Impact of human created environments in the pathogenic potential and antimicrobial resistance of staphylococci from wild neotropical primates in Brazil.

The non-human primate (NHP) Leontopithecus rosalia is an endangered species native of Brazil and lives in forest fragments with different levels of contact with humans (natural, private and urban). Other NHPs - Callithrix spp. - were introduced by humans and co-exist and interact with the native species in these forests. To evaluate if living in or close to human-modified environments could constitute a risk for L. rosalia, we compared the prevalence, genetic background, antibiotic susceptibility and virulence gene content of staphylococci collected from the native and the introduced species from different forest fragments. We found that presence in human-dominated environments increased the colonization rate of L. rosalia with Mammaliicoccus sciuri (former Staphylococcus sciuri) from 18 % to 85 % (p = 0.0001) and of Callithrix spp with Staphylococcus aureus from 6 % to 100 % (p = 0.0001). According to molecular typing data obtained differences probably resulted from dissemination of these bacterial species from the invader NHP species and from humans. Changes in microbiota were paralleled by an increase in the prevalence of Panton-Valentine Leukocidin gene and in resistance to beta-lactams, macrolides and/or lincosamides as exposure to human environment increased. In particular, erythromycin resistance in S. aureus from Callithrix spp. increased from 0 % to 50 % and resistance rate to at least one antibiotic in coagulase-negative staphylococci species from L. rosalia increased from 13 % to 56 % (p = 0.0003). Our results showed that contact of native animal species with human-created environments increased the content of antimicrobial resistant and pathogenic bacteria on their commensal microbiota, which ultimately can impact on their health. IMPORTANCE: Endangered animal species are vulnerable to environmental alterations and human activities have been repeatedly identified as factors driving drastic changes in the natural landscape. It is extremely important to monitor changes in the environment surrounding protected species, because this could lead to early detection of any potential threats. In this study, we found that the contact of L. rosalia - a protected non-human primate from Brazil - with human environments is related to changes in their commensal microbiota. These included an increase in the number of pathogenic and antibiotic resistant bacteria, which have a higher potential to cause infections that are more difficult to treat. We provided evidence for the harmful impact human contact has on L. rosalia. Also, our results suggest that monitoring of commensal microbiota of protected animal species might be a useful way of sensing the risks of protected species to human exposure.

Metagenomic analysis reveals novel dietary-related viruses in the gut virome of marmosets hybrids (Callithrix jacchus x Callithrix penicillata), Brazil.

Viral metagenomics has contributed enormously to the characterization of a wide range of viruses infecting animals of all phyla in the last decades. Among Neotropical primates, especially those introduced, knowledge about viral diversity remains poorly studied. Therefore, using metagenomics based on virus enrichment, we explored the viral microbiota present in the feces of introduced common marmosets (Callithrix sp.) in three locations from the Silva Jardim region in the State of Rio de Janeiro, Brazil. Fecal samples were collected from nine marmosets, pooled into three sample pools, and sequenced on Illumina MiSeq platform. Sequence reads were analyzed using a viral metagenomic analysis pipeline and two novel insect viruses belonging to the Parvoviridae and Baculoviridae families were identified. The complete genome of a densovirus (Parvoviridae family) of 5,309 nucleotides (nt) was obtained. The NS1 and VP1 proteins share lower than 32% sequence identity with the corresponding proteins of known members of the subfamily Densovirinae. Phylogenetic analysis suggests that this virus represents a new genus, provisionally named Afoambidensovirus due to its discovery in the Brazilian Atlantic Forest. The novel species received the name Afoambidensovirus incertum 1. The complete circular genome of a baculovirus of 107,191 nt was also obtained, showing 60.8% sequence identity with the most closely related member of the Baculoviridae family. Phylogenetic analysis suggests that this virus represents a new species in the Betabaculovirus genus, provisionally named Betabaculovirus incertum 1. In addition, sequences from several families of arthropods in the three pools evaluated were characterized (contigs ranging from 244 to 6,750 nt), corroborating the presence of possible insect hosts with which these new viruses may be associated. Our study expands the knowledge about two viral families known to infect insects, an important component of the marmosets' diet. This identification in hosts' feces samples demonstrates one of the many uses of this type of data and could serve as a basis for future research characterizing viruses in wildlife using noninvasive samples.

Sloth's giant tick (Amblyomma varium) parasitizing free-ranging maned sloth (Bradypus torquatus) in the Atlantic Forest biome, Brazil.

Amblyomma varium is a neotropical tick popularly known as the sloth's giant tick, during the adult stage is found almost exclusively on mammals of the Bradypodidae and Megalonychidae families of the superorder Xenarthra. The maned sloth (Bradypus torquatus) is the only sloth species in danger of extinction in Brazil. This species is arboreal and descends to the ground only to perform defecation behavior. This behavior can be the determining factor for infestation and infection by different parasitic agents, including A. varium. This study reported the occurrence of A. varium found parasitizing free-living maned sloths (B. torquatus) in the Atlantic Forest biome of Northeast and Southeast Brazil. A total of 36 individuals were evaluated and more than 50% of the individuals were parasitized by ticks of the species A. varium. This study confirmed the importance of B. torquatus as a host for adults and nymphs of A. varium and reported the variation in length of the spurs on coxa IV of A. varium males.

O Amblyomma varium é um carrapato neotropical popularmente conhecido como carrapato gigante da preguiça, durante a fase adulta é encontrado quase que exclusivamente em mamíferos das famílias Bradypodidae e Megalonychidae da superordem Xenarthra. A preguiça-de-coleira (Bradypus torquatus) é a única espécie de preguiça ameaçada de extinção no Brasil. Esta espécie é arbórea e desce ao solo apenas para realizar comportamentos de defecação. Esse comportamento pode ser o fator determinante para infestação e infecção por diferentes agentes parasitários, incluindo A. varium. Este estudo relatou a ocorrência de A. varium encontrado parasitando preguiças-de-coleira (B. torquatus) de vida livre no bioma Mata Atlântica do Nordeste e Sudeste do Brasil. Foram avaliados 36 indivíduos e mais de 50% dos indivíduos estavam parasitados por carrapatos da espécie A. varium. Este estudo confirmou a importância de B. torquatus como hospedeiro para adultos e ninfas de A. varium e relatou a variação no comprimento dos espinhos da coxa IV de machos de A. varium.

5meCpG epigenetic marks neighboring a primate-conserved core promoter short tandem repeat indicate X-chromosome inactivation.

X-chromosome inactivation (XCI) is the epigenetic transcriptional silencing of an X-chromosome during the early stages of embryonic development in female eutherian mammals. XCI assures monoallelic expression in each cell and compensation for dosage-sensitive X-linked genes between females (XX) and males (XY). DNA methylation at the carbon-5 position of the cytosine pyrimidine ring in the context of a CpG dinucleotide sequence (5meCpG) in promoter regions is a key epigenetic marker for transcriptional gene silencing. Using computational analysis, we revealed an extragenic tandem GAAA repeat 230-bp from the landmark CpG island of the human X-linked retinitis pigmentosa 2 RP2 promoter whose 5meCpG status correlates with XCI. We used this RP2 onshore tandem GAAA repeat to develop an allele-specific 5meCpG-based PCR assay that is highly concordant with the human androgen receptor (AR) exonic tandem CAG repeat-based standard HUMARA assay in discriminating active (Xa) from inactive (Xi) X-chromosomes. The RP2 onshore tandem GAAA repeat contains neutral features that are lacking in the AR disease-linked tandem CAG repeat, is highly polymorphic (heterozygosity rates approximately 0.8) and shows minimal variation in the Xa/Xi ratio. The combined informativeness of RP2/AR is approximately 0.97, and this assay excels at determining the 5meCpG status of alleles at the Xp (RP2) and Xq (AR) chromosome arms in a single reaction. These findings are relevant and directly translatable to nonhuman primate models of XCI in which the AR CAG-repeat is monomorphic. We conducted the RP2 onshore tandem GAAA repeat assay in the naturally occurring chimeric New World monkey marmoset (Callitrichidae) and found it to be informative. The RP2 onshore tandem GAAA repeat will facilitate studies on the variable phenotypic expression of dominant and recessive X-linked diseases, epigenetic changes in twins, the physiology of aging hematopoiesis, the pathogenesis of age-related hematopoietic malignancies and the clonality of cancers in human and nonhuman primates.

Helminths found in marmosets (Callithrix penicillata and Callithrixjacchus) introduced to the region of occurrence of golden lion tamarins (Leontopithecusrosalia) in Brazil.

The introduction of exotic species can increase the risk of extinction of native species through the introduction of new diseases, predation or resource competition. The marmosets Callithrix penicillata and Callithrix jacchus and hybrids of these two species have been introduced to privately owned forests in the lowland Atlantic forest of the Rio de Janeiro State in Brazil, the region of occurrence of the endangered golden lion tamarin, Leontopithecus rosalia. Because the ecology and biology of the marmosets and tamarins is similar, there is a reasonable risk that the marmosets would transmit pathogens such as endo parasites. The objective of this study was to identify the helminth fauna present in the introduced marmosets through an analysis of fecal samples of wild caught animals, and to evaluate the parasitological profile according to age, sex and geographical location. Eggs belonging to the Acanthocephala and Nematoda were found in the feces. One nematode egg type was identified as being Primasubulura jacchi. The ocurrence of nematodes was higher in males, acanthocephala were found in all age groups and sexes and P. jacchi was found with higher frequency in adult females. The geographic distribution analysis revealed that some of the forests had a higher predominance of parasites. Little is known about the pathology of parasites in free living Neotropical mammals, and this knowledge would be necessary to infer about the risk (form a parasitological standpoint) that the marmoset presence represents for the survival of the endangered golden lion tamarin.

Behavioral and ecological interactions between reintroduced golden lion tamarins ( Leontopithecus rosalia Linnaeus, 1766 ) and introduced marmosets ( Callithrix spp, Linnaeus, 1758 ) in Brazil's Atlantic Coast forest fragments

Callithrix jacchus e spp (micos estrela) tem sido introduzido no estado do Rio de Janeiro em áreas designadas para a reintrodução do Leontopithecus rosalia (mico leão dourado). Os objetivos deste estudo foram estimar a população de sagüis em dois fragmentos particulares, e caracterizar a interação entre as espécies. A densidade de sagüis (0.09 a/ha) foi maior que a de mico-leão (0.06 a/ha). O grau de associação entre mico-leão e o sagüi variou entre grupos e estações do ano (inverno= 61%, verão =35%). Foram observados sinais de competição durante o inverno quando a presença do mico estrela está associada a mudanças no forrageio do mico-leão e aumento de comportamentos territoriais. No verão, sinais de benefícios foram uma diminuição da vigilância por parte dos adultos de micos leões. Além disso, os micos leões foram observados se alimentando de goma proveniente dos buracos feitos pelos micos estrela. Estes resultados sugerem que o mico estrela é uma ameaça para a conservação do mico-leão, no entanto.