Mechanical loading activates the YAP/TAZ pathway and chemokine expression in the MLO-Y4 osteocyte-like cell line.

Osteocytes are mechanosensitive cells that control bone remodeling in response to mechanical loading. To date, specific signaling pathways modulated by mechanical loading in osteocytes are not well understood. Yes associated protein (YAP) and transcriptional coactivator with PDZ-binding motif (TAZ), the main effectors of the Hippo pathway, are reported to play a role in mechanotransduction and during osteoblastogenesis. Here, we hypothesized that YAP/TAZ signaling mediates osteocyte mechanosensing to target genes of the bone remodeling process. We aimed to investigate the contribution of YAP/TAZ in modulating the gene expression in an osteocyte-like cell line MLO-Y4. We developed a 3D osteocyte compression culture model from an MLO-Y4 osteocyte cell line embedded in concentrated collagen hydrogel. 3D-mechanical loading led to the increased expression of mechanosensitive genes and a subset of chemokines, including M-csf, Cxcl1, Cxcl2, Cxcl3, Cxcl9, and Cxcl10. The transcription regulators YAP and TAZ translocated to the nucleus and upregulated their target genes and proteins. RNAseq analysis revealed that YAP/TAZ knockdown mediated the regulation of several genes including osteocyte dendrite formation. Use of YAP/TAZ knockdown partially blunted the increase in M-csf and Cxcl3 levels in response to MLO-Y4 compression. These findings demonstrate that YAP/TAZ signaling is required for osteocyte-like cell mechano-transduction, regulates the gene expression profiles and controls chemokine expression.

Hand preference in unimanual and bimanual coordinated tasks in wild western lowland gorillas (Gorilla gorilla gorilla) feeding on African ginger (Zingiberaceae).

OBJECTIVES: Bimanual coordinated behaviors are critical for detecting robust individual hand preference in nonhuman primates but are particularly challenging to observe in the wild. This study focuses on spontaneous feeding behavior on African ginger (Aframomum sp. and Renealmia sp.), which involves a unimanual task (reaching and pulling out a ginger stem) and a bimanual coordinated task (extracting pith from a ginger stem) by wild western lowland gorillas. MATERIALS AND METHODS: Study subjects were 21 gorillas in the Moukalaba-Doudou National Park, Gabon. We examined whether they exhibit significant hand preference at the individual and group levels for both tasks. RESULTS: Sixteen gorillas showed significant hand preference in the unimanual task, whereas all 21 individuals showed significant hand preference in the bimanual coordinated task. Hand preference was significantly stronger in the bimanual coordinated task than in the unimanual task. It is noteworthy that gorillas showed a significant right-hand preference at the group level for the bimanual task (roughly 70% of the subjects). DISCUSSION: This study confirmed that bimanual coordinated tasks are more sensitive in detecting hand preferences in nonhuman primates. In addition to the bimanual nature of the task, the precision grip for processing and the importance of African ginger as a food resource might influence the expression of hand preference. Evidence of a group-level right-hand preference may support the "postural origins theory." Because all wild African great apes feed on the pith of African ginger, comparing this task and its hand preferences can contribute toward a better understanding of the evolution of handedness in Hominidae.

Hydrogel films and coatings by swelling-induced gelation.

Hydrogel films used as membranes or coatings are essential components of devices interfaced with biological systems. Their design is greatly challenged by the need to find mild synthesis and processing conditions that preserve their biocompatibility and the integrity of encapsulated compounds. Here, we report an approach to produce hydrogel films spontaneously in aqueous polymer solutions. This method uses the solvent depletion created at the surface of swelling polymer substrates to induce the gelation of a thin layer of polymer solution. Using a biocompatible polymer that self-assembles at high concentration [poly(vinyl alcohol)], hydrogel films were produced within minutes to hours with thicknesses ranging from tens to hundreds of micrometers. A simple model and numerical simulations of mass transport during swelling capture the experiments and predict how film growth depends on the solution composition, substrate geometry, and swelling properties. The versatility of the approach was verified with a variety of swelling substrates and hydrogel-forming solutions. We also demonstrate the potential of this technique by incorporating other solutes such as inorganic particles to fabricate ceramic-hydrogel coatings for bone anchoring and cells to fabricate cell-laden membranes for cell culture or tissue engineering.

Estimation of chimpanzee community size and genetic diversity in Kahuzi-Biega National Park, Democratic Republic of Congo.

A small chimpanzee habitat in the montane forest of Kahuzi-Biega National Park, Democratic Republic of Congo, is connected with the lowland forest of this park through a corridor, which is affected by human encroachment. To assess the conservation status of the chimpanzee population in this small habitat, we estimated the size of the community and evaluated its genetic diversity by using 279 fecal samples collected in the montane forest of Kahuzi. Using autosomal microsatellite (or short tandem repeat, STR) loci, we identified 32 individuals, comprising 19 females and 13 males. Samples from 24 individuals were collected at least twice and a genetic mark-recapture analysis estimated that the community size was 36 (range: 32-42). Data on nest site sharing confirmed that all the samples belonged to the same community. Nest site sharing information may be useful in population studies of unhabituated chimpanzees. The genetic structure and diversity of the 32 genotyped individuals was assessed using Y-chromosomal short tandem repeat (Y-STR) loci and mitochondrial D-loops. One dominant Y-STR haplotype was found, whereas there was no dominant haplotype in the mitochondrial region, reflecting a female-biased dispersal pattern, which is typical of chimpanzees. The genetic diversity for three markers in Kahuzi chimpanzees was comparable to that in other eastern chimpanzee populations. A relatively high heterozygosity and negative inbreeding coefficient (FIS ) for STR loci suggests that the study community belongs to an outbreeding chimpanzee population. These findings suggest that individuals of the study community may have reproductive contact with other chimpanzee individuals from neighboring communities in Kahuzi-Biega National Park, at least in the recent past. Am. J. Primatol. 77:1015-1025, 2015. © 2015 Wiley Periodicals, Inc.

Death of a one-armed blackback male due to severe injuries in a group of wild western lowland gorillas (Gorilla gorilla gorilla) in Moukalaba-Doudou National Park, Gabon.

The survival of limb-disabled primates in the wild has been widely reported. Nevertheless, their ultimate fate is little documented. It is important to understand the influence of limb disability on primate survival from a conservation perspective, as many African great apes suffer from limb injuries caused by entrapment in snares. Here, we report the death of a one-armed blackback male in a large one-male group of wild western lowland gorillas. The subject was a blackback male (14 years old) named Dodo, who lost his right forearm in August 2008. On 8 December 2019, Dodo was found to have suffered serious bleeding injuries to the front of his body, including large lacerations and puncture wounds. On 14 December his corpse was found in the forest. We provide evidence to suggest that his injuries were more likely caused by intraspecific aggression, though a predatory attack by a leopard could not be completely ruled out. His one-armed disability could have made him more vulnerable to attack from either a gorilla or leopard and led to his fatal injury. This report shows that a gorilla who had previously overcome a disability in one arm in childhood may die prematurely, in part, owing to this disability in young adulthood.

Local people enhance our understanding of Afrotropical frugivory networks.

Afrotropical forests are undergoing massive change caused by defaunation, i.e., the human-induced decline of animal species,1 most of which are frugivorous species.1,2,3 Frugivores' depletion and their functional disappearance are expected to cascade on tree dispersal and forest structure via interaction networks,4,5,6,7 as the majority of tree species depend on frugivores for their dispersal.8 However, frugivory networks remain largely unknown, especially in Afrotropical areas,9,10,11 which considerably limits our ability to predict changes in forest dynamics and structures using network analysis.12,13,14,15 While the academic workforce may be inadequate to fill this knowledge gap before it is too late, local ecological knowledge appears as a valuable source of ecological information and could significantly contribute to our understanding of such crucial interactions for tropical forests.16,17,18,19,20,21 To investigate potential synergies between local ecological knowledge and academic knowledge,20,21 we compiled frugivory interactions linking 286 trees to 100 frugivore species from the academic literature and local ecological knowledge coming from interviews of Gabonese forest-dependent people. Here, we showed that local ecological knowledge on frugivory interactions was substantial and original, with 39% of these interactions unknown by science. We demonstrated that combining academic and local ecological knowledge affects the functional relationship linking frugivore body mass to seed size, as well as the network structure. Our results highlight the benefits of bridging knowledge systems between academics and local communities for a better understanding of the functioning and response to perturbations of Afrotropical forests.

Male genetic structure and paternity in western lowland gorillas (Gorilla gorilla gorilla).

The male dispersal patterns of western lowland gorillas (WLGs, Gorilla gorilla gorilla) are not well understood. To determine whether most silverbacks stay close to their relatives, we analyzed autosomal and Y-chromosomal microsatellites (STRs) in wild WLGs at Moukalaba, Gabon. We obtained STR genotypes for 38 individuals, including eight silverbacks and 12 adult females in an approximately 40 km(2) area. Among them, 20 individuals were members of one identified group (Group Gentil; GG), including one silverback and six adult females. The silverback sired all 13 of the offspring in GG and no Y-STR polymorphism within GG was found, as expected in a one-male group structure. Over all silverbacks sampled, Y-STR diversity was high considering the limited sampling area, and silverbacks with similar Y-STR haplotypes were not always located in nearby areas. Although the misclassification rate of kinship estimates in this study was not negligible, there were no kin dyads among all silverbacks sampled. These results suggest that silverbacks born in the same group do not stay close to each other after maturation. The Y-STR diversity in this study was similar to that of a previous study conducted in an area that was approximately 150 times larger than our study area. Similarity of WLG Y-STR diversity between studies at different sampling scales suggests that male gene flow may not be geographically limited. These results suggest that WLG males normally disperse from their natal areas after maturation, at least, in Moukalaba.

Assessment of landscape-scale distribution of sympatric great apes in African rainforests: concurrent use of nest and camera-trap surveys.

Information on the distribution and abundance of sympatric great apes (Pan troglodytes troglodytes and Gorilla gorilla gorilla) are important for effective conservation and management. Although much research has been done to improve the precision of nest-surveys, trade-offs between data-reliability and research-efficiency have not been solved. In this study, we used different approaches to assess the landscape-scale distribution patterns of great apes. We conducted a conventional nest survey and a camera-trap survey concurrently, and checked the consistency of the estimates. We divided the study area (ca. 500 km²), containing various types of vegetation and topography, into thirty 16-km² grids (4 km × 4 km) and performed both methods along 2-km transects centered in each grid. We determined the nest creator species according to the definitions by Tutin & Fernandez [Tutin & Fernandez, 1984, Am J Primatol 6:313-336] and estimated nest-site densities of each species by using the conventional distance-sampling approach. We calculated the mean capture rate of 3 camera traps left for 3 months at each grid as the abundance index. Our analyses showed that both methods provided roughly consistent results for the distribution patterns of the species; chimpanzee groups (parties) were more abundant in the montane forest, and gorilla groups were relatively homogeneously distributed across vegetation types. The line-transect survey also showed that the number of nests per nest site did not vary among vegetation types for either species. These spatial patterns seemed to reflect the ecological and sociological features of each species. Although the consistent results may be largely dependent on site-specific conditions (e.g., high density of each species, distinct distribution pattern between the two species), conventional nest-surveys and a subsequent check of their consistency with independent estimates may be a reasonable approach to obtain certain information on the species distribution patterns. Further analytical improvement is necessary for camera-traps to be considered a stand-alone method.

Pharyngeal Communities and Antimicrobial Resistance in Pangolins in Gabon.

Wildlife can be a reservoir and source of zoonotic pathogens for humans. For instance, pangolins were considered one of the potential animal reservoirs of SARS-CoV-2. The aim of this study was to assess the prevalence of antimicrobial-resistant species (e.g., extended-spectrum ß-lactamase [ESBL]-producing Enterobacterales) and Staphylococcus aureus-related complex and to describe the bacterial community in wild Gabonese pangolins. The pharyngeal colonization of pangolins sold in Gabon (n = 89, 2021 to 2022) was analyzed using culture media selective for ESBL-producing Enterobacterales, S. aureus-related complex, Gram-positive bacteria and nonfermenters. Phylogenetic analyses of ESBL-producing Enterobacterales was done using core-genome multilocus sequence typing (cgMLST) and compared with publicly available genomes. Patterns of cooccurring species were detected by network analysis. Of the 439 bacterial isolates, the majority of species belonged to the genus Pseudomonas (n = 170), followed by Stenotrophomonas (n = 113) and Achromobacter (n = 37). Three Klebsiella pneumoniae isolates and one Escherichia coli isolate were ESBL-producers, which clustered with human isolates from Nigeria (MLST sequence type 1788 [ST1788]) and Gabon (ST38), respectively. Network analysis revealed a frequent cooccurrence of Stenotrophomonas maltophilia with Pseudomonas putida and Pseudomonas aeruginosa. In conclusion, pangolins can be colonized with human-related ESBL-producing K. pneumoniae and E. coli. Unlike in other African wildlife, S. aureus-related complex was not detected in pangolins. IMPORTANCE There is an ongoing debate if pangolins are a relevant reservoir for viruses such as SARS-CoV-2. Here, we wanted to know if African pangolins are colonized with bacteria that are relevant for human health. A wildlife reservoir of antimicrobial resistance would be of medical relevance in regions were consumption of so-called bushmeat is common. In 89 pangolins, we found three ESBL-producing Klebsiella pneumoniae strains and one ESBL-producing Escherichia coli strains, which were closely related to isolates from humans in Africa. This points toward either a transmission between pangolins and humans or a common source from which both humans and pangolins became colonized.

Netrin-1 Secreted by Human Osteoarthritic Articular Chondrocytes Promotes Angiogenesis in Vitro.

OBJECTIVE: Netrin-1 expression in articular cartilage is correlated with osteoarthritic changes. We aimed to investigate the contribution of Netrin-1 secreted by human osteoarthritic articular chondrocytes to angiogenesis process in vitro. DESIGN: Human articular chondrocytes were extracted from non-osteoarthritic (n = 10) and osteoarthritic (n = 22) joints obtained from surgical specimens and incubated for 24 hours. Medium conditioned by non-osteoarthritic and osteoarthritic articular chondrocytes were collected. Human umbilical vein endothelial cells (HUVEC) were treated with control and conditioned medium and assessed using assays for cell adherence, migration, and tube formation. Netrin-1 expression and secretion was compared between non-osteoarthritic and osteoarthritic chondrocytes by qPCR, Western blot, and ELISA. The role of chondrocyte-secreted Netrin-1 on HUVEC functions was assessed by immunological neutralization using an anti-Netrin-1 monoclonal antibody. RESULTS: As compared with medium conditioned by non-osteoarthritic chondrocytes, medium conditioned by osteoarthritic chondrocytes permitted tube formation by HUVEC. Both non-osteoarthritic and osteoarthritic chondrocytes expressed Netrin-1 at the RNA and protein levels. At the RNA level, Netrin-1 expression did not differ between non-osteoarthritic and osteoarthritic chondrocytes. At the protein level, Netrin-1 appeared as a full protein of 64 kDa in non-osteoarthritic chondrocytes and as two cleaved proteins of 55 kDa and 64 kDa in osteoarthritic chondrocytes. Immunological neutralization of endogenous Netrin-1 reduced the pro-angiogenic and pro-inflammatory transcriptional profile of HUVEC treated with the medium conditioned by osteoarthritic chondrocytes, as well as their capacities to form tubes. CONCLUSIONS: Medium conditioned by osteoarthritic chondrocytes permits tube formation by HUVEC in vitro. This permissive effect is mediated by Netrin-1.

Seasonality in daily movement patterns of mandrills revealed by combining direct tracking and camera traps.

Movement is a fundamental characteristic of animals, but challenging to measure noninvasively. Noninvasive methods for measuring travel have different weaknesses, so multiple techniques need to be applied multiple techniques for reliable inferences. We used two methods, direct tracking and camera trapping, to examine the variation in time and seasonal differences in movement rates of mandrills (Mandrillus sphinx), an elusive primate that lives in large groups in central Africa. In a 400-km2 rainforest area in Moukalaba-Doudou National Park, Gabon, we tracked unidentified groups 46 times from 2009 to 2013. We systematically placed 157 terrestrial camera traps in the same area from 2012 to 2014 and recorded groups 309 times. Generalized additive mixed models (GAMMs) of the tracking data indicated that the group travel speed varied with time and season. In the fruiting season, the movement rate fluctuated with time in a bimodal pattern, whereas in the nonfruiting season, it increased monotonously with time. The predicted day range was longer in the fruiting season (6.98 km) than in the nonfruiting season (6.06 km). These seasonal differences suggest responses to changes in food resources and temperature. Camera-trap detection rates showed similar temporal and seasonal patterns to the tracking data, allowing us to generalize our findings to the population level. Moreover, cameras never detected mandrills at night, and we observed that they slept high in trees and hardly moved until the next morning, all suggesting their strict avoidance of nighttime movement. This study demonstrated the significance of the multiple-method approach in drawing robust conclusions on temporal patterns of animal movement.

Le mouvement est une caractéristique fondamentale des animaux, mais il est difficile de le mesurer de manière non invasive. Les méthodes non invasives de mesure des déplacements présentent des faiblesses différentes, de sorte que nous devons appliquer plusieurs techniques à un même sujet pour obtenir des déductions fiables. Nous avons utilisé deux méthodes, le suivi direct et le piégeage photographique, pour examiner les variations temporelles et la différence saisonnière des taux de déplacement des mandrills (Mandrillus sphinx), un primate insaisissable vivant en grands groupes. Dans une zone de forêt tropicale humide de 400 km2 du parc national de Moukalaba-Doudou au Gabon, nous avons suivi des groupes non identifiés 46 fois entre 2009 et 2013. Dans la même zone, nous avons systématiquement placé 157 pièges photographiques terrestres de 2012 à 2014 et enregistré des groupes 309 fois. Les modèles mixtes additifs généralisés (GAMM) des données de suivi ont indiqué que la vitesse de déplacement des groupes variait en fonction du temps et des saison. Pendant la saison de fructification, la vitesse de déplacement fluctuait dans le temps selon un schéma bimodal, tandis que pendant la saison de non-fructification, elle augmentait de façon monotone avec le temps. Les distances journalières prédites étaient plus longues pendant la saison de fructification (6,98 km) que pendant la saison de non-fructification (6,06 km). Ces différences saisonnières suggèrent des réponses aux changements de ressources alimentaires et de température. Les taux de détection des pièges photographiques ont montré des tendances temporelles et saisonnières similaires à celles des données de suivi, ce qui nous permet de généraliser nos conclusions au niveau de la population. De plus, les caméras n'ont jamais détecté les mandrills la nuit, et nous avons observé qu'ils dormaient haut dans les arbres et ne bougeaient pratiquement pas jusqu'au lendemain matin, ce qui suggère qu'ils évitent strictement de se déplacer la nuit. Cette étude a démontré l'importance de l'approche à méthodes multiples pour tirer des conclusions solides sur les modes temporels de déplacement des animaux.

Phylogenetic Groups, Pathotypes and Antimicrobial Resistance of Escherichia coli Isolated from Western Lowland Gorilla Faeces (Gorilla gorilla gorilla) of Moukalaba-Doudou National Park (MDNP).

(1) Background: Terrestrial mammals in protected areas have been identified as a potential source of antimicrobial-resistant bacteria. Studies on antimicrobial resistance in gorillas have already been conducted. Thus, this study aimed to describe the phylogroups, pathotypes and prevalence of antimicrobial resistance of Escherichia coli isolated from western lowland gorilla's faeces living in MDNP. (2) Materials and Methods: Ninety-six faecal samples were collected from western lowland gorillas (Gorilla gorilla gorilla) during daily monitoring in the MDNP. Sixty-four E. coli isolates were obtained and screened for phylogenetic and pathotype group genes by polymerase chain reaction (PCR) after DNA extraction. In addition, antimicrobial susceptibility was determined by the disk diffusion method on Mueller Hinton agar. (3) Results: Sixty-four (64%) isolates of E. coli were obtained from samples. A high level of resistance to the beta-lactam family, a moderate rate for fluoroquinolone and a low rate for aminoglycoside was obtained. All E. coli isolates were positive in phylogroup PCR with a predominance of A (69% ± 11.36%), followed by B2 (20% ± 19.89%) and B1 (10% ± 8.90%) and low prevalence for D (1% ± 3.04%). In addition, twenty E. coli isolates (31%) were positive for pathotype PCR, such as EPEC (85% ± 10.82%) and EPEC/EHEC (15% ± 5.18%) that were obtained in this study. The majority of these MDR E. coli (DECs) belonged to phylogenetic group A, followed by MDR E. coli (DECs) belonging to group B2. (4) Conclusion: This study is the first description of MDR E. coli (DECs) assigned to phylogroup A in western lowland gorillas from the MDNP in Gabon. Thus, wild gorillas in MDNP could be considered as asymptomatic carriers of potential pathogenic MDR E. coli (DECs) that may present a potential risk to human health.

Aging Cartilage in Wild-Type Mice: An Observational Study.

OBJECTIVE: To describe the spontaneous evolution of age-related changes affecting knee joint articular cartilage, walking speed and a serum biomarker of cartilage remodeling in C57BL/6-JRj wild-type male mice. DESIGN: Histological changes were assessed by the Osteoarthritis Research Society International (OARSI) score (0=normal, 6=vertical clefts/erosion to the calcified cartilage extending >75% of the articular surface) in newborn, 1-week- and 1-, 3-, 6-, 9- and 12-month-old C57BL/6-JRj wild-type male mice, walking speed by the Locotronic system, and serum C-terminal telopeptide of type II collagen (CTX-II) content by ELISA in 1-, 3-, 6-, and 9-month-old C57BL/6-JRj wild-type male mice. RESULTS: Mean (SD) OARSI score significantly increased from 0.2 (0.3) to 1.3 (0.6) (p=0.03) between 1 and 3 months of age and from 1.3 (0.6) to 3.3 (0.6) (p=0.04) between 3 and 6 months of age. Mean walking speed was stable between 1 and 6 months of age but significantly decreased from 11.4 (1.8) to 3.2 (0.8) cm.s-1 (p=0.03) between 6 and 9 months of age. Serum CTX-II content was maximal at 1 month of age, then decreased from 12.2 (8.5) to 2.4 (8.4) pg/ml (p=0.02) between 1 and 3 months of age, remaining low and stable thereafter. CONCLUSIONS: C57BL/6-JRj wild-type male mice showed continuously increasing osteoarthritic changes but delayed decreasing walking speed with age. These variations were maximal between 3 and 9 months of age. Maximal serum CTX-II content preceded these changes.

Mechanical Stretch of High Magnitude Provokes Axonal Injury, Elongation of Paranodal Junctions, and Signaling Alterations in Oligodendrocytes.

Increasing findings suggest that demyelination may play an important role in the pathophysiology of brain injury, but the exact mechanisms underlying such damage are not well known. Mechanical tensile strain of brain tissue occurs during traumatic brain injury. Several studies have investigated the cellular and molecular events following a static tensile strain of physiological magnitude on individual cells such as oligodendrocytes. However, the pathobiological impact of high-magnitude mechanical strain on oligodendrocytes and myelinated fibers remains under investigated. In this study, we reported that an applied mechanical tensile strain of 30% on mouse organotypic culture of cerebellar slices induced axonal injury and elongation of paranodal junctions, two hallmarks of brain trauma. It was also able to activate MAPK-ERK1/2 signaling, a stretch-induced responsive pathway. The same tensile strain applied to mouse oligodendrocytes in primary culture induced a profound damage to cell morphology, partial cell loss, and a decrease of myelin protein expression. The lower tensile strain of 20% also caused cell loss and the remaining oligodendrocytes appeared retracted with decreased myelin protein expression. Finally, high-magnitude tensile strain applied to 158N oligodendroglial cells altered myelin protein expression, dampened MAPK-ERK1/2 and MAPK-p38 signaling, and enhanced the production of reactive oxygen species. The latter was accompanied by increased protein oxidation and an alteration of anti-oxidant defense that was strain magnitude-dependent. In conclusion, mechanical stretch of high magnitude provokes axonal injury with significant alterations in oligodendrocyte biology that could initiate demyelination.

Improving the standards for gut microbiome analysis of fecal samples: insights from the field biology of Japanese macaques on Yakushima Island.

Fecal DNA-based 16S ribosomal RNA (rRNA) gene sequencing using next-generation sequencers allows us to understand the dynamic gut microbiome adaptation of animals to their specific habitats. Conventional techniques of fecal microbiome analysis have been developed within the broad contexts defined by human biology; hence, many of these techniques are not immediately applicable to wild nonhuman primates. In order to establish a standard experimental protocol for the analysis of the gut microbiomes of wild animals, we selected the Japanese macaques (Macaca fuscata yakui) on Yakushima Island. We tested different protocols for each stage of fecal sample processing: storage, DNA extraction, and choice of the sequencing region in the bacterial 16S rRNA gene. We also analyzed the gut microbiome of captive Japanese macaques as the control. The comparison of samples obtained from identical macaques but subjected to different protocols showed that the tested storage methods (RNAlater and lysis buffer) produced effectively the same composition of bacterial operational taxonomic units (OTUs) as the standard frozen storage method, although the relative abundance of each OTU was quantitatively affected. Taxonomic assignment of the detected bacterial groups was also significantly affected by the region being sequenced, indicating that sequencing regions and the corresponding polymerase chain reaction (PCR) primer pairs for the 16S rRNA gene should be carefully selected. This study improves the current standard methods for microbiome analysis in wild nonhuman primates. Japanese macaques were shown to be a suitable model for understanding microbiome adaptation to various environments.

Identification and molecular characterization of novel primate bocaparvoviruses from wild western lowland gorillas of Moukalaba-Doudou National Park, Gabon.

Bocaparvoviruses have been studied extensively owing to their ability to cause respiratory illness or gastroenteritis in humans. Some bocaparvoviruses have been detected in non-human primates (gorillas and chimpanzees), but the diversity and evolution of these viruses are not fully understood. In this study, we collected 107 fecal samples from wild western lowland gorillas in Moukalaba-Doudou National Park in Gabon to investigate the presence of bocaparvoviruses. Using a combination of pan-bocaparvovirus PCR and individual identification by microsatellite genotyping, we found that two samples from two apparently healthy infant gorillas were positive for bocaparvovirus. Sequencing and phylogenetic analyses revealed that the two gorilla bocaparvovirus strains are nearly identical and are closely related to viruses in the species Primate bocaparvovirus 2 (with 86.0% nucleotide identity to a human bocavirus 2 isolate). To our knowledge, this is the first report showing the presence of a non-human primate bocaparovirus within Primate bocaparvovirus 2. Our findings provide novel insights into the diversity and evolution of bocaparvoviruses and highlight the importance of surveying these viruses for the safe management of gorilla-based ecotourism.

Interaction of the alpha-helical H6 peptide from the pro-apoptotic protein tBid with cardiolipin.

BH3 interacting domain death agonist (Bid), a pro-apoptotic member of the Bcl-2 family of proteins, is activated through cleavage by caspase-8. The active C-terminal fragment of Bid (tBid) translocates to the mitochondria where it interacts with cardiolipins at contact sites and induces the release of cytochrome c by a mechanism that is not yet fully understood. It has been shown that the alpha-helices alphaH6 and alphaH7 (which create the hairpin-forming domain of tBid) mediate the insertion of Bid into mitochondrial membranes and are essential for the cytochrome c-releasing activity. In the present study, we focused on the interaction between the alphaH6 and the mitochondrial membrane. By the use of single-cell electropermeabilization associated with flow cytometric analysis of intact cells, we demonstrated that H6 is able to induce cell death when used in the micromolar range. We also studied the interactions of the alphaH6 with artificial monolayers. We showed that the presence of negatively charged cardiolipins greatly enhances the insertion of alphaH6 into the phospholipid monolayer. The modification of two charged amino acid residues in alphaH6 abolished its insertion and also its in vivo effects. Furthermore, the negative values of the excess areas of mixing indicate that attractive interactions between cardiolipins and alphaH6 occur in the mixed monolayers. Fluorescence microscopy observations revealed that alphaH6 significantly disrupts cardiolipin packing and stabilizes the fluid lipid phase. These results suggest that cardiolipins at the contact sites between the two mitochondrial membranes could mediate the binding of tBid via alphaH6.