Health impacts of poor water quality on an endangered shorebird breeding programme in Aotearoa New Zealand.

CASE HISTORY: Two clusters of mortality among endangered tuturuatu/tchuriwat'/shore plover (Thinornis novaeseelandiae) have occurred at captive breeding facilities around New Zealand in recent years. In the first, four chicks died at Pukaha National Wildlife Centre (Mount Bruce, NZ) in February 2016, and in the second five adult birds at the Cape Sanctuary (Cape Kidnappers, NZ) died in 2022. CLINICAL FINDINGS: In 2016, four chicks were noted to become weak, have increased vocalisations and closed eyes prior to death. The remaining chicks were treated for 5 days with amoxycillin/clavulanate orally twice daily. Water containers and brooders were cleaned and disinfected with chlorhexidine. No further mortality was seen.In the 2022 cluster, three adult breeding birds died acutely and five others showed inappetence, weight loss and diarrhoea approximately 10 days after heavy rains flooded the local river. The five birds were treated with amoxycillin/clavulanate orally twice daily and oral fluids for 5 days. Two birds died and three survived. No breeding occurred in the aviaries in the following season. PATHOLOGICAL FINDINGS: In 2016, the chicks showed pulmonary changes ranging from congestion and oedema to heterophilic inflammation consistent with septicaemia.In 2022, the adult birds showed proliferation of bacteria in the distal small intestine associated with mucosal ulceration and heterophilic infiltration. Acid-fast staining of the caecal contents in one bird showed organisms consistent with Cryptosporidium spp. LABORATORY FINDINGS: Aerobic bacterial cultures of the lung and liver of two affected chicks carried out in 2016 showed heavy growth of Plesiomonas shigelloides. The same organism was cultured from water trays and holding tanks containing water boatmen (Sigara arguta) on which the chicks were fed.In 2022, cultures from the livers of three dead birds each showed a mixed bacterial growth with differing dominant organisms (Aeromonas sobria, Hafnia alvei, Citrobacter freundii and an Enterococcus sp.). PCR and sequencing confirmed Cryptosporidium parvum in the caecum of one bird. Fresh faeces from 24 breeding birds from the captive breeding facilities were negative by PCR for Cryptosporidium spp.The captive breeding facilities obtain water for the aviaries and aquatic invertebrates to feed to the chicks from local freshwater sources. Water quality testing at the Cape Sanctuary revealed concentrations of faecal indicator bacteria in excess of safe drinking water guidelines, with peaks following heavy rainfall. CLINICAL RELEVANCE: Fluctuations in water quality associated with mammalian faecal bacteria can adversely affect bird health and impact on captive rearing of endangered wildlife.

Cardiac morphology of North Island brown kiwi (Apteryx mantelli).

AIMS: To investigate the cardiac anatomy of North Island brown kiwi (Apteryx mantelli) through heart morphometric parameters measured at post-mortem examination. METHODS: Morphometric cardiac parameters were established at post-mortem examination of 20 North Island brown kiwi. Birds were classified by gender and age (chicks vs. adults). Measurements included: body mass, heart mass, sternal length, midpoint thickness of left ventricular free wall, midpoint thickness of right ventricular free wall and ratios of heart mass to body mass, left ventricular length to sternal length, right ventricular length to sternal length, length of left ventricle to right ventricle, interventricular septal thickness relative to the sternal length and interventricular septal thickness relative to the left ventricular length. Unadjusted estimates of the median difference and their 95% CI were then reported at each age and sex for all the cardiac morphometric parameters and their ratios. RESULTS: The small sample size led to wide 95% CI for the median difference between gender and age for the cardiac morphometric measurements. Nevertheless, between adult female and male kiwi, the estimated population median differences for heart mass (2.2 (95% CI = -2.9-5.6) g), length (1.2 (95% CI = -2.2-5.6) mm), width (6.1 (95% CI = -1.0-8.2) mm), left ventricular free wall length (5.5 (95% CI = -0.5-8.8) mm) and right ventricular free wall length (2.6 (95% CI = -3.7-6.9) mm) were established. In adult North Island brown kiwi, the heart mass is 0.8 (95% CI = 0.7-0.8)% of the body mass. CONCLUSIONS: The precision of the differences noted in heart measurements recorded between male and female kiwi at each age was limited by the low sample size available for this study. This led to wide CI and an inability to adjust differences observed for gender by differences in other confounders such as body size. With this caveat, there is weak evidence that adult female kiwi have a larger heart size and mass than the adult males. CLINICAL RELEVANCE: These results can be used to improve the diagnosis of cardiac disease in kiwi at post-mortem examination and aid in interpretation of the results of echocardiography in live birds for the antemortem diagnosis of cardiac disorders.

A retrospective study of native wild birds and reptiles admitted to three New Zealand wildlife hospitals due to predation by cats.

AIMS: To investigate the types of wildlife affected and outcomes of hospitalisation for native wild birds and reptiles admitted to three wildlife hospitals in New Zealand following predation by cats. METHODS: We reviewed hospital records of three wildlife hospitals (Wildbase Hospital (WBH) in Palmerston North; The Wildlife Hospital - Dunedin (DWH); and The Nest Te Kohanga (TNTK) in Wellington) for cases of native species with a submission history or diagnosis of predation by cats over the period 2006-2022. Frequency analysis and cross-tabulation of the data were carried out using the factors wildlife hospital, animal order, species, and case outcome. RESULTS: Native wildlife (n = 639) presented following predation by cats to the three wildlife hospitals comprised 598 (93.6%) birds from 31 species and 41 (6.4%) reptiles from eight species. The mortality rate of these patients combined was 61.8% (395/639), with the other 38.2% (244/639) being released from hospital to rehabilitation facilities, released to the wild, or, for a small number, put into permanent captive placements. The most common species of birds admitted due to predation by cats were tui (Prosthemadera novaeseelandiae; n = 173), silvereyes (Zosterops lateralis; n = 142), sacred kingfishers (Todiramphus sanctus; n = 100) and kereru (Hemiphaga novaeseelandiae; n = 70). There was an overall trend to increasing numbers of wildlife admitted for predation by cats over the period of this review, but the different operating periods of the three hospitals should be considered. Each of the three hospitals saw a different mix of species, with Wellington's TNTK seeing the widest diversity of species affected. CONCLUSIONS: Predation by cats was a common cause of native birds and reptiles being admitted to all three wildlife hospitals and a diversity of urban and rural wildlife were affected. CLINICAL RELEVANCE: Predation by cats causes significant mortality in wildlife patients admitted to wildlife hospitals despite best current treatment. Cat management strategies in Aotearoa New Zealand should consider the welfare and health of wildlife.

Morphological characterisation of a novel Eimeria sp. parasite in South Island takahe (Porphyrio hochstetteri).

AIMS: A novel Eimeria sp. is described from the endangered South Island takahe (Porphyrio hochstetteri). METHODS: Faeces were collected from South Island takahe in managed populations across two geographic locations within New Zealand, linked by extensive bird translocation. Faecal flotations were performed and oocysts collected were sporulated in sterile water at 25°C for 5 days. Morphometric measurements including length:width (L:W) ratios of oocysts and sporocysts were made of 65 oocysts using light microscopy under oil immersion. A retrospective histological review of archived post-mortem material from takahe was performed. RESULTS: Oocysts were ovoid (mean L:W 1.2 (SD 0.1)) with mean dimensions of 18.0 (SD 0.8) × 15.2 (SD 0.7) µm, and walls 0.9 (SD 0.1) µm thick at the equator. A micropyle was absent and the oocyst residuum was present as a granular spherical mass. Four sporocysts were present and were elongate ovoid in shape (mean L:W ratio 1.5), with mean dimensions of 9.7 (SD 1.1) × 6.3 (SD 0.7) µm and each containing two sporozoites with anterior and posterior refractile bodies. This morphological description is distinct from other Eimeria spp. described in hosts of the family Rallidae. The retrospective review of tissue was hindered by the poor quality of the preserved gut tissue, but coccidial organisms were able to be identified in the caudal ileum and caecum of a single bird. CONCLUSIONS: A novel Eimeria sp. was identified in the faeces of takahe. We propose this species be named Eimeria takahe. CLINICAL RELEVANCE: Conservation of the endangered South Island takahe is reliant on human management of populations under varying degrees of captive management. An understanding of diseases affecting this host, including parasites and their identification as unique to the takahe or introduced from interactions with other hosts, is essential to the maintenance of overall bird health and informs management decisions in this recovery programme.Abbreviations: L:W: Length:width.

Presence and shedding of Chlamydia psittaci in waterfowl in a rehabilitation facility and in the wild in New Zealand.

AIMS: To determine the frequency of Chlamydia psittaci infection, shedding dynamics of C. psittaci, and C. psittaci genotype diversity in waterfowl temporarily resident in a rehabilitation facility and in mallards in the wild. METHODS: Conjunctival-choanal-cloacal swabs were collected from apparently healthy captive wild mallards (Anas platyrhynchos; n = 114) and paradise shelducks (Tadorna variegata; n = 10) temporarily housed at a waterfowl breeding and rehabilitation facility (Wellington, NZ) and from wild mallards in Palmerston North (n = 50), and Southland (n = 50). DNA extracted from the swabs was analysed using quantitative PCR (qPCR) high-resolution melt curve (HRM) analysis, targeting the ompA gene of C. psittaci. RESULTS: Of the captive waterfowl, 39/114 (34%) mallards and 6/10 (60%) paradise shelducks were positive for C. psittaci as were 24/100 (24%) wild mallards. All wild mallards and paradise shelducks carried only C. psittaci genotype C. In captive wild mallards, genotypes A and C, and a mixed infection of both genotypes were found. Captive wild mallards and paradise shelducks were found to be shedding 4 to 5 × 104 and 1 × 105 to 4 × 105 copies of C. psittaci DNA per swab, respectively, with wild mallards shedding 4-677 DNA copies/swab. CONCLUSIONS: Based on qPCR-HRM analysis, a high proportion of wild mallards were infected with C. psittaci but these birds were shedding only a small amount of bacterial DNA. The proportion of sampled ducks that were infected and the extent of bacterial shedding were higher in the birds in a wildlife rehabilitation facility. The major C. psittaci genotype found in the mallards and paradise shelducks was genotype C. This is the first detection of C. psittaci genotype A and co-infection of genotype A and C in ducks. CLINICAL RELEVANCE: These results indicate that mallards are a reservoir of C. psittaci and therefore may pose a zoonotic risk to people involved in duck hunting, wildlife care and recreational duck feeding. Mallards may also pose a transmission risk to native birds, especially in captive facilities and this has conservation implications for the management of endangered native birds.

Surgical repair of a meningoencephalocoele in a kakapo (Strigops habroptilus).

CASE HISTORY: A kakapo (Strigops habroptilus) chick hatched on an off-shore island of New Zealand with a small white mass protruding through the cranial skin of the head. The chick's growth followed a normal pattern for kakapo but at 3 weeks of age the cranium mass was non-reducible and fixed in place and the chick was removed from the island for diagnostic imaging and hand-rearing. CLINICAL FINDINGS AND TREATMENT: A computed tomography (CT) examination revealed a full-thickness circular defect in the central cranium with suspected herniation of brain and dura. Surgery was performed at 37 days of age, and the herniated dura was dissected from the open fontanelle. Attempts to reduce the herniated tissue were unsuccessful, so the herniated dura and cortex were clamped and resected. The dura was closed and the periosteum of the skull was scarified and monofilament polypropylene mesh was secured tautly over the fontanelle. The mesh graft was infused with autologous bone marrow harvested from the ulna in an attempt to stimulate osteogenesis in the mesh repair. The skin flap was then closed. Post-operative recovery and healing were without complication. A CT examination 4 weeks after surgery showed no recurrence of the hernia, and a composite of mesh and scar over the open fontanelle which had reduced in diameter. The chick was released back onto an off-shore island with a radio transmitter and it continues to be monitored regularly. PATHOLOGICAL FINDINGS: The tissue resected at surgery consisted of a cylindrical core of cerebral parenchyma overlain by a mildly hyperplastic epidermis, and large amounts of oedematous fibrovascular tissue arising from the leptomeninges. DIAGNOSIS: Rostral parietal meningoencephalocoele. CLINICAL RELEVANCE: This is the first report of successful surgical resolution of a meningoencephalocoele in any bird. Techniques from human neurosurgery were adapted for the unique anatomical features of the avian skull. The risks of the procedure included increased intra-cranial pressure resulting in anaesthetic complications or death, cerebrospinal fluid leakage, meningitis or recurrence of the meningoencephalocoele. In the longer term, there was a risk of developmental deficits in cognition or behaviour. None of these complications eventuated in the short to medium term, probably due to the small size of the meningoencephalocoele.

Prevalence and characterisation of wounds in sheep attributed to attacks by kea (Nestor notabilis) on high country farms in New Zealand.

Aims: To characterise and classify wounds in sheep suspected to have been caused by attacks by kea (Nestor notabilis) (kea strike), and to report the prevalence of these wounds on five high country farms in the South Island of New Zealand.Methods: Data were collected from farms between 28 August 2012 and 20 September 2013. Sheep were examined opportunistically immediately after shearing for signs of wounds caused by kea. The age and sex of sheep were also recorded. Wounds were measured and characterised as recent, healing, or healed, and the estimated true prevalence was calculated for each farm.Results: Injuries consistent with kea strike wounds were identified in 70/13,978 (0.5%) sheep examined. The estimated true prevalence varied between farms, from 0 (95% CI = 0-0.16) to 1.25 (95% CI = 0.97-1.61)%. Of the 76 wounds identified, 61 (80%) were located in the lumbar region, and 74 (97%) consisted of full-thickness ulceration of the skin, one showed evidence of injury to muscle and one to bone. The median length of the 63 wounds measured was 6 (min 1, max 23.5) cm, and 10/63 (13%) were categorised as recently healed, 47/63 (62%) as healing, and 17/63 (22%) as recent wounds.Conclusions: The results of this study show that kea strike on sheep was occurring at a low prevalence on the high country farms surveyed. The wounds identified were survivable, but the welfare impact of kea strike on sheep should be considered in balance with the conservation status of kea. There was clear variation in the prevalence of wounds attributed to kea strike between the farms but we were not able to identify the risk factors contributing to these differences. Future studies of kea strike should examine variables such as altitude, local kea density and distribution, and differences in kea strike management and husbandry practices, and should include high country farms without a history of kea strike.

Thermal burns of the spectacle associated with supplementary heating in native New Zealand geckos.

Case history: Gradual onset of ocular opacity was observed in three gold-striped geckos (Woodworthia chrysosiretica), and five Pacific geckos (Dactylocnemis pacificus) held in two adjacent terrariums in a zoological institution located in the North Island of New Zealand. Ultraviolet light and heat had been provided for the previous 3-4 years by a fluorescent bulb, but in the last 4 weeks of winter a ceramic heat bulb had been added, situated 10 cm above the upper mesh of the cageClinical findings: All eight geckos presented with mostly bilateral lesions of varying severity confined to the central or upper quadrant of the spectacles. These lesions ranged from variable areas of opacity within the stroma of the spectacle to similarly distributed ulcers of the surface epithelium of both spectacles. The spectacle lesions in the Pacific geckos responded well to treatment with topical combined antimicrobial therapy, within 18-29 days. The gold-striped geckos suffered complications including dysecdysis, severe spectacle ulceration and perforation, mycotic spectaculitis, and widespread mycotic dermatitis resulting in death or leading to euthanasia.Pathological findings: In the three gold-striped geckos, there were extensive areas of deep ulceration and replacement of the spectacle with a thick serocellular crust containing large numbers of fungal elements. The affected areas of the stroma were expanded by large deposits of proteinaceous and mucinous material, pyknotic cellular debris and moderate numbers of heterophils and macrophages as well as infiltrating fungal hyphae.Diagnosis: Mycotic spectaculitis with ulceration and perforation, and disseminated mycotic dermatitis likely secondary to thermal burns.Clinical relevance: This is the first report of thermal burns of the spectacle in any reptile. There was species variation in the burn severity with gold-striped geckos showing more severe lesions, possibly due to a mix of behavioural and anatomical factors. The thermal burns to the spectacles in three cases were complicated by delayed healing, perforation, dysecdysis and severe mycotic infection.

Untargeted metabolic profiling of dogs with a suspected toxic mitochondrial myopathy using liquid chromatography-mass spectrometry.

'Go Slow myopathy' (GSM) is a suspected toxic myopathy in dogs that primarily occurs in the North Island of New Zealand, and affected dogs usually have a history of consuming meat, offal or bones from wild pigs (including previously frozen and/or cooked meat). Previous epidemiological and pathological studies on GSM have demonstrated that changes in mitochondrial structure and function are most likely caused by an environmental toxin that dogs are exposed to through the ingestion of wild pig. The disease has clinical, histological and biochemical similarities to poisoning in people and animals from the plant Ageratina altissima (white snakeroot). Aqueous and lipid extracts were prepared from liver samples of 24 clinically normal dogs and 15 dogs with GSM for untargeted liquid chromatography-mass spectrometry. Group-wise comparisons of mass spectral data revealed 38 features that were significantly different (FDR<0.05) between normal dogs and those with GSM in aqueous extracts, and 316 significantly different features in lipid extracts. No definitive cause of the myopathy was identified, but alkaloids derived from several plant species were among the possible identities of features that were more abundant in liver samples from affected dogs compared to normal dogs. Mass spectral data also revealed that dogs with GSM have reduced hepatic phospholipid and sphingolipid concentrations relative to normal dogs. In addition, affected dogs had changes in the abundance of kynurenic acid, various dicarboxylic acids and N-acetylated branch chain amino acids, suggestive of mitochondrial dysfunction.

Causes of adult mortality in two populations of New Zealand sea lions (Phocarctos hookeri).

The New Zealand sea lion is an endangered species endemic to New Zealand. While causes of death are well described for pups of this species, mortality in adults is poorly characterised. This study investigated causes of death in 136 New Zealand sea lions in two different populations: a major breeding site on remote, uninhabited Enderby Island in the sub-Antarctic, and a slowly increasing recolonising population on the inhabited mainland. For animals with at least a partial diagnostic investigation (n = 112), the most frequently diagnosed causes of mortality were infectious disease (41/112; 37%), particularly tuberculosis due to M. pinnipedii (20/112; 18%), and conspecific trauma (27/112; 24%). Anthropogenic trauma was an important cause of death in mainland sea lions (9/33; 26%). Deliberate anthropogenic mortality has previously been identified as the greatest potential threat to population recovery for mainland sea lions, and as human and pinniped populations increase, managing interactions between these species will become increasingly important.

Wildlife diseases in New Zealand: recent findings and future challenges.

Our knowledge of diseases in New Zealand wildlife has expanded rapidly in the last two decades. Much of this is due to a greater awareness of disease as a cause of mortality in some of our highly threatened species or as a limiting factor to the successful captive rearing of intensely managed species such as hihi (Notiomystis cincta), kiwi (Apteryx spp.) and kakapo (Strigops habroptilus). An important factor contributing to the increase of our knowledge has been the development of new diagnostic techniques in the fields of molecular biology and immunohistochemistry, particularly for the diagnosis and epidemiology of viral and protozoan diseases. Although New Zealand remains free of serious exotic viruses there has been much work on understanding the taxonomy and epidemiology of local strains of avipox virus and circoviruses. Bacterial diseases such as salmonellosis, erysipelas and tuberculosis have also been closely investigated in wildlife and opportunist mycotic infections such as aspergillosis remain a major problem in many species. Nutritional diseases such as hyperplastic goitre due to iodine deficiency and metabolic bone disease due to Ca:P imbalance have made significant impacts on some captive reared birds, while lead poisoning is a problem in some localities. The increasing use of wildlife translocations to avoid the extinction of threatened species has highlighted the need for improved methods to assess the disease risks inherent in these operations and other intensive conservation management strategies such as creching young animals. We have also become more aware of the likelihood of inbreeding suppression as populations of many species decrease or pass through a genetic bottleneck. Climate change and habitat loss, however, remain the greatest threats to biodiversity and wildlife health worldwide. Temperature changes will affect our wildlife habitats, alter the distribution of disease vectors and wildlife predators, or directly harm threatened species in vulnerable localities.

Lead concentrations in the blood and eggs of backyard laying hens.

AIMS: To investigate the prevalence of lead exposure in hens and eggs from backyard poultry in a sample of Auckland households, the relationship between concentrations of lead in the blood of the hens and in the shells and yolks of eggs from the same household, and to examine associations with measures of hen health, environment and husbandry factors. METHODS: Thirty households participated in the study from August to November 2016, each providing one adult hen for sampling, an egg from the household if available, and completing a questionnaire on hen husbandry. Concentrations of lead in blood were determined using a portable lead analyser. Eggs were analysed for concentrations of lead in the yolk and shell using inductively coupled plasma mass spectrometry after biological digestion with a mixture of nitric and hydrochloric acid. RESULTS: Twenty three of 30 hens (77%) showed evidence of lead exposure, with median concentrations of lead in blood of 0.77 (min <0.16, max 8.02) µmol/L. All eggs showed evidence of lead exposure, with concentrations of lead in the yolk ranging from 0.003-1.07 mg/kg, and concentrations of lead in the eggshell ranging from <0.1-0.82 mg/kg. A positive correlation existed between concentrations of lead in the blood of a hen and concentrations of lead in egg yolk from the same hen (R2=0.97), and both the yolk (R2=0.58) and shell (R2=0.30) of an egg from her flock. No association was found between concentrations of lead in blood and hen health indices measured in this study. Concentrations of lead in blood were higher in hens from properties with homes built before 1941 than between 1941-1960 (p=0.03), and in hens from properties with weatherboard homes than brick homes (p=0.049). CONCLUSIONS AND CLINICAL RELEVANCE: There was a high prevalence of lead exposure in this sample of Auckland backyard chickens, with the majority of hens being sub-clinically affected. Associations were found between concentrations of lead in the blood of the hens, and properties with homes built before 1941 and clad in weatherboard. Concentrations of lead in over half the egg yolks sampled were at levels sufficient to warrant human health concern. The assessment of concentrations of lead in backyard poultry and eggs intended for human consumption is recommended to protect human and bird health.

An epidemiological investigation of an idiopathic myopathy in hunting dogs in New Zealand.

AIM: To conduct an epidemiological investigation of an idiopathic myopathy, known as "Go Slow" (GSM), which was initially recognised in dogs used for pig hunting. A secondary aim was to describe the hunting activities, diet and health of dogs used for pig hunting in New Zealand. METHODS: A retrospective cohort study was conducted between June 2014-June 2017. Cases of GSM in dogs were diagnosed by veterinarians using a combination of clinical history, physical examination findings, serum biochemistry and/or skeletal muscle histology. A telephone interview was conducted with the owner or primary veterinarian to provide information regarding the dog's diet and exercise over the 7 days preceding the onset of clinical signs. In August 2015, a separate online survey of owners of dogs used for pig hunting was conducted to characterise the normal hunting activities, diet and health of these dogs. RESULTS: A total of 86 cases of GSM were recruited, of which 58 (67%) were pig hunting dogs, 16 (19%) pet dogs and 12 (14%) working farm dogs. Cases were most commonly reported in the upper North Island, and 65 (76 (95% CI=67-85)%) were from the Northland region. Processed commercial dog food had been fed to 93 (95% CI=88-98)% of affected dogs. Ingestion of raw, frozen or cooked wild pig in the preceding week was reported for 76 (88 (95% CI=82-95)%) dogs with the myopathy. In the survey of owners of healthy pig hunting dogs, 203 eligible responses were received; pig hunting was reported to most commonly occur in Northland (20.2%), Waikato (22.3%) and Bay of Plenty (23.2%) regions. Commercial dog food was fed to 172 (85 (95% CI=80-90)%) of the dogs included in this survey, and 55 (27 (95% CI=20-33)%) had eaten wild pig in the preceding week. The most common reported health problem in pig hunting dogs was traumatic wounds. CONCLUSIONS: Cases of GSM were most commonly recognised in dogs used for pig hunting, but also occurred in pet and working farm dogs. The disease was most frequently reported in the upper North Island of New Zealand and ingestion of wild pig was a consistent feature in cases of this myopathy. CLINICAL RELEVANCE: To minimise the risk of dogs developing this myopathy, it would seem prudent to avoid feeding any tissues from wild pigs to dogs in areas where the disease is known to occur.

A preliminary survey of Chlamydia psittaci genotypes from native and introduced birds in New Zealand.

AIM: To describe the Chlamydia psittaci genotypes in samples from native and introduced birds from New Zealand by analysis of the sequence variation of the ompA gene. METHODS: DNA was extracted from samples collected from a non-random sample of birds; either swabs from live asymptomatic birds or birds with clinical signs, or formalin-fixed, paraffin-embedded (FFPE) samples from historical post-mortem cases. The presence of C. psittaci in all samples had been confirmed using a quantitative PCR assay. The C. psittaci ompA gene was amplified and sequenced from samples from 26 native and introduced infected birds comprising 12 different species. These sequences were compared to published available C. psittaci genotypes. RESULTS: Genotypes A and C of C. psittaci were identified in the samples. Genotype A was identified in samples from nine birds, including various native and introduced species. Genotype C was identified in samples from 16 different waterfowl species, and a mixed infection of both genotypes was found in a kaka (Nestor meridionalis). In native birds, C. psittaci infection was confirmed in seven new host species. CONCLUSIONS AND CLINICAL RELEVANCE: Two genotypes (A and C) of C. psittaci were found in samples from a wider range of both native and introduced species of birds in New Zealand than previously reported. Both genotypes have been globally associated with significant disease in birds and humans. These initial results suggest the host range of C. psittaci in New Zealand birds is under-reported. However, the prevalence of C. psittaci infection in New Zealand, and the associated impact on avian and public health, remains to be determined. There are biosecurity implications associated with the importation of birds to New Zealand if there is a limited diversity of C. psittaci genotypes present.

Cases of mortality in little penguins (Eudyptula minor) in New Zealand associated with avian malaria.

CASE HISTORY A little penguin (Eudyptula minor) of wild origin, in captivity at Wellington Zoo, became inappetent and lethargic in March 2013. Despite supportive care in the zoo's wildlife hospital, the bird died within 24 hours. CLINICAL FINDINGS Weight loss, dehydration, pale mucous membranes, weakness, increased respiratory effort and biliverdinuria were apparent on physical examination. Microscopic evaluation of blood smears revealed intra-erythrocytic stages of Plasmodium spp. and a regenerative reticulocytosis in the absence of anaemia. PATHOLOGICAL FINDINGS Post-mortem findings included reduced body condition, dehydration, pulmonary congestion and oedema, hepatomegaly, splenomegaly, hydropericardium and subcutaneous oedema. Histopathological findings included protozoal organisms in sections of lung, liver and spleen. A marked, diffuse, sub-acute interstitial histiocytic pneumonia was present. Accumulation of haemosiderin was noted in the Kupffer cells of the liver and in histiocytic-type cells in the spleen. MOLECULAR TESTING DNA was extracted from frozen portions of the liver. Nested PCR results and DNA sequencing confirmed infection of the deceased little penguin with Plasmodium (Huffia) elongatum lineage GRW06. DIAGNOSIS Avian malaria due to Plasmodium (Huffia) elongatum GRW06 RETROSPECTIVE INVESTIGATION A retrospective analysis of 294 little penguin cases in the Massey University post-mortem database revealed three other potential avian malaria cases. Analysis of archived tissues using a nested PCR for Plasmodium spp. followed by DNA sequencing revealed that a little penguin which died at Auckland Zoo was infected with P. elongatum GRW06 and two wild little penguins found dead on New Zealand beaches were infected with P. relictum SGS1 and Plasmodium. sp. lineage LINN1. Therefore, the overall frequency of deaths in little penguins associated with avian malaria was 4/295 (1.36%). CLINICAL RELEVANCE Our results suggest that avian malaria is associated with sporadic mortality in New Zealand's little penguins both in the wild and in captivity, but there is no evidence of mass mortality events due to Plasmodium spp. infection.

Use of a real-time PCR to explore the intensity of Plasmodium spp. infections in native, endemic and introduced New Zealand birds.

Avian malaria, caused by Plasmodium spp., is an emerging disease in New Zealand (NZ). To detect Plasmodium spp. infection and quantify parasite load in NZ birds, a real-time polymerase chain reaction (PCR) (qPCR) protocol was used and compared with a nested PCR (nPCR) assay. A total of 202 blood samples from 14 bird species with known nPCR results were tested. The qPCR prevalences for introduced, native and endemic species groups were 70, 11 and 21%, respectively, with a sensitivity and specificity of 96·7 and 98%, respectively, for the qPCR, while a sensitivity and specificity of 80·9 and 85·4% were determined for the nPCR. The qPCR appeared to be more sensitive in detecting lower levels of parasitaemia. The mean parasite load was significantly higher in introduced bird species (2245 parasites per 10 000 erythrocytes) compared with endemic species (31·5 parasites per 10 000 erythrocytes). In NZ robins (Petroica longipes), a significantly lower packed cell volume was found in birds that were positive for Plasmodium spp. compared with birds that were negative. Our data suggest that introduced bird species, such as blackbirds (Turdus merula), have a higher tolerance for circulating parasite stages of Plasmodium spp., indicating that introduced species are an important reservoir of avian malaria due to a high infection prevalence and parasite load.

A retrospective survey into the presence of Plasmodium spp. and Toxoplasma gondii in archived tissue samples from New Zealand raptors: New Zealand falcons (Falco novaeseelandiae), Australasian harriers (Circus approximans) and moreporks (Ninox novaeseelandiae).

Human colonisation of New Zealand has resulted in the introduction of emerging diseases, such as avian malaria and toxoplasmosis, which arrived with their exotic avian and mammalian hosts. Plasmodium spp. and Toxoplasma gondii have a wide host range, and several species of endemic New Zealand birds have developed a fatal disease following infection with either pathogen. However, no reports of either toxoplasmosis or avian malaria in New Zealand raptors, namely, the New Zealand falcons (Falco novaeseelandiae), Australasian harriers (Circus approximans) and moreporks (Ninox novaeseelandiae) exist in the literature. Therefore, this study was designed to determine if these two pathogens are present in these raptors through a retrospective analysis of archived tissue samples. Detection and isolate identification of these pathogens was determined using established histological and molecular techniques. All three species of New Zealand raptors tested positive for the presence of Plasmodium spp. (10/117; 8.5%) and an atypical genotype of T. gondii (9/117; 7.7%). Plasmodium lineages identified include P. elongatum GRW6, P. relictum SGS1, P. relictum PADOM02 and Plasmodium sp. LINN1. Two Australasian harriers and one morepork tested positive for the presence of both Plasmodium spp. and T. gondii. However, the pathogenicity of these organisms to the raptors is unclear as none of the tissues showed histological evidence of clinical disease associated with Plasmodium spp. and T. gondii infections. Thus, these results demonstrate for the first time that these two potential pathogens are present in New Zealand's raptors; however, further research is required to determine the prevalence and pathogenicity of these organisms among the living populations of these birds in the country.

Efficacy of anthelmintic treatment in captive-reared black stilts (Himantopus novaezelandiae) released to the wild.

AIMS: To assess the efficacy of praziquantel (PZQ) administered to control helminths in captive-reared critically endangered black stilts (Himantopus novaezelandiae) before their release into the wild and determine the effect on their survival shortly after release. METHODS: Of 77 captive black stilts, 37 were treated with PZQ prior to release into the wild in South Canterbury, New Zealand, in August and September 2007. Faecal helminth egg counts (FEC) were measured before and after anthelmintic treatment, and before and after release to the wild using modified faecal flotation and sedimentation methods. In addition, total helminth counts were determined in 11 of the birds that died following release, as well as four captive and 11 other wild stilts. RESULTS: The efficacy of PZQ against trematodes was 92% and against Capillaria spp. was 34%. No trematode or Capillaria spp. eggs were detected in treated birds 1 day after treatment, but FEC increased 3-5 days after treatment. There were no differences in the total helminth counts for trematodes, cestodes or Capillaria spp. in control or treated birds (p>0.2). Survival did not differ between treatment groups in the August or September releases (p>0.4). Of control and treated birds, 11/17 (65%) and 8/14 (57%) survived to 31 days, respectively, in the August release, and 16/20 (80%) and 10/15 (67%) survived to 84 days, respectively, in the September release. CONCLUSION: Overall, the results suggest that PZQ treatment may be an unnecessary cost and the risks of producing anthelmintic resistance, injuring the birds during processing or producing an unnecessary stress response at the time of release could exceed any likely benefits. CLINICAL RELEVANCE: It is recommended that pre-release anthelmintic treatment of black stilts should be used only if indicated by health screening. Any treatment should incorporate annual efficacy testing to monitor the emergence of anthelmintic resistance.