Molecular detection of Sarcocystis sp. in a kept under human care black capuchin monkey (Sapajus nigritus., Goldfuss 1809) with necrotizing encephalitis.

A senile male black capuchin monkey (Sapajus nigritus) kept under human care in a Zoo was found dead after 2 weeks presenting signals of weight loss and hyporexia. Histopathological revealed a necrotizing encephalitis. Although it was not observed microscopically, Sarcocystis sp infection was detected in brain tissue from molecular assays. These infections have been rarely described in neotropical primates, particularly associated with tissue lesions.

Safety and immunogenicity of an adjuvanted recombinant spike protein-based severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) vaccine, SpikeVet™, in selected Carnivora, Primates and Artiodactyla in Australian zoos.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) can infect a broad range of animal species and has been associated with severe disease in some taxa. Few studies have evaluated optimal strategies to mitigate the risk to susceptible zoo animals. This study evaluated the safety and immunogenicity of a protein-based veterinary SARS-CoV-2 vaccine (SpikeVet™) in zoo animals. Two to three doses of SpikeVet™ were administered intramuscularly or subcutaneously 3-4 weeks apart to 354 zoo animals representing 38 species. SpikeVet™ was very well tolerated across all species. Minor adverse effects were observed in 1.69% of animals vaccinated, or 1.04% of vaccine doses administered. Preliminary immunogenicity analyses in representative carnivores (meerkats, lions) and an artiodactylid (domestic goat) showed SpikeVet™-immunized animals developed serum antibodies able to neutralize a range of SARS-CoV-2 variants, including the vaccine-homologous Wuhan and Mu variants, as well as vaccine-heterologous Omicron BA.2 and XBB.1 strains. Prior to vaccination, all eight lions were seropositive for Wuhan strain by surrogate viral neutralization testing, suggesting past infection with SARS-CoV-2 or cross-reactive antibodies generated by another closely related coronavirus. These results from a range of zoo species support the ongoing development of SpikeVet™ as a safe and effective veterinary SARS-CoV-2 vaccine.

Identification of potentially zoonotic parasites in captive orangutans and semi-captive mandrills: Phylogeny and morphological comparison.

Cysts and trophozoites of vestibuliferid ciliates and larvae of Strongyloides were found in fecal samples from captive orangutans Pongo pygmaeus and P. abelii from Czech and Slovak zoological gardens. As comparative material, ciliates from semi-captive mandrills Mandrillus sphinx from Gabon were included in the study. Phylogenetic analysis of the detected vestibuliferid ciliates using ITS1-5.8s-rRNA-ITS2 and partial 18S ribosomal deoxyribonucleic acid (rDNA) revealed that the ciliates from orangutans are conspecific with Balantioides coli lineage A, while the ciliates from mandrills clustered with Buxtonella-like ciliates from other primates. Morphological examination of the cysts and trophozoites using light microscopy did not reveal differences robust enough to identify the genera of the ciliates. Phylogenetic analysis of detected L1 larvae of Strongyloides using partial cox1 revealed Strongyloides stercoralis clustering within the cox1 lineage A infecting dogs, humans, and other primates. The sequences of 18S rDNA support these results. As both B. coli and S. stercoralis are zoonotic parasites and the conditions in captive and semi-captive settings may facilitate transmission to humans, prophylactic measures should reflect the findings.

Investigation of Toxoplasma infection in zoo animals using multispecies ELISA and GRA7 nested PCR.

BACKGROUND: Toxoplasma is an obligate intracellular protozoan that causes an important zoonotic disease with a worldwide distribution. Felids are the definitive hosts of this parasite, while virtually all warm-blooded animals, including birds, serve as intermediate hosts. Four ring-tailed lemurs (Lemur catta) in the Taipei Zoo died of acute Toxoplasma infection in June 2019. Since then, Toxoplasma has occasionally been identified in this Zoo during necropsy of dead animals and PCR of animal blood samples. Therefore, a general survey of Toxoplasma infection in animals in the Zoo seems to be needed. METHODS AND RESULTS: An indirect multispecies ELISA was used for the first time to screen for Toxoplasma infection in 326 serum samples collected from 75 species of animals. The infection rate of Toxoplasma was 27% (88/326). A commercial latex agglutination (LAT) assay was used to re-examine the samples with doubtful and uncertain ELISA results (151 samples from 42 species). The infection rate increased to 36.2% (118/326), and the indirect multispecies ELISA appeared to be applicable to 31 of 75 species animals included in this study. Nested PCR assays targeting the dense granule protein 7 (GRA7) gene and B1 gene were also used to detect Toxoplasma in DNA samples extracted from 10 liver or blood specimens from 8 animals. GRA7 gene fragments were amplified from 8 samples from 7 animals, while B1 gene fragments were amplified from only 4 samples from 4 animals. From the B1 nested PCR and the sequence data of GRA7 fragments amplified from infectious specimens, the animals in the Zoo were speculated to have been infected by at least three different Toxoplasma variants. CONCLUSIONS: According to the serological investigation, we speculated that over one-third (36.2%) of animals in Taipei Zoo presented the infection of Toxoplasma, and the indirect multispecies ELISA we used can be applied to detect Toxoplasma infection in 31 animal species included in this study. Sequence analysis revealed that at least three Toxoplasma variants were infecting the animals of Taipei Zoo.

Prevalence and multilocus genotyping of Giardia duodenalis in zoo animals in three cities in China.

Giardia duodenalis is a flagellated parasitic microorganism that parasitizes in the intestines of humans and animals. Although asymptomatic infections commonly exist in both humans and animals, some enteric symptoms have been reported in immunocompromised individuals, posing a threat to public health. Children could be infected with G. duodenalis through an environment contaminated by infective animals. Thus, the investigation of the prevalence and genotypes of G. duodenalis in zoo animals is important. In this study, 672 fecal samples of 113 species of animals, including non-human primates, artiodactyla, perissodactyla, proboscidian, marsupial, birds, carnivora, and rodents, were collected from three zoos in Hangzhou city, Dalian city, and Suzhou city in China. The samples were screened for the positivity of G. duodenalis by nested PCR based on the ß-giardin (bg), triosephosphate isomerase (tpi), and glutamate dehydrogenase (gdh) gene loci. The overall G. duodenalis prevalence was 10.6% (71/672). The prevalence in non-human primates, artiodactyla, perissodactyla, proboscidian, marsupial, birds, carnivora, and rodent was 6.9% (10/144), 9.0% (12/133), 17.1% (6/35), 0% (0/6), 8.7% (2/23), 13.3% (28/211), 6.7% (7/105), and 40.0% (6/15), respectively. The region and category were considered risk factors for G. duodenalis infection in zoo animals (p < 0.001). Additionally, four genotypes of G. duodenalis were identified in zoo animals, including assemblage E (n = 46), assemblage A (n = 18), assemblage B (n = 6), and assemblage D (n = 1). The assemblages A, B, D, and E are also genotypes observed in humans and other animals. Among the sequences obtained in our study, one multilocus genotype (MLG) of the sub-assemblage AI was observed within assemblage A. Furthermore, three MLGs were detected within assemblage B. These findings reveal G. duodenalis genetic variability in zoo animals in three cities in China and suggest that zoo animals could be a potential source of human infection with G. duodenalis.

Analysis of Yersinia pseudotuberculosis Isolates Recovered from Deceased Mammals of a German Zoo Animal Collection.

Yersinia pseudotuberculosis is an important pathogen for both humans and animals. It can infect livestock, as well as pets and wild animals. During recent years, a number of reports have described the isolation of Y. pseudotuberculosis from zoo animals, mainly birds and mammals, for which the infection was mostly lethal. Between 2005 and 2019, there were at least 17 cases of deceased mammals, belonging to five different species, which suffered from a Y. pseudotuberculosis infection at the Zoo Wuppertal, Germany. Since only scarce information exists on the properties of Y. pseudotuberculosis from zoo animals, we characterized eight isolates, covering all infected species, in detail. All isolates were members of biotype 1, but belonged to five serotypes, five sequence types (STs), and seven core-genome multilocus sequence types (cgMLSTs). Using pulsed-field gel electrophoresis (PFGE) analysis and whole-genome sequencing (WGS), the seven isolates could be discriminated from each other. They differed significantly regarding their virulence genes and mobile genetic elements. While the virulence plasmid pYV existed in all serotypes (five isolates), a complete high-pathogenicity island (HPI) was detected only in the serotypes O:1a, O:1b, and O:13 (four isolates), but not in O:2a and O:2b. Similarly, the content of other plasmids and prophages varied greatly between the isolates. The data demonstrate that the deceased mammals were infected by seven individual isolates and not by a single type predominating in the zoo animals.

Amino Acid Nutrition for Optimum Growth, Development, Reproduction, and Health of Zoo Animals.

Proteins are large polymers of amino acids (AAs) linked via peptide bonds, and major components for the growth and development of tissues in zoo animals (including mammals, birds, and fish). The proteinogenic AAs are alanine, arginine, aspartate, asparagine, cysteine, glutamate, glutamine, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, and valine. Except for glycine, they are all present in the L-isoform. Some carnivores may also need taurine (a nonproteinogenic AA) in their diet. Adequate dietary intakes of AAs are necessary for the growth, development, reproduction, health and longevity of zoo animals. Extensive research has established dietary nutrient requirements for humans, domestic livestock and companion animals. However, this is not true for many exotic or endangered species found in zoos due to the obstacles that accompany working with these species. Information on diets and nutrient profiles of free-ranging animals is needed. Even with adequate dietary intake of crude protein, dietary AAs may still be unbalanced, which can lead to nutrition-related diseases and disorders commonly observed in captive zoo species, such as dilated cardiomyopathy, urolithiasis, gut dysbiosis, and hormonal imbalances. There are differences in AA metabolism among carnivores, herbivores and omnivores. It is imperative to consider these idiosyncrasies when formulating diets based on established nutritional requirements of domestic species. With optimal health, populations of zoo animals will have a vastly greater chance of thriving in captivity. For endangered species especially, maintaining stable captive populations is crucial for conservation. Thus, adequate provision of AAs in diets plays a crucial role in the management, sustainability and expansion of healthy zoo animals.

Avian anaesthesia related mortality and the associated risk factors in a UK zoological collection.

OBJECTIVE: To analyse avian anaesthesia-related mortality in a UK zoological collection over a 5-year period and identify risk factors for mortality. STUDY DESIGN: Retrospective cohort study. ANIMALS: A total of 135 individual birds across 37 species, anaesthetized during 206 events in a UK zoological collection between 1 January 2014 and 30 June 2019 (inclusive). METHODS: Anaesthesia records were reviewed and variables such as age, body condition, weight, sex, duration of anaesthesia and health status were collated. Anaesthesia-related mortality was defined as those deaths occurring during anaesthesia and up to 7 days postanaesthesia. Outcome was analysed using multivariable conditional logistic regression. Overall mortality was defined and included birds euthanised during anaesthesia for non-anaesthesia related reasons. Data were summarised as median (range). A value of p < 0.05 was considered significant. Relative risks and 95% confidence intervals (95% CI) were calculated for the association between risk factors and anaesthetic death where a statistically significant difference was found. RESULTS: The overall mortality rate was 10.19% (95% CI = 6.06-14.3%), while anaesthesia-related mortality was 3.88% (95% CI = 1.69-7.51%). Birds with an abnormal health status had a 15.53-fold (95% CI = 1.95-123.63) increased risk of death compared with those with a normal health status. The duration of anaesthesia was also a statistically significant risk factor (p = 0.021) in the univariable analysis, but not when combined with health status. No other variables were associated with anaesthesia-related mortality. CONCLUSIONS AND CLINICAL RELEVANCE: Abnormal health status and longer anaesthetic procedures were associated with a significantly increased risk of anaesthesia-related death in this population of birds. It is recommended that anaesthetic duration is minimized, and pre-existing diseases are diagnosed where possible prior to general anaesthesia of birds. Anaesthetizing healthy birds was associated with a low risk of mortality.

Serological survey of Coxiella burnetii infections in dairy cattle, sheep, goats and zoo animals in Hungary - Short communication.

Q fever is a disease of high zoonotic potential, but interest in its causative agent is rather low although it causes some public health problems in Hungary. The prevalence of Q fever is highly variable by country. The main reservoirs of the disease are the same domestic ruminant species everywhere, but the epidemiological profile depends on the features of the specific reservoir. The aim of this large-scale study was to demonstrate the importance of Q fever in different species as a possible source for human infection in most regions of Hungary. A total of 851 serum samples from 44 dairy farms, 16 sheep flocks, 4 goat farms and 3 zoos located in different parts of Hungary were tested. The presence of antibodies to Coxiella burnetii was surveyed in dairy cattle (n = 547), goats (n = 71), sheep (n = 200) and zoo animals (n = 33). The animal species tested in Hungary showed different seroprevalence values of C. burnetii infection. Seropositivity by the enzyme-linked immunosorbent assay was found in 258 out of 547 (47.2%) cows and in 69 out of 271 (25.5%) small ruminants, among them in 47 out of 200 (23.5%) sheep and in 22 out of 71 (31.0%) goats. Antibodies to C. burnetii were not detected in zoo animals. Seropositivity was demonstrated in 44 out of 44 (100%) dairy cattle farms, with at least one serum sample found to be positive on each farm. The seropositivity rate of small ruminant farms was 55.0% (11 positive out of 20 tested), with 9 out of 16 (56.3%) sheep flocks and 2 out of 4 (50.0%) goat herds showing seropositivity.

Assessing preferences of two zoo-housed Aldabran giant tortoises (Aldabrachelys gigantea) for three stimuli using a novel preference test.

Recent research has uncovered many complex cognitive traits and affective processes in many reptile species, such as the ability to make choices that are rewarding or pleasurable. The investigation herein was initiated after many years of anecdotal reports, and preliminary observations, that the two male Aldabran giant tortoises (Aldabrachelys gigantea), Wilbur and Little John, at Melbourne Zoo (Melbourne, Australia) would approach the fence-line of their enclosure, sometimes multiple times daily, to "solicit" interactions from zoo visitors and/or zoo staff. Following other research with Galapagos giant tortoises (Geochelone nigra), this experiment was a novel preference test with two male Aldabran giant tortoises to assess their individual preferences for three stimuli: (1) human interaction (shell and neck rubs provided using hands); (2) food (highly palatable carrot pieces); and (3) an "enrichment" ball (a red "boomer ball"). The results found that Wilbur highly preferred human interaction and food over the ball; whereas Little John preferred food, and showed some initial interest in the ball, but did not approach the human on any of his 13 human interaction trials. These results reflect previous published findings, that human interaction is a preferred "enrichment," but is dependent upon a multitude of other contextual factors, and is not preferred at all times by all individuals. Individual variation may be very important to understand to have greater positive effect on the welfare-related needs of each giant tortoise housed in captivity. Further research is clearly required to enhance the welfare and well-being of many types of zoo-housed reptiles.

Natural Canine Distemper Virus Infection in Linnaeus's 2-Toed Sloths (Choloepus didactylus).

An outbreak of canine distemper virus in a private zoo in eastern Tennessee in July 2016 led to fatal clinical disease in 5 adult, wild-caught Linnaeus's 2-toed sloths (Choloepus didactylus). Clinical signs included hyporexia, lethargy, mucopurulent nasal discharge, and oral and facial ulcers. At necropsy, affected animals had crusts and ulcers on the lips, nose, tongue, and oral cavity. Microscopically, all sloths had widespread, random, hepatic necrosis; lymphoid depletion; and bronchointerstitial pneumonia. The central nervous system did not contain gross or histopathologic lesions in any of the 5 sloths, although immunoreactivity for viral antigen was present within vessel walls. Epithelial cells and histiocytes within numerous organs contained intranuclear and intracytoplasmic inclusions and occasional syncytial cells. Canine distemper virus was confirmed with immunohistochemistry and virus isolation. Viral sequencing identified the novel American-4 strain prevalent in eastern Tennessee wildlife. This is the first pathologic characterization of canine distemper virus infection in sloths (family Choloepodidae, order Pilosa) and emphasizes the significant morbidity and mortality in this species.

Prevalence and subtypes of Blastocystis sp. infection in zoo animals in three cities in China.

Blastocystis is a highly prevalent eukaryotic parasite of many animals and humans worldwide. It can compromise the gastrointestinal tract and cause gastrointestinal symptoms, constituting a serious threat to human health and animal growth. Many animals are potential sources of Blastocystis infection in humans. However, limited data are available regarding the prevalence and subtype distribution of Blastocystis infection among zoo animals in China. Therefore, the present study examined the prevalence and subtypes of Blastocystis in zoo animals in Hangzhou, Dalian, and Suzhou cities, China. Of 450 fecal samples from zoo animals, 27 (6.0%) were PCR-positive for Blastocystis, with 7.7% (8/104), 11.3% (7/62), 16.7% (3/18), 1.8% (2/114), 6.3% (1/16), 9.5% (2/21), and 3.6% (4/109) in artiodactyla, aves, rodentia, nonhuman primates, perissodactyla, marsupialia, and carnivora, respectively. Significant differences in the prevalence of Blastocystis were found among different animal groups (P < 0.05). Sequence analysis showed 7 known subtypes (ST2, ST4, ST5, ST7, ST8, ST10, and ST14) of Blastocystis in the present study, with ST10 (10/27) as the predominant subtype in all three of the examined zoos. To our knowledge, this is the first report of Blastocystis infection in Damaliscus dorcas, Cervus elaphus, Macropus rufogriseus, Grus japonensis, Trichoglossus haematodus, Panthera tigris ssp. tigris (white), Panthera tigris ssp. altaica, Lycaon pictus, Suricata suricatta, and Dolichotis patagonum in China. These results demonstrate the presence of Blastocystis infection in zoo animals and provided baseline data for preventing and controlling Blastocystis infection in zoo animals and humans in China.

MOLECULAR CHARACTERIZATION OF FECAL ESCHERICHIA COLI ISOLATED FROM ZOO ANIMALS.

Accredited zoos and animal parks play an important role in animal health research and conservation, providing important insights on matters of public health including zoonotic infectious diseases and antimicrobial resistance (AMR). The emergence and spread of AMR is a complex phenomenon that jeopardizes human and animal health and also threatens the long-term survival of endangered species. The presence of ß-lactamases in clinical isolates is particularly significant as they can jeopardize the efficacy of critically important antimicrobials. Although the presence of ß-lactamases and extended-spectrum ß-lactamases (ESBLs) producing Enterobacteriaceae in zoo animals has been reported, data are not available for northern European countries. In addition, few data are available on phylogenetic grouping of Escherichia coli isolated from zoo animals that can provide additional information on the host-bacterium relationship and on the pathogenicity of isolates. This study aimed to characterize fecal E. coli isolated from 33 healthy zoo animals from 22 species in Ireland, using conventional and molecular microbiological methods. All E. coli isolates were ampicillin resistant, but combined resistance to amoxicillin and clavulanic acid was not detected. Three E. coli isolates sampled from one Amur tiger, one Bornean orangutan, and one Southern white rhino were multidrug resistant, and blaTEM was detected in E. coli recovered from the Amur tiger and the Bornean orangutan. Other ß-lactamases, including ESBLs and AmpCs and plasmid-mediated mcr-1 and mcr-2, were not detected. Overall, E. coli isolates investigated were susceptible to the majority of the antimicrobials tested, and only two animals shed E. coli carrying ß-lactamase-encoding genes. The majority of isolates belonged to phylogenetic group B1. The screening of the AMR phenotype and genotype of zoo animal E. coli provides useful data that is relevant to antimicrobial stewardship in the zoo veterinary services and relevant to the bank of knowledge needed for tackling AMR.

Effect of paternal age on the birth sex ratio in captive populations of aye-aye (Daubentonia madagascariensis (Gmelin)).

For the management of captive populations of zoo animals, it is important to elucidate factors that affect the offspring birth sex ratio. On the basis of the sex allocation theory, the Trivers-Willard and mate attractive/quality hypotheses predict that maternal and paternal conditions affect offspring birth sex ratios. We examined these predictions for the birth sex ratio of aye-aye Daubentonia madagascariensis (Gmelin) by analyzing the pedigree information in the International Studbook. We found that the birth sex ratio of the aye-aye was affected by the paternal age, but not maternal age and other environmental factors (birth year, season, and institution). The younger the sire, the more the offspring sex ratio was biased toward males. These results are useful for the effective population management of captive aye-aye and illustrated the usefulness of the sex allocation theory in the sex ratio management of zoo animals.

The effects of zoo visitors on Quokka (Setonix brachyurus) avoidance behavior in a walk-through exhibit.

The behavior of zoo animals may be influenced by visitors, with possible implications on animal welfare. We examined the effects of the presence of visitors on the presence and visibility of free-ranging quokkas (Setonix brachyurus) in preferred areas of a walk-through enclosure at Melbourne Zoo, Australia. In a controlled experiment, two visitor treatments were randomly imposed: (1) enclosure open to visitors as normal ("Open") and (2) enclosure closed to visitors ("Closed"). Treatments were imposed for 2-day periods, with five replicates of each treatment (10 2-day periods in total). Instantaneous point sampling of camera footage at 1-min intervals from 09:00 to 16:00 hr each day recorded the location and visibility of quokkas. Data were analyzed using a two-factor ANOVA (visitor treatment and study area), with 2-day summary values as the unit of analysis. While the presence of quokkas in the study areas was not affected by treatment, fewer (p < 0.05) quokkas were visible from the visitor paths and more (p < 0.05) quokkas were obscured from the visitor paths in the "Open" treatment than the "Closed" treatment. Number of quokkas differed between study areas across treatments (p < 0.01), but there was no interaction (p > 0.05) between visitor treatment and study area. The reduction in quokka visibility when visitors were present indicates that visitors were at least moderately fear provoking for the quokkas in this experiment, but further research is required to examine the effects of visitor presence and behaviors on the behavior and stress physiology of quokkas in walk-through enclosures, as there may be possible welfare implications.

RETROSPECTIVE EVALUATION OF A NOVEL SUSTAINED-RELEASE IVERMECTIN VARNISH FOR TREATMENT OF WOUND MYIASIS IN ZOO-HOUSED ANIMALS.

Myiasis is a major disease condition in human and veterinary medicine. Domestic, free-ranging, and zoo-housed animals can be severely affected by myiasis. Depending on case severity, multiple treatment episodes may be indicated and can lead to recurrent capturing, handling stress, and anesthetics, all of which increase the risk of adverse responses (including death) individually and also in the herd. As an insecticide, ivermectin is often used for larval control. A total of 28 individual myiasis cases were retrospectively evaluated, out of which 11 cases were also treated using an ivermectin sustained-release varnish (SRV). The clinical outcome of all cases was assessed and the results suggest that the use of a topical ivermectin SRV (with or without concurrent injectable ivermectin) can reduce handling and treatments, has no adverse effects, and has minimal recurrence of the disease when compared with cases treated without it.

An eight-year survey of the intestinal parasites of carnivores, hoofed mammals, primates, ratites and reptiles in the Ljubljana zoo in Slovenia.

Problems with parasitic infections and their interspecies transmissions are common in zoological gardens and could pose serious health damage to captive animals. This study presents results of eight-year monitoring of intestinal parasites in animals from Zoo Ljubljana, Slovenia. A total of 741 faecal samples from 40 animal species were collected two to four times per year and examined microscopically. Intestinal parasites were detected in 45% of samples, with detection of helminths (Cestoda, Nematoda - Ascaridida, Enoplida, Strongylida, Oxyurida, Rhabditida and Trichurida) and protists (Apicomplexa and Ciliophora) in 25% and 13% of samples, respectively; mixed infection was found in 7% of samples. The mostly infected were ungulates (61%), followed by reptiles (44%), ratites (29%), primates (22%) and carnivores (7%). During the observation period, the number of infected animal species increased from 8 to 25. This is the first long-term monitoring study of intestinal parasites in zoo animals from Slovenia. Routine monitoring of parasitic infection and regular deworming and hygienic measures are necessary to prevent gastrointestinal infections in captive animals.

Effects of visitor numbers on captive European red squirrels (Sciurus vulgaris) and impacts on visitor experience.

Visitors to zoological collections can have substantial effects on captive animals that vary according to species, enclosure design, visitor proximity, and husbandry methods. One particularly intense form of visitor interaction occurs in immersive exhibits such as walk-through enclosures. Such enclosures are increasingly common but effects on animal behavior are currently understudied. Here, the behavior of captive European red squirrels (Sciurus vulgaris) is studied in relation to visitor numbers in a walk-through enclosure. We also quantify the correlation between squirrel encounters and visitor experience. Interaction with humans increased significantly as the number of visitors inside the enclosure increased. The number of children present significantly increased locomotion and decreased eating, possibly due to disturbance and squirrels moving away from busy areas. By contrast, the number of adults significantly increased eating and decreased inactivity due to squirrels approaching visitors. The positive reinforcement training used by the keepers (offering food rewards to the squirrels for coming to them to allow routine medical checks) meant that squirrels associated adults with food opportunities. Squirrel encounter rate (number of squirrels seen by each group of visitors) was significantly affected by the number of adults and visitor duration (positive relationships) and noise as perceived by visitors (negative relationship). Encounter rate was positively correlated with overall visitor experience. Our results indicate that visitors affect behavior but this effect is influenced by husbandry methods. It is vital that visitors, especially children, minimize noise, and move slowly in the enclosure, both for the sake of the animals and their own experience.

Prevalence and characterization of Chlamydia DNA in zoo animals in Japan.

Because many people visit zoos, prevention of zoonoses is important from the standpoint of public health. This study examined the prevalence of Chlamydia among zoo animals in Japan by PCR and characterized these bacteria by performing phylogenetic analyses of the sequences of the variable domain (VD) 2 and VD4 regions of the ompA gene, which encodes the Chlamydia major outer membrane protein. Fecal samples were collected from 1150 zoo animals in five zoos and examined for Chlamydia DNA. Chlamydia psittaci DNA was found in 3.9% of mammals, 7.2% of birds and 8.1% of reptiles. The prevalence of Chlamydia pneumoniae DNA was significantly higher in reptiles (5.8%) than in mammals (0.3%) and birds (0.3%). Phylogenetic analysis of the ompA VD2 region from 18 samples showed that nine were in three different clusters containing C. psittaci strains, six were in a cluster containing C. pneumoniae strains and three each formed a distinct branch. Furthermore, phylogenetic analysis of the ompA VD4 region showed that C. pneumoniae DNAs from reptiles were close to those from human patients. The C. pneumoniae DNAs from the European glass lizard, Emerald tree boa, and Panther chameleon were classified in clusters that were distinct from other strains, suggesting that these reptiles had each been infected with a specific C. pneumoniae genotype. This study showed that diverse Chlamydia strains have been prevalent among a variety of zoo animals.

Tuberculosis in elephants-a reemergent disease: diagnostic dilemmas, the natural history of infection, and new immunological tools.

Tuberculosis (TB) in elephants has been described since ancient times. However, it was not until 1996 when infection with Mycobacterium tuberculosis was identified in a herd of circus elephants that significant research into this disease began. The epidemiology and natural history of TB were unknown in elephants since there had been no comprehensive screening programs, and diagnostic techniques developed for cervidae and bovidae were of unknown value. And, while precepts of test and slaughter were the norm for cattle and deer, this was considered untenable for an endangered species. With no precedent for the treatment of TB in animals, treatment regimens for elephants were extrapolated from human protocols, which guided changes to the Guidelines for the Control of Tuberculosis in Elephants. In the absence of diagnostic testing to confirm cure in elephants, the efficacy of these treatment regimens is only beginning to be understood as treated elephants die and are examined postmortem. However, because of pressures arising from public relations related to elephant husbandry and the added considerations of TB infection in animals (whether real or imagined), sharing of information to aid in research and treatment has been problematic. Here we review the challenges and successes of the diagnosis of tuberculosis in elephants and discuss the natural history of the disease to put the work of Landolfi et al on the immunological response to tuberculosis in elephants in perspective.