Lead in Ammunition: A Persistent Threat to Health and Conservation

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 Novel Psittacine Adenovirus Identified During an Outbreak of Avian Chlamydiosis and Human Psittacosis: Zoonosis Associated with Virus-Bacterium Coinfection in Birds

Chlamydophila psittaci is found worldwide, but is particularly common among psittacine birds in tropical and subtropical regions. While investigating a human psittacosis outbreak that was associated with avian chlamydiosis in Hong Kong, we identified a novel adenovirus in epidemiologically linked Mealy Parrots, which was not present in healthy birds unrelated to the outbreak or in other animals. The novel adenovirus (tentatively named Psittacine adenovirus HKU1) was most closely related to Duck adenovirus A in the Atadenovirus genus. Sequencing showed that the Psittacine adenovirus HKU1 genome consists of 31,735 nucleotides. Comparative genome analysis showed that the Psittacine adenovirus HKU1 genome contains 23 open reading frames (ORFs) with sequence similarity to known adenoviral genes, and six additional ORFs at the 3′ end of the genome. Similar to Duck adenovirus A, the novel adenovirus lacks LH1, LH2 and LH3, which distinguishes it from other viruses in the Atadenovirus genus. Notably, fiber-2 protein, which is present in Aviadenovirus but not Atadenovirus, is also present in Psittacine adenovirus HKU1. Psittacine adenovirus HKU1 had pairwise amino acid sequence identities of 50.3–54.0% for the DNA polymerase, 64.6–70.7% for the penton protein, and 66.1–74.0% for the hexon protein with other Atadenovirus. The C. psittaci bacterial load was positively correlated with adenovirus viral load in the lung. Immunostaining for fiber protein expression was positive in lung and liver tissue cells of affected parrots, confirming active viral replication. No other viruses were found. This is the first documentation of an adenovirus-C. psittaci co-infection in an avian species that was associated with a human outbreak of psittacosis. Viral-bacterial co-infection often increases disease severity in both humans and animals. The role of viral-bacterial co-infection in animal-to-human transmission of infectious agents has not received sufficient attention and should be emphasized in the investigation of disease outbreaks in human and animals.     

Towards the Conservation of Endangered Avian Species: A Recombinant West Nile Virus Vaccine Results in Increased Humoral and Cellular Immune Responses in Japanese Quail (Coturnix japonica)

West Nile Virus (WNV) arrived in North America in 1999 and is now endemic. Many families of birds, especially corvids, are highly susceptible to WNV and infection often results in fatality. Avian species susceptible to WNV infection also include endangered species, such as the Greater Sage-Grouse (Centrocercus uropbasianuts) and the Eastern Loggerhead Shrike (Lanius ludovicianus migrans). The virus has been shown to contribute towards the likelihood of their extinction. Although a clear and present threat, there exists no avian WNV vaccine available to combat this lethal menace. As a first step in establishing an avian model for testing candidate WNV vaccines, avian antibody based reagents were assessed for cross-reactivity with Japanese quail (Coturnix japonica) T cell markers CD4 and CD8; the most reactive were found to be the anti-duck CD8 antibody, clone Du-CD8-1, and the anti-chicken/turkey CD4 antibody, clone CT4. These reagents were then used to assess vaccine performance as well as to establish T cell populations in quail, with a novel population of CD4/CD8 double positive T cells being identified in Japanese quail. Concurrently, non-replicating recombinant adenoviruses, expressing either the WNV envelope or NS3 ‘genes’ were constructed and assessed for effectiveness as avian vaccines. Japanese Quail were selected for testing the vaccines, as they provide an avian model that parallels the population diversity of bird species in the wild. Both the level of WNV specific antibodies and the number of T cells in vaccinated birds were increased compared to unvaccinated controls. The results indicate the vaccines to be effective in increasing both humoral and cellular immune responses. These recombinant vaccines therefore may find utility as tools to protect and maintain domestic and wild avian populations. Their implementation may also arrest the progression towards extinction of endangered avian species and reduce the viral reservoir that potentiates infection in humans.     

Prevalence of Chlamydophila psittaci in Fecal Droppings from Feral Pigeons in Amsterdam, The Netherlands

In many cities, the feral rock dove is an abundant bird species that can harbor Chlamydophila psittaci. We determined the prevalence and genotype of C. psittaci in fresh fecal samples from feral pigeons in Amsterdam, The Netherlands. The prevalence was 7.9% overall (26/331; 95% confidence interval, 5 to 11). Ten genotyped PCR-positive samples were all genotype B.     

Andean and California condors possess dissimilar genetic composition but exhibit similar demographic histories

While genetic diversity of threatened species is a major concern of conservation biologists, historic patterns of genetic variation are often unknown. A powerful approach to assess patterns and processes of genetic erosion is via ancient DNA techniques. Herein, we analyzed mtDNA from historical samples (1800s to present) of Andean Condors (Vultur gryphus) to investigate whether contemporary low genetic variability is the result of recent human expansion and persecution, and compared this genetic history to that of California condors (Gymnogyps californianus).We then explored historic demographies for both species via coalescent simulations. We found that Andean condors have lost at least 17% of their genetic variation in the early 20th century. Unlike California condors, however, low mtDNA diversity in the Andean condor was mostly ancient, before European arrival. However, we found that both condor species shared similar demographies in that population bottlenecks were recent and co‐occurred with the introduction of livestock to the Americas and the global collapse of marine mammals. Given the combined information on genetic and demographic processes, we suggest that the protection of key habitats should be targeted for conserving extant genetic diversity and facilitate the natural recolonization of lost territories, while nuclear genomic data should be used to inform translocation plans.     

The immunobiology of the innate response to Toxoplasma gondii

Toxoplasma gondii is a unique intracellular parasite. It can infect a variety of cells in virtually all warm-blooded animals. It has a worldwide distribution and, overall, around one-third of people are seropositive for the parasite, with essentially the entire human population being at risk of infection. For most people, T. gondii causes asymptomatic infection but the parasite can cause serious disease in the immunocompromised and, if contracted for the first time during pregnancy, can cause spontaneous abortion or congenital defects, which have a substantial emotional, social and economic impact. Toxoplasma gondii provokes one of the most potent innate, pro-inflammatory responses of all infectious disease agents. It is also a supreme manipulator of the immune response so that innate immunity to T. gondii is a delicate balance between the parasite and its host involving a coordinated series of cellular interactions involving enterocytes, neutrophils, dendritic cells, macrophages and natural killer cells. Underpinning these interactions is the regulation of complex molecular reactions involving Toll-like receptors, activation of signalling pathways, cytokine production and activation of anti-microbial effector mechanisms including generation of reactive nitrogen and oxygen intermediates.     

Experimental lead poisoning in turkey vultures (Cathartes aura)

Lead-induced mortality appears to have been a major factor in the decline of the California condor (Gymnogyps californianus). We orally dosed turkey vultures (Cathartes aura) with BB-sized lead shot from January 1988 through July 1988 to determine physiologic response (delta-aminolevulinic acid dehydratase inhibition, erythrocyte protoporphyrin levels, anemia), diagnostic tissue lead concentrations (blood, liver, and kidney), and comparative sensitivity of this species. Two turkey vultures died and two became so intoxicated they were euthanized. Overall, responses of measured parameters were comparable to other species exposed to lead although there was considerable individual variation. Survival time (143-211 days), even with the large numbers of shot and constant redosing, was much longer than reported for other species of birds, suggesting considerable tolerance by turkey vultures to the deleterious effects of lead ingestion. Based on these observations, turkey vultures appear to be poor models for assessing the risk of lead poisoning to California condors or predicting their physiologic response.     

Detection and genetic characterisation of Toxoplasma gondii circulating in free-range chickens,pigs and seropositive pregnant women in Benue state,Nigeria

Toxoplasma gondii parasites present strong but geographically varied signatures of population structure. Populations sampled from Europe and North America have commonly been defined by over-representation of a small number of clonal types, in contrast to greater diversity in South America. The occurrence and extent of genetic diversity in African T. gondii populations remains understudied, undermining assessments of risk and transmission. The present study was designed to establish the occurrence, genotype and phylogeny of T. gondii in meat samples collected from livestock produced for human consumption (free-range chickens, n = 173; pigs, n = 211), comparing with T. gondii detected in blood samples collected from seropositive pregnant women (n = 91) in Benue state, Nigeria. The presence of T. gondii DNA was determined using a published nested polymerase chain reaction, targeting the 529 bp multicopy gene element. Samples with the highest parasite load (assessed using quantitative PCR) were selected for PCR-restriction fragment length polymorphism (PCR-RFLP) targeting the surface antigen 3 (SAG3), SAG2 (5’ and 3’), beta-tubulin (BTUB) and dense granule protein 6 (GRA6) loci, and the apicoplast genome (Apico). Toxoplasma gondii DNA was detected in all three of the populations sampled, presenting 30.6, 31.3 and 25.3% occurrence in free-range chickens, pigs and seropositive pregnant women, respectively. Quantitative-PCR indicated low parasite occurrence in most positive samples, limiting some further molecular analyses. PCR-RFLP results suggested that T. gondii circulating in the sampled populations presented with a type II genetic background, although all included a hybrid type I/II or II/III haplotype. Concatenation of aligned RFLP amplicon sequences revealed limited diversity with nine haplotypes and little indication of host species-specific or spatially distributed sub-populations. Samples collected from humans shared haplotypes with free-range chickens and/or pigs. Africa remains under-explored for T. gondii genetic diversity and this study provides the first detailed definition of haplotypes circulating in human and animal populations in Nigeria.     

Impact of the California lead ammunition ban on reducing lead exposure in golden eagles and turkey vultures

Predatory and scavenging birds may be exposed to high levels of lead when they ingest shot or bullet fragments embedded in the tissues of animals injured or killed with lead ammunition. Lead poisoning was a contributing factor in the decline of the endangered California condor population in the 1980s, and remains one of the primary factors threatening species recovery. In response to this threat, a ban on the use of lead ammunition for most hunting activities in the range of the condor in California was implemented in 2008. Monitoring of lead exposure in predatory and scavenging birds is essential for assessing the effectiveness of the lead ammunition ban in reducing lead exposure in these species. In this study, we assessed the effectiveness of the regulation in decreasing blood lead concentration in two avian sentinels, golden eagles and turkey vultures, within the condor range in California. We compared blood lead concentration in golden eagles and turkey vultures prior to the lead ammunition ban and one year following implementation of the ban. Lead exposure in both golden eagles and turkey vultures declined significantly post-ban. Our findings provide evidence that hunter compliance with lead ammunition regulations was sufficient to reduce lead exposure in predatory and scavenging birds at our study sites.     

Seroprevalence for selected pathogens of zoonotic importance in wild nutria (Myocastor coypus)

Little information is present in the literature on diseases of the nutria (Myocastor coypus) in the wild. Serum samples obtained from 176 trapped animals were tested for antibody reactivity against 11 pathogens of significant zoonoses. Sera were positive against Leptospira (38.0 %, odds relative risk?=?0.03, Taylor series 95 % confidence limits 0.01–0.06), Toxoplasma gondii (27.8 %, odds relative risk?=?2.30, Taylor series 95 % confidence limits 1.23–4.31), Chlamydophila psittaci (21.0 % odds relative risk?=?4.94, Taylor series 95 % confidence limits 2.75–8.89), Streptococcus equi subspecies zooepidemicus (15.9 % odds relative risk?=?1.87, Taylor series 95 % confidence limits 1.04–3.37), and encephalomyocarditis virus (3.4 %, odds relative risk?=?0.05, Taylor series 95 % confidence limits 0.02–0.12). Some of the rodents showed antibodies at high titers, mostly indicating recent exposure. Seroprevalence rates varied among the location and age groups, although not by season or gender. All samples were seronegative for Brucella spp., Francisella tularensis, vesicular stomatitis virus, rabies virus, foot-and-mouth disease virus, and Encephalitozoon cuniculi.     

Survey for Selected Pathogens and Evaluation of Disease Risk Factors for Endangered Hawaiian Monk Seals in the Main Hawaiian Islands

A recently reestablished and increasing population of Hawaiian monk seals in the main Hawaiian Islands (MHI) is encouraging for this endangered species. However, seals in the MHI may be exposed to a broad range of human, pet, livestock, and feral animal pathogens. Our objective was to determine the movement and foraging habitats of Hawaiian monk seals in the MHI relative to the potential exposure of seals to infectious diseases in near-shore marine habitats. We captured 18 monk seals in the MHI between January 27, 2004 and November 29, 2005, tested them for various infectious diseases, and then monitored the foraging movements of 11 of them using satellite-linked radio transmitters for the next 32–167 days. All seals tested negative for canine adenovirus, calicivirus, four morbilliviruses, phocine herpes virus, Leptospira sp., and feline and canine heartworm antigen/antibody. Six of the seals tested positive on complement fixation for Chlamydophila abortus (formerly Chlamydia psittaci). Four seals demonstrated positive titers to Sarcocystis neurona, two to Neospora caninum, and two to Toxoplasma gondii. Fecal cultures showed approximately half (n = 6) positive for E. coli 0157, no Salmonella sp., and only one with Campylobacter sp. Satellite monitored seals spent considerable time foraging, traveling, and resting in neritic waters close to human population centers, agricultural activity, and livestock ranges, and sources of land-based water runoff and sewage dispersal. Consequently, Hawaiian monk seals in the MHI may be at risk of exposure to several infectious disease agents associated with terrestrial animals that can contaminate marine habitats from runoff along drainages and that are known to cause disease in marine mammals. Further, some seals overlapped substantially in their use of coastal habitats and several moved among islands while foraging and were seen on beaches near each other. This suggests that diseased seals could infect healthy conspecifics throughout the MHI.     

Molecular genetic characterization of avian <Emphasis Type="Italic">Chlamydophila psittaci</Emphasis> isolates

RFLP analysis of the omp1 gene was used to characterize 51 avian Chlamydophila psittaci isolates. The analysis confirmed the predominance of genotype A in parrot C. psittaci isolates and revealed new omp1 genotypes in corvid C. psittaci isolates. The corvid isolates proved to lack an extrachromosomal plasmid. The omp1 and rRNA IGS sequences were determined for the new isolates. Phylogenetic analysis showed that isolate 1V, obtained from a crow, is intermediate in several characters between C. abortus and C. psittaci. The results were compared with data on the phylogenetic relationships of earlier chlamydium isolates.     

Need and Seek for Dietary Micronutrients: Endogenous Regulation,External Signalling and Food Sources of Carotenoids in New World Vultures

Among birds, vultures show low concentrations of plasma carotenoids due to the combination of their large size, general dull colouration and a diet based on carrion. We recorded the concentration of each carotenoid type present in plasma of the Andean condor (Vultur gryphus) according to age and sex, that determine colour signalling and dominance hierarchies in the carcasses. We compared the carotenoid profile in wild condors with that of captive condors fed with a controlled diet of flesh to test the hypothesis that wild individuals could acquire extra carotenoids from vegetal matter contained in carcass viscera and fresh vegetation. Wild American black vultures (Coragyps atratus) were also sampled to evaluate the potential influence of colouration in the integument on absorption and accumulation patterns of plasma carotenoids. A remarkably higher concentration of lutein than β-carotene was found in wild condors, while the contrary pattern was recorded in American black vultures and captive condors. We found a consistent decrease in all plasma carotenoids with age, and a lower concentration of most xanthophylls in male compared to female wild condors. Positive correlations of all carotenoids indicated general common absorption and accumulation strategies or a single dietary source containing all pigments found in plasma. The comparatively low total concentration of carotenoids, and especially of lutein rather than β-carotene, found in captive condors fed with a diet restricted to flesh supports the hypothesis that Andean condors can efficiently acquire carotenoids from vegetal matter in the wild. Andean condors seem to be physiologically more competent in the uptake or accumulation of xanthophylls than American black vultures, which agrees with the use of colour-signalling strategies in sexual and competitive contexts in the Andean condor. This study suggests that vultures may use dietary vegetal supplements that provide pigments and micronutrients that are scarce or missing in carrion.     

Seroprevalence of Toxoplasma gondii Infection in Police Dogs in Shenyang,Northeastern China

In recent years, worldwide surveys of Toxoplasma gondii infection in dogs have been reported. However, only limited surveys of T. gondii infection in police dogs have been available, including China. In the present study, we report the seroprevalence of T. gondii in police dogs in Shenyang, northeastern China. Sera from 291 police dogs were examined for T. gondii antibodies with the modified agglutination test (MAT), and 30.9% animals were tested seropositive. The results of the present study indicated a relatively high prevalence of T. gondii infection in police dogs in Shenyang, China.     

The effects of rearing method on social behaviors of mentored,captive‐reared juvenile California condors

Puppet‐reared and parent‐reared captive‐bred California condor (Gymnogyps californianus) juveniles were studied before their release into the wild. Behavioral data were collected during social interactions within two cohorts of juveniles (N = 11) and their adult mentors (N = 5). The purposes of this study were to (1) document the social behaviors of mentored juvenile California condors, and (2) compare social behaviors for two different rearing methods (puppet‐reared versus parent‐reared) during two phases of the mentoring process (San Diego Wild Animal Park versus release sites). Of the 17 behaviors examined by 2 × 2 analyses of variance (ANOVAs), two significant interactions between the rearing method and mentoring phase were found: pulls feathers and feeds alone. For both behaviors, parent‐reared condors engaged in these activities more often at the zoo and less often at the release pens than did the puppet‐reared condors. The main effect of rearing was also significant for two behaviors: near others, and receives contact aggression from other. Parent‐reared birds were more likely to be near another bird and receive contact aggression, regardless of mentoring phase, than puppet‐reared birds. The effect size for 16 of the 17 behaviors was greater for the rearing method than for mentoring phase. Rearing method differences may persist long‐term, as parent‐reared adult mentors were significantly more aggressive than puppet‐reared adult mentors. Dominance relations were examined for both cohorts, with the first cohort exhibiting a strong linear relationship (h′ = 0.86, P = 0.018), whereas the second cohort exhibited a moderate but non‐significant linear hierarchy (h′ = 0.63, P = 0.21). The rearing method had no effect on dominance among the juveniles, but adults were probably dominant to juveniles (P = 0.052; the difference was nearly significant). Although social behaviors between the two rearing groups were similar in most respects, this study is the first to document measurable differences between puppet‐ and parent‐reared captive‐bred California condor juveniles. Zoo Biol 27:1–18, 2008. © 2007 Wiley‐Liss, Inc.     

Health evaluation of Galapagos Hawks (Buteo galapagoensis) on Santiago Island, Galapagos

Galapagos Hawks (Buteo galapagoensis), the only endemic, diurnal raptor species in Galapagos, are currently distributed on eight Galapagos Islands having been extirpated from three of the human-inhabited islands. In January 2009, we performed health assessments of 89 Galapagos Hawks on Santiago Island, Galapagos. Four of the 89 Galapagos Hawks (4%) evaluated had physical abnormalities. Blood parameters did not differ between males and females, except for aspartate transaminase values, which were significantly higher in females than males. No Galapagos Hawks tested positive for antibodies to avian encephalitis virus, Marek virus, and paramyxovirus-1 or to haemosporidian antigen. Chlamydophila psittaci antigen was detected in 2 of 86 Galapagos Hawks (2%), with 24 of 43 Galapagos Hawks (56%) antibody-positive for avian adenovirus-1 and 1 of 48 Galapagos Hawks (2%) antibody positive for Toxoplasma gondii. There were no significant differences in infectious disease results based on sex. This study contributes to the understanding of the health status of the Galapagos Hawk and to the establishment of baseline information for the species.     

Seroprevalence of Toxoplasma gondii and Trichinella spiralis infections in wild boars (Sus scrofa) in Korea

Toxoplasma gondii and Trichinella spiralis are important zoonotic pathogens with worldwide distributions. In Korea, several outbreaks of human toxoplasmosis and trichinellosis due to the consumption of infected wild animals have been reported. The purpose of this study was to determine the seroprevalence of T. gondii and T. spiralis infections in wild boars killed in Korea from December 2009 to October 2011. A total of 521 wild boars hunted in eight provinces were examined for antibodies to T. gondii and T. spiralis by using commercial ELISA kits. Overall, 25.1% of serum samples from individual boars were seropositive for T. gondii and 1.7% were seropositive for T. spiralis. Seropositive for T. gondii was found in the boars in all the eight provinces investigated and for T. spiralis in four provinces. This is the first report on the seroprevalence of T. gondii and T. spiralis infections in wild boars in Korea. The consumption of undercooked wild boar meat may expose humans to a high risk of infection.     

Effectiveness of Action to Reduce Exposure of Free-Ranging California Condors in Arizona and Utah to Lead from Spent Ammunition

California condors (Gymnogyps californianus) released into the wild in Arizona ranged widely in Arizona and Utah. Previous studies have shown that the blood lead concentrations of many of the birds rise because of ingestion of spent lead ammunition. Condors were routinely recaptured and treated to reduce their lead levels as necessary but, even so, several died from lead poisoning. We used tracking data from VHF and satellite tags, together with the results of routine testing of blood lead concentrations, to estimate daily changes in blood lead level in relation to the location of each bird. The mean daily increment in blood lead concentration depended upon both the location of the bird and the time of year. Birds that spent time during the deer hunting season in two areas in which deer were shot with lead ammunition (Kaibab Plateau (Arizona) and Zion (Utah)) were especially likely to have high blood lead levels. The influence upon blood lead level of presence in a particular area declined with time elapsed since the bird was last there. We estimated the daily blood lead level for each bird and its influence upon daily mortality rate from lead poisoning. Condors with high blood lead over a protracted period were much more likely to die than birds with low blood lead or short-term elevation. We simulated the effect of ending the existing lead exposure reduction measures at Kaibab Plateau, which encourage the voluntary use of non-lead ammunition and removal of gut piles of deer and elk killed using lead ammunition. The estimated mortality rate due to lead in the absence of this program was sufficiently high that the condor population would be expected to decline rapidly. The extension of the existing lead reduction program to cover Zion (Utah), as well as the Kaibab plateau, would be expected to reduce mortality caused by lead substantially and allow the condor population to increase.     

Morbillivirus and <Emphasis Type="Italic">Toxoplasma</Emphasis> Exposure and Association with Hematological Parameters for Southern Beaufort Sea Polar Bears: Potential Response to Infectious Agents in a Sentinel Species

Arctic temperatures are increasing in response to greenhouse gas forcing and polar bears have already responded to changing conditions. Declines in body stature and vital rates have been linked to warming-induced loss of sea-ice. As food webs change and human activities respond to a milder Arctic, exposure of polar bears and other arctic marine organisms to infectious agents may increase. Because of the polar bear’s status as arctic ecosystem sentinel, polar bear health could provide an index of changing pathogen occurrence throughout the Arctic, however, exposure and monitoring protocols have yet to be established. We examine prevalence of antibodies to Toxoplasma gondii, and four morbilliviruses (canine distemper [CDV], phocine distemper [PDV], dolphin morbillivirus [DMV], porpoise morbillivirus [PMV]) including risk factors for exposure. We also examine the relationships between antibody levels and hematologic values established in the previous companion article. Antibodies to Toxoplasma gondii and morbilliviruses were found in both sample years. We found a significant inverse relationship between CDV titer and total leukocytes, neutrophils, monocytes, and eosinophils, and a significant positive relationship between eosinophils and Toxoplasma gondii antibodies. Morbilliviral prevalence varied significantly among age cohorts, with 1–2 year olds least likely to be seropositive and bears aged 5–7 most likely. Data suggest that the presence of CDV and Toxoplasma gondii antibodies is associated with polar bear hematologic values. We conclude that exposure to CDV-like antigen is not randomly distributed among age classes and suggest that differing behaviors among life history stages may drive probability of specific antibody presence.