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Introduction: The future of human space missions relies on the ability to provide adequate food resources for astronauts and also to reduce stress due to the environment (microgravity and cosmic radiation). In this context, microgreens have been proposed for the astronaut diet because of their fast-growing time and their high levels of bioactive compounds and nutrients (vitamins, antioxidants, minerals, etc.), which are even higher than mature plants, and are usually consumed as ready-to-eat vegetables., Methods: Our study aimed to identify the best light recipe for the soilless cultivation of two cultivars of radish microgreens (Raphanus sativus, green daikon, and rioja improved) harvested eight days after sowing that could be used for space farming. The effects on plant metabolism of three different light emitting diodes (LED) light recipes (L1-20% red, 20% green, 60% blue; L2-40% red, 20% green, 40% blue; L3-60% red, 20% green, 20% blue) were tested on radish microgreens hydroponically grown. A fluorimetric-based technique was used for a real-time non-destructive screening to characterize plant methabolism. The adopted sensors allowed us to quantitatively estimate the fluorescence of flavonols, anthocyanins, and chlorophyll via specific indices verified by standardized spectrophotometric methods. To assess plant growth, morphometric parameters (fresh and dry weight, cotyledon area and weight, hypocotyl length) were analyzed., Results: We observed a statistically significant positive effect on biomass accumulation and productivity for both cultivars grown under the same light recipe (40% blue, 20% green, 40% red). We further investigated how the addition of UV and/or far-red LED lights could have a positive effect on plant metabolite accumulation (anthocyanins and flavonols)., Discussion: These results can help design plant-based bioregenerative life-support systems for long-duration human space exploration, by integrating fluorescence-based non-destructive techniques to monitor the accumulation of metabolites with nutraceutical properties in soilless cultivated microgreens., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Garegnani, Sandri, Pacelli, Ferranti, Bennici, Desiderio, Nardi and Villani.)

The effect of visitor presence on animal behavior needs to be evaluated to ensure the welfare and improve the husbandry of the individuals in zoos. This study aims to assess the effect of visitor presence on the behavior and welfare of pairs of Amur tiger, snow leopard, and Eurasian lynx at Parco Natura Viva, Italy. The study was made of two periods: the baseline, in which the zoo was closed, and the visitor presence period, in which the zoo was opened. Per period and per subject 12 30-min observations were done. Continuous focal animal sampling method was used to collect behavior duration of the big cats. The main results of the study highlighted that, when visitors were present, all felids except for the female lynx were significantly more inactive than in the baseline. Moreover, despite interindividual and inter-species variability in the significance level of findings, natural behaviors such as attentive behavior, exploration/marking, locomotion, and positive social interactions were performed more in the baseline than in the visitor presence period. Finally, when visitors were present, as the studied subjects experienced daily longer exposure to visitors, inactivity increased whereas individual species-typical behaviors (e.g., locomotion) and positive social interactions decreased. Thus, visitors seem to partially alter the behavioral time-budgets of the study big cats, increasing inactivity and decreasing the performance of species-specific behavior, at least in some individuals., (© 2023 The Authors. Zoo Biology published by Wiley Periodicals LLC.)

Aims: Indri indri is a lemur of Madagascar which is critically endangered. The analysis of the microbial ecology of the intestine offers tools to improve conservation efforts. This study aimed to achieve a functional genomic analysis of three Lactiplantibacillus plantarum isolates from indris., Methods and Results: Samples were obtained from 18 indri; 3 isolates of Lp. plantarum were obtained from two individuals. The three isolates were closely related to each other, with <10 single nucleotide polymorphisms, suggesting that the two individuals shared diet-associated microbes. The genomes of the three isolates were compared to 96 reference strains of Lp. plantarum. The three isolates of Lp. plantarum were not phenotypically resistant to antibiotics but shared all 17 genes related to antimicrobial resistance that are part of the core genome of Lp. plantarum. The genomes of the three indri isolates of Lp. plantarum also encoded for the 6 core genome genes coding for enzymes related to metabolism of hydroxybenzoic and hydroxycinnamic acids. The phenotype for metabolism of hydroxycinnamic acids by indri isolates of Lp. plantarum matched the genotype., Conclusions: Multiple antimicrobial resistance genes and gene coding for metabolism of phenolic compounds were identified in the genomes of the indri isolates, suggesting that Lp. plantarum maintains antimicrobial resistance in defense of antimicrobial plant secondary pathogens and that their metabolism by intestinal bacteria aids digestion of plant material by primate hosts., (© The Author(s) 2023. Published by Oxford University Press on behalf of Applied Microbiology International.)

Intestinal bacteria establish a specific relationship with the host animal, which causes the acquisition of gut microbiota with a unique composition classified as the enterotype. As the name suggests, the Red River Hog is a wild member of the pig family living in Africa, in particular through the West and Central African rainforest. To date, very few studies have analysed the gut microbiota of Red River Hogs (RRHs) both housed under controlled conditions and in wild habitats. This study analysed the intestinal microbiota and the distribution of Bifidobacterium species in five Red River Hog (RRH) individuals (four adults and one juvenile), hosted in two different modern zoological gardens (Parco Natura Viva, Verona, and Bioparco, Rome) with the aim of disentangling the possible effects of captive different lifestyle and host genetics. Faecal samples were collected and studied both for bifidobacterial counts and isolation by means of culture-dependent method and for total microbiota analysis through the high-quality sequences of the V3-V4 region of bacterial 16S rRNA. Results showed a host-specific bifidobacterial species distribution. Indeed, B. boum and B. thermoacidophilum were found only in Verona RRHs, whereas B. porcinum species were isolated only in Rome RRHs. These bifidobacterial species are also typical of pigs. Bifidobacterial counts were about 10 6 CFU/g in faecal samples of all the individuals, with the only exception for the juvenile subject, showing 10 7 CFU/g. As in human beings, in RRHs a higher count of bifidobacteria was also found in the young subject compared with adults. Furthermore, the microbiota of RRHs showed qualitative differences. Indeed, Firmicutes was found to be the dominant phylum in Verona RRHs whereas Bacteroidetes was the most represented in Roma RRHs. At order level, Oscillospirales and Spirochaetales were the most represented in Verona RRHs compared with Rome RRHs, where Bacteroidales dominated over the other taxa. Finally, at the family level, RRHs from the two sites showed the presence of the same families, but with different levels of abundance. Our results highlight that the intestinal microbiota seems to reflect the lifestyle (i.e., the diet), whereas age and host genetics are the driving factors for the bifidobacterial population.

A comparative quali-quantitative study of the peel extracts of eight Punica granatum cultivars obtained from underexploited areas of South Italy was carried out in order to valorize them as health-promoting by-products. The results showed that all of the samples possessed 45 ellagitannins, consisting mainly of polyhydroxyphenoyls; 10 flavonoids, belonging to flavonol, flavone, and catechin classes; and 2 anthocyanins. The most representative compounds underwent quantification through a LC-MS/MS multiple reaction monitoring (MRM)-based method; their qualitative profile was almost superimposable, while variability in the quantitative phenolic content was observed. The antioxidant activity was investigated using cell-free and cell-based assays. The in vitro anti-inflammatory potential was also studied by monitoring three typical markers of inflammation (IL-1β, IL-6, and TNF-α). Moderate differences in both activities were observed between the cultivars. Results showed that all of the investigated peels have a potential use as healthy bioactive phytocomplexes due to the interesting antioxidant and anti-inflammatory activities; in particular from the bioinformatic approaches a series of compounds, including galloyl-, pedunculagin- and ellagic acid-based, were found to be highly correlated with bioactivity of the extracts. Finally, the bioactivities showed by a Campanian local cultivar, 'Granato di Aiello del Sabato', could promote its cultivation by local farmers and germplasm conservation.

Although some studies investigated lateralization in reptiles, little research has been done on chelonians, focusing only on few behaviours such as righting response and escape preference. The aim of this study was to investigate lateralization in Aldabra giant tortoises (Aldabrachelys gigantea), focusing on asymmetrical positioning of the limbs and the head during resting behaviour, called sleep-like behaviour, involving both wild tortoises and individuals under human care. Subjects of the study were 67 adult Aldabra tortoises (54 free ranging on Curieuse, 13 under human care in Mahè Botanical Garden). For each tortoise observed during sleep-like behaviour, we recorded the position of the head (on the left, on the right or in line with the body midline) and we collected which forelimb and hindlimb were kept forward. Moreover, the number of subjects in which limbs were in a symmetrical position during the sleep-like behaviour was recorded. Based on our results, the number of tortoises with asymmetrical position of head and limb was higher (head: 63%; forelimbs: 88%; hindlimbs: 70%) than the number of tortoises with symmetrical position of the head and the limb. Regarding the head, throughout the subjects found with the asymmetrical position of the head during sleep-like behaviour, tortoises positioning the head on the right (42%) were more than those sleeping with the head on the left (21%). We found a relationship between the position of the forelimbs and hindlimbs during sleep-like behaviour. We reported no differences between Mahè (under human care) and Curieuse (wild) tortoises. Findings of this preliminary study underlined traces of group-level lateralization in head positioning during the sleep-like behaviour, possibly due to a left-eye/right-hemisphere involvement in anti-predatory responses and threatening stimuli as reported in reptiles and other vertebrates. This study aims at adding data on brain lateralization, often linked to lateralized behaviours, in reptiles, especially in chelonians., (© 2021. The Author(s).)

Dynamic modulation of endothelial cell-to-cell and cell-to-extracellular matrix (ECM) adhesion is essential for blood vessel patterning and functioning. Yet the molecular mechanisms involved in this process have not been completely deciphered. We identify the adhesion G protein-coupled receptor (ADGR) Latrophilin 2 (LPHN2) as a novel determinant of endothelial cell (EC) adhesion and barrier function. In cultured ECs, endogenous LPHN2 localizes at ECM contacts, signals through cAMP/Rap1, and inhibits focal adhesion (FA) formation and nuclear localization of YAP/TAZ transcriptional regulators, while promoting tight junction (TJ) assembly. ECs also express an endogenous LPHN2 ligand, fibronectin leucine-rich transmembrane 2 (FLRT2), that prevents ECM-elicited EC behaviors in an LPHN2-dependent manner. Vascular ECs of lphn2a knock-out zebrafish embryos become abnormally stretched, display a hyperactive YAP/TAZ pathway, and lack proper intercellular TJs. Consistently, blood vessels are hyperpermeable, and intravascularly injected cancer cells extravasate more easily in lphn2a null animals. Thus, LPHN2 ligands, such as FLRT2, may be therapeutically exploited to interfere with cancer metastatic dissemination., (© 2021 Camillo et al.)

Here, we investigated the possible linkages among geophagy, soil characteristics, and gut mycobiome of indri (Indri indri), an endangered lemur species able to survive only in wild conditions. The soil eaten by indri resulted in enriched secondary oxide-hydroxides and clays, together with a high concentration of specific essential micronutrients. This could partially explain the role of the soil in detoxification and as a nutrient supply. Besides, we found that soil subject to geophagy and indris' faeces shared about 8.9% of the fungal OTUs. Also, several genera (e.g. Fusarium, Aspergillus and Penicillium) commonly associated with soil and plant material were found in both geophagic soil and indri samples. On the contrary, some taxa with pathogenic potentials, such as Cryptococcus, were only found in indri samples. Further, many saprotrophs and plant-associated fungal taxa were detected in the indri faeces. These fungal species may be involved in the digestion processes of leaves and could have a beneficial role in their health. In conclusion, we found an intimate connection between gut mycobiome and soil, highlighting, once again, the potential consequent impacts on the wider habitat., (© 2021. The Author(s).)

Many studies have highlighted evidence of lateralized behaviours in vertebrates and invertebrates, indicating that cerebral lateralization might not be uniquely human. Flamingos, as highly social species, might represent an interesting model in the study of lateralization, as this trait appears to be useful in gregarious animals. This study aims to investigate the presence of lateralized behaviours in zoo flamingos. The subjects of this study were 41 greater flamingos ( Phoenicopterus roseus ) and 29 Chilean flamingos ( Phoenicopterus chilensis ). Ten 20-minute observation sessions per subject were carried out, using focal animal sampling method. Bouts of side preferences were recorded for foraging, neck resting and preening. Moreover, bouts and duration of leg use for postural support during unipedal standing (leg stance) were also collected. Results highlighted a group-level right-side preference for foraging in the whole sample as well as within each species. Within greater flamingos, group-level right-side preferences were also reported for neck resting and leg stance. Differences between the two flocks were found, with greater flamingos displaying right preferences to a greater extent than Chilean flamingos. Males showed a more pronounced right preference than females for some categories. This study adds to previous literature highlighting the presence of group-level behavioural lateralization in flamingos.

Early-life experiences may considerably affect the behavioural patterns of adult primates. Particularly, atypical rearing practices might lead to abnormal behaviours and social-sexual deficiencies in captive, adult non-human primates. We conducted behavioural observations of mother-reared (n = 5) and hand-reared (n = 6) adult chimpanzees in a social group at Parco Natura Viva, Italy. We used continuous focal animal sampling to collect behavioural data focusing on individual and social behaviours. We found that all study subjects performed individual and social species-specific behaviours. However, mother-reared chimpanzees performed locomotion and affiliative behaviours significantly more than hand-reared subjects. In addition to these species-typical behaviours, hand-reared chimpanzees showed significantly more abnormal behaviours than mother-reared subjects. Therefore, these findings suggest that hand-rearing could have wide-reaching effects on the behavioural repertoire in adult zoo-housed chimpanzees. Hence, even if sometimes human intervention in rearing may be necessary to ensure the survival of captive infant chimpanzees, our results suggest that zoo-housed chimpanzees might benefit from minimised human-animal interactions and exposure to conspecifics throughout their development. These suggestions should be implemented in regular husbandry practices., (© 2021 S. Karger AG, Basel.)

Urinalysis could be helpful to investigate the health status of giraffes held in captivity using noninvasive methods to avoid animal handling or anesthesia. We collected 52 voided urine samples from 20 giraffes of different ages, sexes, and subspecies from the ground. To evaluate potential interference by soil contaminants, a pilot study was performed using 20 urine samples obtained from 10 cows. All bovine and 29 giraffe samples were subjected to routine urinalysis including urine specific gravity (USG). All samples were analyzed for urine total protein (uTP), urine creatinine (uCrea) concentration, and urine protein-to-urine creatinine ratio (UPC). Urinary proteins were separated by SDS-PAGE electrophoresis. No significant differences were determined between free-catch and urine sampled from the ground in cows. Giraffe urine was pale-yellow, with alkaline pH (>8.0) and a mean USG of 1.035 ± 0.013. The uTP, uCrea, and UPC expressed as median (range) were 0.20 (0.08-0.47) g/L, 2.36 (0.62-5.2) g/L, and 0.08 (0.05-0.15), respectively. SDS-PAGE allowed the separation of protein bands with different molecular masses, including putative uromodulin at 90 kD, putative albumin at 64 kD, and putative immunoglobulin heavy and light chains at 49 kD and 25 kD, respectively. Urine collection from the ground appears to be a reliable technique for urinalysis and urine electrophoresis in giraffes.

Although subcellular positioning of endosomes significantly impacts on their functions, the molecular mechanisms governing the different steady-state distribution of early endosomes (EEs) and late endosomes (LEs)/lysosomes (LYs) in peripheral and perinuclear eukaryotic cell areas, respectively, are still unsolved. We unveil that such differences arise because, while LE retrograde transport depends on the dynein microtubule (MT) motor only, the one of EEs requires the cooperative antagonism of dynein and kinesin-14 KIFC1, a MT minus end-directed motor involved in cancer progression. Mechanistically, the Ser-x-Ile-Pro (SxIP) motif-mediated interaction of the endoplasmic reticulum transmembrane protein stromal interaction molecule 1 (STIM1) with the MT plus end-binding protein 1 (EB1) promotes its association with the p150Glued subunit of the dynein activator complex dynactin and the distinct location of EEs and LEs/LYs. The peripheral distribution of EEs requires their p150Glued-mediated simultaneous engagement with dynein and SxIP motif-containing KIFC1, via HOOK1 and HOOK3 adaptors, respectively. In sum, we provide evidence that distinct minus end-directed MT motor systems drive the differential transport and subcellular distribution of EEs and LEs in mammalian cells., (© 2020 The Authors. Published under the terms of the CC BY 4.0 license.)

Seven bifidobacterial strains were isolated from the faeces of two adult males of the two-toed sloth ( Choloepus didactylus ) housed in Parco Natura Viva, in Italy. Comparative sequence analysis of 16S rRNA and of five housekeeping ( hsp 60, rpo B, clp C, dna J, dna G) genes revealed that these strains were classified into two clusters. On the basis of 16S rRNA gene sequence similarity, the type strain of Bifidobacterium catenulatum subsp. kashiwanohense DSM 21854 T (95.4 %) was the closest neighbour to strain in Cluster I (BRDM 6 T ), whereas the type strain of Bifidobacterium dentium DSM 20436 T (values were in the range of 98‒99.8 %) was the closest neighbour to the other six strains in Cluster II. The average nucleotide identity (ANI) values of BRDM 6 T and of strains in Cluster II with the closely related type strains were 76.0 and 98.9 % (mean value) respectively. Therefore, genotyping based on the genome sequence of the strain BRDM 6 T combined with phenotypic analyses clearly revealed that the strain BRDM 6 T represents a novel species for which the names Bifidobacterium choloepi sp. nov. (BRDM 6 T =NBRC 114053 T =BCRC 81222 T ) is proposed.

A microbiome is defined as a complex collection of microorganisms and their genetic material. Studies regarding gut microbiomes of different animals have provided ecological and evolutionary information showing a strong link between health and disease. Very few studies have compared the gut microbiota of animals housed under controlled conditions and those in wild habitats. Little research has been performed on the reptile gut microbiota, and what studies do exist are mainly focused on carnivorous reptiles. The aim of this study was first to describe the overall microbiota structure of Aldabra giant tortoises ( Aldabrachelys gigantea ) and, second, to compare the microbiota of tortoises living under natural conditions and tortoises living in controlled environments, such as zoological and botanical parks, in Italy and in the Seychelles. Seventeen fecal samples were collected from giant tortoises located on Curieuse Island (CI, n = 8), at the Botanical Garden (BG, n = 3) in Mahé (Seychelles Islands) and at Parco Natura Viva-Garda Zoological Park (PNV, n = 6) in Verona (Italy). The V3-V4 region of the 16S rRNA gene was amplified in order to characterize the gut microbiota profile. Overall, the major phyla identified were Bacteroidetes 42%, Firmicutes 32%, and Spirochaetes 9%. A higher microbial diversity (alpha indices) was observed for the BG samples as compared to the PNV samples (Shannon: 5.39 vs. 4.43; InvSimpson: 80.7 vs. 25; Chao1: 584 vs. 377 p < 0.05). The results in the present study showed a significant difference in beta diversity between the samples from CI, BG, and PNV ( p = 0.001), suggesting a different bacterial fecal profile of giant tortoises at the different habitats. This study provided novel insights into the effects of different environmental conditions on the gut microbial communities of giant tortoises. In particular, differences were reported regarding the bacterial gut community structure between tortoises in natural and in controlled environments. These results could help to improve the management of giant tortoises under human care, thus enhancing ex-situ conservation efforts far from the species geographic range., (Copyright © 2020 Sandri, Correa, Spiezio, Trevisi, Luise, Modesto, Remy, Muzungaile, Checcucci, Zaborra and Mattarelli.)

Three bifidobacterial Gram-stain-positive, non-spore forming and fructose-6-phosphate phosphoketolase-positive strains, SMA1 T , SMB2 and SMA15 T were isolated from the faeces of two adult males of the squirrel monkey (Saimiri sciureus). On the basis of 16S rRNA gene sequence similarities, the type strain of Bifidobacterium primatium DSM 100687 T (99.3%; similarity) was the closest neighbour to strains SMA1 T and SMB2, whereas the type strain of Bifidobacterium stellenboschense DSM 23968 T (96.5%) was the closest neighbour to strain SMA15 T . The average nucleotide identity (ANI) values of SMA1 T and SAM15 T with the closely related type strains were 93.7% and 88.1%, respectively. The in silico DNA‒DNA hybridization values with the closest neighbours were 53.1% and 36.9%, respectively. GC contents of strains SMA1 T and SMA15 T were 63.6 and 66.4 mol%, respectively. Based on the phylogenetic, genotypic and phenotypic data obtained, the strains SMA1 T and SMA15 T clearly represent two novel taxa within the genus Bifidobacterium for which the names Bifidobacterium saimiriisciurei sp. nov. (type strain SMA1 T  = BCRC 81223 T  = NBRC 114049 T  = DSM 106020 T ) and Bifidobacterium platyrrhinorum sp. nov. (type strain SMA15 T  = BCRC 81224 T  = NBRC 114051 T  = DSM 106029 T ) are proposed., (Copyright © 2020 Elsevier GmbH. All rights reserved.)

Fifteen bifidobacterial strains were obtained from faeces of Rousettus aegyptiacus; after grouping them by RAPD PCR only eight were selected and characterized. Analysis of 16S rRNA and of five housekeeping (hsp60, rpoB, clpC, dnaJ, dna G) genes revealed that these eight strains were classified into five clusters: Cluster I (RST 8 and RST 16 T ), Cluster II (RST 9 T and RST 27), Cluster III (RST 7 and RST 11), Cluster IV (RST 19), Cluster V (RST 17) were closest to Bifidobacterium avesanii DSM 100685 T (96.3%), Bifidobacterium callitrichos DSM 23973 T (99.2% and 99.7%), Bifidobacterium tissieri DSM 100201 T (99.7 and 99.2%), Bifidobacterium reuteri DSM 23975 T (98.9%) and Bifidobacterium myosotis DSM 100196 T (99.3%), respectively. Strains in Cluster I and strain RST 9 in Cluster II could not be placed within any recognized species while the other ones were identified as known species. The average nucleotide identity values between two novel strains, RST 16 T and RST 9 T and their closest relatives were lower than 79% and 89%, respectively. In silico DNA-DNA hybridization values for those closest relatives were 32.5 and 42.1%, respectively. Phenotypic and genotypic tests demonstrated that strains in Cluster I and RST 9 T in Cluster II represent two novel species for which the names Bifidobacterium vespertilionis sp. nov. (RST 16 T =BCRC 81138 T =NBRC 113380 T =DSM 106025 T ; RST 8=BCRC 81135=NBRC 113377) and Bifidobacterium rousetti sp. nov. (RST 9 T =BCRC 81136 T =NBRC 113378 T =DSM 106027 T ) are proposed., (Copyright © 2019 Elsevier GmbH. All rights reserved.)

A novel irregularly shaped and slightly curved rod bacterial strain, GLDI4/2 T , showing activity of fructose 6-phosphate phosphoketolase was isolated from a faecal sample of an adult gelada baboon ( Theropithecus gelada ). Phylogenetic analyses based on 16S rRNA genes as well as multilocus sequences (representing fusA , gyrB and xfp genes) and the core genome revealed that GLDI4/2 T exhibited phylogenetic relatedness to Alloscardovia omnicolens DSM 21503 T and to Alloscardovia macacae DSM 24762 T . Comparative analysis of 16S rRNA gene sequences confirmed the phylogenetic results showing the highest gene sequence identity with strain A. omnicolens DSM 21503 T (96.0 %). Activities of α- and β-gluco(galacto)sidases were detected in strain GLDI4/2 T , which is characteristic for almost all members of the family Bifidobacteriaceae . Compared to other Alloscardovia species its DNA G+C content (43.8 mol%) was very low. Phylogenetic studies and the evaluation of phenotypic characteristics, including the results of biochemical, physiological and chemotaxonomic analyses, confirmed the novel species status for strain GLDI4/2 T , for which the name Alloscardoviatheropitheci sp. nov. is proposed. The type strain is GLDI4/2 T (=DSM 106019 T =JCM 32430 T ).

Four novel Gram-stain-positive, non spore forming and fructose-6-phosphate phosphoketolase-positive strains were isolated from the faeces of a cotton top tamarin (Saguinus oedipus) and an emperor tamarin (Saguinus imperator). Phylogenetic analyses based on 16S rRNA revealed that bifidobacterial strains TRE 1 T exhibit close phylogenetic relatedness to Bifidobacterium catulorum DSM 103154 (96.0%) and Bifidobacterium tissieri DSM 100201 (96.0%); TRE D T and TRE H T were closely related to Bifidobacterium longum subsp. longum ATCC 15708 T with similarity values of 97.4% and 97.5%, respectively; TRI 7 T was closely related to Bifidobacterium tissieri DSM 100201 (96.0%). The Average Nucleotide Identity (ANI) and in silico DDH (isDDH) analysis with closest neighbour supported an independent phylogenetic position of all strains with values ranged from 74 to 85% for ANI and from 24 to 28% for isDDH. DNA base composition of the four strains was in the range of 58.3-63.5mol% G+C. Based on the phylogenetic, genotypic and phenotypic data, the strains TRE 1 T , TRE D T , TRE H T and TRI 7 T clearly represent four novel taxa within the genus Bifidobacterium for which the names Bifidobacterium primatium sp. nov. (type strain TRE 1 T =DSM 100687 T =JCM 30945 T ), Bifidobacterium scaligerum sp. nov. (type strain TRE D T =DSM 103140 T =JCM 31792 T ), Bifidobacterium felsineum sp. nov. (type strain TRE H T =DSM 103139 T =JCM 31789 T ) and Bifidobacterium simiarum sp. nov. (type strain TRI 7 T =DSM 103153 T =JCM 31793) are proposed., (Copyright © 2018 Elsevier GmbH. All rights reserved.)

The Northern bald ibis ( Geronticus eremita ) (NBI) is one of the most threatened birds in the world. Intense conservation efforts have been undertaken and several research projects on the species are being done in Morocco and in Europe. Observing animal behaviour has been proved to be an efficient and non-invasive technique to assess the animal welfare, with the performance of a wide array of natural behaviours being one of the mostly used indicators of good mental and physical well-being. The aim of this study was to investigate the behaviour of a flock of 14 zoo-living NBI of different ages. The study focused on the variety of species-specific individual and social behaviours, in the light of reintroduction of the study juveniles in the wild. Per subject, 20 10-min. sessions were done. A continuous focal animal sampling method was used to collect individual and social behaviours. Behavioural data have been compared between adults and juveniles. Moreover, a Behavioural Variety Index (BVI) has been proposed and calculated based on previous literature describing natural ibis behaviours. The BVI might help in the evaluation of the variety of behaviours performed by each individual and the monitoring of the diversity of the behavioural repertoire of zoo animals. Our results showed that the birds performed species-specific behaviours and no abnormal behaviour was reported. Moreover, the BVI highlighted a good behavioural variety as each bird performed approximately 78% of the natural behaviours described in the Northern bald ibis and in close relative species. Our findings seem to suggest the presence of qualitative and quantitative similarities between the behavioural repertoires of the study ibises and those described in wild conspecifics, suggesting a good welfare of the colony. Finally, the BVI proposed in the current study seems to be a useful and practical tool to test behavioural diversity in zoo animals., Competing Interests: Caterina Spiezio is employed by Parco Natura Viva as head of the Research and Conservation Department. Camillo Sandri is employed by Parco Natura Viva as general curator and veterinarian. Barbara Regaiolli is employed by Parco Natura Viva as researcher in the Research and Conservation Department.

Same-sex pair bonds have been documented in several animal species and they are widespread in birds. However, little is known about the evolutionary origin and the adaptive value of such behaviour. The aim of this study was to investigate the parental behaviour of four zoo female greater flamingos involved in two breeding pairs, housed in a flock at Parco Natura Viva, Italy. Further, the behaviour of the study females was compared with that of male and female flamingos in heterosexual pairs described in a previous published work on this same flock. For each pair, the behaviour of both birds during the incubation period was recorded and twenty 10-minute sessions were run within the incubation period. A continuous focal animal sampling method was used to collect data on location (on the nest or not on the nest) and the parental care behaviour (e.g.: agonistic behaviours toward disturbing conspecifics, egg-care, nest-building, self-comfort behaviour, sleeping) of the two pairs. Data of the current study females were compared with those of females and males involved in heterosexual pairs of this same flock. Results showed that within each pair the egg-layer female stayed away from the nest more than the other female. In addition, the female that did not lay an egg was more involved in agonistic behaviour compared to other females, particularly when in specific locations. In heterosexual pairs, male flamingos were more involved in the incubation and in nest protection. Moreover, no significant differences in the time spent on the nest and away from the nest between the heterosexual male and the non-layer females of same-sex pairs were found. The same findings were reported when comparing heterosexual females and the egg-layer females of the same-sex pairs. Therefore, our findings suggest that in greater flamingos the behaviour of the female-female pairs seems to be equivalent to that of male-female bonds. Such research provides more insight into flamingo social behaviour, and their reproductive cycle, and provides information on why pair bonds may form and how these affect the wider breeding behaviour of the flock., Competing Interests: The authors declare there are no competing interests.

Due to the great number of greater flamingos in captivity and their long life span, studying their behavior and welfare might be useful to improve the husbandry and breeding of this species in zoos. This study aimed to investigate factors affecting the breeding activity of captive greater flamingos (Phoenicopterus roseus) from 2012 to 2016. To estimate reproductive success, the number of pairs, eggs laid, and hatchlings were recorded. In addition, information on age, egg-laying history of the partners, and pair composition was collected. An increase in the number of pairs, eggs, and hatchlings was reported over the years. For each breeding season, there were pairs who laid more than once, especially in 2014 and 2015. Approximately 50% of pairs were monogamous between consecutive years; however, the percentage dropped gradually when comparing nonconsecutive years. Senescence and previous experience seemed to affect the reproductive success of the study flamingos. In conclusion, different factors can influence the reproduction of greater flamingos in zoos. All these factors are related to nonhuman animal welfare and need to be considered in developing and improving management practices.

Bifidobacterium sp. strains TRE 1, TRE D, TRE H, and TRI 7 were isolated from two tamarins housed in Parco Natura Viva, Garda Zoological Park S.r.l. (Bussolengo, Verona, Italy). These strains belong to four putative novel species of the genus Bifidobacterium The genome sizes were 2.7 Mb for TRE 1, 2.7 Mb for TRE D, 2.4 Mb for TRE H, and 2.7 Mb for TRI 7. The average GC contents were 63.18% for TRE 1, 58.27% for TRE D, 57.11% for TRE H, and 63.79% for TRI 7., (Copyright © 2018 Puglisi et al.)

Three Gram-stain-positive, non-spore-forming, microaerophilic and fructose-6-phosphate phosphoketolase positive strains were isolated from a faecal sample of an adult subject of the emperor tamarin (Saguinus imperator). Given that the isolates revealed identical BOX PCR profiles, strain TRI 5 T was selected as a representative and characterized further. Comparative analysis of 16S rRNA gene sequence similarity revealed that strain TRI 5 T was closely related to Bifidobacterium saguini DSM 23967 T (96.4 %) and to Bifidobacterium longum subsp. longum ATCC 15708 (96.2 %). Multilocus sequence analyses of five housekeeping genes showed the close phylogenetic relatedness of this strain to Bifidobacterium breve DSM 20213 T (hsp60 94.1 %), Bifidobacterium saguini DSM 23967 T (clpC 91 %), Bifidobacterium avesanii DSM 100685 T (dnaG 80.3 %), Bifidobacterium longumsubsp. infantis ATCC 15697 T (dnaJ 85.3 %) and Bifidobacterium longumsubsp. longum ATCC 15708 (rpoB 93 %), respectively. The peptidoglycan type was A3β, with an interpeptide bridge comprising l-Orn (Lys) - l-Ser - l-Ala - l-Thr - l-Ala. The DNA G+C content of strain TRI 5 T was 60.9 mol%. Based on the data provided, strain TRI 5 T represents a novel species of the genus Bifidobacterium for which the name Bifidobacteriumcallitrichidarum sp. nov. is proposed. The type strain is TRI 5 T (=DSM 103152 T =JCM 31790 T ).

In the last years, studies on captive greater flamingos have increased. Research on zoo animals is important to improve the knowledge on these species and to improve their ex-situ and in-situ conservation. The aim of the present study was to investigate the parental behaviour of a captive colony of greater flamingo hosted at Parco Natura Viva, an Italian zoological garden, to improve the knowledge on this species in zoos. In particular, the present study investigated and compared the parental care of females and males in 35 breeding pairs of greater flamingos. For each pair, we collected durations of parental care behaviour of both females and males, recording their position in relation to the nest (near the nest, on the nest, away from the nest) and individual and social behaviours performed. First, both partners were involved in parental care and displayed species-specific behaviours reported in the wild. The main results were that males spent more time than females on the nest ( P = 0.010) and near it ( P = 0.0001) and were more aggressive toward other flamingos than females, both when sitting on the nest ( P = 0.003) and when near the nest ( P = 0.0003). Therefore, male flamingos seem to be more involved in incubation duties and nest protection than females. This kind of research is important not only to expand the knowledge on bird species such as flamingos, but also to improve their husbandry and breeding in controlled environment. Indeed, understanding animal behaviour allows us to gain insights into their individual and social needs, addressing potential animal welfare issues., Competing Interests: The authors declare there are no competing interests.

Antimicrobial resistance (AMR) in a wide range of infectious agents is a growing public health threat. Birds of prey are considered indicators of the presence of AMR bacteria in their ecosystem because of their predatory behaviour. Only few data are reported in the literature on AMR strains isolated from animals housed in zoos and none about AMR in raptors housed in zoological gardens. This study investigated the antibiotic sensitivity profile of the isolates obtained from the conjunctival and cloacal bacterial flora of 14 healthy birds of prey, 6 Accipitriformes , 3 Falconiformes and 5 Strigiformes , housed in an Italian zoological garden. Staphylococcus spp. was isolated from 50% of the conjunctival swabs, with S. xylosus as the most common species. From cloacal swabs, Escherichia coli was cultured from all animals, while Klebsiella spp. and Proteus spp. were isolated from a smaller number of birds. Worthy of note is the isolation of Escherichia fergusonii and Serratia odorifera , rarely isolated from raptors. Staphylococci were also isolated. All the isolates were multidrug resistant (MDR). To the author's knowledge, this is the first report regarding the presence of MDR strains within raptors housed in a zoological garden. Since resistance genes can be transferred to other pathogenic bacteria, this represents a potential hazard for the emergence of new MDR pathogens. In conclusion, the obtained data could be useful for ex-situ conservation programmes aimed to preserve the health of the endangered species housed in a zoo.

Forty-five microorganisms were isolated on bifidobacteria selective medium from one faecal sample of an adult subject of the cotton-top tamarin (Saguinus oedipus L.). All isolates were Gram-positive, catalase-negative, anaerobic, fructose-6-phosphate phosphoketolase positive, and asporogenous rod-shaped bacteria. In this study, only eight out of the forty-five strains were characterized more deeply, whereas the others are still currently under investigation. They were grouped by BOX-PCR into three clusters: Cluster I (TRE 17(T), TRE 7, TRE 26, TRE 32, TRE 33, TRE I), Cluster II (TRE C(T)), and Cluster III (TRE M(T)). Comparative analysis of 16S rRNA gene sequences confirmed the results from the cluster analysis and revealed relatively low level similarities to each other (mean value 95%) and to members of the genus Bifidobacterium. All eight isolates showed the highest level of 16S rRNA gene sequence similarities with Bifidobacterium scardovii DSM 13734(T) (mean value 96.6%). Multilocus sequence analysis (MLSA) of five housekeeping genes (hsp60, rpoB, clpC, dnaJ and dnaG) supported their independent phylogenetic position to each other and to related species of Bifidobacterium. The G+C contents were 63.2%, 65.9% and 63.0% for Cluster I, Cluster II and Cluster III, respectively. Peptidoglycan types were A3α l-Lys-l-Thr-l-Ala, A4β l-Orn (Lys)-d-Ser-d-Glu and A3β l-Orn-l-Ser-l-Ala in Clusters I, II and III, respectively. Based on the data provided, each cluster represented a novel taxon for which the names Bifidobacterium aerophilum sp. nov. (TRE 17(T)=DSM 100689=JCM 30941; TRE 26=DSM 100690=JCM 30942), Bifidobacterium avesanii sp. nov. (TRE C(T)=DSM 100685=JCM 30943) and Bifidobacterium ramosum sp. nov. (TRE M=DSM 100688=JCM 30944) are proposed., (Copyright © 2016 Elsevier GmbH. All rights reserved.)

Forty-three strains of bifidobacteria were isolated from the faeces of two adult black lemurs, Eulemur macaco . Thirty-four were identified as Bifidobacterium lemurum , recently described in Lemur catta . The nine remaining isolates were Gram-positive-staining, non-spore-forming, fructose-6-phosphate phosphoketolase-positive, microaerophilic, irregular rod-shaped bacteria that often presented Y- or V-shaped cells. Typing techniques revealed that these isolates were nearly identical, and strain LMM&#95;E3 T was chosen as a representative and characterized further. Phylogenetic analysis based on 16S rRNA gene sequences clustered this isolate inside the genus Bifidobacterium and showed the highest levels of sequence similarity with B. lemurum DSM 28807 T (99.3 %), with Bifidobacterium pullorum LMG 21816 T and Bifidobacterium longum subsp. infantis ATCC 15697 T (96.4 and 96.3 %, respectively) as the next most similar strains. The hsp60 gene sequence of strain LMM&#95;E3 T showed the highest similarity to that of Bifidobacterium stellenboschense DSM 23968 T (93.3 %), and 91.0 % similarity to that of the type strain of B. lemurum . DNA-DNA reassociation with the closest neighbour B. lemurum DSM 28807 T was found to be 65.4 %. The DNA G+C content was 62.3 mol%. Strain LMM&#95;E3 T showed a peptidoglycan structure that has not been detected in bifidobacteria so far: A3α l-Lys-l-Ser-l-Thr-l-Ala. Based on the phylogenetic, genotypic and phenotypic data, strain LMM&#95;E3 T represents a novel species within the genus Bifidobacterium , for which the name Bifidobacterium eulemuris sp. nov. is proposed; the type strain is LMM&#95;E3 T ( = DSM 100216 T  = JCM 30801 T ).

Four Gram-positive-staining, microaerophilic, non-spore-forming, fructose-6-phosphate phosphoketolase-positive bacterial strains were isolated from a faecal sample of a 5-year-old ring-tailed lemur (Lemur catta). The strains showed a peculiar morphology, resembling a small coiled snake, a ring shape, or forming a little 'Y' shape. The isolated strains appeared identical, and LMC 13T was chosen as a representative strain and characterized further. Strain LMC 13T showed an A3β peptidoglycan type, similar to that found in Bifidobacterium longum. The DNA base composition was 57.2 mol% G+C. Almost-complete 16S rRNA, hsp60, rpoB, dnaJ, dnaG, purF, clpC and rpoC gene sequences were obtained, and phylogenetic relationships were determined. Comparative analysis of 16S rRNA gene sequences showed that strain LMC 13T showed the highest similarity to B. longum subsp. suis ATCC 27533T (96.65 %) and Bifidobacterium saguini DSM 23967T (96.64 %). Strain LMC 13T was located in an actinobacterial cluster and was more closely related to the genus Bifidobacteriumthan to other genera in the Bifidobacteriaceae. On the basis of these results, strain LMC 13T represents a novel species within the genus Bifidobacterium, for which the name Bifidobacterium lemurum sp. nov. is proposed; the type strain is LMC 13T ( = DSM 28807T = JCM 30168T).