 0.68 0.105 − Creatinine (μmol L 1) 41.7 ± 1.78 57.1 ± 1.78 , 0.001* 50.4 ± 2.72 68.7 ± 3.43 0.002* − Glucose (U L 1) 11.33 ± 0.67 17.80 ± 0.67 , 0.001* 14.62 ± 1.2 23.29 ± 1.42 , 0.001* − Bilirubin (U L 1) 1.03 ± 0.08 1.13 ± 0.08 0.409 1.20 ± 0.11 1.019 ± 0.14 0.473 − AST (U L 1) 176.2 ± 5.91 46.2 ± 5.91 , 0.001* 50.1 ± 9.27 40.1 ± 11.63 0.572 − ALT (U L 1) 84.7 ± 3.82 32.6 ± 3.82 , 0.001* 34.3 ± 5.41 29.7 ± 6.87 0.689 − GGT (U L 1) 16.47 ± 0.91 14.61 ± 0.91 0.021* 13.26 ± 1.15 13.37 ± 1.46 0.968 − ALP (U L 1) 1,679 ± 164.1 1,421 ± 164.1 0.275 1,821 ± 392.2 728 ± 443.6 0.033* − Protein (gL 1) 55.13 ± 0.75 56.80 ± 0.75 0.129 56.88 ± 1.21 56.66 ± 1.51 0.923 − Albumin (gL 1) 40.56 ± 0.76 39.78 ± 0.76 0.138 40.13 ± 1.22 37.13 ± 1.52 0.189 − Globulin (gL 1) 14.47 ± 0.66 17.77 ± 0.66 0.002* 16.57 ± 1.04 19.53 ± 1.31 0.166 Albumin/globulin ratio 4.03 ± 0.41 2.37 ± 0.41 0.013* 2.37 ± 0.56 2.07 ± 0.72 0.807 − Calcium (mmol L 1) 2.29 ± 0.02 2.30 ± 0.02 0.793 2.27 ± 0.03 2.36 ± 0.04 0.147 − Phosphate (mmol L 1) 2.51 ± 0.13 2.01 ± 0.13 0.009* 2.214 ± 0.18 1.66 ± 0.21 0.148 − Creatine kinase (U L 1) 24,018 ± 1,463.9 1,005 ± 1,463.9 , 0.001* 118 ± 2,015 2,537 ± 2,565 0.580 − Cholesterol (mmol L 1) 3.894 ± 0.13 3.050 ± 0.13 0.002* 2.94 ± 0.20 2.96 ± 0.25 0.952 − Triglyceride (mmol L 1) 0.867 ± 0.10 1.51 ± 0.10 , 0.001* 1.46 ± 0.13 1.58 ± 0.16 0.704

 AST, aspartate amino transferase; ALT, alanine transaminase; GGT, gamma glutamyl transferase; ALP, alkaline phosphatase *Denotes statistically significant difference between values

creatinine (P , .) and triglycerides (P = .) were Sera from all  bettongs were negative for antibodies positively related to body weight, whereas mean corpuscular to T. gondii at both time points. One female had detectable haemoglobin (P = .), urea (P = .) and globulin antibodies to alphaherpesviruses MaHV- and MaHV-,as (P = .) were not. determined by serum neutralization assay at translocation,

TABLE 3 Ectoparasite assemblages found on eastern bettongs reintroduced from Tasmania to the Australian Capital Territory, Australia, at translocation and post-reintroduction, and accession numbers.

Present at Present Species translocation post-reintroduction Accession number(s) Listrophoridae Paraheterodoxus erinaceus No Yes AR 1594 Paraheterodoxus? n. sp. Yes Yes AR 1572, 1574 Hetrodoxus cf. ualabati No Yes AR 1590 Trombiculidae Eutrombicula macropus No Yes AR 1593 Guntheria cf. pertinax Yes Yes AR 1579 Guntheria cf. shareli No Yes AR 1614 Laelapidae Haemolaelaps hatteni Yes No AR 1575 Thadeua greeni Yes Yes AR 1585 Atopomelidae Cytostethum (Metacytostethum) intermedium Yes Yes AR 1576 Cytostethum (Metacytostethum) tasmaniense Yes Yes AR 1576 Cytostethum (Metacytostethum) thetis Yes Yes AR 1576 Cytostethum (Metacytostethum) wallabia Yes Yes AR 1576 Ixodidae Ixodes cornuatus Yes No AR 1586 Ixodes tasmani Yes No AR 1587 Ixodes trichosuri Yes Yes AR 1571 Pygiopsyllidae Pygiopsylla zethi Yes No AR 1573 Stephanocercidae Stephanocircus harrisoni Yes No AR 1588

but had no detectable antibodies  months later. The other TABLE 4 Prevalence of parasites on a sample of  eastern bettongs  individuals were seronegative at translocation and post- reintroduced from Tasmania to the Australian Capital Territory, reintroduction. A pooled nasal/conjunctival/urogenital tract Australia, at translocation and post-reintroduction. swab from a single male was positive for herpesvirus DNA At translocation Post-reintroduction (potoroid herpesvirus , PotHV-), using PCR at transloca-   No. of individuals No. of individuals tion, but was negative months later. The other were Parasite affected (%) affected (%) negative for herpesvirus DNA, using PCR at both time Endoparasites points. Strongylid eggs 13 (43.3) 22 (73.3) Fleas, lice, mites and ticks recovered from bettongs were Strongylid larvae 2 (6.7) 6 (20) deposited in the Australian National Wildlife Collection, Capillariid-like eggs 1 (3.3) 0 (0) CSIRO, Canberra. Five ectoparasite species (Haemolaelaps Eimeria gaimardi 3 (10) 5 (16.7) hatteni, Ixodes cornuatus, I. tasmani, Pygiopsylla zethi, Ectoparasites Stephanocircus harrisoni) present at translocation were not Ticks 23 (76.7) 2 (6.7) Lice 7 (23.3) 11 (36.7) recovered post-reintroduction; however, an additional four Fleas 13 (43.3) 0 (0) species (Paraheterodoxus erinaceus, Hetrodoxus cf. ualabti, Mites 28 (93.3) 13 (43.3) Eutrombicula macropus, Guntheria cf. shareli) were recovered post-reintroduction (Table ). Details of the prevalence of endo- and ectoparasites recovered from bettongs at both time points are in Table . reintroduced populations. Despite recommendations for ongoing health monitoring in reintroduction programmes there are few reports of continued monitoring beyond trans-  Discussion location and establishment (Kock et al., ; Work et al., ). Furthermore, post-reintroduction health monitoring Our data demonstrate changing health parameters in east- is often limited to direct observations of morbidity and mor- ern bettongs following reintroduction to the Australian tality, with changes at a physiological level receiving scant mainland and are suggestive of improved health in the attention. Our data demonstrate the value of ongoing

comprehensive health evaluations for assessing the response Western Australia, suggesting post-reintroduction haemo- of individuals and populations to reintroduction, and can be globin concentration may be more representative of normal used as an adjunct to traditional measures for assessing re- values for the eastern bettong in optimal environments introduction outcomes. (Pacioni et al., ). Lower haemoglobin concentrations Body condition index, usually calculated as the residuals in response to seasonal declines in diet quality have been de- of a linear regression of body weight against a linear mor- monstrated in western grey kangaroos Macropus fuliginosus, phometric measure, has been used to assess body condition common wallaroos M. robustus and quokkas Setonix bra- in a number of macropodoid species (Stirrat, ; Robert & chyurus (Ealey & Main, ; Shield, ; Algar et al., Schwanz, ). Despite previous validation of the use of ). In woylies, haemoglobin concentration was also body weight and pes length for calculating body condition shown to have a positive association with rainfall, presum- index in eastern bettongs, through calculating total body ably mediated via changing nutritional content of forage water and hence proportion of body fat via isotope dilution, species (Pacioni et al., ). In contrast, seasonal changes we found no relationship in this study (Johnson, ). The in nutrition did not influence haemoglobin concentration reasons for this were not readily evident but the assump- in agile wallabies Macropus agilis or allied rock wallabies tions that underlie the calculation of body condition index Petrogale assimilis and there was no significant difference using this method have been questioned previously (Green, in haemoglobin concentration between three subpopula- ). Consequently we assessed changes in body weight in tions of tammar wallabies Macropus eugenii living in separ- response to reintroduction. Despite the fact that bettongs at ate habitats of variable nutritional quality (Spencer & Tidbinbilla Nature Reserve received supplemental food Speare, ; Stirrat, ; Robert & Schwanz, ). there was no difference in mean body weight between the Increases in red cell count and haematocrit in eastern two reintroduced populations, and body weight increased bettongs post-reintroduction may also be explained by im- at both sites. This, coupled with haematological and proved nutrition at the reintroduction sites. In woylies, red biochemical data, suggests the source populations in cell count and haematocrit have also been shown to have a Tasmania were experiencing suboptimal nutrition. positive association with rainfall (Pacioni et al., ). In Body condition has been positively correlated with agile wallabies from the wet–dry tropics, haematocrit (but the availability of hypogeous fungi in both eastern and nor- not red cell count) was lower during the dry season, and thern bettongs Bettongia tropica (Johnson, ; Johnson & poor nutrition was postulated as the cause (Stirrat, ). McIlwee, ). We made no attempt to quantify dietary hy- Haematocrit can increase in response to haemoconcentra- pogeous fungi intake or assess the nutritional quality of the tion but eastern bettongs at the source locations and the re- diet at the source habitat or reintroduction sites. However, introduction sites had free access to water, making bettongs were collected from agricultural land and disturbed haemoconcentration secondary to dehydration unlikely. and fragmented habitat in Tasmania; sites which may have Additionally, total protein, albumin and haematocrit all in- represented suboptimal habitat and afforded poorer quality crease in dehydrated individuals, and in this study neither diets. Alternatively, lower population densities and lack of total protein nor albumin values increased significantly competition at the reintroduction sites may have resulted post-reintroduction. in greater resource availability, improved nutrition and We interpreted the neutrophilia, lymphocytosis and hence higher body weights for reintroduced bettongs. monocytosis observed in bettongs at translocation as a Comparisons of haematological and biochemical vari- physiological leucocytosis in response to the prolonged per- ables at translocation and post-reintroduction revealed iod of confinement and transportation prior to sampling significant differences in a range of variables. Some species- (Stockham & Scott, ). The higher white blood cell, neu- specific variability in the response of haematological and trophil and monocyte count observed in the Tidbinbilla biochemical parameters to changing environmental condi- population compared with the Mulligans Flat population tions has been found in other macropodoids (Ealey & Main, may reflect variable levels of physiological stress at the two ; Shield, ; Algar et al., ; Stirrat, ; Pacioni sites. Eastern bettongs at Tidbinbilla have higher faecal cor- et al., ; Robert & Schwanz, ). Of particular interest ticosteroid metabolites (W. Batson, unpubl. data), which in eastern bettongs were increases in haematocrit, haemo- could support the possibility of an unknown environmental globin, red cell count, creatinine and triglycerides post- stressor. reintroduction. These variables were positively correlated Creatinine is influenced by muscle mass and renal func- with weight (rather than reintroduction per se), suggesting tion (Stockham & Scott, ). We attributed the increased they may be useful measures of nutritional status in eastern creatinine in bettongs post-reintroduction to the greater bettongs. body mass of the animals rather than dehydration. Haemoglobin concentration in eastern bettongs post- Reduced creatinine concentrations have been observed in reintroduction was comparable to reference values reported malnourished white-tailed deer Odocoileus virginianus for the closely related woylie Bettongia penicillata in and in tammar wallabies under nutritional stress

(Delgiudice et al., ; Robert & Schwanz, ). In con- host–parasite factors that resulted in five ectoparasites fail- trast, creatinine concentrations were not significantly differ- ing to persist on reintroduced bettongs are unknown but ent, despite significant changes in body mass, in agile could include environmental factors, changes to host physi- wallabies experiencing seasonal nutritional fluctuations ology and immune function, host density, transmission ef- (Stirrat, ). Urea was also significantly higher in bettongs ficiency or the lack of intermediate hosts (MacLeod et al., post-reintroduction, and previous studies of wallaroos, west- ). Twelve of the bettongs were treated with ivermectin – ern grey kangaroos, agile wallabies and tammar wallabies ( μgkg ) subcutaneously at translocation to reduce (but have demonstrated a relationship between poor-quality not eliminate) gastrointestinal nematode burdens (Portas diets or reduced protein intake and low urea concentrations et al., ). However, the ectoparasites that failed to persist (Ealey & Main, ; Algar et al., ; Stirrat, ; Robert & on these individuals also failed to persist on untreated Schwanz, ). However, urea was not positively related to conspecifics at translocation, making treatment with iver- weight in this study, and its usefulness as a potential measure mectin an unlikely explanation for their disappearance. of diet quality or protein intake in eastern bettongs is unclear. Furthermore, two of the ectoparasites that failed to persist An association between triglycerides and kidney fat, a were fleas against which ivermectin is ineffective. traditional measure of body condition in ungulate species, Of the four novel parasites detected post-reintroduction, has been demonstrated in Iberian wild goats Capra pyrenaica Paraheterodoxus erinaceus has been reported previously (Serrano et al., ). In macropodoids, triglycerides have from long-nosed potoroos Potorous tridactylus in been positively correlated with body condition index in Tasmania; Hetrodoxus ualabati has been reported from tammar wallabies but not in agile wallabies (Stirrat, ; the swamp wallaby Wallabia bicolor in Victoria, New Robert & Schwanz, ). The positive correlation between South Wales and Queensland; Eutrombicula macropus has triglycerides and body weight in this study suggests that tri- been reported from macropodoids in the Northern glycerides may be a useful indicator of body condition in Territory, Queensland, Victoria and South Australia; and eastern bettongs. Guntheria shareli has been reported from the red-legged Somewhat unexpectedly, given the observed changes in pademelon Thylogale stigmatica and the bush rat Rattus fus- other parameters, neither total protein nor albumin differed cipes in north Queensland (von Kéler, ; Domrow & significantly in eastern bettongs post-reintroduction. How- Lester, ; Portas et al., ). Sympatric macropodoids ever, globulins were significantly higher post-reintroduction were the most likely source of Paraheterodoxus erinaceus but were not positively correlated with weight. Total protein and Eutrombicula macropus; the macropodoid host species and albumin have been used as indicators of body condition for these parasites are present in the Australian Capital in ungulates (Bahnak et al., ; Caldeira et al., ). Albu- Territory, including a population of long-nosed potoroos min is positively correlated with body condition index in tam- at Tidbinbilla Nature Reserve. Of the two previously re- mar wallabies (Robert & Schwanz, ) and total protein and ported hosts for Guntheria shareli only the bush rat occurs albumin are associated with protein intake in agile wallabies in the Australian Capital Territory, and thus represents the (Stirrat, ). The relationship between total proteins, globu- most likely source of this ectoparasite. However, identifica- lins and albumin and body weight in eastern bettongs requires tion was based solely on morphological features of the scu- further investigation. tum, which may have been damaged in preparation of skin Creatine kinase and aspartate amino transferase values scrapings, hence our designation G. cf. shareli. Similarly, were approximately - and four-fold lower, respectively, variation in the distribution of setae on females of the post-reintroduction. These differences can be explained by Heterodoxus species recovered compared with that de- the prolonged period of confinement and transport (up to  scribed in the published key for the genus (von Kéler, hours) prior to sampling at translocation compared with the ) may or may not represent normal anatomical vari- relatively short period of confinement in traps for sampling ation, hence our designation H. cf. ualabati. The swamp post-reintroduction. The initial values probably reflect wallaby, sympatric at both reintroduction sites, is the most exertional myopathy, and the values obtained post- likely source of this louse species. reintroduction probably reflect more normal values for Serum-virus neutralization assays were used to detect this species (Portas et al., ). Post-reintroduction creatine antibodies against MaHV- and MaHV- (alphaherpes- kinase values are comparable with those obtained for another viruses) in serum samples, whereas PCR was used to detect potoroid marsupial, Gilbert’spotorooPotorous gilbertii any herpesvirus DNA (including the gammaherpesvirus, (Vaughan et al., ). Significant changes in a range of PotHV-) in swab samples. The changing antibody and other biochemical parameters, including electrolytes, are PCR status observed in a small number of bettongs is con- less readily explained and are of unknown clinical sistent with observations in other species, where herpesvirus significance. shedding has been detected intermittently following period- Parasite species are frequently lost when host species ic reactivation from latency (Roizman & Pellett, ). colonize a new environment (Torchin et al., ). The Similarly, one individual was seropositive at translocation

but seronegative post-reintroduction. Serum antibodies to (ethics protocol A/). Funding was provided by the herpesviruses generally persist over time but antibody levels Australian Capital Territory Government and an Australian can decrease in some individuals and fall below detectable Research Council Linkage Grant (LP). ADM was levels, particularly in the absence of reinfection or reactiva- supported by an Australian Research Council Future tion (Van Der Poel et al., ; Kaashoek et al., ; Fellowship (FT). We thank the Tasmanian Whitley, ). Reactivation of latent herpesvirus infections Department of Primary Industries, Parks, Water and often occurs during periods of host immune-compromise or Environment for assistance with the original reintroduction. stress; a study in Australian marsupials identified poor body We acknowledge Don Fletcher, Andrea Reiss, Peter Holz, condition score as a risk factor for herpesvirus shedding Kate Grarock, Elyce Fraser, Ani Kunz, David Dobroszczyk, (Stalder et al., ). The higher body weights and changes Scott Ryan, Kym Birgan, Helen Crisp, Claire Wimpenny, to haematological and biochemical parameters suggestive of Mel Snape, Nicola Munro and Jenny Newport for logistical improved nutritional status could therefore help to explain support. We thank Saul Cunningham, Iain Gordon, Daniel why these viruses were not detected post-reintroduction. Iglesias, Margaret Kitchin and Sue McIntyre for advice and However, further studies are required to investigate this hy- support. We acknowledge Mulligans Flat–Goorooyaroo pothesis, particularly as the rate of herpesvirus infection de- Woodland Experiment and Mulligans Flat Woodland tected at translocation was low. Sanctuary for supporting the translocation. In conclusion, we found that the body weight of eastern bettongs increased significantly post-reintroduction but was not significantly different between reintroduction sites or References between the sexes in response to reintroduction. A range ALGAR, D., ARNOLD, G.W. & GRASSIA,A.() Effects of nitrogen of haematological and biochemical parameters changed and season on western grey kangaroo hematology. The Journal of post-reintroduction but there were few differences between Wildlife Management, , –. reintroduction sites or between the sexes in response to re- BAHNAK, B.R., HOLLAND, J.C., VERME, L.J. & OZOGA, J.J. () introduction. Specifically, haemoglobin, red cell count, Seasonal and nutritional effects on serum nitrogen constituents in  – haematocrit, creatinine and triglycerides increased and white-tailed deer. The Journal of Wildlife Management, , . BATSON, W., FLETCHER, D., PORTAS, T., CRISP, H., RYAN, S., were positively related to weight, providing potentially use- WIMPENNY, C. et al. (in press) Reintroduction of eastern bettong to ful proxies for assessing the nutritional status of eastern bet- a critically endangered woodland habitat in the Australian Capital tongs, and hence the suitability of their habitat. Given that Territory, Australia. IUCN Global Re-introduction Perspectives all individuals studied survived post-reintroduction, we Series.  were unable to establish if any of the variables might be use- BATSON, W.G., GORDON, I.J., FLETCHER, D.B. & MANNING,A.( ) The effect of pre-release captivity on post-release performance in ful predictors of survival. Of the infectious diseases and reintroduced eastern bettongs Bettongia gaimardi. Oryx, http://dx. parasites considered, none impacted overall translocation doi.org/./S. success or individual survival. However, ongoing monitor- CABEZAS, S., CALVETE,C.&MORENO,S.() Survival of ing is recommended, as these diseases and parasites could be translocated wild rabbits: importance of habitat, physiological and  – expected to be of greater importance in suboptimal habitats. immune condition. Animal Conservation, , . CALDEIRA, R.M., BELO, A.T., SANTOS, C.C., VAZQUES, M.I. & The health of eastern bettongs appeared to improve follow- PORTUGAL, A.V. () The effect of long-term feed restriction and ing reintroduction from Tasmania to the Australian Capital over-nutrition on body condition score, blood metabolites and Territory. This study demonstrates the value of comprehen- hormonal profiles in ewes. Small Ruminant Research, , –. sive and ongoing health evaluation during reintroduction CLARIDGE, A.W., SEEBECK,J.&ROSE, R.W. () Bettongs, Potoroos programmes. By assessing these health parameters we and the Musky Rat-kangaroo. CSIRO Publishing, Collingwood, Australia. were able to record the physiological response of eastern CLARKE, J., WARREN, K., CALVER, M., DE TORES, P., MILLS,J.& bettongs to reintroduction, and gain additional information ROBERTSON,I.() Hematologic and serum biochemical not attained with standard measures of reintroduction suc- reference ranges and an assessment of exposure to infectious cess, such as survival and fecundity. We recommend that diseases prior to translocation of the threatened western ringtail comprehensive and ongoing health evaluations be incorpo- possum (Pseudocheirus occidentalis). Journal of Wildlife Diseases,  – rated into future reintroductions of the eastern bettong and , . 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VON KÉLER,S.() A revision of the Australasian Boopiidae Biographical sketches (Insecta: Phthiraptera), with notes on the Trimenoponidae. Australian Journal of Zoology Supplementary TIMOTHY PORTAS is a wildlife veterinarian whose work encompasses Series, , –. reintroduction programmes and diseases of free-ranging wildlife. ROSS WEBSTER, K.N., HILL, N.J., BURNETT,L.&DEANE, E.M. () CUNNINGHAM is a statistician who works in the field of conservation Ectoparasite infestation patterns, haematology and serum and landscape ecology. DAVID SPRATT is a parasitologist who studies biochemistry of urban-dwelling common brushtail possums. host–parasite relationships in Australian fauna. JOANNE DEVLIN’s re- Wildlife Biology, , –. search focuses on the pathogenesis and epidemiology of viral diseases WHITLEY, R.J. () Herpes simplex viruses. In Fields Virology, th in wildlife and domestic animals. PETER HOLZ is a wildlife veterinar- edition (eds D.M. Knipe & P.M. Howley), pp. –. Lippincott ian whose research includes Australian marsupials and bats. WILLIAM Williams & Wilkins, Philadelphia, USA. BATSON specializes in reintroduction biology, including the tactical ’ WORK, T.M., KLAVITTER, J.L., REYNOLDS, M.H. & BLEHERT,D. manipulation of applied processes. JANE OWENS s work includes the () Avian botulism: a case study in translocated endangered epidemiology of infectious disease in wild and domestic animals. Laysan ducks (Anas laysanensis) on Midway Atoll. Journal of ADRIAN MANNING conducts research in the field of landscape ecol- Wildlife Diseases, , –. ogy, conservation biology and restoration ecology.