A new standardization for the use of chicken embryo: selection of target from the phage display library and infection.

The filamentous bacteriophage M13KO7 (M13) is the most used in phage display (PD) technology and, like other phages, has been applied in several areas of medicine, agriculture, and in the food industry. One of the advantages is that they can modulate the immune response in the presence of pathogenic microorganisms, such as bacteria and viruses. This study evaluated the use of phage M13 in the chicken embryos model. We inoculated 13-day-old chicken embryos with Salmonella Pullorum (SP) and then evaluated survival for the presence of phage M13 or E. coli ER2738 (ECR) infected with M13. We found that the ECR bacterium inhibits SP multiplication in 0.32 (M13-infected ECR) or 0.44 log UFC/mL (M13-uninfected ECR) and that the ECR-free phage M13 from the PD library can be used in chicken embryo models. This work provides the use of the chicken embryo as a model to study systemic infection and can be employed as an analysis tool for various peptides that M13 can express from PD selection. KEY POINTS: • SP-infected chicken embryo can be a helpful model of systemic infection for different tests. • Phage M13 does not lead to embryonic mortality or cause serious injury to embryos. • Phage M13 from the PD library can be used in chicken embryo model tests.

The inoculation eschar of Rickettsia parkeri rickettsiosis in Brazil: Importance and cautions.

Two well characterized tick-borne rickettsioses occur in Brazil. Rickettsia rickettsii caused spotted-fever, transmitted by Amblyomma sculptum and Amblyomma aureolatum, is a severe disease with a high case-fatality rate in the southeastern region of the country. Rickettsia parkeri strain Atlantic rainforest infections transmitted by adult Amblyomma ovale ticks cause a milder non-lethal febrile disease with an eschar (necrosis) at the tick bite site. Clinical diagnosis of rickettsiosis is challenging, particularly during the early stages of the illness when signs and symptoms are non-specific. Since eschar at the tick bite site has emerged as the main clinical feature of mild R. parkeri infections and used to differentiate it from severe R. rickettsii infection, its proper recognition, distinction from other tick bite lesions, and boundaries as a clinical tool must be highlighted. Of importance, eschars induced by Rickettsia must be differentiated from dermatoses caused by other tick-borne skin infections as well from lesions caused by the tick bite itself. We herein highlight information on eschar in rickettsial diseases in Brazil and discuss the need for further research on its clinical relevance and application in the diagnosis of spotted fever caused by R. parkeri strain Atlantic rainforest. In particular, we draw attention to diagnosis of other febrile diseases in the presence of concomitant tick bites.

Author Correction: Clinical and serological evaluation of capybaras (Hydrochoerus hydrochaeris) successively exposed to an Amblyomma sculptum-derived strain of Rickettsia rickettsii.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

Clinical and serological evaluation of capybaras (Hydrochoerus hydrochaeris) successively exposed to an Amblyomma sculptum-derived strain of Rickettsia rickettsii.

Brazilian spotted fever (BSF), caused by Rickettsia rickettsii, is the most lethal tick-borne disease in the western hemisphere. In Brazil, Amblyomma sculptum ticks are the main vector. Capybaras (Hydrochoerus hydrochaeris), the largest living rodents of the world (adults weighing up to 100 Kg), have been recognized as amplifying hosts of R. rickettsii for A. sculptum in BSF-endemic areas; i.e., once primarily infected, capybaras develop bacteremia for a few days, when feeding ticks acquire rickettsial infection. We conducted experimental infections of five capybaras with an A. sculptum-derived strain of R. rickettsii and performed clinical and bacteremia evaluation during primary and subsequent infections. Bacteremia was detected in all capybaras during primary infection, but not in subsequent infections. All animals seroconverted to R. rickettsii (titres range: 64-32,768), and remained seropositive throughout the study. Primary infection resulted in clinical spotted fever illness in four capybaras, of which two had a fatal outcome. Subsequent infections in seropositive capybaras resulted in no clinical signs. Capybaras developed a sustained immune response that prevented a second bacteremia. This condition may imply a high reproduction rate of capybaras in BSF-endemic areas, in order to continuously generate capybaras susceptible to bacteremia during primary infection.

Amblyomma sculptum (Amblyomma cajennense complex) tick population maintained solely by domestic pigs.

In Brazil Amblyomma sculptum from Amblyomma cajennense complex is a tick species with a rather broad distribution encompassing the Cerrado Biome and devastated areas of the Atlantic rainforest. This species is also the major vector to humans of Rickettsia rickettsii, agent of the Brazilian spotted fever. Main primary hosts for the adult stage and frequently associated to high environmental infestation levels are the native capybaras and tapirs and a domestic host, the horse. Recent, however, solely circumstantial evidence, has shown that pigs (Sus scrofa) domestic, feral or exotic (wild boars) as well as wild suidae may maintain A. sculptum populations as well. We herein describe the maintenance of an A. sculptum tick population by domestic pigs for three years when raised in paddocks. Observations herein described occurred in the facilities of an intensive outdoor pig production system within a farm in Uberaba municipality southeast Brazil. Initial evaluation was done upon request due to human tick-bite complains in June 2013. Later tick infestations were evaluated at each of eight consecutive seasons from August 2014 to May 2016. In the first evaluation, 104 nymphs and one female A. sculptum were collected from four pigs and 269 nymphs of A. sculptum and 12 Amblyomma spp larva clusters from the paddocks. Only one A. sculptum nymph was found in each of the surrounding bovine pasture and a nearby riparian forest. Seven Rhipicephalus spp and three Amblyomma spp larva were found in the former as well. Infestations were greatly reduced with elimination of the two most infested paddocks and mowing of vegetation. Still along the inspection of eight consecutive seasons A sculptum ticks were collected overwhelmingly adults from pigs. Thus, observations herein described show that outdoor pig raising may provide both a permissive environment and adequate host allowing for a minimal threshold for the maintenance of A. sculptum populations. Since raising domestic pigs in paddocks to increase animal welfare is an increasing practice, it may indicate a new trend in this parasitism, with yet an unknown effect upon animal and human health.