mRNA vaccination of rabbits alters the fecundity, but not the attachment, of adult Ixodes scapularis.

19ISP is a nucleoside-modified mRNA-lipid nanoparticle vaccine that targets 19 Ixodes scapularis proteins. We demonstrate that adult I. scapularis have impaired fecundity when allowed to engorge on 19ISP-immunized rabbits. 19ISP, therefore, has the potential to interrupt the tick reproductive cycle, without triggering some of the other effects associated with acquired tick resistance. This may lead to the development of new strategies to reduce I. scapularis populations in endemic areas.

Metarhizium pingshaense photolyase expression and virulence to Rhipicephalus microplus after UV-B exposure.

The current study examined the impact of ultraviolet (UV)-B radiation in Metarhizium pingshaense blastospores' photolyase expression and their virulence against Rhipicephalus microplus. Blastospores were exposed to UV under laboratory and field conditions. Ticks were treated topically with fungal suspension and exposed to UV-B in the laboratory for three consecutive days. The expression of cyclobutane pyrimidine dimmers (CPDs)-photolyase gene maphr1-2 in blastospores after UV exposure followed by white light exposure was accessed after 0, 8, 12, 24, 36, and 48 h. Average relative germination of blastospores 24 h after in vitro UV exposure was 8.4% lower than 48 h. Despite this, the relative germination of blastospores exposed to UV in the field 18 h (95.7 ± 0.3%) and 28 h (97.3 ± 0.8%) after exposure were not different (p > 0.05). Ticks treated with fungus and not exposed to UV exhibited 0% survival 10 days after the treatment, while fungus-treated ticks exposed to UV exhibited 50 ± 11.2% survival. Expression levels of maphr1-2 8, 12, and 24 h after UV-B exposure were not different from time zero. Maphr1-2 expression peak in M. pingshaense blastospores occurred 36 h after UV-B exposure, in the proposed conditions and times analyzed, suggesting repair mechanisms other than CPD-mediated-photoreactivation might be leading blastospores' germination from 0 to 24 h.

Entomopathogenic fungus treatment changes the gut bacterial diversity of Rhipicephalus microplus ticks.

BACKGROUND: Ticks are obligate bloodsucking parasites responsible for significant economic losses and concerns with human and animal health, mainly due to the transmission of pathogens. Entomopathogenic fungi have been intensively studied as an alternative strategy for tick control that can be used in combination with synthetic acaricides in the integrated management of ticks. Here, we investigated how the gut bacterial community of Rhipicephalus microplus is shaped after Metarhizium anisopliae treatment and how the tick susceptibility to the fungus is affected after disrupting gut bacterial microbiota. METHODS: Partially engorged tick females were artificially fed with pure bovine blood or blood plus tetracycline. Two other groups received the same diet and were topically treated with M. anisopliae. The guts were dissected, and the genomic DNA was extracted 3 days after the treatment; the V3-V4 variable region of the bacterial 16S rRNA gene was amplified. RESULTS: The gut of ticks that received no antibiotic but were treated with M. anisopliae exhibited lower bacterial diversity and a higher occurrence of Coxiella species. The Simpson diversity index and Pielou equability coefficient were higher in the gut bacterial community when R. microplus were fed with tetracycline and fungus-treated. Ticks from fungus-treated groups (with or without tetracycline) exhibited lower survival than untreated females. Previous feeding of ticks with the antibiotic did not change their susceptibility to the fungus. Ehrlichia spp. were not detected in the gueated groups. CONCLUSIONS: These findings suggest that myco-acaricidal action would not be impacted if the calf hosting these ticks is under antibiotic therapy. Moreover, the hypothesis that entomopathogenic fungi can affect the bacterial community in the gut of R. microplus engorged females is endorsed by the fact that ticks exposed to M. anisopliae exhibited a dramatic reduction in bacterial diversity. This is the first report of an entomopathogenic fungus affecting the tick gut microbiota.

Encapsulation of entomopathogenic fungal conidia: evaluation of stability and control potential of Rhipicephalus microplus.

The use of chemical acaricides is the primary strategy to control tick infestations. Nonetheless, chemical resistance in ticks has been reported. Thus, complementary methods such as biological control using entomopathogenic fungi (EPF) have been investigated. EPF, although efficient, have their viability compromised when applied under natural conditions, which indicates that formulation development is essential. Some researchers have demonstrated the efficacy of ionic gelation in protecting EPF against deleterious abiotic factors. In the present study, we conducted the ionic gelation technique to encapsulate Metarhizium anisopliae (Metschn.) Sorokin (Hypocreales: Clavicipitaceae) conidia in 2% (EC 2%) and 3% (EC 3%) sodium alginate. Next, the quantity and viability of encapsulated conidia (EC) were determined. The morphology of particles was characterized by using Scanning Electron Microscopy (SEM). EC and non-encapsulated conidia (NEC) were stored at room temperature (26.8 °C) and in the freezer (-11.9 °C) to shelf-life testing. For UV-B irradiance tolerance and thermotolerance tests, EC and NEC were exposed to UV-B (6.0 or 8.0 kJ m - 2) and heat (42 ºC). In addition, biological parameters of Rhipicephalus microplus Canestrini (Acari: Ixodidae) engorged females exposed to EC were evaluated. The particles presented a spherical shape, more homogeneous (EC 2%) or heterogeneous (EC 3%). Encapsulation decreased (4.8×) the conidial concentration and did not affect their viability. On the other hand, encapsulation increased the shelf life of conidia at room temperature as well as their UV-B tolerance and thermotolerance (6 h). The fungal particles decreased the biological parameters of females more significantly than the NEC. As far as we know, we reported for the first time the use of the ionic gelation to encapsulate entomopathogenic fungi toward controlling R. microplus.

Utilization of Metarhizium as an insect biocontrol agent and a plant bioinoculant with special reference to Brazil.

Brazil has a long history of using biological control and has the largest program in sugarcane agriculture to which a biocontrol program has been applied. This achievement is at least partly due to the utilization of the entomopathogenic fungus Metarhizium. This well-known fungal genus exhibits pathogenicity against a broad range of arthropod hosts and has been used globally as a biocontrol agent. This fungus is also a root symbiont, and in this capacity, it is a plant growth promoter. However, this feature (i.e., as a plant symbiont) has yet to be fully explored and implemented in Brazil, although the number of reports demonstrating Metarhizium's utility as a plant bioinoculant is increasing. The Brazilian bioproduct industry targets agricultural pests, and is limited to two Metarhizium species represented by four fungal isolates as active ingredients. Entomopathogenic fungi have also been successful in controlling arthropods of public health concern, as shown in their control of mosquitoes, which are vectors of diseases. The isolation of new indigenous Metarhizium isolates from a variety of substrates such as soil, insects, and plants shows the wide genetic diversity within this fungal genus. In this review, we emphasize the significance of Metarhizium spp. for the biological control of insects in Brazil. We also suggest that the experience and success of biological control with fungi in Brazil is an important resource for developing integrated pest management and sustainable strategies for pest control worldwide. Moreover, the future implementation prospects of species of Metarhizium being used as bioinoculants and possible new advances in the utility of this fungus are discussed.

Inhibition of Dopamine Activity and Response of Rhipicephalus microplus Challenged with Metarhizium anisopliae.

Dopamine modulates ticks and insect hemocytes and links these arthropods' nervous and immune systems. For the first time, the present study analyzed the effect of a dopamine receptor antagonist on the survival, biological parameters, phagocytic index, and dopamine detection in the hemocytes of ticks challenged by Metarhizium anisopliae. The survival and egg production index of Rhipicephalus microplus were negatively impacted when ticks were inoculated with the antagonist and fungus. Five days after the treatment, the survival of ticks treated only with fungus was 2.2 times higher than ticks treated with the antagonist (highest concentration) and fungus. A reduction in the phagocytic index of hemocytes of 68.4% was observed in the group inoculated with the highest concentration of the antagonist and fungus compared to ticks treated only with fungus. No changes were detected in the R. microplus levels of intrahemocytic dopamine or hemocytic quantification. Our results support the hypothesis that dopamine is crucial for tick immune defense, changing the phagocytic capacity of hemocytes and the susceptibility of ticks to entomopathogenic fungi.

Compatibility of different Metarhizium spp. propagules with synthetic acaricides for controlling Rhipicephalus microplus.

The inappropriate use of synthetic acaricides has selected resistant Rhipicephalus microplus populations. The present study evaluated the compatibility of different Metarhizium spp. propagules (conidia, blastospores, and microsclerotia) by incubating them with synthetic acaricides (amitraz, deltamethrin, and a combination of cypermethrin, chlorpyrifos, and citronellal) for 1 h, 5 h, 10 h, and 24 h. Conidia and microsclerotia of the tested isolates were usually more tolerant to synthetic acaricides than blastospores. Our study also analyzed the in vitro effect of deltamethrin associated with fungal propagules for controlling a population of R. microplus females that were not susceptible to this synthetic acaricide. The use of entomopathogenic fungi in association with deltamethrin in this tick population caused a greater tick control than did the use of the fungus or the synthetic acaricide separately.

How Dopamine Influences Survival and Cellular Immune Response of Rhipicephalus microplus Inoculated with Metarhizium anisopliae.

Dopamine (DA) is a biogenic monoamine reported to modulate insect hemocytes. Although the immune functions of DA are known in insects, there is a lack of knowledge of DA's role in the immune system of ticks. The use of Metarhizium anisopliae has been considered for tick control, driving studies on the immune response of these arthropods challenged with fungi. The present study evaluated the effect of DA on the cellular immune response and survival of Rhipicephalus microplus inoculated with M. anisopliae blastospores. Exogenous DA increased both ticks' survival 72 h after M. anisopliae inoculation and the number of circulating hemocytes compared to the control group, 24 h after the treatment. The phagocytic index of tick hemocytes challenged with M. anisopliae did not change upon injection of exogenous DA. Phenoloxidase activity in the hemolymph of ticks injected with DA and the fungus or exclusively with DA was higher than in untreated ticks or ticks inoculated with the fungus alone, 72 h after treatment. DA was detected in the hemocytes of fungus-treated and untreated ticks. Unveiling the cellular immune response in ticks challenged with entomopathogenic fungi is important to improve strategies for the biological control of these ectoparasites.

UV-B tolerances of conidia, blastospores, and microsclerotia of Metarhizium spp. entomopathogenic fungi.

The aim of the present study was to analyze ten native Metarhizium spp. isolates as to their UV-B tolerances. Comparisons included: different fungal propagules (conidia, blastospores, or microsclerotia [MS]); conidia in aqueous suspensions or in 10% mineral oil-in-water emulsions; and conidia mixed with different types of soil. The UV-B effect was expressed as the germination of conidia or culturability of blastospores and MS relative to nongerminated propagules. Metarhizium anisopliae LCM S05 exhibited high tolerance as blastospores and/or MS, but not as conidia; LCM S10 and LCM S08 had positive results with MS or conidia but not blastospores. The formulations with 10% mineral oil did not always protect Metarhizium conidia against UV-B. Conidia of LCM S07, LCM S08, and LCM S10 exhibited the best results when in aqueous suspensions, 24 h after UV-B exposure. In general, conidia mixed with soil and exposed to UV-B yielded similar number of colony forming units as conidia from unexposed soil, regardless the soil type. It was not possible to predict which type of propagule would be the most UV-B tolerant for each fungal isolate; in conclusion, many formulations and propagule types should be investigated early in the development of new fungal biocontrol products.

Larvicidal activity, route of interaction and ultrastructural changes in Aedes aegypti exposed to entomopathogenic fungi.

Blastospores or conidia (formulated or not) of entomopathogenic fungi were assessed against Aedes aegypti larvae. Larvae (L2) were exposed to 105, 106, 107, and 108 propagules mL-1 water suspension. Mineral oil at 0.1%, 0.5%, or 1.0% (v/v) was employed to observe the effect on larval survival. The 0.1% mineral oil did not affect larval survival. Accordingly, 107 propagules mL-1 and 0.1% mineral oil were used to prepare all fungal emulsions. The fungal suspension or formulation was prepared as follows: 107 propagules mL-1 on 0.03% TweenⓇ 80 (v/v) aqueous solution or 107 propagules mL-1 on 0.03% TweenⓇ 80 plus 0.1% mineral oil; larval survival rates were evaluated for 7 days, and median survival time (S50) was also determined. The presence of fungi in larvae was examined both histologically and by scanning electron microscopy 24 h or 48 h after exposure. To evaluate the larval growth, larvae were exposed to 107 propagules mL-1 for 48 hours and their length measured using a digital caliper. Here, propagules had similar results in reducing the larvae survival rate and time. The treatment with Beauveria bassiana s.l. at 108 propagules mL-1 or with Metarhizium anisopliae s.l. at 108 blastopores mL-1 reduced the larval survival time to two days. M. anisopliae s.l. at 108 conidia mL-1 reduced the survival time to three days. The survival time of larvae submitted to the other treatments ranged from 6 days to over 7 days. M. anisopliae s.l. or B. bassiana s.l. oil-in-water emulsions at 107 propagules mL-1 yielded better results than the water suspensions, the larvae survival rate was 2 days for both propagules in oil-in-water emulsion. Larvae exposed to blastospores from both isolates or M. anisopliae conidia were longer than in the other treatments. Scanning electron microscopy and histology analyzes found fungi predominantly in the gut, mouthparts, and perispiracular lobes of larvae. Formulated fungus yielded better results than the aqueous suspensions for control of mosquito larvae. Thus, for the first time, the effect of mineral oil on the fungal interaction on A. aegypti larvae was observed as well as the effect of entomopathogenic fungi in the growth of larvae, supporting the search for strategies to control this arthropod.

Atividades da intervenção de enfermagem “cuidados com a ostomia” / Nursing intervention activities \"care for ostomy”

Objetivo: avaliar as evidências sobre as atividades propostas pela intervenção da NIC “Cuidados coma ostomia”. Método: revisão integrativa, realizada em março de 2016 nas bases de dados MEDLINE, CINAHL,Web of Science, SCOPUS e LILACS. Os estudos foram categorizados a partir da identificação das intervençõescondizentes com a taxonomia da intervenção na NIC >, bem como as atividadespropostas. Resultados: a coleta resultou em 23 artigos, de maioria qualitativos (52%), com nível de evidênciaIV (60,9%), identificando-se, nestes, 23 das 25 atividades de Enfermagem propostas na NIC, sendo a atividade“Orientar o paciente/pessoa importante quanto ao uso de dispositivo/cuidados de ostomia” a mais frequente(60,9%). Conclusão: verificou-se, portanto, que as ações mais frequentes realizadas pelos enfermeiros sereferem às orientações de manejo do ostoma com a inclusão do cuidador e que há necessidade de maisestudos com melhores níveis de evidência nessa temática.