QUARANTINE AND HEALTH SCREENING
4.1 Quarantine
Quarantine is important in ex situ-breeding programs, both at establishment with the arrival of founder stock and for inter-institutional transfers. A period of 30 days has been recommended as a minimum standard duration (Woodford 2000); however, the DRA will generally guide the duration and level of isolation required for a given CT.
Where possible and ideally, animals in ex situ-breeding programs specifically for CTs should be held in permanent quarantine.During the quarantine period animals typically undergo anaesthesia for comprehensive health evaluation, disease screening and application of a unique identifier such as a microchip transponder or ear tag. For CTs comprising direct wild to wild translocations, a period of in situ or ex situ quarantine maybe implemented. The duration of quarantine maybe determined by the length of time required for diagnostic testing to be completed or by the incubation period of specific diseases of concern. In addition to its obvious role as a disease mitigation strategy, quarantine also provides a period of acclimation before release, which may be particularly important in assisted colonisation programs. Given that quarantine has been associated with elevated FGMCs, for example in eastern bettongs (Batson et al. 2017), welfare considerations are also important in determining the length of the quarantine period.
4.2 Health and disease screening
Baseline health and disease data are unknown for many native mammal species. Performing comprehensive health evaluations serves several functions. It allows for baseline health and disease parameters to be established, which in turn can be used for the conservation management of the species. This data can then be used to inform disease risk analysis and mitigation strategies and forms the basis for longitudinal monitoring. Finally, health evaluations can be used as a basis for selecting candidates of an appropriate health status for CTs, thus maximising both conservation and welfare outcomes.
Health screening of conspecifics and sympatric species at the donor and recipient sites has been advocated to gain a broader understanding of potential disease effects. However, in reality logistical and resource constraints often dictate that health screening is limited to individuals under direct consideration for CTs. Sympatric macropods were screened for a range of parasites and pathogens before reintroduction of brush-tailed rock-wallabies in the Grampians National Park, Vic. but there are few other examples of targeted surveillance of native mammal species at the recipient site before CT (Schultz et al. 2011).
Standardised assessment protocols and accurate record keeping are vital to ensure consistency throughout the process and for accurate comparisons during experimental evaluation of CT techniques. Anaesthesia is recommended for performing a complete physical examination and sample collection. Frequently, health and disease screening is carried out in remote locations, often at the point of capture if the donor population is free-ranging, necessitating substantial forward planning to ensure successful field work (Fig. 2.2). Effective biosecurity protocols are also an important component of trapping, handling and sampling procedures to reduce the spread of pathogens and parasites and the possibility of cross-contamination of samples collected for disease screening (Hillman et al. 2016).
Deciding which diseases to screen for in CT programs will be influenced by the outcome of the DRA, the availability of validated testing methodologies for a given pathogen and species, the volume of blood and other biological samples that can be safely collected and logistical and cost limitations. Some disease processes with multifactorial aetiologies may be difficult to detect despite health screening. For example, despite comprehensive pre-release oral cavity examinations in brush-tailed rock-wallabies several reintroduced animals succumbed to macropod progressive periodontal disease within 3 mo of release, suggesting early signs of the disease were not detected during health screening (Schultz et al.
2011).Table 2.2. Selected diseases that may warrant consideration in disease risk analysis or disease investigation for macropod conservation translocations
| Pathogen class | Specific pathogens | Disease screening and investigation testing options |
| Viruses | Macropodid herpesviruses (MaHV-1, MaHV-2, MaHV-3, MaHV-4, MaHV-5) Potoroid herpesviruses (PotHV-1 and an incompletely described herpesvirus from brush-tailed bettongs [Bettongia penicillata]) | Serum - serum-virus neutralisation assay (MaHV-1, MaHV-2 only, although may cross-react with other alphaherpesviruses) Conjunctival, nasal, oropharyngeal and urogenital swabs - panherpesvirus PCR Fresh tissue - virus isolation, PCR |
| Orbiviruses (Wallal, Warrego, Eubenangee serogroups) | Serum - serum-virus neutralisation assay (Wallal and Warrego serogroups only) Formalin-fixed tissue samples - PCR, immunohistochemistry Fresh tissue samples - virus isolation, PCR | |
| Papillomaviruses (including B. penicillata papillomavirus type 1 [BpPV1]) | Fresh or frozen skin samples - PCR | |
| Encephalomyocarditis virus | Serum - serum neutralisation test Tissue samples - virus isolation | |
| Uncharacterised pox viruses | Skin biopsy - histopathology | |
| Bacteria | Bordetella bronchiseptica | Nasal swabs, tracheal wash, bronchoalveolar lavage - bacterial culture |
| Salmonella spp., Campylobacter spp., Yersinia pseudotuberculosis | Tissue samples, faeces - bacterial culture | |
| Treponema spp. - Gilbert's potoroo (Potorous gilbertii) | Urogenital swabs - bacterial culture, dark-field microscopy | |
| Coxiella burnetii | Faecal sample - qPCR Serum - complement fixation test, ELISA | |
| Erysipelothrix rhusiopathiae | Tissue samples - bacterial culture | |
| Leptospira spp. | Serum - microscopic agglutination test, indirect haemagglutination assay, microsphere immunoassay, ELISA Urine - dark-field microscopy, culture, PCR | |
| Mycobacterium spp. (typically, managed animals) | Tissue aspirates, bronchoalveolar lavage samples - acid-fast stains, culture and PCR Tissue samples - histopathology, culture and PCR | |
| Clostridium tetani, C. perfringens | Tissue samples - bacterial culture | |
| Burkoholderia pseudomallei | Whole blood and/or serum - culture using selective media, biochemical tests, latex agglutination assays, PCR Tissue samples - culture using selective media, PCR | |
| Fungi | Cryptococcus spp. (potoroids in particular) | Serum - LCAT, cryptococcal antigen lateral flow assay Cerebrospinal fluid - LCAT Tissue samples - culture, PCR |
| Protozoa | Toxoplasma gondii | Serum - modified agglutination test, indirect fluorescent antibody test, ELISA Fresh/frozen tissue - PCR Formalin-fixed tissue - histopathology, immunohistochemistry, PCR |
| Eimeria spp., Isospora spp. | Faeces - faecal flotation | |
| Neospora caninum | Serology -cELISA and N. caninum-agglutination test Tissue - histopathology, immunohistochemistry, PCR |
| Pathogen class | Specific pathogens | Disease screening and investigation testing options |
| Protozoa (cont.) | Trypanosoma spp. (T. copemani, T. noyesi, T. vegrandis, uncharacterised Trypanosoma spp.) | Blood smears - cytology (low sensitivity) Whole blood - PCR combined with Sanger sequencing |
| Babesia macropus | Blood smears, brain and kidney impression smears - cytology Tissue - histopathology, PCR | |
| Leishmania spp. | PCR on tissue or blood samples Serology - indirect fluorescent antigen test and ELISA | |
| Besnoitia spp. | Nasal smears - cytology Tissue samples - histopathology | |
| Theileria spp. (T. gilberti, T. penicillata, T. brachyuri, T. fuliginosa, uncharacterised Theileria spp.) | Blood smears - light microscopy (low sensitivity) Whole blood - PCR targeting the 18S rRNA gene | |
| Helminths | Various GI nematodes Potentially pathogenic spp. include third- stage larvae of Labiomultiplex spp., Labiosimplex spp. and Rugopharynx rosemariae (eastern [Macropusgiganteus] and western [M. fuliginosus] grey kangaroos only); Strongyloides spp.; Globocephaloides spp.; and Hypodontus macropi | Identification of parasites obtained at necropsy; speciation of nematode eggs presents in faeces is generally not possible |
| Fasciola hepatica | Faeces - faecal sedimentation techniques Identification of parasites obtained at necropsy | |
| Echinococcus granulosa | Pulmonary cysts may be identified radiographically Identification of parasites obtained at necropsy | |
| Angiostrongylus cantonensis | Fresh brain or meninges - microscopy Fixed brain - histopathology | |
| Arthropods | Sarcoptes scabiei | Skin scrapings - microscopy |
| Thadeua spp. | Skin scrapings - microscopy | |
| Ixodes spp., Haemaphysalis spp., Amblyomma spp. | Clinical examination | |
| Non-infectious disease | Exertional myopathy | Blood - biochemistry (CK, AST etc.) Formalin-fixed tissue - histopathology |
Table 2.3.
Selected diseases that may warrant consideration in disease risk analysis or disease investigation for koala (Phascolarctos cinereus) conservation translocations| Pathogen class | Specific pathogens | Disease screening and investigation testing options |
| Viruses | Koala retroviruses (KoRV-A, KoRV-B, KoRV-J, and numerous subtypes) Phascolarctid herpesviruses (PhaHV-1, PhaHV-2) Papillomaviruses | Blood, faeces, tissue - PCR Conjunctival, nasal, oropharyngeal and urogenital swabs - pan-herpesvirus PCR Fresh tissue - virus isolation, PCR Tissue - histopathology, PCR |
| Bacteria | Chlamydia pecorum, C. pneumoniae Coxiella burnetii | Conjunctival and urogenital swabs - PCR Formalin-fixed tissue - immunohistochemistry Faecal sample - qPCR Serum - complement fixation test, ELISA |
| Fungi | Cryptococcus gatti, C. neoformans | Serum - LCAT, cryptococcal antigen lateral flow assay Cerebrospinal fluid - LCAT Tissue samples - culture, PCR |
| Protozoa | Trypanosoma irwini, T. gilletti, T. copemani, T. vergrandis | Blood smears - cytology (low sensitivity) Whole blood - PCR combined with Sanger sequencing |
| Helminths | Durikainema phascolarcti | Tissue samples - histopathology |
| Arthropods | Koalachirus perkinsi Sarcoptes scabiei | Fur pluck samples - microscopy Skin scrapings - microscopy |
| Non-infectious disease | Oxalate nephrosis | Urine - urinalysis Ultrasonography |
Fig. 2.2. Field set up for health evaluation of bush rats (Rattus fuscipes) during a reintroduction program. Photo: Larry Vogelnest
Table 2.4. Selected diseases that may warrant consideration in disease risk analysis or disease investigation for wombat conservation translocations
| Pathogen class | Specific pathogens | Disease screening and investigation testing options |
| Viruses | Vombatid herpesviruses (VoHV-1, VoHV-2, VoHV-3) Encephalomyocarditis virus | Conjunctival, nasal, oropharyngeal and urogenital swabs - pan herpesvirus PCR Fresh tissue - virus isolation, PCR Serum - serum neutralisation test Tissue samples - virus isolation |
| Bacteria | Leptospira spp. Including L. interrogans serovar Pomona | Serum - microscopic agglutination test, indirect haemagglutination assay, microsphere immunoassay, ELISA Urine - dark-field microscopy, culture, PCR |
| Fungi | Emmonsia parva | Formalin-fixed tissue - histopathology, PCR Fresh tissue - fungal culture, PCR |
| Protozoa | Toxoplasma gondii Eimeria arundeli, E. wombati, E. ursini Trypanosoma spp. | Serum - modified agglutination test, indirect fluorescent antibody test, ELISA Fresh/frozen tissue - PCR Formalin-fixed tissue - histopathology, immunohistochemistry, PCR Faeces - faecal flotation Blood smears - cytology (low sensitivity) Whole blood - PCR combined with Sanger sequencing |
| Helminths | Echinococcus granulosus Baylisascaris tasmaniensis Strongyloides speari Fasciola hepatica | Typically diagnosed at necropsy Pulmonary cysts may be identified radiographically Identification of parasites obtained at necropsy Faeces - Baermann technique Faeces - faecal sedimentation techniques Identification of parasites obtained at necropsy |
| Arthropods | Sarcoptes scabiei Aponomma auruginans, Ixodes spp., Echidnophaga spp. | Skin scrapings - PCR or microscopic examination Clinical examination |
Table 2.5. Selected diseases that may warrant consideration in disease risk analysis or disease investigation for dasyurid conservation translocations
| Pathogen class | Specific pathogens | Disease screening and investigation testing options |
| Viruses | Herpesviruses (DaHV-1, DaHV-2, DaHV-3 and an undescribed gammaherpesvirus from eastern quolls [Dasyurus viverrinus]) | Conjunctival, nasal, oropharyngeal and urogenital swabs - panherpesvirus PCR |
| Bacteria | Bacillus cereus - managed dibblers (Parantechinus apicalis) | Faeces - bacterial culture |
| Salmonella spp. including S. mississippi, S. potsdam, S. freemantle | Faeces - bacterial culture | |
| Campylobacter jejuni | Faeces - bacterial culture | |
| Mycobacterium spp. (cutaneous disease in managed animals) | Swabs and fresh tissue - mycobacterial culture, acid-fast staining, PCR | |
| Leptospira spp. including L. borgpetersenii serovar Javanica, L. weilli serovar Cellodoni | Serum - microscopic agglutination test, indirect haemagglutination assay, microsphere immunoassay, ELISA Urine - dark-field microscopy, culture, PCR | |
| Coxiella burnetii | Faecal sample -qPCR Serum - complement fixation test, ELISA | |
| Fungi | Cryptococcus spp. | Serum - LCAT, cryptococcal antigen lateral flow assay Cerebrospinal fluid - LCAT Tissue samples - culture, PCR |
| Protozoa | Sarcocystis mucosa | Formalin-fixed tissue - histopathology |
| Haemogregarina dasyuri, Hepatazoon dasyuri | Blood smear - microscopy Whole blood - PCR | |
| Toxoplasma gondii | Serum - modified agglutination test, indirect fluorescent antibody test, ELISA Fresh/frozen tissue - PCR Formalin-fixed tissue - histopathology, immunohistochemistry, PCR | |
| Trypanosoma copemani | Blood smears - cytology (low sensitivity) Whole blood - PCR combined with Sanger sequencing | |
| Giardia spp. | Faeces - wet preparation, centrifugal faecal flotation, immunofluorescence microscopy, PCR | |
| Babesia spp. | Blood smears - cytology Tissue - histopathology, PCR | |
| Helminths | Trichinella pseudospiralis (Tas. only) | Formalin-fixed tissue - histopathology |
| Baylisascaris tasmaniensis | Faeces - faecal flotation | |
| Ophidascaris robertsi | Identification of parasites obtained at necropsy Formalin-fixed tissue - histopathology | |
| Cercopithifilaria johnstoni | Identification of parasites obtained at necropsy Formalin-fixed tissue - histopathology | |
| Arthropods | Ixodes spp., Pygiopsylla hoplia, Uropsylla tasmanica | Clinical examination |
| Dasyurochirus major, Demodex spp., Myocoptes spp. | Skin scrapes, fur pluck - microscopy | |
| Neoplasia | Devil facial tumour disease | Clinical examination Formalin-fixed tissue - histopathology, immunohistochemistry |
Table 2.6. Selected diseases that may warrant consideration in disease risk analysis or disease investigation for possum and glider conservation translocations
| Pathogen class | Specific pathogens | Disease screening and investigation testing options |
| Viruses | Wobbly possum disease - common brush-tailed possum (Trichosurus vulpecula); unknown if a nidovirus is the aetiological agent in Australia | Tissue samples - RT-qPCR |
| Papillomavirus | Fresh or frozen skin samples - PCR | |
| Uncharacterised pox viruses | Skin biopsy - histopathology | |
| Brush-tailed possum adenovirus (PoAdV-1), described from possums in NZ | Intestinal contents - PCR | |
| Gammaherpesviruses in yellow-bellied glider and Leadbeater's possums | Ocular, cloacal, nasal oropharyngeal swabs - universal herpesvirus PCR | |
| Bacteria | Coxiella burnetii | Faecal sample - qPCR Serum - complement fixation test, ELISA |
| Clostridium piliforme | Tissue samples - bacterial culture, histopathology and silver staining | |
| Mycobacterium ulcerans | Tissue samples - histopathology, culture and PCR Faecal samples - PCR | |
| Yersinia pseudotuberculosis | Tissue samples - bacterial culture | |
| Lepstospira spp. including L. interrogans serovar Balcanica | Serum - microscopic agglutination test, indirect haemagglutination assay, microsphere immunoassay, ELISA Urine - dark-field microscopy, culture, PCR | |
| Francisella tularensis ssp. holarctica biovar japonica | Liver - culture, direct fluorescent antigen testing, PCR | |
| Staphylococcus aureus - implicated in exudative dermatitis of common brush-tailed possums (T. vulpecula) and swollen paw syndrome of eastern ring-tailed (Pseudocheirusperegrinus), although both diseases likely have a multifactorial aetiology | Direct swabs - bacterial culture | |
| Listeria monocytogenes | Tissue samples - bacterial culture | |
| Fungi | Emmonsia crescens | Formalin-fixed tissue - histopathology, PCR Fresh tissue - fungal culture, PCR |
| Protozoa | Toxoplasma gondii | Serum - modified agglutination test, indirect fluorescent antibody test, ELISA Fresh/frozen tissue - PCR Formalin-fixed tissue - histopathology, immunohistochemistry, PCR |
| Cryptosporidium parvum | Faeces - acid-fast staining, direct fluorescent antibody test | |
| Giardia intestinalis | Faeces - wet preparation, centrifugal faecal flotation, immunofluorescence microscopy, PCR Serum - ELISA | |
| Plasmodium spp. - Leadbeater's possum (Gymnobelideus leadbeateri) | Blood smear - microscopy Whole blood - PCR | |
| Helminths | Angiostrongylus cantonensis | Fresh brain or meninges - microscopy Fixed brain - histopathology |
| Anoplotaenia dasyuri, Bertiella trichosuri, Marsupostrongylus minesi, Parastrongyloides trichosuri | Identification of parasites obtained at necropsy | |
| Trichinellapseudospiralis (Tasmania only) | Formalin-fixed tissue - histopathology | |
| Arthropods | Trichosurolaelaps crassipes, Echidnophagia myrmecobii, Notoedres muris, Ixodes spp. | Clinical examination or skin scrape samples - microscopy |
Table 2.7. Selected diseases that may warrant consideration in disease risk analysis or disease investigation for bandicoot and greater bilby (Macrotis lagotis) conservation translocations
| Pathogen class | Specific pathogens | Disease screening and investigation testing options |
| Viruses | Bandicoot papillomatosis carcinomatosis viruses (BPCV) BPCV-1 - western barred bandicoot (Perameles bougainville) BPCV-2 - southern brown bandicoot (Isoodon obesulus) | Skin swabs, fresh or frozen biopsy samples - PCR Formalin-fixed biopsies - histopathology and in situ hybridisation |
| Peramelid herpesvirus-1 | Conjunctival, nasal, oropharyngeal and urogenital swabs - pan-herpesvirus PCR | |
| Encephalomyocarditis virus | Serum - serum neutralisation test Tissue samples - virus isolation | |
| Bacteria | Coxiella burnetii | Faecal sample - qPCR Serum - complement fixation test, ELISA |
| Erysipelothrix rhusiopathiae | Tissue samples - bacterial culture | |
| Chlamydia spp. | Conjunctival and urogenital swabs - PCR Formalin-fixed tissue - immunohistochemistry | |
| Mycobacterium spp. M. ulcerans | Direct swabs, tissue samples or faeces - culture and/or PCR | |
| Salmonella spp., Campylobacter spp. | Faeces - bacterial culture | |
| Leptospira spp. L. interrogans serovar Perameles, L. weilii serovar Topaz | Serum - microscopic agglutination test, indirect haemagglutination assay, ELISA Urine - dark-field microscopy, culture, PCR | |
| Fungi | Cryptococcus spp. | Serum - LCAT, cryptococcal antigen lateral flow assay Cerebrospinal fluid - LCAT Tissue samples - culture, PCR |
| Protozoa | Toxoplasma gondii | bgcolor=white>Serum - modified agglutination test, indirect fluorescent antibody test, ELISA|
| Cryptosporidium spp. C. muris - managed greater bilbies (Macrotis lagotis) | Faeces - acid-fast staining, direct fluorescent antibody test | |
| Giardia spp. G. peramelis | Faeces - wet preparation, centrifugal faecal flotation, immunofluorescence microscopy, PCR Serum - ELISA | |
| Eimeria spp. E. kanyana - western barred bandicoot | Faeces - faecal flotation | |
| Hepatozoon spp. | Blood smear - microscopy Whole blood - PCR | |
| Babesia spp. | Blood smears - cytology Tissue - histopathology, PCR | |
| Trypanosoma spp. | Blood smears - cytology Whole blood - PCR | |
| Helminths | Asymmetracantha tasmaniensis, Gnathostoma spp., Capillaria spp., Parastrongyloides australis, Paremelostrongylus spp., Physalaptera peramelis, Moniliformis semoni | Faeces - zinc sulfate faecal flotation to identify capillarid and spirurid ova Recovery of helminths at necropsy for identification |
| Pathogen class | Specific pathogens | Disease screening and investigation testing options |
| Helminths (cont.) | Cercopithifilaria johnstoni | Identification of parasites obtained at necropsy Formalin-fixed tissue - histopathology |
| Arthropods | Ixodes spp., Pygiopsylla hoplia, Pygiopsyllazethi, Haemolaelaps marsupialis, Sarcoptes scabiei, Schoutedenichia emphyla, Stephanocircus daysuri, Petauralges spp., Ornithonyssus bacoti | Clinical examination or skin scrape samples - microscopy |
5.