Ex-situ Conservation Programmes Research Articles

Near extinct Argyreia versicolor and rare Argyreia mekongensis are dependent on carpenter bee pollinators.

Argyreia versicolor and Argyreia mekongensis are extremely rare plant species. The former had not been seen for nearly 100 years until two individuals were found in Thailand in 2018, and only a handful of populations are known for the latter. The aims of this study were to examine the breeding systems of A. versicolor and A. mekongensis using pollination experiments and to determine their potential pollinators via floral observations. Our controlled pollination experiments uncovered the self-incompatibility of both species. Pollinator censuses indicated that females of two carpenter bee species, Xylocopa aestuans and Xylocopa latipes, were the predominant floral visitors for both Argyreia species. Our observations confirmed a harmonious match between the floral shape of both Argyreia species and the body sizes of these pollinators, ensuring effective pollen transfer and validating their role as putative pollinators. In line with the high frequency of pollinator visits observed, our controlled pollination experiments found no evidence of pollen limitation under field conditions. The findings of this study hold significance for the conservation of these endangered species, yet the situation is dire for A. versicolor, with one of the two individuals under study recently lost. Hence, it is crucial to intensify monitoring efforts for the species, aiming to identify additional individuals for potential inclusion in an ex-situ conservation program. Simultaneously, safeguarding the habitat of these plant species and their pollinators will be critical.

Sperm morphology, and macro and microscopic description of male and female reproductive tract in the North African houbara bustard (Chlamydotis undulata)

Ex-situ conservation programs are useful tools for preserving endangered species, and knowledge of their reproductive physiology is essential for achieving conservation objectives in captive breeding programs. The North African houbara bustard (Chlamydotis undulata) is a vulnerable bird species bred in captivity to restore its natural populations. To better understand the reproductive biology of this species, we first described the anatomy and histology of the reproductive tracts of male and female North African houbara bustards. We described and classified the normal and abnormal sperm morphologies for the species. The reproductive tracts of males and females were collected during the breeding season (BS) (21 males and 14 females) and the non-breeding season (NBS) (four males and two females). Pictures and measurements of the testes and oviducts were taken, and testis colour was graded from 1 (black) to 6 (white). Organs were fixed and processed for histological analysis, scanning electron microscopy (SEM), and transmission electron microscopy (TEM). The testis, epididymis, oviduct size, testicular capsule thickness, seminiferous tubule (ST) diameter, and histological layers of the oviduct were compared between the BS and NBS groups. Sperm morphometry was evaluated using fixed ejaculates (n=22, from 22 males). Ejaculates (n=417, 166 males) were collected and fixed during the BS, and their morphology and morphometry were evaluated using phase-contrast microscopy. Ejaculate mixes (n=12) were fixed and analysed using SEM and TEM. Overall, the anatomical structure and histology of the male and female reproductive tracts were similar to those of other avian species. The testes were symmetric, and a regression of testis and oviduct size, the thickness of the mucosa and serosa of the oviduct, and ST diameter (p<0.001) were observed during NBS compared with those during BS. Sperm storage structures are located inside the oviduct at the uterovaginal junction and infundibulum. Melanin granules were observed in the interstitial space of the seminiferous epithelium, which explains the testis pigmentation. Melanin was also found in the ovaries and the uterus. Houbara sperm have an elongated nuclear form and a round-shaped acrosome lacking perforatorium and rostrum. Most analysed samples were limitrophe normospermic (normal= 68.5 ± 14.7%), houbara sperm morphology could be classified into eight categories: normal sperm, acrosome, head, midpiece, tail, and teratogenic defects, swollen head, and cytoplasmic droplet. This study provides the first anatomical and histological descriptions of the male and female reproductive tracts of the North African houbara bustard. Further investigation into the functional role of melanin in the testes is warranted.

The endangered brain: actively preserving ex-situ animal behaviour and cognition will benefit in-situ conservation.

Endangered species have small, unsustainable population sizes that are geographically or genetically restricted. Ex-situ conservation programmes are therefore faced with the challenge of breeding sufficiently sized, genetically diverse populations earmarked for reintroduction that have the behavioural skills to survive and breed in the wild. Yet, maintaining historically beneficial behaviours may be insufficient, as research continues to suggest that certain cognitive-behavioural skills and flexibility are necessary to cope with human-induced rapid environmental change (HIREC). This paper begins by reviewing interdisciplinary studies on the 'captivity effect' in laboratory, farmed, domesticated and feral vertebrates and finds that captivity imposes rapid yet often reversible changes to the brain, cognition and behaviour. However, research on this effect in ex-situ conservation sites is lacking. This paper reveals an apparent mismatch between ex-situ enrichment aims and the cognitive-behavioural skills possessed by animals currently coping with HIREC. After synthesizing literature across neuroscience, behavioural biology, comparative cognition and field conservation, it seems that ex-situ endangered species deemed for reintroduction may have better chances of coping with HIREC if their natural cognition and behavioural repertoires are actively preserved. Evaluating the effects of environmental challenges rather than captivity per se is recommended, in addition to using targeted cognitive enrichment.

Hope is the last thing lost: Colombian captive-bred population of the critically endangered Orinoco crocodile (Crocodylus intermedius) is a genetic reservoir that could help to save the species from extinction

A purpose of ex-situ populations is the preservation of genetic variation, but this is a challenging task since genetic diversity is commonly lost through each generation, and so the establishment of management guidelines should be a high priority. Fifty years ago, the National University of Colombia began a breeding program in the Roberto Franco Tropical Station (in Villavicencio, Meta) to conserve the critically endangered Orinoco crocodile Crocodylus intermedius. Despite the large number of individuals raised and kept in captivity, the Station has not been able to release individuals due to a lack of a complete genetic characterization that could determine whether the population is genetically viable. In this study we used a panel of 17 microsatellite loci to overcome this problem. We estimated from the founder animals and the live crocodiles the inbreeding, heterozygosities, the number of alleles, and their richness, and frequencies to understand the effects of managing a captive breeding program without considering genetic profiles. Our results revealed that the living population maintains much of its founder diversity with high levels of heterozygosity and low overall inbreeding, making it suitable for maintaining captive breeding and for implementing wild releases. We estimated the individual genetic diversity of the living crocodiles, as well as their relationships. This information, combined with the size, sex, and location, allowed us to propose combinations and to restructure the breeding groups. We demonstrated that molecular data could be used to improve the management of ex-situ conservation programs well beyond what could be achieved with pedigree information alone.

Assessing molecular diversity among 87 species of the Quercus L. genus by RAPD markers

Oaks (Quercus sp.) are among the most ecologically and economically important woody Angiosperms of the northern hemisphere. Nowadays, the reduction of Quercus biodiversity is becoming a matter of global concern, and several oak species have been included in the IUCN Red List of Threatened Species. Consequently, characterization and preservation strategies for the oak germplasm are largely promoted. Thus, in this work, the genetic diversity existing among 87 different Quercus species was assessed using the RAPD markers, in order to better typify these specimens, to show the amazing DNA variability of this plant genus, and to confirm or infer new putative molecular correlations. Our data were discussed taking into consideration the phylogenetic and phylogeographic relationships previously proposed by the literature. In general, the obtained results corroborated that the evolutionary pattern of Quercus genus has been extremely intricate and continues to change rapidly, making it difficult to be fully resolved. The evidence collected in the present investigation would confirm the complex evolution of the oaks, due to their high migration capacity, divergence rate, and hybridization propensity. This research, performed on a so large series of species, represents a positive contribution for highlighting the genetic diversity within collections of Quercus germplasm and favouring ex-situ conservation programmes.

Determination of sexual maturity of Indonesian box turtle (Cuora amboinensis couro) based on straight carapace length

Predicting the turtle's reproductive status and sexual maturity is necessary for ex-situ breeding and conservation program. The histology of the seminiferous tubules of the Indonesian box turtle (Cuora amboinensis couro) at various ages has not been studied much. This study aims to develop equations to predict the sexual maturity of Indonesian box turtles based on straight carapace length (SCL). Six turtles with SCL sizes 10-21 cm were sacrificed and their testes were taken for morphometric measurements and histological preparations for Hematoxylin Eosin staining. Pearson’s correlation of SCL with testicular maturity parameters was analyzed, followed by Anova Regression. The results showed that there were differences in the morphometry and topographic anatomy of the Indonesian box turtle testes between SCL below 15 cm and above 15 cm. Six turtles with SCL size of 10-21 cm were sacrificed and their testes were taken for morphometric measurements and histological preparations for Hematoxylin Eosin staining. Pearson's correlation of SCL with testicular maturity parameters was analyzed, followed by Regression Anova. The results showed that there were differences in the morphometry and topographic anatomy of the Indonesian box turtle testes between SCL below 15 cm and above 15 cm.

Urine biochemical profile analysis and transabdominal ultrasonography for pregnancy detection in the endangered lowland anoa (Bubalus depressicornis)

Abstract. Bowolaksono A, Mayasari A, Sundari AM, Simamora ATJ, Suryaningsih R, Suryawan A, Simamora ATJ, Abinawanto, Sjahfirdi L, Hasna A, Supriatna J, Kyes RC. 2022. Urine biochemical profile analysis and transabdominal ultrasonography for pregnancy detection in the endangered lowland anoa (Bubalus depressicornis). Biodiversitas 23: 6056-6061. The population of lowland anoas (Bubalus depressicornis) has decreased significantly due to illegal hunting and habitat loss. As such, the urgency in developing successful ex-situ conservation programs cannot be overstated. Ex-situ breeding programs of anoas, however, are hindered by difficulties in early pregnancy detection. The failure to detect pregnancy at an early stage often results in miscarriage due to repeated breeding attempts of unconfirmed pregnant females. Moreover, frequent false heat during 5 to 6 months of pregnancy corroborated the repeated breeding of pregnant females. Therefore, early detection techniques are needed to improve breeding success. The present study evaluated the use of urine biochemical profiles as a minimally invasive approach for early pregnancy detection and transabdominal ultrasonography as a confirmation tool of false heat in four lowland anoas. Statistical analysis showed that glucose and protein level were significantly elevated during pregnancy since the 0 days of post-mating (p&lt;0.05) and consistently higher compared to the non-pregnant period. Meanwhile, transabdominal ultrasonography successfully identified organ structures and development by the 155th, 176th, 180th, and 191st days of pregnancy. The present study suggests that urine biochemical data provide a promising, minimally invasive approach to early pregnancy detection in anoas. Incorporating urine data and ultrasonography can significantly help to maintain pregnancy by validating false heat.

What affects the desiccation tolerance threshold of Brazilian Eugenia (Myrtaceae) seeds?

Desiccation sensitive (DS) seeds are shed at high water contents (WC) and metabolically active, but WC thresholds vary broadly among species even in the same genus. Eugenia is an important ecological genus that has high occurrence in several Brazilian morphoclimatic domains. In this study, we assessed seed desiccation tolerance of five Eugenia species collected in specific meteorological conditions. We reported the species geographical ranges and verified the rainfall and temperature of species sites in the year prior to seed collection. We also assessed initial WC, seed germination and vigor and seedling growth upon desiccation. Eugenia uniflora was the widest spread among the five species, while E. astringens was the most restricted. In this specific study, widespread species showed a higher WC threshold than restricted species. In the same way, the WC of fresh seeds was not correlated to the desiccation tolerance threshold. Seed desiccation tolerance was species dependent and correlated with the environmental status of seed collection sites. Wetter and warmer conditions were correlated to the E. uniflora higher DS threshold. Low rainfall and temperature corresponded to a lower desiccation sensitivity of E. astringens seeds. Seeds of the five species lost half viability between 0.44 and 0.25g H2O g DW- 1 and after 65-270h of desiccation. Our results indicate that abiotic factors impact plant populations during the seed production season and can drive seed desiccation tolerance threshold and physiological behavior. These results should be taken into account in ex-situ plant conservation programs and tropical species management.

Genetic Diversity of the Endangered Endemic Anoa (Bubalus spp): Implication for Conservation

Anoa is an endemic ungulate in Sulawesi and its status now is endangered because the population continues to decline. Conservation genetics is one of the crucial issues in the anoa conservation strategy and action plan 2013-2022 document, but this genetic data is not yet available. To investigate and provide valuable information for conservation genetics measures, thirteen polymorphic microsatellites were used to analyze 20 adult anoa. Anoa has relatively low genetic diversity within populations (HO = 0.58), and high genetic differentiation among populations (FST = 0157). Although the anoa population has a bottleneck signal (T.P.M: 0.019; P0.05), the bottleneck simulation results show that the loss of genetic diversity is being slow over the next 100 years (9.5%). We provide some recommendations for conservation genetics based on the findings in this paper, including monitoring and genetically mapping for other anoa populations due to bottleneck signals, establishing the founder of the ex-situ population by examining their genetic diversity status, maintaining and increasing the number of individuals in the ex-situ population to genetically safe population size, and managing anoa populations by avoiding inbreeding. In-situ and ex-situ conservation programs should be combined to maintain the genetic diversity of anoa.

Genetic purity of a rear-edge population of Carex podogyna Franch. et Sav. (Cyperaceae) maintained under interspecific hybridization.

Interspecific hybridization is a critical issue in conservation biology because it may drive small populations to extinction through direct or indirect processes. In this study, to develop a conservation strategy for an endangered rear-edge population of Carex podogyna in Ashiu, Kyoto, Japan, we performed a molecular genetic analysis of the wild population and an ex-situ population established from wild seeds. Microsatellite genotypic data revealed a complete loss of genetic diversity in the wild population, suggesting that it has long been prone to genetic drift due to isolation as a small population. In contrast, microsatellite analysis of 13 ex-situ individuals detected multiple alleles that are not harbored in the wild C. podogyna population. Sequence analysis revealed that these individuals are likely natural hybrids between C. podogyna and a co-occurring species, C. curvicollis, although established hybrids have never been found in the natural habitat. Based on our observation of variegated leaves in hybrid individuals, we propose that hybrids have been excluded by natural selection and/or interspecific competition caused by insufficient productivity of photosynthesis, although other genetic and ecological factors may also be influential. Overall, this study indicates that natural mechanisms selectively removing the hybrids have maintained the genetic purity of this rear-edge population of C. podogyna, and also emphasizes the importance of genetic assessment in ex-situ conservation programs.

Ex-situ conservation and diversity index of greened ex-tin mine

The greened ex-tin mine comprises planted timber species of Acacia mangium, Hopea odorata and Khaya ivorensis. The ex-tin mine was formerly a lowland dipterocarps mixed forest. The man-made forest was established at 4 x 5 m spacing or 500 stem/ha on slime tailings. However, high mortality of the timber species due to damages by wild pigs and farm animals have reduced the plantation of 3.2 ha to only 93 stems of Acacia mangium, 92 stems of Hopea odorata and 130 stems of Khaya ivorensis at 20 years after planting. The regeneration covering the planting line was kept and the planting spaces in-between the planting lines were cleared for planting of another 14 species of endemic, endangered and threatened tree species (EETS). The ex-situ conservation program of planting 1500 stems of EETS to the 3 ha site was sponsored by Asian Forest Cooperation Organization (AFoCO). The 100% vegetation survey was carried out 4 years after planting. The species richness, Shannon diversity index and species evenness were computed for the nurse stand, and the nurse stands with EETS. The results obtained for species richness, Shannon Index (H) and Shannon Evenness index (EH) of the nurse stand and nurse stand with EETS was 24, 1.46, 0.46 and 38, 2.30, 0.63, respectively. The nurse stand with EETS has increased in species richness, H and EH of nurse stand to 58%. and 36.9%, respectively. More importantly, none of the introduced EETS was found in the composition of the natural regeneration of the 20 years old nurse stand. The 21 species naturally regenerated in the nurse stand were an only pioneer and late pioneer species. This shows that ex-tin mine after the reclamation efforts of planting timber species and the natural regeneration will not be able to regain the initial species composition of a lowland dipterocarps forest. The domestication technology of EETS developed in this project is reproducible. The project has successfully preserved the 14 EETS and the site shall be a demonstration plot for the future seed production area.

Genetic variability and population structure of Ethiopian chickpea (Cicer arietinum L.) germplasm.

Evaluation of the genetic diversity and an understanding of the genetic structure and relationships of chickpea genotypes are valuable to design efficient germplasm conservation strategies and crop breeding programs. Information is limited, in these regards, for Ethiopian chickpea germplasms. Therefore, the present study was carried out to estimate the genetic diversity, population structure, and relationships of 152 chickpea genotypes using simple sequence repeats (SSR) markers. Twenty three SSR markers exhibited polymorphism producing a total of 133 alleles, with a mean of 5.8 alleles per locus. Analyses utilizing various genetic-based statistics included pairwise population Nei's genetic distance, heterozygosity, Shannon's information index, polymorphic information content, and percent polymorphism. These analyses exemplified the existence of high genetic variation within and among chickpea genotypes. The 152 genotypes were divided into two major clusters based on Nei's genetic distances. The exotic genotypes were grouped in one cluster exclusively showing that these genotypes are distinct to Ethiopian genotypes, while the patterns of clustering of Ethiopian chickpea genotypes based on their geographic region were not consistent because of the seed exchange across regions. Model-based population structure clustering identified two discrete populations. These finding provides useful insight for chickpea collections and ex-situ conservation and national breeding programs for widening the genetic base of chickpea.

Conservation biology of the critically endangered salamander, Paradactylodon persicus gorganensis (Clergue-Gazeau &amp; Thorn, 1979) (Amphibia: Hynobiidae) in northeastern Iran

Abstract Paradactylodon persicus gorganensis is a critically endangered salamander found in northeastern Iran which faces increasing major anthropogenic threats such as agriculture, industry and mining. The aim of the current study is to identify key environmental variables associated with the species’ presence. Also, measuring and comparing the water quality parameters in habitats where P. p. gorganensis is present and absent are the other aims of the study. The results indicate that high pH and dissolved oxygen values have a negative effect on the species’ presence, while its presence was strongly correlated with low mean water temperatures. The species’ range in response to the climate change will shift to the south and east, where areas of high elevation occur, because the species favors cold habitats. This study indicates that it will be particularly important to evaluate the whole range where P. p. gorganensis exists when preparing conservation action plans. It is recommended that ex-situ conservation programs are incorporated into conservation planning for the species, because the number of individuals in the wild is so low and their future viability is fragile.

Genetic diversity and population structure of Eurycoma apiculata in Eastern Sumatra, Indonesia

Abstract. Zulfahmi, Parjanto, Purwanto E, Yunus A. 2021. Genetic diversity and population structure of Eurycoma apiculata in Eastern Sumatra, Indonesia. Biodiversitas 22: 4431-4439. Information on genetic variation within and among populations of Eurycoma apiculata plants is important to develop strategies for their conservation, sustainable use, and genetic improvement. To date, no information on genetic variation within and among populations of the E. apiculata has been reported. This study aims to assess genetic diversity within and among populations of E. apiculata based on RAPD markers, and to determine populations to collect E. apiculata genetic material for conservation and breeding programs. Young leaves of E. apiculata were collected from six natural populations. Fifteen RAPD primers were used to assess the genetic diversity of each population. The data obtained were analyzed with POPGEN and Arlequin software. The amplification results of 15 selected primers produced 3-16 loci with all primers 100% polymorphic. At the species level, the mean allele per locus (Na), number of effective alleles (Ne), percentage of polymorphic loci (PPL), Nei’s gene diversity index (He) and Shannon information index (I) were 2.000, 1.244, 100%, 0.167, and 0.286, respectively. At the population level, the mean values for Na, Ne, PPL, He and I were 1.393, 1.312, 39.27%, 0.119, and 0.186, respectively. The highest value of gene diversity within population (He) was found in the Lingga-1 population and the lowest value was found in the Rumbio population. The value of genetic differentiation among populations (GST) of E. apiculata is 0.284, consistent with the results of the AMOVA analysis which found that genetic variation among populations was 23.14%, indicates that the genetic variation of E. apiculata was more stored within populations than among populations. The gene flow (Nm) value of E. apiculata was 1.259 migrants per generation among populations. The Nm value of this species was high category, and could inhibit genetic differentiation among populations. The clustering of E. apiculata population based on the UPGMA dendrogram and PCA was inconsistent with its geographic distribution, reflecting the possibility that genes migration occurred between islands in the past. The main finding of this study was the genetic variation of the E. apiculata mostly stored within the population. Therefore, the population with the highest genetic diversity is a priority for in-situ conservation, and collection of E. apiculata genetic material for ex-situ conservation and breeding programs should be carried out minimum from Lingga-1 and Pokomo populations.

Landscape-level habitat management plan through geometric reserve design for critically endangered Hangul (Cervus hanglu hanglu).

Hangul (Cervus hanglu hanglu), the only red deer subspecies surviving in the Indian subcontinent, is of top conservation priority with global importance. Unfortunately, it has lost much of its historical distribution range, and it is now confined to Dachigam landscape within the Kashmir valley of India. The Government of India initiated a recovery plan in 2008 to augment their numbers through ex-situ conservation programs. However, it was necessary to identify potential hangul habitats in Kashmir valley for adopting landscape-level conservation planning for the species. Based on geometric aspects of reserve design, we modeled hangul habitat using an ensemble approach to identify hangul habitats. The present model indicates that the conifer and broadleaf mixed forests were the most suitable habitats. Only 9% of the total study landscape was found suitable for the species. We identified corridors among the suitable habitat blocks, which may be vital for the species' long-term genetic viability. We suggest reorganizing the existing management of Dachigam National Park (NP) following the landscape level and habitat block-level management planning based on the core principles of geometric reserve design. We recommend that the identified patch (PID-6) in the southern region of the landscape to be converted into a Conservation Reserve or merged with the Overa-Aru Wildlife Sanctuary. This habitat patch PID-6 may be a stepping stone habitat and vital for maintaining the species landscape connectivity and metapopulation dynamics.

White-naped mangabeys\u2019 viable insurance population within European Zoo Network

The success and viability of an ex-situ conservation program lie in the establishment and potential maintenance of a demographically and genetically viable insurance population. Such population reserve may support reintroduction and reinforcement activities of wild populations. White-naped mangabeys are endangered restricted-range African primates which have experienced a dramatic population decrease in their natural habitats over the last few decades. Since 2001, some European zoos singularly monitor an ex-situ population aiming to seek the recovery of the current wild population. The aim of the present paper is to evaluate the genetic status and population demographics of European zoo-captive white-naped mangabeys based on pedigree data. The captive population is gradually growing and preserves specific reproductive and demographic parameters linked to the species. The intensive management program that is implemented has brought about the minimization of inbreeding and average relatedness levels, thus maintaining high levels of genetic diversity despite the existence of fragmented populations. This finding suggests white-naped mangabey ex-situ preservation actions may be a good example of multifaceted conservation throughout studbook management which could be used as a model for other ex-situ live-animal populations.

Studbook and molecular analyses for the endangered black-lion-tamarin; an integrative approach for assessing genetic diversity and driving management in captivity

Breeding strategies based on molecular markers have been adopted by ex-situ conservation programs to assess alternative parameters for the genetic diversity estimates. In this work we evaluated molecular and studbook data for captive populations of black-lion-tamarin (BLT), an endangered primate endemic to Brazil’s Atlantic Forest. Pedigree analyses were performed using BLT studbook information collected from 1973 to 2018. We analyzed the whole captive population since its foundation; the current captive population (CCP); and all extant BLTs in the Brazilian captive population (BCP), separately. Microsatellite analyses were implemented on the BCP individuals from the eighth generation (BCP-F8) only to avoid generation overlap. The expected heterozygosity for BCP-F8, using molecular, data was 0.45, and the initial expected heterozygosity was 0.69. Kinship parameters showed high genetic relationships in both pedigree and molecular analyses. The genealogy-based endogamy evidenced a high inbreeding coefficient, while the molecular analyses suggested a non-inbreeding signature. The Mate Suitability Index showed detrimental values for the majority of potential pairs in the CCP. Nevertheless, some individuals evidenced high individual heterozygosity and allele representation, demonstrating good potential to be used as breeders. Thus, we propose the use of molecular data as a complementary parameter to evaluate mating-pairs and to aid management decision-making.

Effects of personality and rearing-history on the welfare of captive Asiatic lions (Panthera leo persica).

BackgroundThe long-term success of ex-situ conservation programmes depends on species-appropriate husbandry and enrichment practices complemented by an accurate welfare assessment protocol. Zoos and conservation breeding programmes should employ a bottom-up approach to account for intraspecific variations in measures of animal welfare. We studied 35 (14:21) captive Asiatic lions in Sakkarbaug Zoological Garden, Junagadh, India to understand the implications of individual variations on welfare measures. We categorized the subjects based on personality traits (bold or shy), rearing history (wild-rescued or captive-raised), sex, and social-grouping. We explored the association of these categorical variables on welfare indices such as behavioural diversity, latency to approach novel objects, enclosure usage and aberrant repetitive behaviours. Further, we assessed the inter-relationships between different behavioural measures of welfare.ResultsOur results show that intraspecific variations based on rearing-history and personality traits are significantly associated with the welfare states of captive Asiatic lions. Asiatic lions with bold personality traits (M = 0.50, SD = 0.12, N = 21) and those raised in captivity (M = 0.47, SD = 0.12, N = 16) used enclosure space more homogenously compared to shy (M = 0.71, SD = 0.15, N = 14) and wild-rescued (M = 0.67, SD = 0.15, N = 19) animals. Behaviour diversity was significantly higher in captive-raised (M = 1.26, SD = 0.3, N = 16) and bold (M = 1.23, SD = 0.26, N = 21) subjects compared to wild-rescued (M = 0.83, SD = 0.35, N = 19) and shy (M = 0.73, SD = 0.34, N = 14) individuals. Aberrant repetitive behaviours (stereotypy) were significantly lower in bold (M = 7.01, SD = 4, N = 21) and captive-raised (M = 7.74, SD = 5.3) individuals compared to wild-rescued (M = 13.12, SD = 6.25, N = 19) and shy (M = 16.13, SD = 5.4, N = 16) lions. Sex and social-grouping of subjects did not show significant associations with behavioural welfare indices. Interestingly, behaviour diversity was reliably predicted by the enclosure usage patterns and aberrant repetitive behaviours displayed by subjects.DiscussionOur findings underline the importance of individual-centric, behaviour-based, and multi-dimensional welfare assessment approaches in ex-situ conservation programmes. The results suggest that behavioural welfare indices complemented with individual variations can explain inter-individual differences in behavioural welfare measure outcomes of Asiatic lions. These findings also provide zoo managers with a non-invasive tool to reliably assess and improve husbandry practices for Asiatic lions. Understanding the unique welfare requirement of individuals in captivity will be crucial for the survival of the species.

Sexual Behavior of Javan Langur (Trachypithecus auratus) in Taman Safari Indonesia Ex-situ Conservation Facility

Trachypithecus auratus is one of the vulnerable primate species that is declining in number due to illegal hunting and forest degradation. The efforts to increase population can be through ex-situ conservation program that are specific to reproduction. To date, little is known about the reproduction of this species, particularly in the aspect of sexual behavior. The aim of this research was to analyze the sexual behavior of T. auratus at the Taman Safari Indonesia ex-situ conservation facility. This research conducted on five adult individuals from a group of consisting of one male and four females. The group of T. auratus were observed for five months from September 2018 until January 2019. The method used focal animal sampling with BORIS (Behavioral Observation Research Interactive Software). The results showed that the females performed pre-copulation (courtship) behavior such as head shaking (28.90%), hindquarter present (17.33%), raising a forelimb (11.66%), and the male performed pre-copulation (courtship) behavior such as genital inspection (68.46%). Copulation lasts for 60 seconds, starting with male mounting and thrusting as many as 40 seconds and ejaculation which is thought to occur for 20 seconds. At the end of the copulation sequence, a pair of T. auratus showed grooming and foraging together. The conclusion of this research was T. auratus has parameters of sexual behavior with copulation that lasts for 60 seconds and head shaking was sexual behavior that can be used as an signal for mating. This research are expected to be used as basic reproductive biology data to support the breeding and conservation program of T. auratus species.

Captivity and Infection by the Fungal Pathogen Batrachochytrium salamandrivorans Perturb the Amphibian Skin Microbiome.

The emerging fungal pathogen, Batrachochytrium salamandrivorans (Bsal) is responsible for the catastrophic decline of European salamanders and poses a threat to amphibians globally. The amphibian skin microbiome can influence disease outcome for several host-pathogen systems, yet little is known of its role in Bsal infection. In addition, many experimental in-vivo amphibian disease studies to date have relied on specimens that have been kept in captivity for long periods without considering the influence of environment on the microbiome and how this may impact the host response to pathogen exposure. We characterized the impact of captivity and exposure to Bsal on the skin bacterial and fungal communities of two co-occurring European newt species, the smooth newt, Lissotriton vulgaris and the great-crested newt, Triturus cristatus. We show that captivity led to significant losses in bacterial and fungal diversity of amphibian skin, which may be indicative of a decline in microbe-mediated protection. We further demonstrate that in both L. vulgaris and T. cristatus, Bsal infection was associated with changes in the composition of skin bacterial communities with possible negative consequences to host health. Our findings advance current understanding of the role of host-associated microbiota in Bsal infection and highlight important considerations for ex-situ amphibian conservation programmes.