Blue light directly modulates the quorum network in the human pathogen Acinetobacter baumannii.

Quorum sensing modulates bacterial collective behaviors including biofilm formation, motility and virulence in the important human pathogen Acinetobacter baumannii. Disruption of quorum sensing has emerged as a promising strategy with important therapeutic potential. In this work, we show that light modulates the production of acyl-homoserine lactones (AHLs), which were produced in higher levels in the dark than under blue light at environmental temperatures, a response that depends on the AHL synthase, AbaI, and on the photoreceptor BlsA. BlsA interacts with the transcriptional regulator AbaR in the dark at environmental temperatures, inducing abaI expression. Under blue light, BlsA does not interact with AbaR, but induces expression of the lactonase aidA and quorum quenching, consistently with lack of motility at this condition. At temperatures found in warm-blooded hosts, the production of AHLs, quorum quenching as well as abaI and aidA expression were also modulated by light, though in this case higher levels of AHLs were detected under blue light than in the dark, in a BlsA-independent manner. Finally, AbaI reduces A. baumannii's ability to kill C. albicans only in the dark both at environmental as well as at temperatures found in warm-blooded hosts. The overall data indicate that light directly modulates quorum network in A. baumannii.

Small to modest impact of social group on the gut microbiome of wild Costa Rican capuchins in a seasonal forest.

The horizontal transmission of pathogenic and beneficial microbes has implications for health and development of socially living animals. Social group is repeatedly implicated as an important predictor of gut microbiome structure among primates, with individuals in neighboring social groups exhibiting distinct microbiomes. Here we examine whether group membership is a predictor of gut microbiome structure and diversity across three groups of white-faced capuchins (Cebus capucinus imitator) inhabiting a seasonal Costa Rican forest. We collected 62 fecal samples from 18 adult females during four sampling bouts. Sampling bouts spanned the dry-to-wet-to-dry seasonal transitions. To investigate gut microbial composition, we sequenced the V4 region of the 16S rRNA gene. We used the DADA2 pipeline to assign amplicon sequence variants and the RDP database to classify taxa. Our findings are: 1) gut microbiomes of capuchins clustered by social group in the late dry season, but this pattern was less evident in other sampling bouts; 2) social group was a significant variable in a PERMANOVA test of beta diversity, but it accounted for less variation than season; 3) social group was not an important predictor of abundance for the ten most abundant microbial taxa in capuchins; 4) when examining log2-fold abundances of microbes between social groups, there were significant differences in some pairwise comparisons. While this is suggestive of group-wide differences, individual variation may have a strong impact and should be assessed in future studies. Overall, we found a minor impact of social group membership on the gut microbiota of wild white-faced capuchins. Future research including home range overlap and resource use, as well as fine-scale investigation of individual variation, will further elucidate patterns of socially structured microbes.

Oral microbiota and their antibiotic susceptibility in free-living monkeys in Goiás State, Brazil: Repercussions for injuries in humans.

INTRODUCTION: Goiás State, which is in the midwest region of Brazil, has several urban forests. This fact, along with the expansion of urban areas within the limits of Forest Conservation Units, increases the contact between humans and wildlife, such as capuchin monkeys. The impulsive behavior of these animals and the scarcity of food cause them to vigorously search for food, leading to direct encounters with Park visitors, which can result in scratches and bites and making them potential disseminators of pathogenic microorganisms. METHODS: Ten specimens of bearded capuchin monkey (Sapajus libidinosus) were captured at the Onofre Quinan Environmental Park in Anápolis, Goiás, Brazil. Samples were collected from the monkeys, and the bacteria and fungi present in the samples were isolated and identified. Then, the identified microorganisms were subjected to antibiotic susceptibility testing. RESULTS: A total of 111 bacteria and 12 fungi were isolated, including two strict anaerobic bacteria of the genus Peptostreptococcus, 109 facultative anaerobic bacteria, and 12 yeasts. Among the facultative bacteria, enterobacteria and Staphylococcus were common. Resistance to tetracycline and ampicillin antibiotics was detected in the enterobacteria, and resistance to tetracycline, erythromycin, and clindamycin was detected in the Staphylococci. The other strains were sensitive to all tested antimicrobials. Cefoxitin showed 100% efficacy in all isolated bacteria. CONCLUSIONS: For bites from capuchin monkeys, we recommend performing complete hygiene and antibiotic therapy, according to medical recommendations. Given the 100% effectiveness of cefoxitin, it should be considered for this type of injury, especially in the study region.

Seasonality of the gut microbiota of free-ranging white-faced capuchins in a tropical dry forest.

Research on the gut microbiota of free-ranging mammals is offering new insights into dietary ecology. However, for free-ranging primates, little information is available for how microbiomes are influenced by ecological variation through time. Primates inhabiting seasonal tropical dry forests undergo seasonally specific decreases in food abundance and water availability, which have been linked to adverse health effects. Throughout the course of a seasonal transition in 2014, we collected fecal samples from three social groups of free-ranging white-faced capuchin monkeys (Cebus capucinus imitator) in Sector Santa Rosa, Área de Conservación Guanacaste, Costa Rica. 16S rRNA sequencing data reveal that unlike other primates, the white-faced capuchin monkey gut is dominated by Bifidobacterium and Streptococcus. Linear mixed effects models indicate that abundances of these genera are associated with fluctuating availability and consumption of fruit and arthropods, whereas beta diversity clusters by rainfall season. Whole shotgun metagenomics revealed that the capuchin gut is dominated by carbohydrate-binding modules associated with digestion of plant polysaccharides and chitin, matching seasonal dietary patterns. We conclude that rainfall and diet are associated with the diversity, composition, and function of the capuchin gut microbiome. Additionally, microbial fluctuations are likely contributing to nutrient uptake and the health of wild primate populations.

Oral microbiota and their antibiotic susceptibility in free-living monkeys in Goiás State, Brazil: Repercussions for injuries in humans

Abstract INTRODUCTION: Goiás State, which is in the midwest region of Brazil, has several urban forests. This fact, along with the expansion of urban areas within the limits of Forest Conservation Units, increases the contact between humans and wildlife, such as capuchin monkeys. The impulsive behavior of these animals and the scarcity of food cause them to vigorously search for food, leading to direct encounters with Park visitors, which can result in scratches and bites and making them potential disseminators of pathogenic microorganisms. METHODS: Ten specimens of bearded capuchin monkey (Sapajus libidinosus) were captured at the Onofre Quinan Environmental Park in Anápolis, Goiás, Brazil. Samples were collected from the monkeys, and the bacteria and fungi present in the samples were isolated and identified. Then, the identified microorganisms were subjected to antibiotic susceptibility testing. RESULTS: A total of 111 bacteria and 12 fungi were isolated, including two strict anaerobic bacteria of the genus Peptostreptococcus, 109 facultative anaerobic bacteria, and 12 yeasts. Among the facultative bacteria, enterobacteria and Staphylococcus were common. Resistance to tetracycline and ampicillin antibiotics was detected in the enterobacteria, and resistance to tetracycline, erythromycin, and clindamycin was detected in the Staphylococci. The other strains were sensitive to all tested antimicrobials. Cefoxitin showed 100% efficacy in all isolated bacteria. CONCLUSIONS: For bites from capuchin monkeys, we recommend performing complete hygiene and antibiotic therapy, according to medical recommendations. Given the 100% effectiveness of cefoxitin, it should be considered for this type of injury, especially in the study region.

The microbial reproductive ecology of white-faced capuchins (Cebus capucinus).

Changes in reproductive status influence energy and nutrient requirements in female primates. The gut microbiota may buffer changes in energy demands, with shifts in community composition increasing the energy production potential of the gut during pregnancy and lactation. In this study, we examine changes in the gut microbiome of wild, female white-faced capuchins (Cebus capucinus) across different reproductive states. Fecal samples (n = 39) were collected from five adult females over the course of a year. Gut microbial community composition was assessed using 16S rRNA gene sequences, and PICRUSt was used to make metagenomic functional predictions. We found a significant relationship between reproductive state and both the structure and predicted function of the gut microbiome, neither of which were associated with host diet. For example, the relative abundance of Firmicutes was significantly lower in lactating females compared with cycling females; the relative abundance of Actinobacteria was significantly higher in pregnant females compared with lactating females, and there was a trend toward higher relative abundances of Proteobacteria in pregnant females compared with cycling females. The results of this study suggest that, in addition to behavioral and dietary adaptions, the gut microbiota may play a role in allowing female primates to meet their changing energetic needs during reproduction. Further studies of the "microbial reproductive ecology" of primates will help advance our understanding of gut microbial contributions to primate energetics.

Influence of fruit and invertebrate consumption on the gut microbiota of wild white-faced capuchins (Cebus capucinus).

OBJECTIVES: Invertebrate consumption is thought to be an integral part of early hominin diets, and many modern human populations regularly consume insects and other arthropods. This study examines the response of gut microbial community structure and function to changes in diet in wild white-faced capuchins (Cebus capucinus), a primate that incorporates a large proportion of invertebrates in its diet. The goal of the study is to better understand the role of both fruit and invertebrate prey consumption on shaping primate gut microbiomes. MATERIALS AND METHODS: Fecal samples (n = 169) and dietary data were collected over 12 months. The V3-V5 region of microbial 16S rRNA genes was amplified and sequenced. The IM-TORNADO pipeline was used to analyze sequences. RESULTS: White-faced capuchin gut bacterial communities were characterized primarily by Firmicutes (41.6%) and Proteobacteria (39.2%). There was a significant relationship between the invertebrate diet composition of individual capuchins and their gut microbiome composition. However, there was no relationship between the fruit diet composition of individual capuchins and their gut microbiome composition, even when examining multiple timescales. DISCUSSION: The results of our study indicate that there is a stronger relationship between gut microbial community structure and invertebrate diet composition than between gut microbial community structure and fruit consumption. As invertebrates and other animal prey play an important role in the diet of many primates, these results give important insight into the role of faunivory in shaping the evolution of host-microbe interactions in primates.

High-Resolution Melting Curve Analysis of the 16S Ribosomal Gene to Detect and Identify Pathogenic and Saprophytic Leptospira Species in Colombian Isolates.

AbstractIt is important to identify the circulating Leptospira agent to enhance the performance of serodiagnostic tests by incorporating specific antigens of native species, develop vaccines that take into account the species/serovars circulating in different regions, and optimize prevention and control strategies. The objectives of this study were to develop a polymerase chain reaction (PCR)-high-resolution melting (HRM) assay for differentiating between species of the genus Leptospira and to verify its usefulness in identifying unknown samples to species level. A set of primers from the initial region of the 16S ribosomal gene was designed to detect and differentiate the 22 species of Leptospira. Eleven reference strains were used as controls to establish the reference species and differential melting curves. Twenty-five Colombian Leptospira isolates were studied to evaluate the usefulness of the PCR-HRM assay in identifying unknown samples to species level. This identification was confirmed by sequencing and phylogenetic analysis of the 16S ribosomal gene. Eleven Leptospira species were successfully identified, except for Leptospira meyeri/Leptospira yanagawae because the sequences were 100% identical. The 25 isolates from humans, animals, and environmental water sources were identified as Leptospira santarosai (twelve), Leptospira interrogans (nine), and L. meyeri/L. yanagawae (four). The species verification was 100% concordant between PCR-HRM and phylogenetic analysis of the 16S ribosomal gene. The PCR-HRM assay designed in this study is a useful tool for identifying Leptospira species from isolates.

Mycoplasma sp. infection in captive Marcgrave's capuchin monkeys (Sapajus flavius).

Three species of hemoplasmas are known to infect monkeys, 'Candidatus Mycoplasma kahanei', 'Candidatus Mycoplasma haemomacaque' and 'Candidatus Mycoplasma aoti'. Marcgrave's capuchin monkey (Sapajus flavius) is a critically endangered neotropical primate species of the Brazilian Atlantic Forest, and thus, sustained efforts are needed to protect the health and well-being of these animals. This study has aimed to screen 12 captive S. flavius from the State of Paraíba, northeastern Brazil, for hemoplasma infection. Packed cell volume (PCV) was measured and a pan-hemoplasma PCR protocol performed. A total of 8/12 (66.6%) monkeys were positive for the 16S rDNA gene of Mycoplasma sp., showing an identity of 99% to Mycoplasma sp. previously detected in Sapajus apella and Sapajus sp. monkeys from the Brazilian Amazon. Further studies should be conducted in order to elucidate whether this potentially novel Mycoplasma sp. is able to cause anemia in Sapajus sp. monkeys from Brazil.

Conjunctival flora, Schirmer's tear test, intraocular pressure, and conjunctival cytology in neotropical primates from Salvador, Brazil.

BACKGROUND: This study aimed to establish reference values for selected ophthalmic diagnostic tests in healthy neotropical primates from Salvador, Brazil. METHODS: A total of 73 intact adults, including Callithrix jacchus (n = 31), Callithrix penicillata (n = 8), Cebus sp. (n = 22), and Cebus xanthosternos (n = 9) were used to evaluate the normal conjunctival bacterial flora. Cebus xanthosternos (n = 12) were used to evaluate tear production with Schirmer's tear test (STT), intraocular pressure (IOP), and conjunctival cytology. RESULTS: For all animals evaluated, Gram-positive bacteria were predominant. Results of the diagnostic tests in Cebus xanthosternos were as follows: STT: 14.92 ± 5.46 mm/minutes, IOP: 19.62 ± 4.57 mmHg, and conjunctival cytology revealed intermediate squamous epithelial cells in great quantities. CONCLUSIONS: These ophthalmic reference values will be particularly useful to diagnose discrete or unusual pathological changes in the neotropical primates eye.

Skin mycoflora of Cebus primates kept in captivity and semicaptivity.

BACKGROUND: This study was performed to determine the fungal mycoflora in healthy tufted capuchins primates (Cebus sp) kept in captivity and semicaptivity to allow a more realistic interpretation on the basis of fungi isolated from their skin. Furthermore, we aimed at evaluating the potential risk of infection to humans by fungi perpetuated in the tegument of monkeys of this genus. METHODS: For the collection of skin material, the carpet method was used, followed by seeding in Sabouraud dextrose agar culture media, Mycosel agar, Dermatophyte Test Medium and Sabouraud agar supplemented with olive oil. RESULTS: Seventeen genera of fungi were detected, being three of them isolated only in the captivity animals (Acremonium - Cephalosporium, Phoma and Trichosporon). The genera of fungi with the higher frequencies were identified in the semicaptivity capuchins (Fusarium, Aspergillus and Penicillium). CONCLUSIONS: Many of the genera of fungi identified are potential pathogens for immune-compromised monkeys and humans.

Frequency of different human mollicutes species in the mucosa of the oropharynx, conjunctiva, and genitalia of free-ranging and captive capuchin monkeys (Cebus spp.).

This study is the first to evaluate the occurrence of several Mollicutes species in Brazilian capuchin monkeys (Cebus spp.). Mollicutes were detected by culture and polymerase chain reaction (PCR) in samples of the oropharyngeal, conjuctiva, and genital mucosae of 58 monkeys. In the oropharynx, Mollicutes in general (generic PCR to the Class), and those of the genus Ureaplasma (genus PCR), were detected in 72.4% and 43.0% of the samples, respectively. The identified species in this site included: Mycoplasma arginini (43.1%), M. salivarium (41.4%), and M. pneumoniae (19.0%). Both Ureaplasma and Mycoplasma are genera of the order Mycoplasmatales. In the preputial/vaginal mucosa, PCR detected Mollicutes in general in 27.58% of the samples, the genus Ureaplasma in 32.7%, the species M. arginini in 8.6%, and Acholeplasma laidlawii of the order Acholeplasmatales in 1.7% In the conjunctiva, Mollicutes in general were detected in 29.3% of the samples, with 1.7% being identified as A. laidlawii. Culturing was difficult due to contamination, but two isolates were successfully obtained. The Mollicutes species of this study provided new insights into these bacteria in Brazilian Cebus. Studies are lacking of the actual risk of Mollicutes infection or the frequency at which primates serve as permanent or temporary reservoirs for Mollicutes. In the present study, the samples were collected from monkeys without clinical signs of infection. The mere presence of Mollicutes, particularly those also found in humans, nevertheless signals a need for studies to evaluate the impact of these microorganisms on the health of non-human primates (NHPs) and the possibility of cross-species transmission between NHPs and humans.

Genetic diversity of oral Fusobacterium nucleatum isolated from patients with different clinical conditions.

The genetic diversity of 23 oral Fusobacterium nucleatum isolated from 15 periodontal patients, eight from seven healthy subjects, nine from nine AIDS patients and two from two Cebus apella monkeys were analyzed. EcoRI restricted the bacterial DNA and 28 ribotypes grouped from A to J groups were obtained. Isolates formed 24 ribotypes which were contained into A, B, C, D, E and F groups, and three reference strains and two clinical isolates of A. actinomycetemcomitans, and E. coli CDC formed four different ribotypes into the G, H, I and J groups. Moreover, from nine F. nucleatum from AIDS patients, six were ribotyped as group C and three as group D. By using ribotyping we distinguished F. nucleatum recovered from different sources. It is possible that isolates from AIDS patients may contain some phenotypic or genotypic factor did not observed in this study.

Genetic diversity of oral Fusobacterium nucleatum isolated from patients with different clinical conditions

Neste estudo foi avaliada a diversidade genética de 23 amostras de Fusobacterium nucleatum isoladas da cavidade bucal de 15 pacientes com doença periodontal, de oito cepas isoladas de sete indivíduos sadios, de nove isoladas de nove pacientes com AIDS e de duas isoladas de dois macacos Cebus apella. Pela ação da enzima EcoRI sobre o DNA bacteriano foram reconhecidos 28 ribotipos agrupados de A a J. Os isolados testados formaram 24 ribotipos os quais foram contidos nos grupos A, B, C, D, E e F, e as três cepas de referência e dois isolados clínicos de A. actinomycetemcomitans e E. coli CDC formaram quatro diferentes ribotipos contidos nos grupos G, H, I e J. Em adição, as nove cepas de F. nucleatum isoladas de pacientes com AIDS, seis pertenciam ao grupo C e três ao grupo D. Usando-se a ribotipagem foi possível distinguir F. nucleatum isolados de diferentes origens.

Plasmid profile in oral Fusobacterium nucleatum from humans and Cebus apella monkeys.

Fusobacterium nucleatum is a strict anaerobe and is indigenous of the human oral cavity. This organism is commonly recovered from different monomicrobial and mixed infections in humans and animals. In this study, the plasmid profile, the plasmid stability and the penicillin-resistance association in oral F. nucleatum isolated from periodontal patients, healthy subjects and Cebus apella monkeys were evaluated. Forty-five F. nucleatum strains from patients, 38 from healthy subjects and seven from C. apella were identified and analyzed. Plasmid extraction was performed in all the isolated strains. These elements were found in 26.7% strains from patients and one strain from C. apella. Strains from healthy subjects did not show any plasmid. Most of strains showed two plasmid bands ranging from 4 to 16 Kb, but digestions with endonucleases showed that they belonged to a single plasmid. The plasmid profile was similar and stable in human and monkey strains. Also, plasmids were classified into three groups according to size. Two strains were positive to beta-lactamase production and no plasmid DNA-hybridization with a beta-lactamase gene probe was observed, suggesting a chromosomal resistance.

Haemagglutination and haemolysis by oral Fusobacterium nucleatum.

Haemagglutination and haemolytic activity of 80 Fusobacterium nucleatum isolates from human and animal origin, on different human blood types was evaluated. All the isolates were able to agglutinate erythrocytes and the most were either alpha-haemolytic or beta-haemolytic. No specificity between haemolysin or haemagglutinin and blood type was observed. Haemagglutination activity was inhibited when D-galactose, D-lactose or D-raffinose were used. Haemagglutination and haemolysis may be important factors in the pathogenesis of human and animal periodontal diseases.

Failure to detect human T-lymphotropic virus antibody in wild-caught New World primates.

We conducted a study to look for a simian counterpart of human T-lymphotropic virus (HTLV) in wild-caught monkeys in the Republic of Panama. Serum specimens were obtained from 102 monkeys (Ateles fusciceps, n = 75; Alouatta villosa, n = 18; and Cebus capucinus, n = 9) captured in Panama's Darien rain forest in 1979-1980. Specimens were screened for HTLV antibody by enzyme-linked immunosorbent assay, and reactive specimens were further tested by Western blot. None of the 102 specimens were seropositive for HTLV. Our findings provide no evidence for an HTLV-like virus in New World primates from Panama, but the sample size was small, and further studies are warranted.

"Paracoccidioidomycosis-infection" survey in Brazilian captive primates (Cebus apella) / Pesquisa da ocorrência da \"Paracoccidioidomicose-infecçäo\" em Primatas brasileiros, mantidos em cativeiro

O objetivo deste trabalho foi estudar alguns aspectos da epidemiologia desta micose, pesquisando a ocorrência de paracoccidioidomicose-infecçäo em animais da Ordem Primata, uma vez que säo filogeneticamente os mais próximos ao homem, único hospedeiro naturalmente susceptível a esta micose sistêmica, no atual estágio de conhecimento. Foram realizados testes de hipersensibilidade do tipo tardio com paracoccidioidina em 33 exemplares de Cebus apella (macaco prego), obtendo-se 33,33//de positividade. Foram também executadas biópsias de reaçöes intradérmicas para exame histológico, testes sorológicos, de fixaçäo de complemento e precipitaçäo em meio líquido no soro destes animais. Os resultados obtidos permitiram verificar a ocorrência de paracoccidioidomicose-infecçäo em primatas näo humanos, sugerindo a possível participaçäo destes animais na epidemiologia da paracoccidioidomicose

In vitro HIV infection of primate lymphocytes.

Peripheral blood mononuclear cells (PBMC) obtained from four different primate species were tested for their respective ability to support the "in vitro" replication of the human immunodeficiency viruses, HIV-1, and HIV-2. PBMC of Cebus apella, patas (Erythrocebus patas), green (cercopithecus aethiops sabaeus) and rhesus monkeys (Macaca mulatta) were infected "in vitro" with either HIV-1 or HIV-2. Cultures were assayed weekly for particle-associated reverse transcriptase activity. Both viruses were found to be cytolytic for all these monkey's PBMC. Low levels of HIV-1 and HIV-2 infection were observed in Cebus cells. However, productive infection was only detected in HIV-2 infected rhesus PBMC. The capacity of HIV-2 to replicate in rhesus cells may provide a useful model for evaluating antiviral drugs and vaccines.

In vitro HIV infection of primate lymphocytes

El propósito de este estudio fue el de evaluar la capacidad de los virus del SIDA (VIH-1 y VIH-2) para multiplicarse en las células mononuclearres de la sangre periférica (CMSP) de cuatro especies de primates. CMSP de Cebus apella, patas (Erythrocebus patas), monos verdes (cercopithecus aethiops sabeus) y rhesus (Macaca mulatta) fueron infectados "in vitro" con VIH-1 y con VIH-2. La multiplicación de estos virus se determinó midiendo la actividad de la enzima retrotranscriptasa en los cultivos infectados. Ambos virus produjeron efectos citipáticos en dichos cultivos. Se observó un bajo nivel de multiplicación de los virus VIH-1 y VIH-2 en las células provenientes de monos Cebus. Sin embargo, el virus VIH-2 se multiplicó eficientemente en CMSP de monos rhesus. La capacidad que posee el virus de la inmunodeficiencia humana tipo 2, (VIH-2) de multiplicarse en estas células, podría ser utilizada para en la evaluación "in vivo" de productos antivirales y de vacunas