Prague Zoo

The Usutu virus (USUV; Orthoflavivirus, Flaviviridae) is a mosquito-borne pathogen causing fatal neuroinfections in susceptible wild and captive birds, particularly blackbirds, other passerines, and owls. Zoological gardens provide favourable conditions for the circulation of such viruses due to the proximity of diverse species and limited options for prevention. Following the sudden death of several Chilean tinamous kept in the Brno zoological garden, we tested tissues sampled from 22 bird cadavers (from zoos, private owners, and free-living birds) for the presence of USUV and West Nile virus (WNV) RNA using duplex reverse transcription qPCR. Near-complete whole-genome sequences were acquired from positive samples by next-generation sequencing and subjected to phylogenetic analyses. Furthermore, serum samples from additional zoo animals and privately owned birds were screened for anti-flavivirus antibodies using ELISA and subsequently confirmed by the virus neutralization test. We report fatal USUV infections in multiple bird species from three zoological gardens in the Czech Republic. Duplex RT-qPCR targeting USUV and West Nile virus (WNV) detected USUV RNA in tissues from two Boreal owls (Aegolius funereus), one Eurasian pygmy owl (Glaucidium passerinum), two Brahminy starlings (Sturnia pagodarum), and four Chilean tinamous (Nothoprocta perdicaria). Additionally, three randomly found cadavers of free-living blackbirds (Turdus merula) tested positive. Pathological findings ranged from minimal pathological changes to pronounced hepatosplenomegaly with intestinal bleeding. Phylogenetic analysis of near-complete genome sequences assigned all viruses to the Europe 2 genetic lineage, revealing partial geographic clustering among isolates obtained in this study. Serological testing confirmed exposure in additional birds and demonstrated cross-neutralisation between anti-USUV and anti-WNV-positive sera. In zoological gardens, flavivirus infections can cause substantial losses, even among rarely bred or endangered species. Given the zoonotic potential of both USUV and WNV, documenting their occurrence in avian hosts is important not only for animal health but also for human disease surveillance from the One Health perspective.

Cloning from historically cryopreserved cells offers a potential means to restore lost genetic variation or increase the representation of particular lineages within bottlenecked species, provided such biobanked materials are archived for such genetic rescue applications. One species for which cloning can provide genetic management benefits is Przewalski’s horse, Equus przewalskii. All ~1800 living Przewalski’s horses, distributed across ex situ breeding facilities and in situ reintroduction sites, are descended from one or more of the five founder lineages established by the 12 horses captured from the wild between 1898 and 1947. Since the 1970s, the San Diego Zoo Wildlife Alliance Biodiversity Bank’s Frozen Zoo® (Escondido, CA, USA) has biobanked cells or tissues of 575 individuals spanning many generations. A pedigree analysis of a subset of deceased individuals represented in the San Diego Zoo Wildlife Alliance Biodiversity Bank’s Frozen Zoo® revealed an underrepresented male that lived from 1975 to 1998, Studbook Number 615 (also known as Kuporovitch), who would be of high value for breeding if cloned. Here, we report that two healthy clones were produced from this cell line using cross-species somatic cell nuclear transfer from 2020 to 2023. Their identification as clones was verified by a standard horse-pedigree genotyping panel, and, for one clone, a whole genome sequencing comparison to the original donor was performed. This is the first time that multiple healthy clones surviving the perinatal period have been produced for an endangered species.

Abstract The house mouse,Mus musculus, is a widely used animal model in biomedical research, with classical laboratory strains (CLS) being the most frequently employed. However, the limited genetic variability in CLS hinders their applicability in evolutionary studies. Wild-derived strains (WDS), on the other hand, provide a suitable resource for such investigations. This study quantifies genetic and phenotypic data of 101 WDS representing 5 species, 3 subspecies, and 8 natural Y consomic strains and compares them with CLS. Genetic variability was estimated using whole mtDNA sequences, thePrdm9gene, and copy number variation at two sex chromosome-linked genes. WDS exhibit a large natural variation with up to 2173 polymorphic sites in mitogenomes, whereas CLS display 92 sites. Moreover, while CLS have twoPrdm9alleles, WDS harbour 46 different alleles. Although CLS resembleM. m. domesticusandM. m. musculusWDS, they differ from them in 8 and 11 out of 15 phenotypic traits, respectively. The results suggest that WDS can be a useful tool in evolutionary studies and have great potential for medical applications.

Analyzing social networks is a powerful tool for understanding the ecology of social species. While most studies focus on the role of each group member, few compare groups with different characteristics. The current population of Przewalski's horses Equus ferus przewalskii at the Great Gobi B Strictly Protected Area (Mongolia) includes groups of wild-born and captive-bred individuals with different experiences with the area (acclimatizing, long-term reintroduced, and wild-born), therefore serving as an ideal natural behavioral lab. We filmed 11 groups for 141.5 hours in summer 2018 (July), late spring 2019 (May, June), and autumn 2019 (September, October). Affiliative and agonistic interactions were recorded, and social networks were created. We tested the influence of origin, experience, season, sex, age, relative time belonging to the group, relatedness, and dominance rank on different network indices at the individual and group levels. We found that groups with greater experience in the area are generally better connected than members of the newly formed groups. However, these strong networks were created by wild-born individuals with very low interaction rates. On the contrary, inexperienced groups composed of captive-bred individuals displayed many interactions but created weak social networks. The results show a trend of behavioral transition from acclimatizing through long-term reintroduced to wild-born groups, supporting that the newly formed groups of released Przewalski's horses need time to display the typical social behavior patterns of wild-born individuals. Long-term monitoring of released Przewalski's horses is recommended to promote the success of this reintroduction program.

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

Longevity-related genes have been found in humans, mice, dogs and in several other animal species. The goal of this study was to perform genetic analysis of long-lived European bisons with the aim to find genes that are associated with longevity using GWAS and further sequencing of a wider sample panel. European bison has a unique history of near extinction and the recovery of the species from just a few founder individuals. Together with the short medium lifespan, the expected genetic homogeneity makes bison a suitable model for studying longevity. Particular single nucleotide polymorphisms within three genes, BCKDHB, FER1L6 and SERPINI2, were found significantly overrepresented in long-lived European bisons. In SERPINI2, the longevity-associated single nucleotide polymorphism localizes to an exon. In the protein encoded by the SERPINI2 gene, amino acid leucine present in the reference European bisons is replaced by tryptophan in the long-lived animals. This study is the first to determine longevity-associated variants in genes in European bison. Association of the FER1L6 gene with longevity shows a possible sex dependency.

Screening of serum and fecal samples from huemul (Hippocamelus bisulcus) and pudu (Pudu puda) from southern Chile for Mycobacterium bovis and Mycobacterium avium paratuberculosis (MAP) found all but four samples Mycobacterium-negative. The positive sequences showed only 92-93% similarity with MAP and were from remote Isla Riesco populations.

BackgroundLongevity-related genes have been found in several animal species as well as in humans. The goal of this study was to perform genetic analysis of long-lived Cane corso dogs with the aim to find genes that are associated with longevity.ResultsSNPs with particular nucleotides were significantly overrepresented in long-lived dogs in four genes, TDRP, MC2R, FBXO25 and FBXL21. In FBXL21, the longevity-associated SNP localises to the exon. In the FBXL21 protein, tryptophan in long-lived dogs replaced arginine present in reference dogs.ConclusionsFour SNPs associated with longevity in dogs were identified using GWAS and validated by DNA sequencing. We conclude that genes TDRP, MC2R, FBXO25 and FBXL21 are associated with longevity in Cane corso dogs.

The gut microbiome of primates is known to be influenced by both host genetic background and subsistence strategy. However, these inferences have been made mainly based on adaptations in bacterial composition - the bacteriome and have commonly overlooked the fungal fraction - the mycobiome. To further understand the factors that shape the gut mycobiome of primates and mycobiome-bacteriome interactions, we sequenced 16 S rRNA and ITS2 markers in fecal samples of four different nonhuman primate species and three human groups under different subsistence patterns (n = 149). The results show that gut mycobiome composition in primates is still largely unknown but highly plastic and weakly structured by primate phylogeny, compared with the bacteriome. We find significant gut mycobiome overlap between captive apes and human populations living under industrialized subsistence contexts; this is in contrast with contemporary hunter-gatherers and agriculturalists, who share more mycobiome traits with diverse wild-ranging nonhuman primates. In addition, mycobiome-bacteriome interactions were specific to each population, revealing that individual, lifestyle and intrinsic ecological factors affect structural correspondence, number, and kind of interactions between gut bacteria and fungi in primates. Our findings indicate a dominant effect of ecological niche, environmental factors, and diet over the phylogenetic background of the host, in shaping gut mycobiome composition and mycobiome-bacteriome interactions in primates.