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Background Koalas are infected with the koala retrovirus (KoRV) that exists as exogenous or endogenous viruses. KoRV is genetically diverse with co-infection with up to ten envelope subtypes (A-J) possible; KoRV-A is the prototype endogenous form. KoRV-B, first found in a small number of koalas with an increased leukemia prevalence at one US zoo, has been associated with other cancers and increased chlamydial disease. To better understand the molecular epidemiology of KoRV variants and the effect of increased viral loads (VLs) on transmissibility and pathogenicity we developed subtype-specific quantitative PCR (qPCR) assays and tested blood and tissue samples from koalas at US zoos (n = 78), two Australian zoos (n = 27) and wild-caught (n = 21) in Australia. We analyzed PCR results with available clinical, demographic, and pedigree data. Results All koalas were KoRV-A-infected. A small number of koalas (10.3%) at one US zoo were also infected with non-A subtypes, while a higher non-A subtype prevalence (59.3%) was found in koalas at Australian zoos. Wild koalas from one location were only infected with KoRV-A. We observed a significant association of infection and plasma VLs of non-A subtypes in koalas that died of leukemia/lymphoma and other neoplasias and report cancer diagnoses in KoRV-A-positive animals. Infection and VLs of non-A subtypes was not associated with age or sex. Transmission of non-A subtypes occurred from dam-to-offspring and likely following adult-to-adult contact, but associations with contact type were not evaluated. Brief antiretroviral treatment of one leukemic koala infected with high plasma levels of KoRV-A, -B, and -F did not affect VL or disease progression. Conclusions Our results show a significant association of non-A KoRV infection and plasma VLs with leukemia and other cancers. Although we confirm dam-to-offspring transmission of these variants, we also show other routes are possible. Our validated qPCR assays will be useful to further understand KoRV epidemiology and its zoonotic transmission potential for humans exposed to koalas because KoRV can infect human cells.

The Mojave Desert tortoise (Gopherus agassizii), federally listed as threatened, has suffered habitat loss and fragmentation due to human activities. Upper respiratory tract disease (URTD), a documented health threat to desert tortoises, has been detected at the Large-Scale Translocation Study Site (LSTS) in southwestern Nevada, US, a fenced recipient site for translocated animals. Our study aimed to 1) estimate prevalence of URTD and Mycoplasma infection at LSTS and three nearby unfenced sites; 2) assess whether Mycoplasma infection status was associated with developing clinical signs of URTD; and 3) determine whether such an association differed between LSTS and unfenced areas. We sampled 421 tortoises in 2016 to describe the current status of these populations. We evaluated three clinical signs of URTD (nasal discharge, ocular discharge, nasal erosions) and determined individual infection status for Mycoplasma agassizii and Mycoplasma testudineum by quantitative PCR and enzyme-linked immunosorbent assay. In 2016, LSTS had the highest prevalence of M. agassizii (25.0%; 33/132), M. testudineum (3.0%; 4/132), and URTD clinical signs (18.9%; 25/132). Controlling for other factors, clinical sign(s) were positively associated with M. agassizii infection (odds ratio [OR]=7.7, P=0.001), and this effect was similar among study sites (P>0.99). There was no association with M. testudineum status (P=0.360). Of the 196 tortoises in a longitudinal comparison of 2011-14 with 2016, an estimated 3.2% converted from M. agassizii-negative to positive during the study period, and incidence was greater at LSTS (P=0.002). Conversion to positive M. agassizii status was associated with increased incidence of clinical signs in subsequent years (OR=11.1, P=0.018). While M. agassizii and URTD are present outside the LSTS, there is a possibility that incidence of Mycoplasma infection and URTD would increase outside LSTS if these populations were to reconnect. Population-level significance of this risk appears low, and any risk must be evaluated against the potential long-term benefits to population viability through increased connectivity.

Over the past 30 years, the California condor (Gymnogyps californianus) population has rebounded from 22 individuals to over 200 birds living in the wild. Historical impacts to the population have been largely anthropogenic. In this study, we explore mortality and cause of death data from condors that died during the years 2010-2014 and compare these to mortality data described by Rideout et al. in 2012, covering the years 1992-2009. In addition, morphologic and genetic analysis of the contents of the upper gastrointestinal (GI) tract was performed on the 2010-2014 condor mortalities to determine animal origins of the last meal eaten. The maximum population at risk within this time frame was 329 birds. During this time, 88 condors died and underwent post-mortem examination, and 41 birds were lost to tracking efforts and presumed dead (crude mortality rate of 39%; 129/329). A cause of death was determined for 66 of the 88 necropsied birds. Lead toxicosis remained a significant negative factor in condor population recovery, being related to the deaths of 37 adult and juvenile condors (proportional mortality rate 56%). Compared to condors succumbing to other causes of death, cattle were less often part of the last meal of lead-intoxicated condors. Based on these data, continued efforts to mitigate the impact of lead on California condors should be pursued.

This study combined a social network analysis and whole-genome sequencing (WGS) to test for general patterns of contagious spread of a mycobacterial infection for which pathways of disease acquisition are not well understood. Our population included 275 cases diagnosed with avian mycobacteriosis that were nested in a source population of 16,430 birds at San Diego Zoo Wildlife Alliance facilities from 1992 through mid-2014. Mycobacteria species were determined using conventional methods and whole genome sequencing (WGS). Mycobacterium avium avium (MAA) and Mycobacterium genavense were the most common species of mycobacteria identified and were present in different proportions across bird taxa. A social network for the birds was constructed from the source population to identify directly and indirectly connected cases during time periods relevant to disease transmission. Associations between network connectivity and genetic similarity of mycobacteria (as determined by clusters of genotypes separated by few single nucleotide polymorphisms, or SNPs) were then evaluated in observed and randomly generated network permutations. Findings showed that some genotypes clustered along pathways of bird connectivity, while others were dispersed throughout the network. The proportion of directly connected birds having a similar mycobacterial genotype was 0.36 and significant (pM. genavense. Evaluations of SNP distributions also showed genotypes of MAA were more related in connected birds than expected by chance; however, no significant patterns of genetic relatedness were identified for M. genavense, although data were sparse. Integrating the WGS analysis of mycobacteria with a social network analysis of their host birds revealed significant genetic clustering along pathways of connectivity, namely for MAA. These findings are consistent with a contagious process occurring in some, but not all, case clusters.

Disease transmission can be identified in a social network from the structural patterns of contact. However, it is difficult to separate contagious processes from those driven by homophily, and multiple pathways of transmission or inexact information on the timing of infection can obscure the detection of true transmission events. Here, we analyze the dynamic social network of a large, and near-complete population of 16,430 zoo birds tracked daily over 22 years to test a novel "friends-of-friends"strategy for detecting contagion in a social network. The results show that cases of avian mycobacteriosis were significantly clustered among pairs of birds that had been in direct contact. However, since these clusters might result due to correlated traits or a shared environment, we also analyzed pairs of birds that had never been in direct contact but were indirectly connected in the network via other birds. The disease was also significantly clustered among these friends of friends and a reverse-time placebo test shows that homophily could not be causing the clustering. These results provide empirical evidence that at least some avian mycobacteriosis infections are transmitted between birds, and provide new methods for detecting contagious processes in large-scale global network structures with indirect contacts, even when transmission pathways, timing of cases, or etiologic agents are unknown. Published version

The goal of this study was to evaluate longitudinal patterns of avian mycobacteriosis spread through a social network. Specifically, we wanted to determine whether the patterns of connectivity over time can predict future infections, and whether this pattern can distinguish between different sources of infection. The study population included 13,409 individuals nested in a larger population of birds that were closely monitored in zoological facilities for over 22 years (1992-2014). A retrospective cohort study design and social network connectivity were used to estimate the association between exposure to an infected bird, and development of mycobacteriosis. Avian mycobacteriosis was diagnosed from histopathology and network connectivity was defined by enclosure histories over discrete time periods. Single-variable and multivariable longitudinal, mixed effects logistic regression models examined whether exposure to infected birds, both directly- and indirectly-connected, was associated with development of mycobacteriosis at the next time step. Our adjusted model showed an increased odds of developing mycobacteriosis (odds ratio = 2.15; 95 % CI: 1.48-3.12; p < 0.001) for birds that were directly exposed (i.e., housed in the same aviary) to another infected bird, compared to those with no exposure. Exposure to a positive, indirectly-connected bird at a previous time step was independently associated with an increased risk of mycobacteriosis (odds ratio = 1.56; 95 % CI: 1.11-2.19). This association persisted in adjusted models even when the indirect contacts were housed in distinctly different aviaries and never had contact with the subject of interest or its environment. Adjusted, risk-stratified models further characterized the type of exposure that increased the risk of avian mycobacteriosis. Birds that were exposed in small aviaries were more likely to develop mycobacteriosis than those exposed in larger aviaries and those with no exposure. The lesion distribution and species of the contact (same species versus different species) were also significant predictors of disease risk. Some findings were sensitive to model variation of time divisions and initiation time. Our study shows avian mycobacteriosis spread through the social network in quantifiable and discernable patterns. We provide empirical evidence that a contagious process drives some of the observed infection, but we also show low transmissibility based on sustained patterns of low incidence over time even when large groups of birds are exposed. Targeted risk mitigation efforts based on the characteristics of the exposure may be effective at reducing risk of avian mycobacteriosis while enhancing population sustainability.

OBJECTIVE To determine the incidence of and risk factors for clinical feline herpesvirus (FHV) infection in zoo-housed cheetahs and determine whether dam infection was associated with offspring infection. DESIGN Retrospective cohort study. ANIMALS 144 cheetah cubs born in 6 zoos from 1988 through 2007. PROCEDURES Data were extracted from the health records of cheetahs and their dams to identify incident cases of clinical FHV infection and estimate incidence from birth to 18 months of age. Univariate and multivariable Cox proportional hazards models, controlling for correlations among cheetahs with the same dam, were used to identify risk factors for incident FHV infection. RESULTS Cumulative incidence of FHV infection in cheetah cubs was 35% (50/144). No significant association between dam and offspring infection was identified in any model. Factors identified as significant through multivariable analysis varied by age group. For cheetahs up to 3 months of age, the most important predictor of FHV infection was having a dam that had received a preparturition FHV vaccine regimen that included a modified-live virus vaccine versus a dam that had received no preparturition vaccine. Other risk factors included being from a small litter, being born to a primiparous dam, and male sex. CONCLUSIONS AND CLINICAL RELEVANCE This study provided the first population-level characterization of the incidence of and risk factors for FHV infection in cheetahs, and findings confirmed the importance of this disease. Recognition that clinical FHV infection in the dam was not a significant predictor of disease in cubs and identification of other significant factors have implications for disease management.

A 3-year-old female Bruce's green pigeon (

Gammaherpesvirus infections are ubiquitous in captive and free-ranging ruminants and are associated with a variety of clinical diseases ranging from subclinical or mild inflammatory syndromes to fatal diseases such as malignant catarrhal fever. Gammaherpesvirus infections have been fully characterized in only a few ruminant species, and the overall diversity, host range, and biologic effects of most are not known. This study investigated the presence and host distribution of gammaherpesviruses in ruminant species at two facilities, the San Diego Zoo and San Diego Zoo Safari Park. We tested antemortem (blood, nasal or oropharyngeal swabs) or postmortem (internal organs) samples from 715 healthy or diseased ruminants representing 96 species and subspecies, using a consensus-based herpesvirus PCR for a segment of the DNA polymerase (DPOL) gene. Among the 715 animals tested, 161 (22.5%) were PCR and sequencing positive for herpesvirus, while only 11 (6.83%) of the PCR positive animals showed clinical signs of malignant catarrhal fever. Forty-four DPOL genotypes were identified of which only 10 have been reported in GenBank. The data describe viral diversity within species and individuals, identify host ranges of potential new viruses, and address the proclivity and consequences of interspecies transmission during management practices in zoological parks. The discovery of new viruses with wide host ranges and presence of co-infection within individual animals also suggest that the evolutionary processes influencing Gammaherpesvirus diversity are more complex than previously recognized.

The preshipment examination, with associated transmissible disease testing, has become standard practice in the movement of animals between zoos. An alternative disease risk-based approach, based on a comprehensive surveillance program including necropsy and preventive medicine examination testing and data, has been in practice since 2006 between the San Diego Zoo and San Diego Zoo Safari Park. A retrospective analysis, evaluating comprehensive necropsy data and preshipment testing over a 5-yr study period, was performed to determine the viability of this model for use with sending animals to other institutions. Animals (607 birds, 704 reptiles and amphibians, and 341 mammals) were shipped to 116 Association of Zoos and Aquariums (AZA)-accredited and 29 non-AZA-accredited institutions. The evaluation showed no evidence of the specific transmissible diseases tested for during the preshipment exam being present within the San Diego Zoo collection. We suggest that a risk-based animal and institution-specific approach to transmissible disease preshipment testing is more cost effective and is in the better interest of animal welfare than the current industry standard of dogmatic preshipment testing.

A histopathologic study of free-ranging hummingbirds found in California, US was performed to identify mortality trends. Tissues from 61 wild hummingbirds representing five native California species collected by the San Diego Zoo from 1996 to 2016 or the Lindsay Wildlife Experience from 2015 to 2017 were histologically examined. Birds were either found dead or moribund at the time of submission or were euthanized due to unresolvable health issues. Long-term rehabilitated birds were excluded from this study. Lesions were sorted by organ, etiology, and gender. The most commonly affected organs were the lung (68%, 40/59), followed by the ingluvies (67%, 34/51) and the liver (54%, 33/61). While some birds had minimal or nonspecific lesions, 23% (14/61) had lesions primarily attributable to trauma, 16% (10/61) had lesions associated with bacteria, fungi, or viruses, 11% (7/61) had parasitic lesions, and 13% (8/61) had multifactorial concurrent processes. Infectious disease lesions included those associated with avian poxvirus, intestinal adenovirus, disseminated aspergillosis, bacterial septicemia, malaria ( Haemoproteus spp.), and mycobacteriosis. The most commonly identified parasitic infection was intestinal cestodiasis, for which there was no significant associated intestinal damage, although the large size of these cestodes may have affected digestion. The incidence of traumatic lesions did not vary significantly by sex, age, species, or sampling location. Other significant findings not related to disease or trauma, but not previously documented, were histologic evidence of a gallbladder and the presence of aortic ossification. Our study reported mortality trends at a population level for free-ranging hummingbirds found in California and identified the presence of intestinal adenovirus and two anatomic structures not previously described.

Johne’s disease, caused by Mycobacterium avium subsp. paratuberculosis (MAP), is a chronic, progressive bacterial enteritis of ruminants that can cause serious losses in both livestock and exotic species. Infection risk in exotic ruminants is associated with maternal infection status, but the effect of other herdmates on risk of infection has not been reported, to our knowledge. We conducted a retrospective cohort study to evaluate the association between MAP infection status and early-life contact with infected herdmates. The study population included 3,234 individuals representing 128 species at San Diego Zoo Global facilities between 1991 and 2010. Animal movement, health, and pathology records were used to trace enclosure-sharing contacts between members of the study population and any MAP-infected animal. Contact-days were counted by age of the reference animal and the number of unique infected individuals contacted. Herdmate infection status was stratified by stage of infection (180 d prior to diagnosis), age, and whether relevant lesions were found at autopsy. Having an infected herdmate was a strong risk factor for infection (OR = 4.4; 95% CI: 1.9–10.3), and each method of defining herdmate infection status showed significant differences in infection risk. The best predictor was number of contact-days within the first week of life, with a 2-fold increase in risk associated with each doubling in the number of contact-days (OR = 2.1; 95% CI: 1.1–4.0). We conclude that early contact with infected animals is an important predictor of MAP infection risk, although the effect size is smaller than that previously described for maternal infection status.

Although Salmonella spp. infection has been identified in captive and free-ranging rhinoceros, clinical cases in black rhinoceros ( Diceros bicornis ) calves have not been described. This case series describes clinical salmonellosis in four black rhinoceros calves. Two calves developed self-limiting diarrhea, recovering after treatment. The other two cases were fatal. One of the fatal cases had a short clinical course, whereas the other case was protracted, with signs reflecting multiple organ system involvement. In all cases, diagnosis was by fecal culture and/or quantitative polymerase chain reaction. A variable clinical presentation, which is typical for salmonellosis in domestic hoofstock, was a feature of these rhinoceros cases. Similarly, postmortem pathology in black rhinoceros calves was consistent with domestic neonatal ungulates with salmonellosis. Potential predisposing factors for infection were considered to be primiparity of the dam and failure of passive transfer in the calf. The case investigation included attempts to identify the source of infection, which was aided by organism serotyping. In one case, the patient's dam and another conspecific in the facility were shown to be asymptomatic shedders of the organism strain responsible for disease in the calf. Further surveillance of captive rhinoceros Salmonella spp. carrier status is needed to inform screening recommendations for this taxa.

Mycoplasmas are an important cause of upper respiratory tract disease (URTD) in desert tortoises (Gopherus agassizii) and have been a main focus in attempts to mitigate disease-based population declines. Infection risk can vary with an animal's population of origin, making screening tests popular tools for determining infection status in individuals and populations. To provide additional methods for investigating URTD we developed quantitative PCR (qPCR) assays specific for agents causing clinical signs of URTD: Mycoplasma agassizii, Mycoplasma testudineum, and Testudinid herpesvirus 2 (TeHV2) and tested necropsied desert tortoises housed at the Desert Tortoise Conservation Center in Las Vegas, Nevada, USA, as well as wild desert tortoises (n=3), during 2010. Findings were compared with M. agassizii enzyme-linked immunosorbent assay (ELISA) data. Based on qPCR, the prevalence of M. agassizii was 75% (33/44) and the prevalence of TeHV2 was 48% (20/42) in the evaluated population. Both agents were also present in the wild tortoises. Mycoplasma testudineum was not detected. The M. agassizii ELISA and qPCR results did not always agree. More tortoises were positive for M. agassizii by nasal mucosa testing than by nasal flush. Our findings suggest that mycoplasmas are not the only agents of concern and that a single M. agassizii ELISA or nasal flush qPCR alone failed to identify all potentially infected animals in a population. Caution should be exercised in using these tests for disposition decisions.

The recent classification as threatened status of the northern koala (Phascolarctos cinereus) by the Australian Government highlights the importance of the conservation and health management of this iconic Australian marsupial. This case series describes gastrointestinal torsion and intussusception in six northern koalas (three males, three females, 2-11 yr old) at the San Diego Zoo from 1976 to 2012. Two koalas died shortly after presentation. Diagnoses of ileocecal intussusception, resulting from enteritis in one case and cecal torsion in the other, were made at postmortem examination. One koala died 4 days after an exploratory laparotomy, with negative findings, and an acute double colonic intussusception was diagnosed at postmortem examination. Two small intestinal mesenteric torsion and one proximal colon mesenteric torsion cases were successfully corrected surgically. In the case of colonic mesenteric torsion, the koala had recurrent clinical signs 2 wk later, and a second surgery requiring resection and anastomosis of ischemic jejunum was performed, with the koala dying shortly afterward. One koala with small intestinal torsion had a recurrence of torsion 22 mo later and subsequently died. The koala with the second case of small intestinal torsion remains alive 14 mo postsurgical correction. All six koalas presented with signs of colic that included anorexia, lethargy, depression, acute abdominal distension, abdominal stretching, decreased fecal output, open-mouth gasping, or a combination of symptoms. Abdominal radiographs may show stacked gastrointestinal linear gas patterns and contrast stasis. Prevalence of torsion and intussusception is low at this institution (2%), although recurrence in individuals is common (50%) and overall survival is poor (83%), which emphasizes the importance of timely recognition, surgical correction, and postoperative management. While inciting etiologies were unable to be determined in these cases, monitoring generalized gastrointestinal health and differing Eucalyptus sp. effects on individual koala's gastrointestinal function, parasite control, and stress minimization through standardized husbandry practices are likely important.

Many scavenging bird populations have experienced abrupt declines across the globe, and intensive recovery activities have been necessary to sustain several species, including the critically endangered California condor (Gymnogyps californianus). Exposure to lead from lead-based ammunition is widespread in condors and lead toxicosis presents an immediate threat to condor recovery, accounting for the highest proportion of adult mortality. Lead contamination of carcasses across the landscape remains a serious threat to the health and sustainability of scavenging birds, and here we summarize recent evidence for exposure to lead-based ammunition and health implications across many species. California condors and other scavenging species are sensitive indicators of the occurrence of lead contaminated carcasses in the environment. Transdisciplinary science-based approaches have been critical to managing lead exposure in California condors and paving the way for use of non-lead ammunition in California. Similar transdisciplinary approaches are now needed to translate the science informing on this issue and establish education and outreach efforts that focus on concerns brought forth by key stakeholders.

A 3-year-old female Bruce's green pigeon (Treron waalia) was presented with granulomatous inflammation of the cere and underlying tissues with osteomyelitis and bone proliferation of the dorsal premaxilla. Biopsy and culture revealed the presence of Mycobacterium avium-intracellulare complex, and multi-antimicrobial treatment was initiated with clarithromycin, ethambutol, rifabutin, and enrofloxacin. The cere lesion improved and no evidence of systemic granulomas was observed over 4 months of treatment, although leukocytosis and monocytosis persisted. Five months after discontinuation of antibiotic therapy, the white blood cell count had normalized, but distal beak irregularities and partial recurrence of the mass were present. The bird died 15 months after discontinuation of antibiotic therapy and necropsy revealed no evidence of active mycobacteriosis of the beak or cere. This report documents an unusual clinical presentation of mycobacteriosis, in addition to its successful resolution.

The identification of feline herpesvirus (FHV) infected cheetahs (Acinonyx jubatus) and characterization of shedding episodes is difficult due to nonspecific clinical signs and limitations of diagnostic tests. The goals of this study were to develop a case definition for clinical FHV and describe the distribution of signs. Medical records from six different zoologic institutions were reviewed to identify cheetahs with diagnostic test results confirming FHV. Published literature, expert opinion, and results of a multiple correspondence analysis (MCA) were used to develop a clinical case definition based on 69 episodes in FHV laboratory confirmed (LC) cheetahs. Four groups of signs were identified in the MCA: general ocular signs, serious ocular lesions, respiratory disease, and cutaneous lesions. Ocular disease occurred with respiratory signs alone, with skin lesions alone, and with both respiratory signs and skin lesions. Groups that did not occur together were respiratory signs and skin lesions. The resulting case definition included 1) LC cheetahs; and 2) clinically compatible (CC) cheetahs that exhibited a minimum of 7 day's duration of the clinical sign groupings identified in the MCA or the presence of corneal ulcers or keratitis that occurred alone or in concert with other ocular signs and skin lesions. Exclusion criteria were applied. Application of the case definition to the study population identified an additional 78 clinical episodes, which represented 58 CC cheetahs. In total, 28.8% (93/322) of the population was identified as LC or CC. The distribution of identified clinical signs was similar across LC and CC cheetahs. Corneal ulcers and/or keratitis, and skin lesions were more frequently reported in severe episodes; in mild episodes, there were significantly more cheetahs with ocular-only or respiratory-only disease. Our results provide a better understanding of the clinical presentation of FHV, while presenting a standardized case definition that can both contribute to earlier diagnoses and be used for population-level studies.

A free-ranging, young adult, female American white pelican ( Pelecanus erythrorhynchos), found dead on the grounds of the San Diego Zoo Safari Park in Southern California, had severe multifocal to coalescing necrotizing hepatitis and splenitis on postmortem examination. Histologically, within the large areas of necrosis were myriad pleomorphic, 5–20 µm in diameter, protozoal organisms with 1 to multiple nuclei. Ultrastructurally, the organisms were consistent with a trichomonad flagellate. Polymerase chain reaction and sequencing of the small subunit ribosomal RNA gene identified nucleotide sequences with 99% identity to Tetratrichomonas gallinarum, which is a common inhabitant of the intestinal tract of galliform and anseriform birds that has occasionally been associated with disease, including typhlitis and hepatitis. Damage to the cecal mucosa in the pelican from trematodes and secondary bacterial infection could have allowed invasion and systemic dissemination of the organism. Exposure of the pelican to a variety of native and exotic anseriform and galliform birds at the zoological institution could have led to cross-species infection and severe manifestation of disease in a novel host.

Quarantine is designed primarily to prevent the introduction of transmissible diseases to zoological collections. Improvements in preventive medicine, disease eradication, and comprehensive pathology programs call into question current industry quarantine standards. Disease risk analysis was used at the San Diego Zoo (SDZ) and the SDZ Safari Park to eliminate quarantine isolation and transmissible disease testing for animals transferred between the two institutions. To determine if a risk-based approach might be valid between other institutions and SDZ, we reviewed quarantine data for animals arriving at SDZ from 81 Association of Zoos and Aquariums (AZA)-accredited and 124 other sources (e.g., non-AZA-accredited institutions, private breeders, private dealers, governmental bodies) over a 5-yr period (2009-2013). No mammal or herptile failed quarantine due to transmissible diseases of concern. Approximately 2.5% of incoming birds failed quarantine due to transmissible disease; however, all 14 failed individuals were obtained from three nonaccredited sources (private breeders, confiscation). The results of our study suggest that a risk-based approach could be used to minimize or eliminate quarantine for the transfer of animals from institutions with comprehensive disease surveillance programs and/or preshipment testing practices. Quarantine isolation with testing remains an essential defense against introducing transmissible diseases of concern when there is a lack of health knowledge about the animals being received.

Following field observations of wild Agassiz's desert tortoises (Gopherus agassizii) with oral lesions similar to those seen in captive tortoises with herpesvirus infection, we measured the prevalence of antibodies to Testudinid herpesvirus (TeHV) 3 in wild populations of desert tortoises in California. The survey revealed 30.9% antibody prevalence. In 2009 and 2010, two wild adult male desert tortoises, with gross lesions consistent with trauma and puncture wounds, respectively, were necropsied. Tortoise 1 was from the central Mojave Desert and tortoise 2 was from the northeastern Mojave Desert. We extracted DNA from the tongue of tortoise 1 and from the tongue and nasal mucosa of tortoise 2. Sequencing of polymerase chain reaction products of the herpesviral DNA-dependent DNA polymerase gene and the UL39 gene respectively showed 100% nucleotide identity with TeHV2, which was previously detected in an ill captive desert tortoise in California. Although several cases of herpesvirus infection have been described in captive desert tortoises, our findings represent the first conclusive molecular evidence of TeHV2 infection in wild desert tortoises. The serologic findings support cross-reactivity between TeHV2 and TeHV3. Further studies to determine the ecology, prevalence, and clinical significance of this virus in tortoise populations are needed.

Investigation of all embryo and neonatal mortalities is essential for optimizing productivity in artificial incubation and hand rearing programs. Because artificial incubation is a complex process with many variables, thorough and systematic evaluations are necessary to identify potential problems. Every step of the process from egg lay through incubation and hatching should be evaluated in conjunction with comprehensive data on management of the breeding population. The most common sources of significant problems include nutrition and management of the breeding population, insufficient parental incubation prior to artificial incubation, abnormal egg weight loss during incubation, and infections of the yolk sac or umbilicus.

The objective of the present study was to determine whether offspring of dams infected with Mycobacterium avium subsp. paratuberculosis (Map) have an increased risk for Map infection. Antemortem and postmortem disease surveillance data were used to identify positive and test-negative ruminants born at the Zoological Society of San Diego (ZSSD) between 1991 and 2007 and to estimate cumulative lifetime incidence. A matched case-control study, nested within the population, was conducted and conditional logistic regression analyses were used to quantify the association between infection status of offspring and their dams. Cases (infected ruminants, n = 47) were matched to controls (test-negative ruminants, n = 152) by species, birth date, birth enclosure, and follow-up time to control for confounding factors. The overall cumulative lifetime incidence of infection was estimated at 2.2%, but it decreased over time and varied by species. There was a significant association between infection status of offspring and their dams (odds ratio [OR] = 6.8, P < 0.01), which is consistent with studies in domestic livestock species. The association was stronger for animals whose dam was diagnosed within 2 years of their birth (OR = 9.0, P < 0.01) than for animals whose dam was diagnosed more than 2 years after their birth (OR = 6.0, P < 0.01) compared to animals with test-negative dams. For positive animals born to a positive dam, 85.3% of the Map infections were attributable to having a positive dam. For the entire population of ZSSD ruminants, 36.8% of the cases were attributable to having a positive dam. Findings will help guide future management of Map infection in zoo ruminant populations.

Myxozoans are spore-forming, metazoan parasites common in cold-blooded aquatic vertebrates, especially fishes, with alternate life cycle stages developing in invertebrates. We report nine cases of infection in free-flying native and captive exotic ducks (Anseriformes: Anatidae) from locations across the United States and describe the first myxozoan in birds, Myxidium anatidum n. sp. We found developmental stages and mature spores in the bile ducts of a Pekin duck (domesticated Anas platyrhynchos ). Spores are lens-shaped in sutural view, slightly sigmoidal in valvular view, with two polar capsules, and each valve cell has 14–16 longitudinal surface ridges. Spore dimensions are 23.1 μm × 10.8 μm × 11.2 μm. Phylogenetic analysis of the ssrRNA gene revealed closest affinity with Myxidium species described from chelonids (tortoises). Our novel finding broadens the definition of the Myxozoa to include birds as hosts and has implications for understanding myxozoan evolution, and mechanisms of geographical and host range extension. The number of infection records indicates this is not an incidental occurrence, and the detection of such widely dispersed cases suggests more myxozoans in birds will be encountered with increased surveillance of these hosts for pathogens.

Mycobacterium avium subsp. avium and Mycobacterium intracellulare are primary causes of mycobacteriosis in captive birds throughout the world, but little is known about how they are transmitted. To define the local epidemiology of infection, we strain-typed 70 M. avium subsp. avium and 15 M. intracellulare culture isolates obtained over a 4-year period from captive birds. Typing was performed using randomly amplified polymorphic DNA (RAPD) PCR, amplified fragment length polymorphic (AFLP) fragment analyses, and for a subset of isolates, DNA sequencing of a segment of the 16S-23S rRNA internal transcribed spacer region. Six strain clusters comprising 43 M. avium subsp. avium, isolates were identified; 42 isolates had unique typing patterns, including all M. intracellulare isolates. Phylo-geographical analyses using RAPD and AFLP fingerprints and animal confinement histories showed no correlation between housing of infected birds and mycobacterial strain-type, except for two animals. The diversity of M. avium subsp. avium and M. intracellulare isolates and minimal evidence for bird-to-bird transmission suggest that environmental reservoirs may be important sources of infection in captivity.

Ingestion of foreign anthropogenic material, here called junk, has been documented in many avian taxa but has become especially problematic in some Old World Gyps vulture populations within the last 30 years. Here, we document the effects of ingested junk on a reintroduced population of the Critically Endangered California Condor Gymnogyps californianus in southern California, U.S.A. Of 13 breeding attempts to date (2001–2005), only one has resulted in successful fledging. Of nests where either the nest substrate was sifted (n = 10) or nestlings (n = 8) were examined, all but one held junk. Nine nestlings hatched in the wild between 2002 and 2005; of these six died at or near nests and two were removed from the wild for health reasons. Four dead nestlings and two removed from the wild held substantial quantities of junk. In two cases, junk ingestion was determined to be the cause of death. Five of six dead nestlings had elevated hepatic copper levels (150–531 ppm dry weight) although the significance of this, if any, remains undetermined. In comparison with historic condor nests (n = 69), junk was more prevalent and of greater size and quantity in reintroduced condor nests. To date, junk ingestion has been the primary cause of nest failure in the reintroduced condor population and threatens the reestablishment of a viable breeding population in southern California.

© 2015 Elsevier B.V. We investigated threats to the California condor (Gymnogyps californianus), a flagship endangered species, using individual data on survival during a 20. year period of intensive recovery efforts. Over the two decades of reintroductions, condors in California had an estimated median survival time of 7.8. years suggesting that 50% of condors are expected to survive in the wild long enough to contribute to recruitment. In general, annual mortality rates exceeded levels necessary for a stable population; however, mortality declined, reaching levels needed for population stability, during the second decade of re-establishment. Intensive management practices, including utility pole aversion training and clinical interventions to prevent lead-related deaths likely contributed to the decrease in mortality rates. Utility line collision and/or electrocution was an important factor causing mortality over the two decades; though, this threat has largely been mitigated through management and targeted efforts in high-risk areas. In the past, wildfires were not considered a major threat to survival of free-flying condors. However, our analyses suggest that forest fires are significantly linked to the hazard of death, and increased wildfire activity in California highlights this population's vulnerability to catastrophic losses from forest fire. Lead poisoning, which was a major driver in the population's decline, was a leading cause of death accounting for the greatest adult mortality, and lead exposure remains the most significant threat. Recent lead ammunition reduction efforts in the condor range in California hold promise for improving the recovery potential for this population.

© 2015 Straub et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Throughout the world, populations of scavenger birds are declining rapidly with some populations already on the brink of extinction. Much of the current research into the factors contributing to these declines has focused on exposure to drug residues, lead, and other toxins. Despite increased monitoring of these declining populations, little is known about infectious diseases affecting scavenger bird species. To assess potential infectious disease risks to both obligate and facultative scavenger bird species, we performed a serosurvey for eleven potential pathogens in three species of scavenging birds in California: the California condor (Gymnogyps californianus), Turkey vulture (Cathartes aura) and golden eagle (Aquila chrysaetos). California condors were seropositive for avian adenovirus, infectious bronchitis virus, Mycoplasma gallisepticum, avian paramyxovirus-2, West Nile virus (WNV) and Toxoplasma gondii. Golden eagles were seropositive for avian adenovirus, Chlamydophila psittaci and Toxoplasma gondii, and Turkey vultures were seropositive for avian adenovirus, Chlamydophila psittaci, avian paramyxovirus-1, Toxoplasma gondii and WNV. Risk factor analyses indicated that rearing site and original release location were significantly associated with a positive serologic titer to WNV among free-flying condors. This study provides preliminary baseline data on infectious disease exposure in these populations for aiding in early disease detection and provides potentially critical information for conservation of the endangered California condor as it continues to expand its range and encounter new infectious disease threats.

The author measured endorsement of the New Ecological Paradigm (NEP) in college students following their involvement in a 2- to 3-week environmental problem module focused on global environmental problems and energy issues. The module included readings, discussion, and a writing exercise, and was presented during 3 sequential semesters within a course on research design. Students from the 3 groups were all tested subsequently in spring 2001. When compared with control data, data from students receiving the educational module showed significant increases in NEP endorsement that declined somewhat with time but remained significantly greater after 3 semesters. In contrast, retention of knowledge from the module showed a predictable decrease over time.

In 1996, a disease outbreak occurred at a captive breeding facility in Idaho, causing anorexia, dehydration, and diarrhea or sudden death in 72 of 110 Northern aplomado falcons ( Falco femoralis septentrionalis ) from 9 to 35 days of age and in 6 of 102 peregrine falcons ( Falco peregrinus ) from 14 to 25 days of age. Sixty-two Northern aplomado and six peregrine falcons died. Epidemiologic analyses indicated a point source epizootic, horizontal transmission, and increased relative risk associated with cross-species brooding of eggs. Primary lesions in affected birds were inclusion body hepatitis, splenomegaly, and enteritis. The etiology in all mortalities was determined by molecular analyses to be a new species of adenovirus distantly related to the group I avian viruses, serotypes 1 and 4, Aviadenovirus . In situ hybridization and PCR demonstrated that the virus was epitheliotropic and lymphotropic and that infection was systemic in the majority of animals. Adeno-associated virus was also detected by PCR in most affected falcons, but no other infectious agents or predisposing factors were found in any birds. Subsequent to the 1996 epizootic, a similar disease caused by the same adenovirus was found over a 5-year period in orange-breasted falcons ( Falco deiroleucus ), teita falcons ( Falco fasciinucha ), a merlin ( Falco columbarius ), a Vanuatu peregrine falcon ( Falco peregrinus nesiotes ), and gyrfalcon × peregrine falcon hybrids ( Falco rusticolus / peregrinus ) that died in Wyoming, Oklahoma, Minnesota, and California. These findings indicate that this newly recognized adenovirus is widespread in western and midwestern North America and can be a primary pathogen in different falcon species.

Prevalence and disease caused by isosporoid coccidia in passerine birds are well recognized, but confusion about the life cycles of the parasites has led to taxonomic inconsistencies. In this study, we characterized segments of the chromosomal small and large-subunit ribosomal RNA (rRNA) genes of coccidial parasites from 23 species of passerine birds, as well as heat shock protein 70, apicoplast rRNA, and chromosomal 5.8s rRNA genes from a subgroup of these animals, and we correlated genetic data with morphologic findings for different parasite developmental stages, host phylogeny, and overall taxonomic relations within the phylum Apicomplexa. Our findings indicate that isosporoid coccidia of passerine birds are monophyletic but exhibit substantial diversity, with most avian species having one or several unique parasite lineages that underwent synchronous speciation with their hosts, interrupted by sporadic episodes of lateral transmission across species and families. Molecular analyses support a homoxenous life cycle, with sexual forms occurring chiefly in the intestines and asexual merozoites present systemically. Rarely, extraintestinal sexual stages can occur. The passerine coccidia are genetically most closely related to species of Eimeria rather than Isospora. We suggest that these parasites, whether identified from blood merozoite stages or fecal oocysts, be provisionally grouped as a homogeneous clade of individual species in a single taxon and formally named when reliable criteria allowing reclassification of related genera in the suborder Eimeriina are clarified.

As the initial phase of a longitudinal study of environmental perspective in college students, resident student opinion was sampled using the New Ecological Paradigm (NEP) scale administered through systematic alphabetical sampling. Sampling was also carried out by a blanket e-mail distribution of surveys for voluntary response. Results showed positive but weak endorsement of the NEP worldview. Reliably greater endorsement was shown in e-mail than in systematic samples, and by women.

Transitional cell carcinomas (TCC) of the urinary bladder were diagnosed in four related fishing cats (Prionailurus viverrinus). The major clinical sign in each case was persistent hematuria unresponsive to medical therapy. Cystotomy and biopsy provided an antemortem diagnosis in three of the fishing cats before euthanasia because of progression of clinical signs. The diagnosis was made in the fourth cat after euthanasia because of renal failure. Hematuria improved temporarily in one of the cats diagnosed antemortem and treated with piroxicam and carboplatin. Attempts to isolate a herpesvirus in two of the cats failed. Histopathologic appearance of the TCC was similar to that described for other species. TCC metastasis to the lungs was noted at necropsy in one cat; metastatic disease was not noted in the other fishing cats on gross or histopathologic examination. TCC of the urinary bladder appears to be more prevalent in fishing cats than in other species of domestic or nondomestic felids.

Medical records and necropsy reports of 410 pigeons and doves from the Zoological Society of San Diego were examined to determine if birds treated with fenbendazole (FBZ) or albendazole (ABZ) had a higher incidence of toxic-related signs than untreated control birds. Birds presented for nonmedical reasons (quarantine, relocation, preshipment examinations, and routine parasite screening) were used exclusively to avoid confounding variables. Birds were observed for 45 days after presentation. Compared with a 4–6% mean weight gain in control birds during the treatment period or hospitalization, birds treated with FBZ had a mean weight loss of 16.1% (P < .001), and birds treated with ABZ had a mean weight loss of 13.3% (P = .004). The percentage of birds with marked leukopenia (white blood cell < 1000/μ l) was higher in FBZ-treated birds (87.5%, P < .001) and ABZ-treated birds (100%, P = .013) than in control birds (12.5%). Bone marrow hypoplasia was found in FBZ-treated birds (48.6%, P < .001) and A...

Methods Mycobacteria isolated from more than 100 birds diagnosed with avian mycobacteriosis at the San Diego Zoo and its Safari Park were cultured postmortem and had their whole genomes sequenced. Computational workflows were developed and applied to identify the mycobacterial species in each DNA sample, to find single-nucleotide polymorphisms (SNPs) between samples of the same species, to further differentiate SNPs between as many as three different genotypes within a single sample, and to identify which samples are closely clustered genomically. Results Nine species of mycobacteria were found in 123 samples from 105 birds. The most common species were Mycobacterium avium and Mycobacterium genavense, which were in 49 and 48 birds, respectively. Most birds contained only a single mycobacterial species, but two birds contained a mixture of two species. The M. avium samples represent diverse strains of M. avium avium and M. avium hominissuis, with many pairs of samples differing by hundreds or thousands of SNPs across their common genome. By contrast, the M. genavense samples are much closer genomically; samples from 46 of 48 birds differ from each other by less than 110 SNPs. Some birds contained two, three, or even four genotypes of the same bacterial species. Such infections were found in 4 of 49 birds (8%) with M. avium and in 11 of 48 birds (23%) with M. genavense. Most were mixed infections, in which the bird was infected by multiple mycobacterial strains, but three infections with two genotypes differing by ≤ 10 SNPs were likely the result of within-host evolution. The samples from 31 birds with M. avium can be grouped into nine clusters within which any sample is ≤ 12 SNPs from at least one other sample in the cluster. Similarly, the samples from 40 birds with M. genavense can be grouped into ten such clusters. Information about these genomic clusters is being used in an ongoing, companion study of mycobacterial transmission to help inform management of bird collections.

To the Editor: The influenza A pandemic (H1N1) 2009 virus rapidly created a global pandemic among humans and also appears to have strong infectivity for a broad range of animal species (1–3). The virus has been found repeatedly in swine and has been detected in a dog, cats, turkeys, and domestic ferrets and in nondomestic animals, including skunks, cheetahs, and giant anteaters (2–4). In some cases, animal-to-animal transmission may have occurred, raising concern about the development of new wildlife reservoirs (2). In 2009, the first recognized occurrence of pandemic (H1N1) 2009 in southern California in April was followed by a surge of cases during October through November (4). During this time, respiratory illness developed in a 12-year-old male American badger (Taxidea taxus taxus), a 19-year-old female Bornean binturong (Arctictis binturong penicillatus), and a 7-year-old black-footed ferret (Mustela nigripes) housed in a San Diego zoological garden. The 3 affected animals had clinical signs that included lethargy, inappetance, dyspnea, nasal discharge, and coughing. The severity of disease in the badger and binturong necessitated euthanasia; the ferret recovered with antibiotic and fluid therapy. Postmortem examination revealed bronchopneumonia with diffuse alveolar damage in the badger and interstitial pneumonia with diffuse alveolar damage in the binturong. Bacterial cultures and Gram stains of affected lung samples were negative. Molecular analyses for several groups of viruses, including Herpesviridae, Paramyxoviridae, Adenoviridae, and all influenza A viruses, were performed on frozen lung samples from the badger and binturong and on frozen conjunctival and pharyngeal swabs from the ferret. Results of PCRs specific for segments of influenza A nucleoprotein, matrix protein, hemagglutinin, and neuraminidase genes were positive in samples from all 3 animals, and DNA sequencing of amplicons identified the viruses as pandemic (H1N1) 2009. Influenza A virus was not detected in samples from the ferret after it recovered. Results of PCRs for all other viruses were negative. Immunohistochemical evaluation of lung samples from the badger for antigens of influenza A virus (5) showed rare staining in bronchiolar epithelial cells (Figure). Figure Lung section from an American badger showing immunohistochemical staining (red chromogen) specific for the pandemic (H1N1) 2009 virus within the nucleus and cytoplasm of bronchiolar epithelial cells and concurrent inflammatory cell infiltrates; hematoxylin ... Respiratory disease in all 3 affected animals seemed to be caused by pandemic (H1N1) 2009 virus. The badger and binturong were generally healthy, no other pathogens were detected, and pulmonary lesions were consistent with influenza pneumonia. In these animals, pandemic (H1N1) 2009 infection was especially aggressive, resulting in irreversible disease. Reports of pandemic (H1N1) 2009 virus in skunks and anteaters also describe severe disease in those species (2,3). In contrast, the infected black-footed ferret in our study had relatively mild clinical illness, consisting only of lethargy. This finding was surprising given recent experimental studies that reported the current pandemic (H1N1) 2009 virus was more pathogenic in domestic ferrets (Mustela putorius furo) than typical seasonal influenza viruses (6). However, several factors could have resulted in the low level of disease in this animal, such as prior immunity to influenza viruses or a low exposure dose. It is also possible black-footed ferrets are innately more resistant to influenza infection than domestic ferrets. The origin of infection in these cases was not determined but was most likely an infected human. All animals had some level of contact with caretakers or veterinarians and were housed separately from other wildlife species. None of the potential human sources of virus had clinical signs before the animals became ill; however, influenza infections in humans can often be mild (7). Wild animals, such as opossums and skunks, that occasionally enter the zoological garden, represent another possible source. Good hygiene and husbandry practices used within the enclosures of the badger, binturong, and ferret failed to prevent infection, which suggests pandemic (H1N1) 2009 is efficiently transmitted to these species. Descriptions of infection in giant anteaters and cheetahs kept under similar conditions also support high transmissibility of influenza A viruses to animals, as do ongoing findings for swine (3,4,8). Although ferrets are known to be susceptible to influenza A virus, to our knowledge, influenza in badgers and binturongs has not been reported. Badgers and binturong have been housed in zoological gardens for decades without incidence of influenza. Increased surveillance for influenza by the scientific community during the pandemic may have resulted in the novel recognition of infection in these species. Alternatively, the current pandemic (H1N1) 2009 virus may have a broader host range and stronger virulence than viruses in the past. Pandemic (H1N1) 2009 was first detected in humans in March 2009 and reached pandemic levels by June of that year, rapidly establishing a rich pool for the development of genetic variants. Naturally acquired disease has now been described in 10 animal species, and experimental infection has been reported in an additional 2 animals (mice and cynomolgus macaques) (9). The ubiquity of pandemic (H1N1) 2009 and its ability to infect a diverse range of hosts is worrisome for the health of wildlife and for the possibility of creating additional reservoirs that could alter the evolution of subtype H1N1 viruses by applying varied selection pressures and establishing new ways of generating unique reassortant strains.

A 10-month-old, female African pygmy falcon ( Polihierax semitorquatus) hatched and housed at the San Diego Zoo developed neurologic signs and died from a cerebral infection with the rat lungworm Angiostrongylus cantonensis. There was an associated mild nonsuppurative meningoencephalitis. This infection was diagnosed on histology and confirmed by detection of species-specific A. cantonensis DNA in formalin-fixed and frozen brain tissue by a polymerase chain reaction assay. To the authors’ knowledge, this infection has not previously been reported in a bird in the United States and has not been known to be naturally acquired in any species in this region of the world. The source of the infection was not definitively determined but was possibly feeder geckos ( Hemidactylus frenatus) imported from Southeast Asia where the parasite is endemic.

During a preventive medicine examination on a 13-yr-old intact female African spot-necked otter (Lutra maculicollis), radiographs were obtained and a cranial thoracic mass was noted. Cytology from an ultrasound-guided fine-needle aspirate was suggestive of a thymoma. Surgical removal was performed and this diagnosis was confirmed by histopathology. At a recheck examination 6 mo postsurgery, there was no evidence of recurrence. This case highlights the value of a comprehensive preventive medicine program that resulted in the early detection of a subclinical thymoma and its successful removal.

Five out of 169 fertile California condor (Gymnogyps californianus) eggs laid in captivity have exhibited chondrodystrophy, a lethal form of dwarfism. Pedigree records indicate that this chondrodystrophy, like similar conditions in chickens, turkeys and quail, is probably inherited as an autosomal, recessive allele. We estimate that the frequency of this putative allele is about 9%. This high frequency is probably due to a founder effect. We consider three management options for the allele: ignoring it, eliminating it by selection and minimizing its phenotypic manifestation by avoiding matings between possible carriers. We recommend minimizing its phenotypic expression because an unacceptably large proportion of condors (up to 78 out of 146) would be prevented from breeding under a selection strategy designed to eliminate the allele. We predict that many captive populations will prove similar to the California condor population in that it will prove inadvisable or impractical to select against one or more deleterious alleles detected in the population.

Lead is one of the most studied toxicants, and overwhelming scientific evidence demonstrates that lead is toxic to several physiological systems in vertebrates, including the nervous, renal, cardiovascular, reproductive, immune, and hematologic systems (Health Risks from Lead-Based Ammunition in the Environment—A Consensus Statement of Scientists 2013). Furthermore, there is no level of lead exposure in children known to be without adverse effects [Centers for Disease Control and Prevention (CDC) 2012a, 2012b]. In light of this evidence, there is an urgent need to end a major source of lead for animals and humans: spent lead bullets and shotgun pellets. Notably, production of lead-based ammunition in the United States accounted for > 69,000 metric tons consumed in 2012; this is second only to the amount of lead used to manufacture storage batteries (U.S. Geological Survey 2013). However, there are few regulations regarding the release of lead into the environment through discharge of lead-based ammunition. For other major categories of lead consumption, such as lead batteries and sheet lead/lead pipes, environmental discharge and disposal are regulated. Therefore, lead-based ammunition is likely the greatest largely unregulated source of lead that is knowingly discharged into the environment in the United States. In contrast, the release or distribution of other major sources of environmental lead contamination (e.g., leaded gasoline, lead-based paint, lead solder) have been substantially regulated and reduced since the mid-1970s (Health Risks from Lead-Based Ammunition in the Environment—A Consensus Statement of Scientists 2013). There is a national discussion—polarized at times—of the health risks posed to humans and wildlife from the discharge of lead-based ammunition. To inform this discussion, a group of 30 nationally and internationally recognized scientists with expertise regarding lead and environmental health recently collaborated to create an evidence-based consensus statement (Health Risks from Lead-Based Ammunition in the Environment—A Consensus Statement of Scientists 2013) supporting the reduction and eventual elimination of lead released to the environment through the discharge of lead-based ammunition. The discharge of lead bullets and shotgun pellets into the environment poses significant health risks to humans and wildlife. The best available scientific evidence demonstrates that the discharge of lead-based ammunition substantially increases environmental lead levels, especially in areas with higher shooting activity (U.S. Environmetal Protection Agency 2012) and that the discharge of lead-based ammunition poses risks of elevated lead exposure to gun users (National Research Council 2012). When lead-containing bullets are used to shoot wildlife, they can fragment into hundreds of small pieces, many of which are small enough to be easily ingested by scavenging animals or to be retained in meat prepared for human consumption (Hunt et al. 2009; Knott et al. 2010; Pauli and Burkirk 2007). Consequently, lead-based ammunition may be a significant source of lead exposure in humans that regularly ingest wild game (Hanning et al. 2003; Johansen et al. 2006; Levesque et al. 2003; Tsuji et al. 2008). In addition, lead pellets and fragments have been reported in gastrointestinal tracts of hunters who consume meat from animals shot with lead-based ammunition (Carey 1977; Reddy 1985). The use of lead pellets in shotgun shells for hunting waterfowl posed a serious threat to wetland birds, and secondarily to bald eagles, in the United States, leading to the U.S. Fish and Wildlife Service’s 1991 nationwide regulations requiring use of nontoxic shotgun pellets for hunting waterfowl (Anderson 1992). However, lead poisoning from ingestion of spent lead-based ammunition fragments continues to pose a particularly serious health threat for scavenging species. These lead-containing fragments remain the principal source of lead exposure to endangered California condors and continue to prevent the successful recovery of these birds in the wild (Church et al. 2006; Finkelstein et al. 2012; Green et al. 2008; Parish et al. 2009; Rideout et al. 2012; Woods et al. 2007). Other wildlife species, such as golden eagles, bald eagles, ravens, turkey vultures, and pumas, are also exposed to the fragments of spent lead ammunition (Burco et al. 2012; Clark and Scheuhammer 2003; Craighead and Bedrosian 2008; Cruz-Martinez et al. 2012; Fisher et al. 2006; Kelly and Johnson 2011; Stauber et al. 2010; Wayland and Bollinger 1999). No rational deliberation about the use of lead-based ammunition can ignore the overwhelming evidence for the toxic effects of lead, or that the discharge of lead bullets and shot into the environment poses significant risks of lead exposure to humans and wildlife. Given the availability of non-lead ammunition for shooting and hunting (Thomas 2013), the use of lead-based ammunition that introduces lead into the environment can be reduced and eventually eliminated. This seems to be a reasonable and equitable action to protect the health of humans and wildlife.

In order to test the effectiveness of captive-rearing and release strategies for future restoration of birds in Hawai'i, this pilot study was conducted in forests where introduced avian disease and mammalian predators were present. Methodology used resulted in the first successful hatching of Drepanidinae eggs in an incubator and subsequent hand-rearing of chicks from hatch. Sixteen Common 'Amakihi (Hemignathus virens virens) (mean hatch weight = 1.4 g) were hand-reared. Two different reintroduction strategies were evaluated for small honeycreepers. Known mortality in the wild after release was due to mosquito-transmitted disease (avian malaria and pox). This pilot study shows that the techniques necessary to hatch, rear, and release endangered Hawaiian honeycreepers are available. However, restoration efforts will probably not succeed unless mosquito-free, predator-controlled reintroduction sites are available or strategies are developed to decrease mortality in naive honeycreepers exposed to disease after release. © 1996 Wiley-Liss, Inc.

Gout is the only enemy that I do not wish to have at my feet. — Reverend Sydney Smith, 1841 Gout, a disease that has been known since antiquity, is an acute, often recurrent arthritis mediated by t...