Luiz Cezar Machado Pereira

Rickettsia amblyommii Infecting Amblyomma auricularium Ticks in Pernambuco, Northeastern Brazil: Isolation, Transovarial Transmission, and Transstadial Perpetuation

This study investigated rickettsial infection in Amblyomma auricularium ticks from the state of Pernambuco, northeastern Brazil. An engorged female of A. auricularium collected from a skunk (Conepatus semistriatus) was sent alive to the laboratory, where the female was found through molecular analysis to be infected by Rickettsia amblyommii. This engorged female oviposited, and its offspring was reared through three consecutive generations, always using tick-naïve rabbits to feed the ticks. PCR performed on five egg pools, 10 larvae, 10 nymphs, and 10 adults of each of the three generations always yielded rickettsial DNA, indicating maintenance of rickettsial infection in the ticks by transstadial and transovarial passages. DNA sequences of random PCR products from eggs, larvae, nymphs, and adults were identified as R. amblyommii. All infested rabbits seroconverted to R. amblyommii antigens at the 21(st) day after infestation, indicating that larvae, nymphs, and adults transmitted R. amblyommii through parasitism. However, no infested rabbit presented fever or any clinical alteration during the experimental period. Rickettsiae were successfully isolated from the two A. auricularium females, and the isolates were established in Vero cell culture. Molecular characterization of the isolates confirmed R. amblyommii by sequencing partial gltA, ompA, and ompB genes. From another sample of 15 A. auricularium adult ticks collected from two armadillos (Euphractus sexcinctus), eight (53.3%) were infected by R. amblyommii. This study reports R. amblyommii infecting the tick A. auricularium for the first time. This is also the first report of rickettsia infecting ticks in the northeastern region of Brazil.

Natural and Anthropogenic Hybridization in Two Species of Eastern Brazilian Marmosets (Callithrix jacchus and C. penicillata).

Animal hybridization is well documented, but evolutionary outcomes and conservation priorities often differ for natural and anthropogenic hybrids. Among primates, an order with many endangered species, the two contexts can be hard to disentangle from one another, which carries important conservation implications. Callithrix marmosets give us a unique glimpse of genetic hybridization effects under distinct natural and human-induced contexts. Here, we use a 44 autosomal microsatellite marker panel to examine genome-wide admixture levels and introgression at a natural C. jacchus and C. penicillata species border along the São Francisco River in NE Brazil and in an area of Rio de Janeiro state where humans introduced these species exotically. Additionally, we describe for the first time autosomal genetic diversity in wild C. penicillata and expand previous C. jacchus genetic data. We characterize admixture within the natural zone as bimodal where hybrid ancestry is biased toward one parental species or the other. We also show evidence that São Francisco River islands are gateways for bidirectional gene flow across the species border. In the anthropogenic zone, marmosets essentially form a hybrid swarm with intermediate levels of admixture, likely from the absence of strong physical barriers to interspecific breeding. Our data show that while hybridization can occur naturally, the presence of physical, even if leaky, barriers to hybridization is important for maintaining species genetic integrity. Thus, we suggest further study of hybridization under different contexts to set well informed conservation guidelines for hybrid populations that often fit somewhere between “natural” and “man-made.”

Hybridization effects and genetic diversity of the common and black-tufted marmoset (Callithrix jacchus and Callithrix penicillata) mitochondrial control region

Hybridization is continually documented in primates, but effects of natural and anthropogenic hybridization on biodiversity are still unclear and differentiating between these contexts remains challenging in regards to primate evolution and conservation. Here, we examine hybridization effects on the mitochondrial DNA (mtDNA) control region of Callithrix marmosets, which provide a unique glimpse into interspecific mating under distinct anthropogenic and natural conditions. DNA was sampled from 40 marmosets along a 50-km transect from a previously uncharacterized hybrid zone in NE Brazil between the ranges of Callithrix jacchus and Callithrix penicillata. DNA was also collected from 46 marmosets along a 30-km transect in a hybrid zone in Rio de Janeiro state, Brazil, where exotic marmosets appeared in the 1980s. Combining Callithrix DNA sampled inside and outside of these hybrid zones, phylogenetic and network analyses show C. jacchus and C. penicillata being parental species to sampled hybrids. We expand limited Callithrix population genetics work by describing mtDNA diversity and demographic history of these parental species. We show ancient population expansion in C. jacchus and historically constant population size in C. penicillata, with the latter being more genetically diverse than the former. The natural hybrid zone contained higher genetic diversity relative to the anthropogenic zone. While our data suggest hybrid swarm formation within the anthropogenic zone due to removed physical reproductive barriers, this pattern is not seen in the natural hybrid zone. These results suggest different genetic dynamics within natural and anthropogenic hybridization contexts that carry important implications for primate evolution and conservation.

Validation of qPCR Methods for the Detection of Mycobacterium in New World Animal Reservoirs

Zoonotic pathogens that cause leprosy (Mycobacterium leprae) and tuberculosis (Mycobacterium tuberculosis complex, MTBC) continue to impact modern human populations. Therefore, methods able to survey mycobacterial infection in potential animal hosts are necessary for proper evaluation of human exposure threats. Here we tested for mycobacterial-specific single- and multi-copy loci using qPCR. In a trial study in which armadillos were artificially infected with M. leprae, these techniques were specific and sensitive to pathogen detection, while more traditional ELISAs were only specific. These assays were then employed in a case study to detect M. leprae as well as MTBC in wild marmosets. All marmosets were negative for M. leprae DNA, but 14 were positive for the mycobacterial rpoB gene assay. Targeted capture and sequencing of rpoB and other MTBC genes validated the presence of mycobacterial DNA in these samples and revealed that qPCR is useful for identifying mycobacterial-infected animal hosts.

New Record of Leopardus pardalis (Linnaeus, 1758) (Carnivora: Felidae) in the Caatinga of the state of Pernambuco, northeastern Brazil

The ocelot ( Leopardus pardalis ) has a wide geographic distribution. However, in Brazil, there is still a lack of information on their population status and, in the Caatinga, little is known about its occurrence. This paper aims to register the new occurrence of Leopardus pardalis in the State of Pernambuco, Northeast Brazil.

Range extension, new state record and geographic distribution map of Acratosaura mentalis (Amaral, 1933) (Squamata: Gymnophthalmidae)

We provide new records of Acratosaura mentalis from three localities in the state of Pernambuco, Brazil, these records extending the known geographical distribution of the species to the municipalities of Sertânia, Custodia and Floresta. We also report A. mentalis from the municipality of Mauriti, which is the first record for the state of Ceara, Brazil.

Correction: Validation of qPCR Methods for the Detection of Mycobacterium in New World Animal Reservoirs

[This corrects the article DOI: 10.1371/journal.pntd.0004198.].

A New Distributon Record of the Threatened Megasoma gyas rumbucheri Fischer, 1968 (Coleoptera: Scarabaeidae: Dynastinae) Northward to the Rio São Francisco, Pernambuco, Brazil

The genus Megasoma Kirby is composed of 17 species of medium to large sized beetles that occur from the southern United States to northern Argentina (Morón 2005; Ratcliffe and Morón 2005; Abadie et al. 2008). In Brazil, seven species and subspecies are known: Megasoma actaeon (L., 1758); Megasoma anubis Chevrolat, 1836; Megasoma gyas gyas (Herbst, 1775); Megasoma gyas porioni Nagai, 2003; Megasoma gyas rumbucheri Fischer, 1968; Megasoma janus janus Felsche, 1906; and Megasoma janus fujitai Nagai, 2003 (Morón 2005; Abadie et al. 2008). Males of Megasoma species have a single bifurcated head horn and, in most species, there are one discal and two lateral horns on the pronotum (Endrödi 1985). In males of M. gyas, the pronotal projections are elongated, acute, and anteriorly directed (Moore 2007). There are three subspecies of M. gyas, all of which occur in Brazil: M. gyas gyas in the southeastern Atlantic Rainforest; M. gyas rumbucheri in the northeastern Caatinga and Caatinga-Cerrado transitions; and M. gyas porioni restricted to the state of Bahia (Morón 2005). Megasoma gyas rumbucheri is considered a rare subspecies and differs from other conspecific subspecies by having a large frontal horn with a strongly bifurcated apex and elytral pubescence considerably longer (Endrodi 1985; Grossi et al. 2008). Megasoma gyas rumbucheri suffers from severe habitat loss in both humid and semi-arid environments of Brazil, where it is considered endangered (“vulnerable” status – Monteiro et al. 2008). Together with the imperative conservation of its remaining habitat, Grossi et al. (2008) proposed the following conservation measures: (1) searching for new populations, (2) studying its population dynamics and life cycle, and (3) captive rearing in order to reintroduce animals in nature. In 2010, M. gyas rumbucheri was recorded at latitudes above the São Francisco River, which also constituted the first record of the subspecies in a Brazilian Conservation Unit (Santos et al. 2013). We now report M. gyas rumbucheri in the state of Pernambuco, where it was last recorded in 1972 (Grossi et al. 2008), and provide biometrical data of the collected specimens. During monitoring activities by the “Projeto de Integração do Rio São Francisco com Bacias Hidrográficas do Nordeste Setentrional”, three males of M. gyas rumbucheri were collected in a locality known as Samambaia (8°19′25′′S