Novel Ehrlichia sp. detected in Magellanic penguins (Sphenicus magellanicus) and in the seabird tick Ixodes uriae from Magdalena Island, southern Chile.

Ehrlichia spp. are obligatory intracellular microorganisms that infect hematopoietic, endothelial or blood cells of mammals. Ticks are the only vectors of these agents in nature. To date, the role of birds and their associated ticks as reservoirs of ehrlichiae remains almost unexplored. In this study, we performed a molecular screening for bacteria of Anaplasmataceae family in samples of spleen (n = 72) and lung (n = 17), recovered from 72 carcasses of Magellanic penguins (Spheniscus magellanicus) in Brazil and Chile. One apparently unengorged tick (Ixodes uriae) was also collected while wandering upon one of the carcasses and submitted to molecular analyses as well. Through conventional and nested PCR protocols three genes (16S rRNA, dsb and groEL) of a new Ehrlichia sp. were partially characterized upon organs of three penguins and in the tick coming from Magdalena Island (Chile). First matches after BLASTn comparisons showed that our sequences share 99.4% (16S rRNA), 94.6% (groEL) and 79.3% (dsb) of identity with "Candidatus Ehrlichia ornithorhynchi", Ehrlichia sp. NS101 and Ehrlichia canis CCZ, respectively. Matrixes of genetic distance including other representatives of the Ehrlichia genus point a 99.4%, 94.0%, and 80.0% of identity with 16S rRNA, groEL and dsb genes from Ehrlichia sp. It25, Ehrlichia sp. NS101, and Ehrlichia chaffeensis San Louis, respectively. A Bayesian phylogenetic analysis of Anaplasmataceae 16S rRNA gene places the detected Ehrlichia sp. into a group with Ehrlichia sp. BAT and Ehrlichia sp. Natal. Although depicting different topologies, Bayesian unrooted phylogenetic trees constructed for groEL and dsb genes position this Ehrlichia sp. into well-supported branches, which reinforces the finding of a new taxon. For the moment, any pathogenic effect of this new Ehrlichia sp. on penguins is still unknown. However, this fact becomes important to assess from a conservation point of view since populations of Magellanic penguins are currently threatened and in an ongoing decrease.

Anaplasmataceae, Borrelia and Hepatozoon agents in ticks (Acari: Argasidae, Ixodidae) from Chile.

Microorganisms harbored by Chilean autochthonous ticks have been scarcely studied and current knowledge is restricted to three species of hard ticks only. The current study aimed to assess the presence of Anaplasmataceae, Borrelia and Hepatozoon agents in ticks collected directly from the environment, on reptiles, birds and mammals in twelve localities from northern, central and southern regions of the country and Antarctica. Ticks were identified by means of a morphological and molecular approach. PCR detections point the occurrence of an Anaplasma-like agent and a relapsing fever Borrelia sp. in Ornithodoros spheniscus; an Ehrlichia sp. and a Hepatozoon sp. in Ornithodoros atacamensis; "Candidatus Neoehrlichia chilensis", Borrelia burgdorferi sensu lato (s. l.), and Hepatozoon in Ixodes ticks morphologically related to the Ixodes sigelos group; and B. burgdorferi s. l. in Ixodes auritulus. Supported by phylogenetic analyses of characterized microorganisms, this study introduces putative vector roles and initial evidence on possible new agents detected in Chilean ticks.

Ticks, rickettsial and erlichial infection in small mammals from Atlantic forest remnants in northeastern Brazil.

We evaluated infection by Rickettsia spp. and Ehrlichia spp in small mammals and their ticks from two Atlantic forest conservation areas in the state of Rio Grande do Norte, northeastern Brazil. A total of 39 small mammals were captured during 2012-2013, encompassing 33 marsupials (29 Didelphis albiventris, four Monodelphis domestica), three Cricetidae rodents (two Necromys lasiurus, one Rattus rattus), one Caviomorpha rodent (Thrichomys apereoides) and two armadillos (Euphractus sexcinctus). The ticks Amblyomma auricularium, Ixodes loricatus, and Ornithodoros mimon were collected from D. albiventris, whereas only A. auricularium was collected from armadillos. Through immunofluorescence assay with Rickettsia spp. antigens, 6/28 (21%) D. albiventris and the single R. rattus specimen reacted to at least one rickettsial antigen, with highest seroprevalence and endpoint titers to Rickettsia amblyommatis. A total of 150 ticks (126 A. auricularium, nine I. loricatus, 15 O. mimon) was tested for rickettsial infection by PCR, which detected only R. amblyommatis in most of the A. auricularium ticks. Lung and spleen samples were collected from small mammals (two N. lasiurus, six D. albiventris, three M. domestica, one T. apereoides, one R. rattus) and were tested by PCR for Anaplasmataceae agents. The spleen from one D. albiventris contained a new ehrlichial agent, here named as Ehrlichia sp. strain Natal. Phylogenetic analysis inferred from the dsb gene of Ehrlichia spp. indicates that this novel agent is potentially a new species. Future studies should monitor the possible role of rickettsial and/or ehrlichial microorganisms as agents of emerging diseases in these degraded areas of Atlantic forest, just as has occurred with other agents in degraded areas of this biome in southeastern Brazil.

Infestação por carrapatos Argasidae e Ixodidae em pequenos mamíferos silvestres da Estação Experimental Rafael Fernandes, Mossoró/RN / Infestation by ticks Argasidae and Ixodidae on small wild mammals at the Experimental Station Rafael Fernandes, Mossoró/RN, Brazil

Poucos estudos avaliaram a diversidade de ectoparasitos e a associação deles com seus hospedeiros que ocorrem no bioma Caatinga, Nordeste do Brasil. Considerando-se essa falta de conhecimento, este estudo objetivou identificar e determinar a ocorrência de carrapatos coletados de pequenos mamíferos da Estação Experimental Rafael Fernandes, no Rio Grande do Norte, Brasil. De janeiro de 2014 a fevereiro de 2015 foram capturados 52 marsupiais (38 Gracilinanus agilis e 14 Monodelphis domestica) e 10 roedores (5 Wiedomys sp., 4 Thrichomys sp. e 1 Rattus norvegicus). Foram identificados os carrapatos Amblyomma auricularium, Amblyomma parvum, Amblyomma sp., Ornithodoros mimon e Ornithodoros sp., empregando estudo morfológico, chaves taxonômicas e sequenciamento parcial do gene mitocondrial 16S rDNA de carrapatos. Todas as associações carrapato-hospedeiro encontradas neste estudo são relatadas pela primeira vez no Rio Grande do Norte e constituem novos dados ecológicos aplicáveis aos ectoparasitos de pequenos mamíferos no nordeste do Brasil.(AU)

Few studies have assessed the diversity of ectoparasites and their associated hosts occurring within the Caatinga biome in northeastern Brazil. Considering this lack of knowledge, in this study we aimed to identify and determine the occurrence of ticks collected from small mammals at the Estação Experimental Rafael Fernandes, in Rio Grande do Norte state, Brazil. From January 2014 to February 2015, we captured 52 marsupials (38 Gracilinanus agilis and 14 Monodelphis domestica) and 10 rodents (5 Wiedomys sp., 4 Thrichomys sp. and 1 Rattus norvegicus). We identified the ticks Amblyomma auricularium, Amblyomma parvum, Amblyomma sp., Ornithodoros mimon and Ornithodoros sp. by a morphological study, the use of taxonomic keys, and the partial sequencing of the tick mitochondrial 16S rDNA gene. All the tick-host associations found in this study are reported for the first time in Rio Grande do Norte and constitute new ecological data concerning ectoparasites of small mammals in northeastern Brazil.(AU)

A new species of Ornithodoros (Acari: Argasidae) from desert areas of northern Chile.

Ornithodoros atacamensis n. sp. is described from larvae collected on the lizard Liolaemus bisignatus and from free-living adults collected in desert areas from the Pan de Azúcar and Llanos de Challe National Parks, in Northern Chile. Additionally, unengorged larvae were obtained from fertilized females, which laid eggs in the laboratory. Morphological and mitochondrial 16S rDNA sequence analyses were performed in order to compare this new soft tick species with other congeneric Neotropical representatives. Larvae of O. atacamensis are morphologically closely related to Ornithodoros talaje sensu stricto, Ornithodoros puertoricensis, Ornithodoros rioplatensis, Ornithodoros guaporensis and Ornithodoros hasei, all belonging to the O. talaje species group. The larval diagnostic characters for this species are a combination of a large pyriform dorsal plate with a length of approximately 300µm, 17 pairs of dorsal setae with five central pairs, hypostome with apex pointed and dental formula 2/2 in most rows, 3/3 apically, and capsule of the Haller's organ oval in shape without reticulations. Phylogenetic analyses inferred from the mitochondrial 16S rRNA gene and a Principal Component Analysis based on morphometric characters provide additional support to the description of O. atacamensis as an independent lineage within the genus clustering within the O. talaje species group.

Ticks and rickettsiae from wildlife in Belize, Central America.

BACKGROUND: The agents of spotted fevers in Latin America are Rickettsia rickettsii, R. parkeri, Rickettsia sp. strain Atlantic rainforest, and R. massiliae. In Continental Central America, R. rickettsii remains the only known pathogenic tick-borne rickettsia. In the present study, ticks were collected from wild mammals in natural areas of Belize. Besides providing new data of ticks from Belize, we investigated rickettsial infection in some of these ticks. Our results provide ticks harboring rickettsial agents for the first time in Central America. METHODS: Between 2010 and 2015, wild mammals were lived-trapped in the tropical broadleaf moist forests of central and southern Belize. Ticks were collected from the animals and identified to species by morphological and molecular analysis (DNA sequence of the tick mitochondrial 16S RNA gene). Some of the ticks were tested for rickettsial infection by molecular methods (DNA sequences of the rickettsial gltA and ompA genes). RESULTS: A total of 84 ticks were collected from 8 individual hosts, as follows: Amblyomma pacae from 3 Cuniculus paca; Amblyomma ovale and Amblyomma coelebs from a Nasua narica; A. ovale from an Eira Barbara; A. ovale, Amblyomma cf. oblongoguttatum, and Ixodes affinis from a Puma concolor; and A. ovale, A. coelebs, A. cf. oblongoguttatum, and I. affinis from two Panthera onca. Three rickettsial agents were detected: Rickettsia amblyommii in A. pacae, Rickettsia sp. strain Atlantic rainforest in A. ovale, and Rickettsia sp. endosymbiont in Ixodes affinis. CONCLUSIONS: The present study provides unprecedented records of ticks harboring rickettsial agents in the New World. An emerging rickettsial pathogen of South America, Rickettsia sp. strain Atlantic rainforest, is reported for the first time in Central America. Besides expanding the distribution of 3 rickettsial agents in Central America, our results highlight the possible occurrence of Rickettsia sp. strain Atlantic rainforest-caused spotted fever human cases in Belize, since its possible vector, A. ovale, is recognized as one of the most important human-biting ticks in the Neotropical region.

Occurrence of antibodies against Toxoplasma gondii and its isolation and genotyping in donkeys, mules, and horses in Brazil.

The occurrence of antibodies against Toxoplasma gondii was determined in donkeys, mules, and horses from different regions of Brazil. Serum samples from 304 donkeys (67.11%), 118 horses (26.05%), and 31 mules (6.84%) were analyzed by means of the indirect fluorescent antibody test (cutoff=64). Antibodies against T. gondii were detected in 129 equids (28.47%) (82 donkeys, 32 horses, and 15 mules). Tissue samples from 19 seropositive and 50 seronegative animals were obtained in order to isolate the parasite by means of mouse bioassay, and T. gondii was isolated from a donkey. Through genotypic characterization of the isolate, by means of polymerase chain reaction (PCR)-restriction fragment length polymorphism (RFLP) using 11 genotypic markers, the genotype #163 (TgCkBr220), which has already been described in chickens in Brazil, was identified.

Defense reaction mediated by NMDA mechanisms in the inferior colliculus is modulated by GABAergic nigro-collicular pathways.

Electrical stimulation of the inferior colliculus (IC) causes a behavioral activation together with autonomic responses similar to fear reactions to threatening situations. GABAergic mechanisms exert a tonic inhibitory control on the neural substrates of aversion in the IC insofar as local injections of GABA agonists or antagonists inhibit or mimic these defensive behaviors, respectively. Recently, we have shown that systemic injections of the GABA-A receptor agonist muscimol unexpectedly enhanced the freezing and escape responses provoked by gradual increases in the intensity of the electrical stimulation of the IC. Taking into account that the neural circuits mediated by excitatory amino acids (EAA) in the IC may be responsible for the integration of fear states, in the present study we examined whether the defensive behavior induced by local injections of NMDA into the IC is influenced by prior treatment with systemic muscimol and also whether this GABAergic control could be exerted by GABAergic fibers that project to the inferior colliculus from the substantia nigra pars reticulata (SNpr). Rats were implanted with two guide-cannulae aimed at the IC and SNpr through which drug microinfusions with glass micropipette could be made with reduced brain damage. One week after surgery, the animals received either NMDA (7 nmol/0.2 microl) or saline into the IC and were placed into the middle of an enclosure where behavioral responses such as freezing, crossings, jumping, rearing, and turnings could be measured as an indirect index of unconditioned fear. These animals were pretreated either with saline or muscimol (0.5 mg/kg, IP) or with brain injections of saline or muscimol (1 nmol/0.2 ìl into SNpr). NMDA applied into the IC produced a behavioral activation with significant increases in all behavioral measures. IP injections of muscimol or into the SNpr enhanced the defense reaction caused by microinjections of NMDA into the IC. These findings give support to the idea that unconditioned defensive responses generated in the IC may be mediated by NMDA mechanisms. Additionally, a reduction of the inhibitory control exerted by nigrocollicular GABAergic neurons seems to be responsible for the unexpected pro-aversive action of systemic injections of muscimol on the neural substrates of aversion mediated by NMDA in the IC.