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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.

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.

Three bifidobacterial Gram-stain-positive, non-spore forming and fructose-6-phosphate phosphoketolase-positive strains, SMA1T, SMB2 and SMA15T 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 100687T (99.3%; similarity) was the closest neighbour to strains SMA1T and SMB2, whereas the type strain of Bifidobacterium stellenboschense DSM 23968T (96.5%) was the closest neighbour to strain SMA15T. The average nucleotide identity (ANI) values of SMA1T and SAM15T 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 SMA1T and SMA15T were 63.6 and 66.4 mol%, respectively. Based on the phylogenetic, genotypic and phenotypic data obtained, the strains SMA1T and SMA15T clearly represent two novel taxa within the genus Bifidobacterium for which the names Bifidobacterium saimiriisciurei sp. nov. (type strain SMA1T = BCRC 81223T = NBRC 114049T = DSM 106020T) and Bifidobacterium platyrrhinorum sp. nov. (type strain SMA15T = BCRC 81224T = NBRC 114051T = DSM 106029T) are proposed.

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 16T), Cluster II (RST 9T and RST 27), Cluster III (RST 7 and RST 11), Cluster IV (RST 19), Cluster V (RST 17) were closest to Bifidobacterium avesanii DSM 100685T (96.3%), Bifidobacterium callitrichos DSM 23973T (99.2% and 99.7%), Bifidobacterium tissieri DSM 100201T (99.7 and 99.2%), Bifidobacterium reuteri DSM 23975 T (98.9%) and Bifidobacterium myosotis DSM 100196T (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 16T and RST 9T 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 9T in Cluster II represent two novel species for which the names Bifidobacterium vespertilionis sp. nov. (RST 16T = BCRC 81138T = NBRC 113380T = DSM 106025T ; RST 8 = BCRC 81135 = NBRC 113377) and Bifidobacterium rousetti sp. nov. (RST 9T = BCRC 81136T = NBRC 113378T = DSM 106027T) are proposed.

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 Bifidobacterium than 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).

Bifidobacteria represent one of the main bacterial groups occurring in the gut of primates and other animals and are well known for their positive role on host health. Moreover, bifidobacteriaceae are used in coevolution studies since they are maintained within host lineages across hundreds of thousands of host generations. Therefore, the discovery of bifidobacterial diversity in the gastrointestinal tract together with the isolation and characterization of novel bacterial taxa in different hosts is essential. Until now no evidence has been reported about the occurrence of bifidobacteria in bats which are a crucial part of the ecosystem and are ecologically and economically advantageous to humans especially frugivorous ones. This study aimed at the investigation of bifidobacteria in bats belonging to Rousettus aegyptiacus species which were born in Egypt and hosted in Natura Viva Garda Zoological Park (Bussolengo, Verona, Italy). Faecal samples were serially diluted and plated on selective medium MRS supplemented with 0.05 % (w/v) cysteine, 100 mg/l norfloxacin, 2 mg/l cycloheximide, 50 mg/l mupirocin and 1 ml/l acetic acid. Strains were typed by BOX-PCR. Representative isolates of each cluster were identified sequencing 16S rRNA gene and studied for phylogenetical relationship with other Bifidobacteriaceae. Similarities among profiles were calculated by clustering the Pearson‟s correlation matrix using UPGMA algorithm. A total of 13 bifidobacteria were isolated and two of them belonged to new bifidobacterial species according to 16S rRNA gene analysis. The majority of strains revealed a phylogenetic relationship with bifidobacteria isolated from non-human primates. This evolutive link of bifidobacterial microbiotas between bats and non-human primates will be further investigated studying bifidobacteria characterizing Rousettus aegyptiacus bats from Egypt in order to offer new insights about bats origin.