Invasion and intracellular survival of Campylobacter jejuni in HEp-2 cells were analyzed by transmission electron microscopy and by viable counts after killing of extracellular bacteria by gentamicin. During the first 30 min after challenge, no bacteria were seen in association with the host cell. After 1 h, campylobacters apparently attached to the cell membrane, with areas of close appositions. In these areas, an intracellular network of actin-like filaments was seen beneath the plasma membrane. Other bacteria were included into endocytic vacuoles. After 3 h, an intense lysosomal response was observed in the host cells, as determined by the presence of myelinic forms and acid phosphatase activity. After 9 h, bacteria still contained in vacuoles showed signs of degradation with a change from spiral to coccal forms. Morphological evidence of phagosome-lysosome fusion was also seen, and these observations by transmission electron microscopy correlated well with a decrease in bacteria viability 9 h after challenge, as determined from separate kinetics studies. Inhibitors of phagocytosis were observed to reduce markedly the entry of C. jejuni into the cells at concentrations which apparently did not affect bacterial viability. These results suggest that the campylobacters were successively attached to the HEp-2 cell membrane, internalized by a phagocytic-like mechanism, and digested after phagosome-lysosome fusion.

The objective of this paper was to report the isolation of two fluoroacetate degrading bacteria from the rumen of goats. The animals were adult goats, males, crossbred, with rumen fistula, fed with hay, and native pasture. The rumen fluid was obtained through the rumen fistula and immediately was inoculated 100 μL in mineral medium added with 20 mmol L−1 sodium fluoroacetate (SF), incubated at 39°C in an orbital shaker. Pseudomonas fluorescens (strain DSM 8341) was used as positive control for fluoroacetate dehalogenase activity. Two isolates were identified by 16S rRNA gene sequencing as Pigmentiphaga kullae (ECPB08) and Ancylobacter dichloromethanicus (ECPB09). These bacteria degraded sodium fluoroacetate, releasing 20 mmol L−1 of fluoride ion after 32 hours of incubation in Brunner medium containing 20 mmol L−1 of SF. There are no previous reports of fluoroacetate dehalogenase activity for P. kullae and A. dichloromethanicus. Control measures to prevent plant intoxication, including use of fences, herbicides, or other methods of eliminating poisonous plants, have been unsuccessful to avoid poisoning by fluoroacetate containing plants in Brazil. In this way, P. kullae and A. dichloromethanicus may be used to colonize the rumen of susceptible animals to avoid intoxication by fluoroacetate containing plants.

RESUMO.-Nos assentamentos rurais há maiores concentrações de reservatórios e vetores da leishmaniose visceral canina (LVC) devido às transformações ambientais resultantes da ação antrópica pela ocupação do território, no entanto, não há estudos no Brasil acerca de informações epidemiológicas da LVC em assentamentos rurais. O objetivo do presente trabalho foi determinar a soroprevalência para Leishmania sp. em cães de assentamentos rurais, bem como identificar os fatores de risco associados a infecção. Foram utilizados 306 cães e o diagnóstico sorológico da LVC foi realizado através do ELISA S7. Dos 306 cães investigados, 118 apresentaram anticorpos anti-Leishmania sp., resultando em frequência de 38,6%. Idade de 12 -24 meses (OR=2,97), idade de 24--48 meses (OR=4,83), idade de 4-6 anos (OR=4,40), idade >6 anos (OR=3,62), contato com aves (OR=1,67) e sexo (fêmea) (OR=1,97) foram apontados como fatores de risco para LVC. Cães de assentamentos rurais do semiárido paraibano apresentaram frequência elevada de anticorpos anti-Leishmania sp., o que torna a população desses locais susceptíveis à infecção. Isso evidencia a necessidade de alerta aos órgãos de vigilância epidemiológica para estabelecer medidas de prevenção e controle dessa zoonose, incluindo ações educacionais e sanitárias nesses assentamentos, já que os mesmos estão situados em áreas com características propícias à instalação da LVC. In rural settlements there are higher concentrations of reservoirs and vectors of canine visceral leishmaniasis (CVL) due to environmental changes resulting from human action by occupation of the territory, however, there are no surveys in Brazil on epidemiological information of CVL in these areas. The aim of this study was to determine the seroprevalence of Leishmania sp. in dogs from rural settlements, as well as to identify the risk factors associated with infection. A total of 306 dogs were tested, and the serological diagnosis of CVL was performed by ELISA S7. Of the 306 dogs, 118 presented anti-Leishmania sp. antibodies., resulting in a frequency of 38.6%. The risk factors identified for CVL seropositivity were: age of 12-24 months (OR=2.97), age of 24-48 months (OR=4.83), age of 4-6 years (OR=4.40), age >6 years (OR=3.62), contact with poultry (OR=1.67) and gender (female) (OR=1.97). Dogs from rural settlements in the semiarid of Paraiba showed high anti-Leishmania sp. antibody frequency, what makes the population of those settlements susceptible to infection. This highlights for the need to alert the epidemiological surveillance agencies to stablish prevention and control methods for that zoonosis, including educational and health activities on these settlements, as they are located in areas with favorable characteristics for CVL installation.

Lutzomyia longipalpis s.l. (Diptera: Psychodidae) is the principal vector of Leishmania infantum chagasi in the Americas, and constitutes a complex of species. Various studies have suggested an incipient speciation process based on behavioral isolation driven by the chemotype of male sexual pheromones. It is well known that natural barriers, such as mountains and rivers can directly influence population divergence in several organisms, including insects. In this work we investigated the potential role played by the Sao Francisco River in eastern Brazil in defining the current distribution of Lu. longipalpis s.l. Our studies were based on analyses of polymorphisms of the cytochrome b gene (cyt b) sequences from Lu. longipalpis s.l. available in public databases, and from additional field-caught individuals. Altogether, 9 distinct populations and 89 haplotypes were represented in the analyses. Lu. longipalpis s.l. populations were grouped according to their distribution in regards to the 10°S parallel: north of 10°S (<10°S); and south of 10°S (>10°S). Our results suggest that although no polymorphisms were fixed, moderate genetic divergences were observed between the groups analyzed (i.e., F ST = 0.184; and Nm = 2.22), and were mostly driven by genetic drift. The population divergence time estimated between the sand fly groups was about 0.45 million years (MY), coinciding with the time of the change in the course of the Sao Francisco River, during the Mindel glaciation. Overall, the polymorphisms on the cyt b haplotypes and the current speciation process detected in Lu. longipalpis s.l. with regards to the distribution of male sexual pheromones suggest a role of the Sao Francisco River as a significant geographical barrier in this process.

This article reports a new case of protothecosis by Prototheca wickerhamii in goats. The animal presented severe respiratory difficulty and nodules, sometimes ulcerated, in the nasal vestibule, mucocutaneous junction of the nostrils and skin of the face. Prototheca wickerhamii was isolated from the lesions. The animal had no clinical or haematologiccl evidence of immunodepression. The agent was highly resistant to antimicrobial drugs. The goat was treated unsuccessfully with fluconazole and euthanised 10 months after the diagnosis of the disease. Histological lesions were necrotising pyogranulomatous dermatitis, rhinitis and osteomyelitis with myriads of walled sporangia characteristic of P. wickerhamii. It is suggested that in goats, protothecosis is characterised by a chronic, slowly progressive infection, which affects immunologically competent goats, causing multifocal, ulcerative, pyogranulomatous and necrotising lesions of the mucosa of the nasal vestibule, mucocutaneous junctions of the nostrils and skin of the face.