Zoos In China Research Articles

Viral metagenomic analysis reveals potential biological hazards in non‐human primates in a zoo

AbstractPathogens have been documented to be transmissible between humans and non‐human primates (NHPs), with NHPs demonstrating susceptibility to human viruses. Conducting surveillance for viruses in NHPs to identify potential zoonotic agents that may emerge or pose a high risk of spillover remains a critical strategy for preparing for and responding to future zoonotic events. This study employed viral metagenomic analysis on nine randomly selected NHPs from the Zoo in China to detect potential pathogens within captive environments. The analysis identified the presence of picornavirus Encephalomyocarditis virus (EMCV), poxvirus (BeAn 58,058 virus, BAV), and retroviruses (human endogenous retrovirus, HERV and baboon endogenous virus, BaEV). These viruses exhibited significant genetic homology to established viral strains, with EMCV demonstrating close relatedness to a strain previously detected in the United States, and BAV beling newly identified in NHPs. The study suggests that zoo rodents, particularly rats, serve as the primary reservoirs for the viruses, thereby posing a potential threat to public health. Therefore, this underscores the imperative to enhance rodent control measures within zoological institutions and provides strategic recommendations for mitigating interspecies viral transmission.

Development and Application of a Portable Environmental DNA Test for the Detection of Rafetus swinhoei in Viet Nam

ABSTRACTSwinhoe's (or Yangtze) giant softshell turtle (Rafetus swinhoei) is a large, critically endangered freshwater turtle and considered among the rarest species in the world. As of 2024, only two individuals have been confirmed to remain alive, one at the Suzhou Zoo in China and one in Xuan Khanh Lake, Viet Nam. The only hope for the long‐term survival of R. swinhoei is finding additional, as yet undiscovered, animals that have thus far eluded detection by traditional survey methods. In recent years, numerous studies have been published on the use of environmental DNA (eDNA) for species detection and monitoring. This method takes advantage of the persistence of DNA in the environment, such as in water, soil, and air. An organism's DNA is shed into the environment through urine, feces, and the sloughing of skin. Species‐specific quantitative polymerase chain reaction (qPCR) testing can be used to detect eDNA in samples collected from the environment. eDNA trials to detect R. swinhoei were initiated by the Asian Turtle Program and Washington State University in 2013. To expand the use of eDNA for species detection by conservationists, we developed and validated a first‐of‐its‐kind innovative point‐of‐detection (POD) qPCR platform for the rapid, onsite detection of R. swinhoei from water samples. Here we show that the portable eDNA test kit can be used for the successful detection of R. swinhoei in a large body of water and that pooling filters may be a useful strategy to reduce test costs and improve detection efficiency. Use of this test can expand the search for R. swinhoei in unexplored and understudied lakes, reservoirs, and other bodies of water where this species may be present and could inform field surveys utilizing eDNA for other threatened species that are rare in nature.

Tourism destination social responsibility and the moderating role of self-congruity

Propósitonuestro artículo investiga las interrelaciones de la responsabilidad social del destino (DSR), el apego emocional, la autocongruencia, la satisfacción experiencial y el comportamiento ambientalmente responsable (ERB).Diseño/metodología/enfoqueutilizando una muestra de 294 visitantes del zoológico de Guangzhou en China, nuestro estudio probó las relaciones antes mencionadas a través del modelo de ecuaciones estructurales.Hallazgosel apego emocional media el efecto de DSR en la satisfacción experiencial, mientras que el apego emocional y la satisfacción experiencial median el efecto de DSR en ERB secuencialmente. Además, la autocongruencia modera la relación entre DSR y apego emocional.Implicaciones prácticasla administración de los zoológicos debe utilizar estrategias de comunicación de DSR de manera más proactiva para que los visitantes estén bien informados sobre sus actividades económicas, filantrópicas, ambientales y sociales en la comunidad anfitriona. Esto tendrá muchas consecuencias positivas, incluidos los ERB de los visitantes.Originalidad/valorel estudio se suma a la literatura de DSR al presentar múltiples mecanismos de mediación y caminos que conducen a los ERB de los visitantes.

Occurrence and Molecular Characteristics of Microsporidia in Captive Red Pandas (Ailurus fulgens) in China.

Enterocytozoon bieneusi and Encephalitozoon spp. are microsporidian pathogens with zoonotic potential that pose significant public health concerns. To ascertain the occurrence and genotypes of E. bieneusi and Encephalitozoon spp., we used nested PCR to amplify the internal transcribed spacer (ITS) gene and DNA sequencing to analyze 198 fecal samples from red pandas from 6 zoos in China. The total rate of microsporidial infection was 15.7% (31/198), with 12.1% (24/198), 1.0% (2/198), 2.0% (4/198) and 1.0% (2/198) for infection rate of E. bieneusi, Encephalitozoon cuniculi, Encephalitozoon intestinalis and Encephalitozoon hellem, respectively. One red panda was detected positive for a mixed infection (E. bieneusi and E. intestinalis). Red pandas living in semi-free conditions are more likely to be infected with microsporidia (χ2 = 6.212, df = 1, p < 0.05). Three known (SC02, D, and PL2) and one novel (SCR1) genotypes of E. bieneusi were found. Three genotypes of E. bieneusi (SC02, D, SCR1) were grouped into group 1 with public health importance, while genotype PL2 formed a separate clade associated with group 2. These findings suggest that red pandas may serve as a host reservoir for zoonotic microsporidia, potentially allowing transmission from red pandas to humans and other animals.

Characterization of a Tigecycline-Resistant and blaCTX-M-Bearing Klebsiella pneumoniae Strain from a Peacock in a Chinese Zoo.

In Chinese zoos, there are usually specially designed bird parks, similar to petting zoos, that allow children and adults to interact with diverse birds. However, such behaviors present a risk for the transmission of zoonotic pathogens. Recently, we isolated eight strains of Klebsiella pneumoniae and identified two blaCTX-M-positive strains from 110 birds, including parrots, peacocks, and ostriches, using anal or nasal swabs in a bird park of a zoo in China. There, K. pneumoniae LYS105A was obtained from a diseased peacock with chronic respiratory diseases by a nasal swab, which harbored the blaCTX-M-3 gene and exhibited resistance to amoxicillin, cefotaxime, gentamicin, oxytetracycline, doxycycline, tigecycline, florfenicol, and enrofloxacin. According to an analysis by whole-genome sequencing, K. pneumoniae LYS105A belongs to serotype ST859 (sequence type 859)-K19 (capsular serotype 19) and contains two plasmids, of which pLYS105A-2 can be transferred by electrotransformation and harbors numerous resistance genes such as blaCTX-M-3, aac(6')-Ib-cr5, and qnrB91. The above-mentioned genes are located in a novel mobile composite transposon, Tn7131, which makes horizontal transfer more flexible. Although no known genes were identified in the chromosome, a significant increase in SoxS upregulated the expression levels of phoPQ, acrEF-tolC, and oqxAB, which contributed to strain LYS105A acquiring resistance to tigecycline (MIC = 4 mg/L) and intermediate resistance to colistin (MIC = 2 mg/L). Altogether, our findings show that bird parks in zoos may act as important vehicles for the spread of multidrug-resistant bacteria from birds to humans and vice versa. IMPORTANCE A multidrug-resistant ST859-K19 K. pneumoniae strain, LYS105A, was obtained from a diseased peacock in a Chinese zoo. In addition, multiple resistance genes such as blaCTX-M-3, aac(6')-Ib-cr5, and qnrB91 were located in a novel composite transposon, Tn7131, of a mobile plasmid, implying that most of the resistance genes in strain LYS105A can be moved easily via horizontal gene transfer. Meanwhile, an increase in SoxS can further positively regulate the expression of phoPQ, acrEF-tolC, and oqxAB, which is the key factor for strain LYS105A to develop resistance to tigecycline and colistin. Taken together, these findings enrich our understanding of the horizontal cross-species spread of drug resistance genes, which will help us curb the development of bacterial resistance.

Influence of soundscape on visiting time in zoos

The soundscape is very important for the environmental quality and animal welfare of zoos. This paper explores the relationship between the soundscape and visiting time, for the case of Guangzhou Zoo in China. In this research, we measured the sound indicators in the zoo along with the visiting time of visitors. This included the cases of with or without animal soundscape around the exhibition area, recorded by video equipment. It was found that there was a positive correlation between visiting time and the consistency of animal sound scene vision. The visiting time of tourists will increase significantly When there is animal sound scene in the exhibition area.The visiting time of tourists is significantly increased when the animal sound is consistent with the animals seen by tourists.Within a certain range, the visiting time is not affected by the sound pressure level.The results of this research are expected to guide the soundscape design, construction and management of zoos.

Exploration of cultural norms and behavioural beliefs about zoo animal behaviour, welfare, ethics and husbandry practices in a sample of the international zoo community.

Beliefs influence the intentions of people to behave in certain ways towards animals. This study presents survey responses from 237 people working in zoos in China and Europe and describes their demographic characteristics. It explores their beliefs about zoo animal behaviour, welfare and ethical issues, and zoo practices, using a survey methodology. These beliefs may be influenced by individual demographic or cultural factors such as age, genderand region of employment, as well as experiential or situative 'norms' within the work environment. Beliefs were significantly influenced by the region of employment with Chinese respondents beliefs being significantly different to beliefs from respondents in the United Kingdom or the rest of Europe. Hierarchical cluster analysis of the survey generated clusters of people from both regions who indicated positive beliefs about zoo animal welfare as well as clu sters indicating a lack of understanding of some zoo animal welfare issues. In addition, a cluster suggesting cognitive dissonance between beliefs about animals welfare and zoo practices was generated from Chinese responses. Factor analysis identified that prioritisation of in-situ conservation within good animal welfare was a key feature in Chinese respondents, whereas European beliefs prioritising in-situ conservation were distinct from those on supporting good animal welfare. This paper identifies similarities and differences in beliefs about zoo animal welfare and zoo husbandry practices between Europe and China, and discusses the underlying norms and values that these beliefs may reflect.

An outbreak of tuberculosis in endangered northern pig-tailed macaques (Macaca leonina) and milu deer (Elaphurus davidianus) from a zoo in China.

Tuberculosis (TB) is a zoonotic disease that affects humans and domesticated and wild animals. Animals in zoos are potentially an important source of TB for humans; however they are often neglected in routine disease surveillance programs. This investigation reports an outbreak of TB in milu deer and northern pig-tailed macaques in a zoo in Wuhan, China, which highlighted the need for improved prevention and control of TB in China. Between 24 November and 9 December 2020 two milu deer and a northern pig-tailed macaque that were displaying signs of wasting died. Post-mortem, histopathological diagnosis and acid fast staining were used for the dead animals. Multiple PCR for Mycobacterium tuberculosis complex (MTBC) was performed to identify the bacterial in both milu deer and northern pig-tailed macaque. The serum antibody iELISA for MTBC was then performed for all the surviving milu deer and northern pig-tailed macaques. Six seropositive milu deer and a seropositive northern pig-tailed macaque were subsequently euthanised and, along with two other dead milu deer, necropsied. DNA from these tissue samples was extracted and detected MTBC using PCR and Real-time PCR. Subsequently bacterial isolation was used to confirm the infection. The lungs of the dead animals displayed gross and histological TB-like lesions and changes, and red staining bacilli were detected in smears of the lesions by microscopy after acid fast staining. Mycobacterium bovis (M. bovis) was detected in the two milu deer and Mycobacterium tuberculosis (M. tb) in the northern pig-tailed macaque using multiple PCR for MTBC. 35.3% surviving milu deer and 50% surviving northern pig-tailed macaques MTBC serologically positive. Six of the euthanised milu deer were also positive on a DNA test for M. bovis and the euthanised northern pig-tailed macaque was positive to M. tb. This is the first report of tuberculosis in the endangered species, milu deer and northern pig-tailed macaques, in China, and warrants urgent attention by researchers and conservation authorities. These cases highlight the need for expanding surveillance for MTBC to zoos in China.

Morphological characteristics and phylogenetic evidence reveal two new species of Acremonium (Hypocreales, Sordariomycetes).

Using chicken feathers as bait, Acremoniumglobosisporum sp. nov. and Acremoniumcurvum sp. nov. were collected from the soil of Yuncheng East Garden Wildlife Zoo and Zhengzhou Zoo in China. They were identified by combining the morphological characteristics and the two-locus DNA sequence (LSU and ITS) analyses. In the phylogenetic tree, both new species clustered into separate subclades, respectively. They were different from their allied species in their morphology. The description, illustrations, and phylogenetic tree of the two new species were provided.

Prevalence and Molecular Identification of Entamoeba spp. in Non-human Primates in a Zoological Garden in Nanjing, China.

ObjectiveEntamoeba spp. are globally distributed zoonotic parasites that infect various hosts, among which non-human primates (NHPs) have been identified as one of the most common hosts of these parasites. Consequently, the infections of Entamoeba spp. in captive NHPs from Nanjing Hongshan Forest Zoo in China were investigated in order to assess their zoonotic potential.MethodsA total of 120 fresh fecal samples, including 19 species of NHPs, were collected from four breeding bases of the zoo from May to June 2019. The infections of six species of Entamoeba spp. were detected by PCR using the 16S or 18S rDNA-specific primers, and the positive samples were sequenced and analyzed.ResultsEntamoeba spp. were detected as positive in 59 NHPs fecal samples (49.17%), including five Entamoeba species: Entamoeba histolytica (7.50%), E. dispar (22.50%), E. coli (22.50%), E. chattoni (10.00%) and E. nuttalli (1.67%). Infection with one Entamoeba species was more common (35%) than co-infections (13.33%) or infections with three Entamoeba species (0.83%). There was a significantly higher prevalence rate of Entamoeba spp. in the species Pongo pygmaeus and Macaca mulatta than in Papio sp., Mandrillus sphinx, and Saimiri sciureus.ConclusionEntamoeba spp. are highly prevalent in the NHPs raised in Nanjing Hongshan Forest Zoo. Therefore, attention should be paid to the development of containment strategies of Entamoeba spp. in this zoological garden.

Prevalence and antimicrobial susceptibility of CTX-M-type-producing Escherichia coli from a wildlife zoo in China.

BackgroundWildlife zoos provide the opportunity for children and adults to interact with animals, However, it's unknown that the risk of contact with animals, which carried zoonotic pathogens and antimicrobial resistant bacteria.ObjectivesThis study aimed to investigate the prevalence and antimicrobial susceptibility of extended‐spectrum β‐lactamases Escherichia coli (ESBLs‐EC) from a wildlife zoo in China.MethodsA total of 93 wildlife faecal samples were collected from a wildlife zoo. Agar dilution method was used to determine the resistant phenotype. Whole genomes sequencing and bioinformatic analysis were employed to evaluate the molecular typing and genetic relationships of ESBLs‐EC.ResultsA total of 23 CTX‐M‐positive ESBLs‐EC were isolated from swan (n = 14), squirrel monkey (n = 5), black hat hanging monkey (n = 2), gibbon monkey (n = 1) and phoenicopteridae (n = 1) respectively. All ESBLs‐EC strains were resistant to cefotaxime, tetracycline, ciprofloxacin and trimethoprim‐sulfamethoxazole, but susceptible to colistin, tigecycline, meropenem and amikacin. By screening whole genome sequences, ESBLs‐EC strains main carried bla CTX‐M‐55 (34.8%, 8/23) and bla CTX‐M‐14 (26.0%, 6/23), following by bla CTX‐M‐27 (21.7%, 5/23), bla CTX‐M‐15 (13.0%, 3/23) and bla CTX‐M‐121 (4.3%, 1/23). ESBLs‐EC strains mainly belonged to phylogroup A (60.9%, 14/23), and ST48, ST746 and ST616 (3 strains respectively, 13.0%) were major ST types. Core genome‐based single nucleotide polymorphism (SNP) analysis suggested that strains from the swan, over the phylogenetic tree, have a closer genetic relationship with strains from other animals (black hat hanging monkey, gibbon monkey, phoenicopteridae and squirrel monkey).ConclusionsCTX‐M type ESBLs‐EC can transmit between animals in wildlife zoos, which may be a risk of spread to animal keepers, veterinarians and visitors when contact with animals. Our study provides that the importance of hygiene measures to minimise the risk of transmission of ESBLs‐EC to visitors in wildlife zoos.

Characterization of the Gut Microbiome and Resistomes of Wild and Zoo-Captive Macaques.

Rhesus macaques (Macaca mulatta) are the most widely distributed species of Old World monkey and are frequently used as animal models to study human health and disease. Their gastrointestinal microbial community likely plays a major role in their physiology, ecology and evolution. Herein, we compared the fecal microbiome and antibiotic resistance genes in 15 free-ranging and 81 zoo-captive rhesus macaques sampled from two zoos in China, using both 16S amplicon sequencing and whole genome shotgun DNA sequencing approaches. Our data revealed similar levels of microbial diversity/richness among the three groups, although the composition of each group differed significantly and were particularly marked between the two zoo-captive and one wild groups. Zoo-captive animals also demonstrated a greater abundance and diversity of antibiotic genes. Through whole genome shotgun sequencing we also identified a mammalian (simian) associated adenovirus. Overall, this study provides a comprehensive analysis of resistomes and microbiomes in zoo-captive and free-ranging monkeys, revealing that semi-captive wildlife might harbor a higher diversity of antimicrobial resistant genes.

A survey of Cryptosporidium prevalence among birds in two zoos in China.

BackgroundCryptosporidiosis is an important zoonotic protozoan disease worldwide, but few studies on this disease have been performed in wild birds; thus, our knowledge of this disease is insufficient, even in zoo birds. Animals in zoos are possible zoonotic disease reservoirs, potentially resulting in zoonotic agent spillover to humans; accordingly, our understanding of such phenomena should be improved.MethodsA total of 263 fresh fecal samples from 43 avian species were randomly collected from the Beijing Zoo and Harbin North Forest Zoo and screened for the prevalence of Cryptosporidium by 18S rRNA gene sequencing. Cryptosporidium species were distinguished based on the combined results of phylogenetic tree and genetic distance analyses conducted with the inclusion of seven avian Cryptosporidium species and 13 avian Cryptosporidium genotypes. The genetic diversity of Cryptosporidium parvum among different hosts, including humans, cattle, dogs, and birds, and the genetic diversity of avian C. parvum among avian hosts in China, Iraq and Brazil were determined based on C. parvum 18S rRNA haplotypes.ResultsThe results of PCR targeting the 18S rRNA gene revealed that 1.9% (5/263) of the samples were Cryptosporidium-positive. Four of the five Cryptosporidium-positive samples originated from white cranes (Grus leucogeranus), and one originated from a flamingo (Phoenicopteridae). Avian C. parvum isolates, including the isolates examined in the present study, showed gene flow with other isolates from different types of hosts, including humans, cattle and dogs, indicating that zoo birds potentially pose zoonotic and pathogenic risks to humans and animals. Additionally, gene flow between avian C. parvum isolates from China and Brazil was detected.ConclusionsTo the best of our knowledge, our results demonstrate C. parvum infection in a flamingo (Phoenicopteridae) and white cranes (Grus leucogeranus) for the first time. The results of our study provide an important reference for understanding the host range, biological characteristics, and molecular epidemiology of C. parvum.

Female wanted for the world's rarest turtle: prioritizing areas where Rafetus swinhoei may persist in the wild

AbstractThe Yangtze giant softshell turtle Rafetus swinhoei is the rarest turtle species, with just two individuals known to be surviving, one male in a zoo in China and one individual recently identified as a female in the wild in Viet Nam. As the species is on the brink of extinction, it is an urgent priority to search for additional individuals in the wild, and for areas where it may still be present. Here, we analysed areas where the species may still occur, identified through interview surveys in Viet Nam. In addition, we introduced a novel system for evaluating the potential for occurrence of this rare and elusive freshwater turtle, using a conservation priority index. This index was based on three recorded variables: (1) probability of the species' presence based on interviews with local fishers, (2) degree of habitat alteration, and (3) fishing intensity, with the latter two based on experts’ evaluation of the sites where the species could potentially be present. There were at least 13 independent, reliable sightings in the 2010s, seven of which were in 2018–2019, indicating that R. swinhoei potentially persists in the wild. Although the species was confirmed at only one site, there are at least three more sites where it is likely to be present, and 13 sites of conservation interest. We provide a description of all sites where the species is potentially present, and a summary of relevant interviews. The Da River system has the highest number of recent sightings. We recommend that a research and conservation project be initiated urgently, and outline how such a project could be implemented.

Prevalence and characterization of antibiotic resistance genes and integrons in Escherichia coli isolates from captive non-human primates of 13 zoos in China

Antimicrobial resistance (AMR) has become a public health concern; but antibiotic resistance genes (ARGs) and integrons that link to AMR of Escherichia coli from non-human primates remain largely unknown. This study aimed to investigate antibiotic resistance, emerging environmental pollutants ARGs, and integrons factors (intI1, intI2 and intI3) in 995 E. coli isolates obtained from 50 species of captive non-human primates of 13 zoos in China. Our result showed 83.62% of the E. coli isolates were resistant to at least one antibiotic and 47.94% isolates showed multiple drug resistances (MDR). The E. coli isolates mainly showed resistance to tetracycline (tetracycline 62.71%, doxycycline 61.11%), β-lactams (ampicillin 54.27%, amoxicillin 52.36%), and sulfonamide (trimethoprim-sulfamethoxazole 36.78%). A total of 423 antibiotic resistance patterns were observed, of which DOX/TET (49 isolates, 4.92%) was the most common pattern. Antibiotic resistance rates among 13 zoos had a significant difference (P < 0.01). We further detected 22 ARGs in the 995 E. coli isolates, of which tetA had the highest occurrence (70.55%). The presence of integrons class 1 and 2 were 24.22% and 1.71%, respectively, while no class 3 integron was found. Significant positive associations were observed among integrons and antibiotics, of which the strongest association was observed for integrons / Gentamicin (OR, 2.642) and integrons / Cefotaxime (OR, 2.512). In addition, cassette arrays were detected in 64 strains of class 1 integron-positive isolates (26.56%) and 10 strains of class 2 integron-positive isolates (58.82%). Eighteen cassette arrays were found within 64 class 1 integron isolates, while 3 cassette arrays were identified within 10 class 2 integron isolates. Our results indicate a high diversity of antibiotic resistance phenotypes in non-human primate E. coli isolates, which carry multiple ARGs and integrons. Corresponding preventive measures should be taken to prevent the spread of integron-mediated ARGs in non-human primates and their living environments in zoos.

Molecular identification and biological characterization of Cryptosporidium muris from camels (Camelus bactrianus) in China

BackgroundCryptosporidium is an opportunistic pathogen that infects a wide variety of vertebrates. The aim of the present study was to characterize Cryptosporidium spp. isolates from Bactrian camels and to foster further understanding of the biological characteristics of the pathogen.MethodsFecal specimens were collected from two 4-year-old Bactrian camels resident at the Kaifeng City Zoo in China and examined for Cryptosporidium. Fecal specimens were screened using the floatation method, and then genomic DNA was extracted from the oocysts and identified by nested-PCR amplification of the small subunit ribosomal RNA (SSU rRNA) gene, the actin gene and the Cryptosporidium oocyst wall-protein (COWP) gene. Subtype analysis was performed based on four minisatellite (MS) loci (MS1, MS2, MS3 and MS16) that were aligned and phylogenetically analyzed to determine the species and subtype of Cryptosporidium. We then established a BALB/c mice infection model and further verified the results through clinical status, pattern of oocyst excretion and histological examination.ResultsCryptosporidium oocyst isolates from the two Bactrian camels had an average (± standard deviation) size of 7.49 ± 0.13 × 5.70 ± 0.10 μm (n = 50). The sequencing and phylogenetic analysis confirmed the species as C. muris. Multilocus sequence typing analysis indicated that the subtypes were M13, M4, M1 and M5. Following the inoculation of BALB/c mice, we found that the prepatent period and number of oocysts per gram increased with increasing infective dose. Oocysts were first detected in the feces of BALB/c mice at 7–8 days post-infection (dpi), with levels peaking twice thereafter, at 15–16 dpi and 19–20 dpi. Histology and scanning electron microscopy studies showed that the stomach contained gastric pits filled with Cryptosporidium that adhered to the surface of gastric mucosa gland epithelial cells, causing the latter to deform, swell and become disordered.ConclusionsThe findings of this study indicated that oocysts isolated from Bactrian camels were from C. muris. This is the first report of C. muris isolated from camels in China. More epidemiological data are needed to understand the prevalence and transmission of C. muris in camels in different geographic areas.Graphical abstract

The Genetic Diversity and Population Genetic Structure of the Red Panda, Ailurus fulgens, in Zoos in China.

Simple SummaryRed pandas (Ailurus fulgens) have been listed as one of the endangered species in China and worldwide. In order to protect the populations as well as to avoid the inbreeding depression in wildlife, monitoring the genetic diversity and population genetic structure of red pandas is necessary. By analyzing the captive red pandas among the zoos in China using PCR and genotyping, we found a high genetic diversity among the populations as high as that of the wild population. This demonstrate the current scientifically-based breeding program can keep the high genetic diversity in captive red pandas.In China, red pandas (Ailurus fulgens) have been raised in zoos for 60 years. It is very important to understand the genetic diversity and population genetic structure of the captive red pandas. Based on 19 microsatellite loci, we investigated genetic diversity and population genetic structure of 116 captive red pandas, with samples taken from 11 captive populations in China. Our results revealed a high genetic diversity among the populations, with mean allelic richness varying from 3.505 (Beijing) to 4.026 (Mianning), and expected heterozygosities varying from 0.631 (Huangshan) to 0.782 (Wenling). In particular, significant deviation from Hardy–Weinberg equilibrium was found in populations of Fuzhou and Jiangsu. The genetic differentiation index across all populations was 0.055, indicating a significant genetic differentiation among the 11 populations. These populations could be divided into three genetic clusters using a microsatellite-based Bayesian clustering analysis, which were consistent with the clustering results of wild populations. We conclude that the genetic diversity among captive red pandas is as high as that of the wild population. More attention should be paid to develop a proper and scientifically-based management program to avoid inbreeding and maintain a high genetic diversity in captive red pandas.

Occurrence and multilocus genotyping of Giardia duodenalis in captive non-human primates from 12 zoos in China

Giardia duodenalis is a common enteric protozoan that infects a range of hosts including humans and other mammals. Multilocus genotyping of G. duodenalis in captive non-human primates (NHPs) from zoos in China is limited. In this study, we evaluated 302 NHP fecal samples collected from 32 different NHP species. The primates were from 12 zoos distributed across eight provinces and two municipalities (Chongqing and Beijing) of China. The overall infection rate was 8.3% (25/302). The six G. duodenalis-positive zoos and their infection rates were: Suzhou Zoo (40.0%, 4/10), Yangzhou Zoo (22.2%, 2/9), Dalian Zoo (16.7%, 4/24), Chengdu Zoo (12.8%, 6/47), Guiyang Forest Wildlife Zoo (12.1%, 7/58), and Changsha Zoo (4.7%, 2/43). Molecular analysis of three loci, beta-giardin (bg), triose phosphate isomerase (tpi), and glutamate dehydrogenase (gdh), showed high genetic heterogeneity, and seven novel subtypes (BIII-1, MB10-1, WB8-1, B14-1, MB9-1, DN7-1, and BIV-1) were detected within assemblage B. Additional analysis revealed 12 different assemblage B multilocus genotypes (MLGs), one known MLG and 11 novel MLGs. Based on phylogenetic analysis, 12 assemblage B MLGs formed two main clades, MLG-SW (10-12, 18) and MLG-SW (13, 14, 16, 17), the other four MLG-SW (15, 19, 20, 21) were scattered throughout the phylogenetic tree in this study. Using multilocus genotyping, this study expands our understanding of the occurrence of Giardia infection and genetic variation in Giardia in captive non-human primates from zoos in China.

Genomic characterization of a novel astrovirus identified in Amur tigers from a zoo in China.

Astroviruses (AstVs) have a very wide range of hosts and are associated with enteric and extra-enteric disease in mammals and birds. Cross-species transmission of AstVs has been observed frequently. In the present study, the genome of a novel astrovirus from Amur tigers (Panthera tigris) from a zoo in China was characterized and was found to have the typical genomic features of other mammal AstVs. It showed the highest nucleotide sequence similarity (46.1-87.3% identity) to AstVs from cats, indicating a close phylogenetic relationship and possible cross-species transmission between them. To our knowledge, this is the first identification and characterization of AstV from tigers, and this virus is the third astrovirus identified in hosts of the family Felidae. The results of this study will be helpful for understanding the origin, genetic diversity, and cross-species transmission of AstV.

Toxoplasma gondii in four captive kangaroos (Macropus spp.) in China: Isolation of a strain of a new genotype from an eastern grey kangaroo (Macropus giganteus).

Marsupials are highly susceptible to Toxoplasma gondii infection. Here, we report T. gondii infection in four kangaroos from a zoo in China. Kangaroos were imported into China in 2000 and were since bred in zoo. In 2017–2018, four kangaroos died due to respiratory system disease or injury. The bodies were submitted to the laboratory to test for T. gondii infection. Antibodies to T. gondii were found in 75% (3/4) of the kangaroos via the modified agglutination test with the cut-off 1:25. Cysts were observed in the histopathological sections of tongue and diaphragm or squashes of fresh myocardium in two kangaroos. These cysts were confirmed as T. gondii by immunohistochemical staining and molecular biological analysis. One viable T. gondii strain was isolated from one kangaroo and designated as TgRooCHn1. DNA from T. gondii tachyzoites obtained from cell culture was characterized by 10 PCR-RFLP markers and the virulence genes ROP5 and ROP18. The genotype of this isolate did not match with any known genotypes; it was designated as ToxoDB#292. The virulence of TgRooCHn1 (104 tachyzoites) was non-lethal to mice, and it formed tissue cysts. To our knowledge, the present study is the first isolation of ToxoDB#292 strain from kangaroo. Improvemets for captive settings were initiated, including greater attention being paied to birds and stray cats, fed frozen meat for carnivores.