Taipei Zoo Research Articles

Sex identification of birds in Taipei Zoo.

As a conservation and breeding institution for birds, Taipei Zoo plays an important role in restoring endangered species. As approximately half of all bird species are monomorphic, precisely confirming the sex of individuals is critical for the management of ex-situ conservation breeding populations, as well as for understanding the sex ratio of those in the wild. Generally, PCR is used more reliably for sex determination versus traditional methods such as plumage, behavior or hormone levels. Nevertheless, the various primer sets and annealing temperatures vary between species, and so inaccurate sexing can occasionally happen due to inadequate PCR conditions. To reduce the probability of misidentification, and to establish a PCR condition database for sex determination across the diverse range of avian taxa, we tested multiple primer sets and annealing temperatures for amplification of the bird sex-specific gene fragments (CHD1) for each captive or rescued avian species held at Taipei Zoo since 2014. A total of 162 species across 22 orders were tested using one or two primer sets. One hundred and fifty-five species were successfully sexed by the primer set 2550F/2718R and the success rate of sex typing reached over 90% of species tested in each order. Most species have suitable PCR annealing temperatures between 45°C and 55°C, and the species in the same avian taxa showed similar results in temperature. This indicates that it is possible to select the annealing temperature of other species in the same family when the species had not been tested before. We expect this study will improve the success rate of identifying sex by using applicable PCR conditions and reduce the time for searching references every time before attempts to PCR sex birds.

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

BackgroundToxoplasma 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 resultsAn 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.ConclusionsAccording 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.

Molecular and serological detection of Toxoplasma gondii infection in mammals in the Taipei Zoo.

Toxoplasmosis caused by Toxoplasma gondii affects both conservation and public health efforts. In the Taipei Zoo, toxoplasmosis was diagnosed in ring-tailed lemurs and a meerkat in 2019 while a freeze-thaw meat strategy had been applied to carnivores before the event. To investigate the possible risk factors associated with T.gondii infection in the Taipei Zoo, 179 veterinary visiting mammals from 2019-2021 and six stray cats were included to detect anti-T.gondii IgG and IgM in their serum via ELISA, and T. gondii in their faeces and blood via PCR. Although the overall T.gondii IgG seroprevalence was 33.5% and PCR positivity was 16.2% in the zoo mammals, the correlation between T.gondii PCR and systemic IgG results was low. An omnivorous diet (adjusted OR=0.4; 95% CI: 0.2-1.0), a herbivorous diet (adjusted OR=3.2; 95% CI: 1.1-9.6), and animals in the Conservation Area where stray cats appeared (adjusted OR=18.3; 95% CI: 3.9-85.9) were independent risk factors for T.gondii infection. The low T.gondii-specific IgM positivity (0.6%) suggests that most animals did not have acute T.gondii infection. In conclusion, our findings indirectly support that feeding frozen meat to carnivores, cleaning fresh food, and restricting access to stray cats to prevent faecal contaminants could prevent animals from T.gondii exposure.

Development of the Animal Conservation Digital Learning Aids and Assessments through the Industry-University Collaborative Course

Due to the rapid changes caused by globalization and internationalization, this study focused on achieving Sustainable Development Goals (SDGs) 4 and 15 via specific digital learning materials—animal conservation apps—particularly designed for enlarging primary school pupils’ knowledge of biodiversity and conservation of natural habitats, and promoting sustainable development and lifelong learning abilities. Through a collaboration with Taipei Zoo, this study recruited 37 undergraduates who took the Learning Design and Practice course to develop the digital learning-assisted materials, namely animal conservation apps and assessment tools that suited the digital learning materials. In the initiative stage of the course, the undergraduates were required to work as a team and to learn in a group by observing and experiencing the model apps provided in the class. The provided apps were developed in compliance with the ADDIE model. In the middle stage of the course, each team was asked to develop their team app and assessment tools following the ADDIE model. In the final stage, each team’s design results were evaluated based on the digital learning material scale and core competency test evaluation: art domain. The results show that the undergraduates were able to integrate the expertise they gained in the course to developing high-quality digital learning materials. According to the educational professionals’ evaluation, the assessment tools designed by the undergraduates scored high marks.

WOUND MANAGEMENT WITH AND WITHOUT MODIFIED CHOUKROUN'S PLATELET-RICH FIBRIN IN RESCUED FORMOSAN PANGOLINS (MANIS PENTADACTYLA PENTADACTYLA).

Trauma is a key threat to wild Formosan pangolins (Manis pentadactyla pentadactyla) and the most frequent reason for presentation to the rescue center of the Taipei Zoo from 2008 to 2017. Of the 105 cases received during this time, 72% presented for trauma. Among these 76 cases, 72% survived and were returned to the wild. Traditional wound management resulted in healing times from 45 to 410 d. Improvements in the success and duration of wound healing could increase the success rate of pangolin release back to the wild. This case series describes wound management in five cases using either traditional wound management practices (two cases) or modified Choukroun's platelet-rich fibrin (MC-PRF) in an attempt to accelerate the wound-healing process (three cases). MC-PRF is relatively easily obtainable and widely used to promote tissue healing in other species. MC-PRF was applied to the wounds of three pangolins every 1 to 2 wk with satisfactory results. The healing time with MC-PRF were 21, 45, and 51 d, shorter than the 60-98 d seen in the cases of traditional wound management in this series. Given the scarcity of literature on pangolins, these cases evaluating both traditional and MC-PRF wound management practices can serve as examples for other pangolin rescue centers in their treatment of traumatic wounds.

Clinical Management of an Intranasal Abscess in a Captive Elongate Tortoise (Indotestudo elongata) by Using Modified Choukroun's Platelet-Rich Fibrin

An adult male elongate tortoise (Indotestudo elongata) was presented to the Animal Medical Center of Taipei Zoo with an abscess on the tip of its nose. After 43 days of daily debridement and systemic antibiotic administration, we sought an alternative treatment modality due to poor wound management. We developed a novel treatment using platelet-rich fibrin (PRF). There are few studies documenting the benefits of PRF for the management of wounds in reptiles. Most published protocols require the use of specific anticoagulants and methods to prepare PRF; however, we found that a modified Choukroun's PRF was relatively easy to prepare and use for wound treatment of an elongate tortoise. Here, we describe a successful outcome of a chronic, nonresolving facial wound after treatment with modified Choukroun's PRF. To our knowledge, this is the first report of a Choukroun's PRF preparation derived from reptilian blood. We recommend further studies and detailed trials to evaluate the efficacy of this protocol in reptiles.

Methods to ameliorate heat stress in non-native alpaca, Vicugna pacos

Despite their ability to adapt to a variety of environments, alpacas suffer from heat stress symptoms when raised in warm and humid regions of the world. Hyperthermia in alpacas not only raises animal welfare concerns, but also decreases productivity. To better understand the relationship between heat stress and alpaca mortality, Taiwanese alpaca facilities were surveyed. Additionally, to address the dearth of experimental research on heat stress reducing amenities in alpacas, an experiment was designed to test the effectiveness of different cooling amenities (water pool, frequent mist spraying, and fans) through temperature, behavioral, location and postural observations of alpacas at the Taipei Zoo during manipulations. Experimental result supports the use of mist sprinklers, water pools, and fans as alpaca cooling amenities, with water pools as being the most effective. Survey result shows a significant portion of alpaca deaths in Taiwan occurred during the summer and that heat stress was a leading cause of death. Furthermore, imported alpacas had a higher mortality rate than those born in Taiwan.

DNA barcoding reveals CITES-listed species among Taiwanese government-seized chelonian specimens.

Compared to traditional morphological identification, DNA barcoding-molecular identification based on sequencing of a segment of mitochondrial cytochrome c oxidase subunit I (COI)-provides a shortcut to authenticating chelonian identifications. Here, we selected 63 government-seized chelonian specimens deposited at Taipei Zoo for DNA barcoding analysis. DNA barcoding and subsequent phylogenetic analysis successfully authenticated 36 chelonian species, including five that are listed in CITES Appendix I. Approximately 90% (57/63) of the specimens were successfully authenticated by our molecular approach, but lack or error of BOLD reference sequences, biological processes such as hybridization, and uncertain species delimitation all reduced the accuracy of DNA barcoding. To increase the accuracy of DNA barcoding, Taipei Zoo will continue to enrich the BOLD database and also establish a genetic database, to include additional genetic markers, by using government-seized chelonian specimens. A fast and accurate method to authenticate seized samples could assist law enforcement agencies to prosecute criminals and restrict illegal exploitation of wild chelonian resources.

Diffuse-type Gastric Mucinous and Signet Ring Cell Adenocarcinoma in a Captive California King Snake (Lampropeltis getula californiae)

An adult female California king snake (Lampropeltis getula californiae) housed in Taipei Zoo was presented with a 2-week history of anorexia, fatigue and abdominal swelling. Exploratory laparotomy revealed a gastric mass with two circular perforations and multiple mottled white to beige protuberances along the mucosal surface. Histologically, the gastric mass showed an invasive, transmural growth of epithelial cells arranged in nests, lobules, acini and sheets in the mucosa and submucosa that progressively transformed into signet ring cells in the muscularis externa and subserosa. All of the neoplastic cells expressed pan-cytokeratin immunohistochemically. Based on the World Health Organization histological criteria, a diagnosis of diffuse-type gastric mucinous and signet ring cell adenocarcinoma was made.

Factors affecting yearly and monthly visits to Taipei Zoo

This study investigated factors affecting yearly and monthly numbers of visits to Taipei Zoo. Both linear and nonlinear regression models were used to estimate yearly visits. The results of both models showed that the “opening effect” and “animal star effect” had a significantly positive effect on yearly visits, while a SARS outbreak had a negative effect. The number of years had a significant influence on yearly visits. Results showed that the nonlinear model had better explanatory power and fitted the variations of visits better. Results of monthly model showed that monthly visits were significantly influenced by time fluctuations, weather conditions, and the animal star effect. Chinese New Year, summer vacation, numbers of holidays, and animal star exhibitions increased the number of monthly visits, while the number of days with temperatures at or below 15 °C, the number of days with temperatures at or above 30 °C, and the number of rainy days had significantly negative effects. Furthermore, the model of monthly visits showed that the animal star effect could last for over two quarters. The results of this study clarify the factors affecting visits to an outdoor recreation site and confirm the importance of meteorological factors to recreation use.

A partial least squares structural equation modeling approach for examining multidimensional values of Taipei Zoo

ABSTRACTThe values associated with a tourism product, such as a hospitality facility, an attraction, or a destination, are intangible for independent evaluation. Meanwhile, tangible features of a tourism product can be used as predictor variables in causal research to assess the values as dependent variables. In the meanwhile, the multidimensional values may function independently from each other. Under this condition, consideration should be made in designing the values as a formative model and using partial least squares structural equation modeling to illustrate its relationships with the predictor variables. This paper uses the Taipei Zoo as a case study by conducting a survey of 472 visitors. The results demonstrate how different conceptual model designs can affect the statistical results and ultimate decision-making concerned with the value design of a tourism product.

Mycolactone-producing Mycobacterium marinum infection in captive Hong Kong warty newts and pathological evidence of impaired host immune function.

A mass mortality event of captive Hong Kong warty newts Paramesotriton hongkongensis with non-granulomatous necrotic lesions occurred in Taipei Zoo, Taiwan, in 2014. Clinically, the sick newts were lethargic and often covered with water mold Saprolegnia sp. on the skin of the body trunk or extremities. Predominant pathological findings were multifocal non-granulomatous necrotic lesions in the liver, spleen, and kidneys and severe skin infection with Saprolegnia sp., with deep invasion and involvement of underlying muscles. The possibility of ranavirus infection was ruled out by negative PCR results. Unexpectedly, abundant intralesional acid-fast positive bacilli were found in the necrotic lesions of the liver, spleen, and kidney in all 14 sick newts. PCR targeting the hsp65, ITS region, and partial 16S rRNA genes was performed, and the sequence identity from amplified amplicons of hsp65 and partial 16S rRNA genes was 100% identical to that of the corresponding gene fragment of Mycobacterium marinum. Further molecular investigations demonstrated that the current M. marinum was a mycolactone-producing mycobacterium with the presence of esxA/esxB genes. Mycolactone is a plasmid-encoded, immunosuppressive, and cytotoxic toxin. The possible immunosuppression phenomenon characterized by systemic non-granulomatous necrotic lesions caused by M. marinum and the unusual deep invasive infection caused by water mold might be associated with the immunosuppressive effect of mycolactone. Therefore, it should be noted that non-granulomatous necrotic lesions in amphibians can be caused not only by ranavirus infection but also by mycobacteriosis.

Rail-to-Earth Resistance Assessment: A Medium Capacity Transit System with Continuous Negative Rails by Potential Measurement

The Brown Line of the Taipei Rapid Transit System is the only rubber-tire medium capacity line in the system. The first phase of its construction was launched in 1988, and revenue service began in 1996. It was frequently referred to as the Muzha Line, as the route runs from the Taipei Zoo in the southeast suburb of Muzha to the vicinity of the Taipei Songshan Airport in the northeast side of the downtown commercial district [1]. The two-track line was built on single reinforced-concrete beam structures supported on reinforced-concrete columns [2], except in a short tunnel. The second phase was an extension to the Taipei Songshan Airport terminal and then to the suburb of Neihu, so it was also referred to as the Neihu Line. The section of the second-phase route near Taipei Songshan Airport was built in an underground tunnel, while the rest of the route was built on single reinforced-concrete beam structures supported on reinforced-concrete columns, like the Muzha Line. Figure 1 shows the schematic route map of the Brown Line with labeling of selected stations. Note that the stations of the first phase were labeled as BR2 through BR13, and the stations of the second phase were labeled as B1 through B11, plus BR1 as the Taipei Songshan Airport Station.

CASE REPORT: A CHRONIC B-CELL LEUKEMIA IN A EURASIAN OTTER (Lutra lutra)

A 12-year-old, female, Eurasian otter (Lutra lutra) reared in Taipei zoo was submitted for necropsy due to sudden death without remarkable clinical history. Parameters of the latest hematology were all within normal limits. At necropsy, there were large amounts of abdominal effusion, diffuse splenohepatomegaly, generalized lymphadenopathy, consolidation of the lung, and bilateral enlargement of both adrenal glands. Histological examination revealed large numbers of small to medium-sized neoplastic lymphocytes disrupting the normal architectures of the spleen, liver, lymph nodes, lung, mesentery, and adrenal glands. Immunohistochemistry showed diffuse positivity of tumor cells for CD79a but negativity for CD3. There were scattered positive signals for Ki-67 mainly in the spleen in sites of extramedullary hematopoiesis and lymph nodes. The neoplasm was considered a chronic lymphocytic leukemia of B-cell origin.

Retrospective study of the medical status of 34 Formosan sika deer (Cervus nippon taiouanu) at the Taipei Zoo from 2003 to 2014

Abstract : The Formosan sika deer (Cervus nippon taiouanus) is an endemic subspecies in Taiwan. The original wilddeer has been extinct since the late 1960s. The largest captive population is located at the Taipei Zoo. Except forinfectious disease outbreaks, no systemic medical research has been reported for this subspecies. This study wasconducted to analyze the medical status of the captive Formosan sika deer population, including the hematologicaland serum chemistry characteristics. To accomplish this, medical records for 34 Formosan sika deer from January2003 to January 2014 were acquired and analyzed. The most common illness and cause of death was trauma, followedby gastrointestinal and respiratory disease, respectively. The hematologic and serum chemical values of healthy adultswere quite different from those of sika deer (Cervus nippon yesoensis). This study provides a closer medicalunderstanding of this subspecies and the results will facilitate its management.Keywords : Formosan sika deer, gastrointestinal disease, respiratory disease, Taiwan, trauma

Aspects of digestive anatomy, feed intake and digestion in the Chinese pangolin (Manis pentadactyla) at Taipei zoo.

Pangolins are considered difficult to maintain in zoos, often attributed to problems in feeding management. Taipei Zoo's designation as a wildlife rescue center for Chinese pangolins (Manis pentadactyla) has resulted in long term feeding experience with development of diets that support recovery, maintenance, and reproduction, as well as experimental opportunities to further understand digestive physiology to optimize nutrition. Opportunistic dissection of 10 animals revealed details of the tongue, salivary glands, stomach and gastrointestinal tract (GIT), including confirmation of anatomical differences between Asian and African pangolin species. Length of the total GIT relative to body length (∼ 9:1) was greater than found in domestic carnivores, more similar to omnivorous species. Intake and digestion trials conducted with 4 animals demonstrated that pangolins maintained body weights (BW; 6-9 kg) consuming diets containing 32-40% crude protein, 20-25% crude fat, and 13-28% crude fiber (DM basis). Daily DM intakes ranged from ∼40 to 70 g per animal, with digestible energy intake 51.5-87.5 kcal /kg BW(0.75) /day; pangolins consumed 9.4-15.2 g DM/kg BW(0.75) /day. Dietary energy data support observations of low metabolism and maintenance requirements for this species, similar to values reported for other myrmecophageous species. Addition of 5% ground chitin to diets improved fecal consistency, and decreased digestibility of DM, protein, and energy; addition of chitosan (chitin treated with NaOH) resulted in diet rejection. This information may assist in enhancement of captive diets, as well as in controlling obesity in pangolins.

Hematologic and serum biochemical parameters of apparently healthy rescued formosan pangolins (Manis pentadactyla pentadactyla).

Natural habitats of pangolins are rapidly deteriorating because of extensive farming, logging, and human construction activities. In addition, the illegal trading of pangolins substantially accelerated the decline of the pangolins' population in southeastern Asia. The maintenance of confiscated pangolins in rescue centers is currently a daunting task for veterinarians and conservation biologists. There is limited information in the literature about the reference values regarding the physiology of pangolins. The purpose of this study is to establish reliable hematologic and serum biochemical reference values for the Formosan pangolin (Manis pentadactyla pentadactyla). Blood samples were collected from 51 apparently healthy pangolins from a population of 117 rescued pangolins at the Taipei Zoo. Sex-related differences were observed in platelet count, alanine aminotransferase level, mean corpuscular hemoglobin concentration, and total protein level. Age-related differences were also noted; juveniles have significantly higher platelet counts and alkaline phosphatase levels than their adult counter parts. The hematologic and serum biochemical reference values for the Formosan pangolin presented in this study can be applied in the medical care of this important species during rescue attempts. It is the first systematic report of blood parameters of apparently healthy pangolins and provides a basis for future investigation of this species. The reference values reported in this study may also be applicable to other pangolin species in the genus Manis.

A Retrospective Study of the Medical Status on 63 African Hedgehogs (Atelerix albiventris) at the Taipei Zoo From 2003 to 2011

Inbreeding of African hedgehogs (Atelerix albiventris) is considered a problem as breeders try to meet the demands of the pet trade. Most studies of African hedgehogs are concentrated in the USA. This study investigates and compares the life span and medical conditions of captive African hedgehogs at the Taipei Zoo, Taiwan. The medical records of 63 African hedgehogs at the Taipei Zoo from 2003 to 2011 were collected and analyzed statistically for this retrospective study. The mean life span of the 39 deceased hedgehogs in the study was 3.4 ± 1.1 years. The average weight was 401 ± 98g. Female hedgehogs were heavier than males. The primary cause of death in this group of animals was neoplasia (35.9%). Oral squamous cell carcinoma was the most common form of neoplasia diagnosed (35.7%). Hematologic and serum biochemical tests from 9 healthy adults revealed increased alanine aminotransaminase, aspartate aminotransferase, total bilirubin, and albumin values compared with references. Decreased values for alkaline phosphatase, amylase, and creatinine levels were also found. The results of this study may provide a better understanding of African hedgehog health and may help to determine potential management problems.

A 20-year disease survey of captive formosan serows (Capricornis swinhoej) at the Taipei Zoo (1991-2011).

The Formosan serow (Capricornis swinhoei) is endemic to Taiwan. The wild population has declined dramatically over the past few decades and the species is listed as a "precious and rare species" protected under law in Taiwan. Disease investigations have been rare except for sporadic observations of wild individuals, and no long-term disease survey has been performed on this species. The objective of this study was to identify and report on the most common diseases in captive Formosan serows and determine the potential causes. Medical records of Formosan serows (n = 62) housed at the Taipei Zoo over a 20-yr period (1991-2011) were collected and analyzed for this study. The most common diseases affected the gastrointestinal system and the skin. Parasitic etiologies accounted for greater than 85% of these diseases, and coinfection was common. Coccidia and lice were the most common endo- and ectoparasites, respectively. High mortality was noted in serows less than 1 yr old associated with parasitism. The results from this study could provide vital information on disease prevention and species management, which may greatly help in rehabilitation of captive and wild populations.

Meningothelial meningioma in a Malayan sun bear (Helarctos malayanus)

A 24-year-old, spayed female Malayan sun bear (Helarctos malayanus) in the Taipei Zoo (Taipei, Taiwan) showed clinical signs of slowly progressive anorexia, dullness, compulsive pacing, and circling. The animal subsequently developed acute severe stupor and persistent recumbency. Postcontrast study of computed tomography revealed a spheroid, extra-axial mass with strong but heterogeneous hyperattenuation in the left temporal lobe of the cerebrum. At necropsy, a solitary, well-circumscribed intracranial mass measuring 3 cm × 2.5 cm × 2 cm was attached to the left pyriform lobe with compression of the adjacent neuroparenchyma. Cytological examination obtained from the mass revealed large clumps and sheets of cohesive polyhedral cells with round nuclei, wispy cytoplasm, and indistinct cell borders. Microscopically, the mass was composed of densely packed round to polygonal cells arranged in lobules and small nests. Psammoma bodies, xanthomatous change, and cholesterol deposition were also noted. Immunohistochemical staining of the tumor was positive for vimentin, pancytokeratin, cytokeratin (CK)34BE12, neuron-specific enolase, and epithelial membrane antigen, but negative for glial fibrillary acidic protein and S100 protein. The cytological, histological, and immunohistochemical features were compatible with a meningothelial meningioma.