Physiological and genomic characterization of Lactiplantibacillus plantarum isolated from Indri indri in Madagascar.

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.

Do not disturb: Visitors' effect on the behavior and welfare of female and male big cats in zoos.

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.

The "right" side of sleeping: laterality in resting behaviour of Aldabra giant tortoises (Aldabrachelys gigantea).

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.

Color preference and manual laterality in the emperor tamarin (Saguinus imperator).

The current research focuses on color preference between red and green stimuli and manual laterality in the emperor tamarin (Saguinus imperator). Trichromacy in primates has been related to a foraging advantage allowing frugivore primates to distinguish ripe from unripe fruits as well to socio-sexual communication, as trichromats would be advantaged in recognizing social and sexual signals. As warm colors can affect the emotive state of the subjects, leading to the activation of one hemisphere over the other (e.g., right hemisphere), this could lead to behavioral lateralization. Thus, studying of hand preference may be relevant when testing color preference. Nine adult zoo emperor tamarins were involved and the study aimed to investigate the preference between red, green, and white cones as well as manual laterality. Tamarins were provided with pairs of red-green, red-white, and green-white combinations of cones. Ten 30-min sessions per combination were carried out and data on the interaction with one of the two cones of each apparatus were collected to assess subjects' color preference. We also recorded the hand used by each subject during the interaction with cones of different colors and the position of the apparatus in respect to the tamarin. We found no preferences for colored versus white cones. Similarly, we reported no group-level preferences within different color combinations, whereas individual-level preferences were found when considering all choices. Finally, we found that red cones elicited a left-hand preference, suggesting a right-hemisphere involvement in the presence of red cones. Although we do not have genetic data on trichromat and dichromat females, the tendency to use the left hand when interacting with red stimuli provides further evidence that warm colors can influence the emotive state of the perceiver, affecting their manual lateralization.

I Like the Way You Eat It: Lemur (Indri indri) Gut Mycobiome and Geophagy.

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.

Paw preference in wolves (Canis lupus): A preliminary study using manipulative tasks.

Behavioural and brain lateralization is widespread among non-human vertebrates. Motor lateralization has been investigated in the domestic dog, revealing that "pawedness" in this species seems to be sex and task related; however, few if any studies considered this asymmetry in wolves (Canis lupus). The aim of this study was to investigate the paw preference of seven wolves housed at Parco Natura Viva - Italy, during the interaction with food-related (FD) and olfactory (OLF) environmental enrichment devices. Eleven sessions were done (22 session in total) per condition (FD and OLF), and data about enrichment manipulation were collected. Most of the wolves manipulated the enrichment devices using one paw rather than both paws. At the individual level, all subjects were lateralized in paw use, six were right pawed, one was left pawed (the alpha male) regardless of the enrichment condition. The fact that one paw rather than both was frequently involved in manipulation could indicate a practical advantage for each individual in being lateralized in paw use. Despite the small sample size, our results provide interesting insights about lateralization in wolves, deserving further investigations. More studies are needed considering factors such as temperament, social rank and task complexity on canid motor lateralization.

Behavioural Laterality in two species of flamingos: greater flamingos and Chilean flamingos.

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.

Effects of Rearing on the Behaviour of Zoo-Housed Chimpanzees (Pan troglodytes).

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.

Bifidobacteria in two-toed sloths (Choloepus didactylus): phylogenetic characterization of the novel taxon Bifidobacterium choloepi sp. nov.

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 (hsp60, rpoB, clpC, dnaJ, dnaG) 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 21854T (95.4 %) was the closest neighbour to strain in Cluster I (BRDM 6T), whereas the type strain of Bifidobacterium dentium DSM 20436T (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 6T 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 6T combined with phenotypic analyses clearly revealed that the strain BRDM 6T represents a novel species for which the names Bifidobacterium choloepi sp. nov. (BRDM 6T=NBRC 114053T=BCRC 81222T) is proposed.

Gum feeder as environmental enrichment for zoo marmosets and tamarins.

Tamarins and marmosets are small-bodied social callitrichines. Wild callitrichines feed on exudates, such as sap and gum; particularly, marmosets are mainly gummivores, while tamarins consume gums only occasionally and opportunistically. Zoo marmosets and tamarins are usually provided with gum arabic as an alternative to the exudates normally found in the wild. The aim of this study was to evaluate the effects of a gum feeder on the behavior and well-being of four zoo-managed callitrichines. We studied four cotton-top tamarins (Saguinus oedipus), four red-handed tamarins (S. midas), two pygmy marmosets (Cebuella pygmaea), and three Geoffroy's marmosets (Callithrix geoffroyi) housed at Parco Natura Viva (Italy). We conducted the study over two different periods, a baseline (control, without the gum feeder) and then a gum feeder (when the gum feeder was provided) period. We used continuous focal animal sampling to collect behavioral data, including durations of social and individual behaviors. We collected 240 min of observations per period per study subject, with a total of 3,120 min for all the subjects in the same period and of 6,240 min in both periods. We analyzed data by using nonparametric statistical tests. First, we found that the gum feeder promoted species-specific behaviors, such as exploration, and diminished self-directed behaviors, suggesting an enriching effect on tamarin and marmoset behavior. Moreover, in red-handed tamarins, the provision of the gum feeder reduced the performance of self-directed and abnormal behavior, specifically coprophagy. These results confirm that gum feeders are effective foraging enrichment tools for zoo marmosets and tamarins.

Alloscardovia theropitheci sp. nov., isolated from the faeces of gelada baboon, the 'bleeding heart' monkey (Theropithecus gelada).

A novel irregularly shaped and slightly curved rod bacterial strain, GLDI4/2T, 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 GLDI4/2T exhibited phylogenetic relatedness to Alloscardovia omnicolens DSM 21503T and to Alloscardovia macacae DSM 24762T. Comparative analysis of 16S rRNA gene sequences confirmed the phylogenetic results showing the highest gene sequence identity with strain A. omnicolens DSM 21503T (96.0 %). Activities of α- and ß-gluco(galacto)sidases were detected in strain GLDI4/2T, 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 GLDI4/2T, for which the name Alloscardoviatheropitheci sp. nov. is proposed. The type strain is GLDI4/2T (=DSM 106019T=JCM 32430T).

Bifidobacterium callitrichidarum sp. nov. from the faeces of the emperor tamarin (Saguinus imperator).

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 5T was selected as a representative and characterized further. Comparative analysis of 16S rRNA gene sequence similarity revealed that strain TRI 5T was closely related to Bifidobacterium saguini DSM 23967T (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 20213T (hsp60 94.1 %), Bifidobacterium saguini DSM 23967T (clpC 91 %), Bifidobacterium avesanii DSM 100685T (dnaG 80.3 %), Bifidobacterium longumsubsp. infantis ATCC 15697T (dnaJ 85.3 %) and Bifidobacterium longumsubsp. longum ATCC 15708 (rpoB 93 %), respectively. The peptidoglycan type was A3ß, with an interpeptide bridge comprising l-Orn (Lys) - l-Ser - l-Ala - l-Thr - l-Ala. The DNA G+C content of strain TRI 5T was 60.9 mol%. Based on the data provided, strain TRI 5T represents a novel species of the genus Bifidobacterium for which the name Bifidobacteriumcallitrichidarum sp. nov. is proposed. The type strain is TRI 5T (=DSM 103152T=JCM 31790T).

Hand preference on unimanual and bimanual tasks in Barbary macaques (Macaca sylvanus).

The presence of group-level handedness in non-human primates remains controversial, as different studies have produced inconsistent results. Bimanual coordinated tasks have been found to elicit more pronounced hand preferences than simple unimanual tasks. The aim of this study was to examine manual lateralization in a group of 15 Barbary macaques during unimanual and bimanual tasks. In the unimanual task, data on simple food reaching in a foraging context were collected. During the bimanual task, macaques had to use one hand to hold a tube-shaped apparatus while reaching with the other hand to retrieve the food inside it. Data on the hand use to retrieve food were collected. First, no significant group-level hand preference was found for the unimanual task. However, a significant right hand bias was evident for the bimanual task. At the individual-level, approximately 47% and 67% of the subjects had a significant hand preference in the unimanual and bimanual task respectively. The strength of the hand preference was greater in the bimanual than in the unimanual task. Findings of this study add Macaca sylvanus to the other species showing a right hand preference for coordinated bimanual tasks. Moreover, our results add to the growing body of evidence that bimanual tasks are more suitable and valid measures to investigate handedness in non-human primates.

Yolk testosterone affects growth and promotes individual-level consistency in behavioral lateralization of yellow-legged gull chicks.

Behavioral lateralization is common in animals and may be expressed at the individual- and at the population-level. The ontogenetic processes that control lateralization, however, are largely unknown. Well-established sex-dependence in androgen physiology and sex-dependent variation in lateralization have led to the hypothesis that testosterone (T) has organizational effects on lateralization. The effects of T exposure in early life on lateralization can be efficiently investigated by manipulating T levels in the cleidoic eggs of birds, because the embryo is isolated from maternal and sibling physiological interference, but this approach has been adopted very rarely. In the yellow-legged gull (Larus michahellis) we increased yolk T concentration within the physiological limits and tested the effects on the direction of lateralization in two functionally fundamental behaviors (begging for parental care and escape to cover) of molecularly sexed hatchlings. We also speculated that T may intervene in regulating consistency, rather than direction of lateralization, and therefore tested if T affected the 'repeatability' of lateral preference in consecutive behavioral trials. T treatment had no effect on the direction of lateralization, but enhanced the consistency of lateral preference in escape responses. Sex did not predict lateralization. Neither behavior was lateralized at the population-level. We therefore showed for the first time in any species an effect of egg T on consistency in lateralization. The implications of the effect of T for the evolution of trade-offs in maternal allocation of egg hormones, and the evolutionary interpretations of findings from our studies on lateralization among unmanipulated birds are discussed.

Bifidobacterium eulemuris sp. nov., isolated from faeces of black lemurs (Eulemur macaco).

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_E3T 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 28807T (99.3 %), with Bifidobacterium pullorum LMG 21816T and Bifidobacterium longum subsp. infantis ATCC 15697T (96.4 and 96.3 %, respectively) as the next most similar strains. The hsp60 gene sequence of strain LMM_E3T showed the highest similarity to that of Bifidobacterium stellenboschense DSM 23968T (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 28807T was found to be 65.4 %. The DNA G+C content was 62.3 mol%. Strain LMM_E3T 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_E3T represents a novel species within the genus Bifidobacterium, for which the name Bifidobacterium eulemuris sp. nov. is proposed; the type strain is LMM_E3T ( = DSM 100216T = JCM 30801T).

Hand preference on unimanual and bimanual tasks in strepsirrhines: The case of the ring-tailed lemur (Lemur catta).

Assessing manual lateralization in non-human primates could be an optimal way to understand the adaptive value of this asymmetry in humans. Though many studies have investigated hand preferences in Old and New World monkeys and apes, fewer studies have considered manual lateralization in strepsirrhines, especially in experimental tasks. This study investigated hand preferences for a unimanual and a bimanual task of 17 captive ring-tailed lemurs (Lemur catta), housed at Parco Natura Viva (VR), Italy. The effect of age on handedness has been also investigated. The lemurs were tested on a unimanual task, in which subjects were restricted to using one hand to retrieve the food inside an apparatus, and on a bimanual task, in which lemurs had to use one hand to keep the apparatus door open while reaching with the other hand to retrieve the food inside it. At the population-level, our results revealed an asymmetrical hand use distribution, in particular a bias toward a right hand preference for food reaching in both the unimanual and the bimanual tasks. Furthermore, at the individual-level, the bimanual task seems to elicit a greater hand preference than the unimanual task. Results of this study underline the importance of experimental tasks in determining hand preference in strepsirrhines. Furthermore, as bimanual tasks elicited a stronger degree of lateralization, they appear to be more suited to investigate manual laterality. Finally, findings from this study highlight the presence of a right hand preference in ring-tailed lemurs, shedding new light on the evolution of human right handedness. Am. J. Primatol. 78:851-860, 2016. © 2016 Wiley Periodicals, Inc.

Manual lateralization in macaques: handedness, target laterality and task complexity.

Non-human primates represent models to understand the evolution of handedness in humans. Despite several researches have been investigating non-human primates handedness, few studies examined the relationship between target position, hand preference and task complexity. This study aimed at investigating macaque handedness in relation to target laterality and tastiness, as well as task complexity. Seven pig-tailed macaques (Macaca nemestrina) were involved in three different "two alternative choice" tests: one low-level task and two high-level tasks (HLTs). During the first and the third tests macaques could select a preferred food and a non-preferred food, whereas by modifying the design of the second test, macaques were presented with no-difference alternative per trial. Furthermore, a simple-reaching test was administered to assess hand preference in a social context. Macaques showed hand preference at individual level both in simple and complex tasks, but not in the simple-reaching test. Moreover, target position seemed to affect hand preference in retrieving an object in the low-level task, but not in the HLT. Additionally, individual hand preference seemed to be affected from the tastiness of the item to be retrieved. The results suggest that both target laterality and individual motivation might influence hand preference of macaques, especially in simple tasks.

Bifidobacterium lemurum sp. nov., from faeces of the ring-tailed lemur (Lemur catta).

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 A3ß 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).

Can an Enrichment Programme with Novel Manipulative and Scent Stimuli Change the Behaviour of Zoo-Housed European Wildcats? A Case Study.

Objects and semiochemicals may be used as enrichment in zoos. Domestic cats release Fraction 3 of Facial Pheromone (F3) by rubbing the muzzle to convey relational and territorial information. We aimed to evaluate whether and how the introduction of novel objects and scent stimuli could change the behaviour of one group (N = 5 subjects) of adult European wildcats (Felis silvestris silvestris) hosted at Parco Natura Viva-Garda Zoological Park, Italy. We assessed the behavioural changes following the introduction of novel objects (blocks and rags) and scent (synthetic F3) via observations over four experimental conditions (baseline, rags, F3 rags, blocks) using continuous focal animal sampling. Our results showed that no behavioural differences were found between the different conditions and the baseline, except for the condition with blocks when significantly less exploration was observed. Between conditions, wildcats performed significantly less individual explorative, affiliative, and agonistic behaviours, but more individual inactivity, when exposed to rags after F3 administration. Our findings suggest that the enrichment programme did not substantively affect the behaviour of the zoo-housed wildcats. However, the behavioural differences recorded between conditions suggest that, while novel objects introduced as visual stimuli (blocks) do not affect the wildcat behaviour, novel manipulative objects (rags) might impact their behaviour. Moreover, the changes in affiliative and agonistic behaviours displayed during the condition with exposure to rags sprayed with F3 suggest that such semiochemical could play an appeasement role within this study group.

Faecal Microbiota Characterisation of Potamochoerus porcus Living in a Controlled Environment.

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 106 CFU/g in faecal samples of all the individuals, with the only exception for the juvenile subject, showing 107 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.