Potential Impact of Chemical Fungicides on the Efficacy of Metarhizium rileyi and the Occurrence of Pandora gammae on Caterpillars in Soybean Crops.

Entomopathogenic fungi may play a crucial role in the regulation of caterpillar populations in soybean crops, either through natural occurrences or applied as mycopesticides. In the present work, we reported the naturally occurring entomopathogenic fungus Pandora gammae attacking the caterpillar Chrysodeixis includens, with infection rates in field trials ran in two consecutive years in the 10-35% range. As many chemicals are potentially harmful to entomopathogenic fungi, this work aimed to investigate the potential impact of two chemical fungicides (azoxystrobin + benzovindiflupyr and trifloxistrobina + prothioconazole) used to control soybean rust (Phakopsora pachyrhizi) on the natural occurrence of P. gammae and Metarhizium rileyi, as well as the efficacy of the latter fungus applied as different formulations against the soybean caterpillars Anticarsia gemmatalis and C. includens. Under laboratory conditions, fungicides used at field-recommended rates had a considerable negative impact on the germinability of M. riley on the medium surface, and all tested formulations did not protect conidia from damage by these chemicals. This harmful effect also impacted host infectivity, as the larval mortality owing to this fungus was reduced by 30-40% compared to that of the fungicide-free treatments. In field trials conducted in two subsequent years, unformulated and formulated M. rileyi conidia applied to soybean plants produced primary infection sites in caterpillar populations after a single spray. Spraying unformulated or formulated M. rileyi conidia following fungicide application on plants did not affect host infection rates over time. Moreover, the use of M. rileyi-based formulations or chemical fungicide did not interfere with the natural infection rates by P. gammae on its host, C. includens. Although a higher degree of exposure to non-selective fungicides can negatively affect fungal entomopathogens, a single foliar application of fungicides may be harmless to both M. rileyi and P. gammae in soybean fields. Additionally, this work showed that naturally occurring wasps and tachnids also play an important role in the regulation of A. gemmatalis and, notably, C. includens, with parasitism rates above 40-50% in some cases.

Diversity of anamorphic Cordyceps (formerly Isaria) isolated from Brazilian agricultural sites.

A total of 53 anamorphic strains of Brazilian Cordyceps species currently maintained in a government-owned culture collection, were reassessed for diversity and species identity using multi-loci-based phylogenetic methods. The strains used in this study were originally obtained from soil samples or were isolated from insects of the orders Hemiptera, Lepidoptera, Coleoptera and Diptera, mostly from agricultural sites. A Bayesian phylogenetic tree was constructed based on a concatenation of five loci (ITS, LSU, RPB1, RPB2 and TEF). In a few cases of ambiguity, morphological traits were also considered for species delimitations. Considerable variability within the set of strains was detected and six Cordyceps species were identified: C. amoenerosea, C. fumosorosea, C. javanica, C. tenuipes and, for the first time, C. brevistroma and C. spegazzinii are reported in Brazil. Four other taxonomically equivocal groups, closely related to other known taxa (C. amoenerosea, C. cateniannulata, C. polyarthra and C. spegazzinii), were also recognized, although further studies will be required to confirm their identifications or their descriptions as new species. Cordyceps javanica was the most common species in our dataset, originally isolated from soil and several different insect orders, and includes 17 strains from the whitefly, Bemisia tabaci. Interestingly, strains previously identified as C. fumosorosea based on morphology and growth characteristics, were shown to be C. javanica, including the active ingredients of some commercial mycoinsecticides. Cordyceps farinosa, usually mentioned in the literature as occurring in Brazil, was not found in our study. Since most strains were from insect crop pests, further studies with hosts from non-agricultural settings or from environmental samples would be advisable for a deeper understanding of the occurrence of anamorphic Cordyceps in Brazil.

Akanthomyces diversity in Brazil and their pathogenicity to plant-sucking insects.

Currently, species within the genus Akanthomyces are poorly studied and explored compared to other hypocrealean entomopathogenic fungi employed as commercial biocontrol agents. This study aimed to molecularly identify 23 Brazilian Akanthomyces strains, most originally isolated from aphids and scales (n = 22), and one from the coffee leaf rust, and further investigate their pathogenicity to six plant-sucking insects as a means to better understand their host spectra. We also explored the capacity of A. muscarius CG935 for blastospore production via liquid fermentation. Akanthomyces dipterigenus, A. muscarius, A. lecanii, and two unidentified species were recognized as naturally occurring in Brazil. Akanthomyces dipterigenus CG829 and A. muscarius CG935 were highly virulent to nymphs of Bemisia tabaci (67.5-85.4% confirmed mortality) and the aphid Aphis fabae (74.6-75.3%), but only the first strain was virulent to the mealybug Planococcus sp. (80.9%). Akanthomyces lecanii CG824 was weakly virulent to all tested insects. None of the strains were pathogenic to the thrips Caliothrips phaseoli, and all strains showed low virulence to the wooly whitefly Aleurothrixus floccosus and the scale Duplachionaspis divergens. Submerged liquid fermentation yields varied from 1.72 × 109 (day 2) to 3.90 × 109 (day 5) blastospores mL-1. Blastospores or aerial conidia from A. muscarius CG935, at a single concentration of 1 × 107 viable propagules mL-1, resulted in 67.5-83.1% mortality of B. tabaci nymphs within 8 days post-treatment. Overall, these results encourage additional studies that could lead to the development of new mycopesticides based on Akanthomyces strains.

Genetic diversity and correlation between morphological traits of pequi fruits (Caryocar brasiliense Camb.) with and without thorns at the endocarp.

Morphological characteristics of pequi fruits (Caryocar brasiliense) allows us to obtain information for the conservation, breeding, and detect genetic variability. Thus, the aims of this research were to characterize the genetic diversity of Caryocar brasiliense that produce fruits with and without thorn at the endocarp; determine the important characters in the study of genetic diversity, and to estimate phenotypic and genotypic correlations. 80 fruits with thorns and 90 fruits without thorns were evaluated. Data were used for analysis: Euclidian distance as a measure of dissimilarity and the Tocher method for delimitation of groups; Principal Component Analysis, and genotypic and phenotypic correlations. Genetic dissimilarity of C. brasiliense ranged from 0.066 (B6-D6) to 0.908 (A14-B3). Tocher method allowed dividing the matrices into three groups, the dendrogram into five groups, and the principal components allowed the distribution in 8 groups. We observed 15 significant correlations for characters of fruit with thorns and 31 correlations for characters of fruit without thorns. The weight of the fruit (with thorns) and the pyrene yield (without thorns) are relevant correlations for production. The groupings generated allowed the classification of the matrices into distinct sets. Allowed the identification of potential producers plants that could be used in breeding programs.

Factors associated with latent tuberculosis among international migrants in Brazil: a cross-sectional study (2020).

BACKGROUND: Migrants are a high priority group for TB control measures due to their high exposure to risk factors such as poverty and social vulnerability. The study aimed to identify factors associated with latent TB among international migrants living in four Brazilian state capitals. This was a cross-sectional study conducted in September and October 2020 in a sample of 903 international migrants living in four Brazilian state capitals: Boa Vista/RR (458), Manaus/AM (136), São Paulo/SP (257), and Curitiba/PR (52). Data were collected with a questionnaire consisting of open and closed questions on personal characteristics, information on TB, and use of preventive measures. Tuberculin skin test (TST) was performed, with reading after 72 h by trained nurses and using 5 mm induration as the positive cutoff. Chi-square test (X2) and Fisher's exact test, both two-tailed, were used to compare statistically significant levels of association between the migrants´ sociodemographic characteristics, vulnerability, and latent TB infection (LTBI). Binary logistic regression was applied to calculate odds ratios and respective 95% confidence intervals. For all the tests, type I error of 5% was defined as statistically significant (p < 0.05). RESULTS: Prevalence of LTBI among migrants was 46.1% in Manaus/AM, 33.3% in São Paulo/SP, 28.1% in Curitiba/PR, and 23.5% in Boa Vista/RR. Factors associated with latent infection were age, male gender, and brown or indigenous race. CONCLUSIONS: The study showed high prevalence of latent TB among international migrants.

Assessment of the diversity of Brazilian entomopathogenic fungi in the genus Beauveria.

We combined matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) along with sequencing of the B locus intergenic region (Bloc) to assess the diversity of Brazilian species within the anamorphic genus Beauveria. A total of 121 strains maintained in a government-owned culture collection and isolated from a range of hosts/substrates over a long time span (1981-2015) were assessed. Strains were collected in five of six Brazilian biomes, mostly in the Atlantic Forest (42.2%) and Cerrado (29.8%), primarily from insect pests of crops. All strains were subjected to MS, and those not accurately identified by this technique were genomically analyzed. Among the outcomes of this study, four taxa from the genus Beauveria were recognized, with the great majority of strains belonging to B. bassiana s.str. (93.4%), followed by B. caledonica (2.5%), B. pseudobassiana (2.5%) and B. amorpha (1.6%). B. bassiana s.str. was found in all biomes and isolated from a wide range of hosts/substrates. Due to low numbers, associations of the remaining Beauveria species with specific hosts or habitats/biomes were not clear, except that all three B. caledonica strains were found only in the Cerrado biome and were associated with adults of the banana weevil, Cosmopolites sordidus (Col.:Curculionidae). B. pseudobassiana is reported for the first time on the South American continent, in a subtropical region and from two insect orders not yet associated with this taxon. We also showed that some strains previously ascribed to B. brongniartii were misidentifications. The biodiversity of Beauveria analyzed in our study was comparatively low. The geographic origins of strains used in our study were biased towards biomes with intense human interventions. Future surveys on more conserved, less environmentally disturbed biomes, such as Caatinga, Pampa, Pantanal, and Amazon are needed for a more comprehensive picture of the diversity of Beauveria and related genera in Brazil.

Within-host interactions of Metarhizium rileyi strains and nucleopolyhedroviruses in Spodoptera frugiperda and Anticarsia gemmatalis (Lepidoptera: Noctuidae).

Members of the family Baculoviridae have been quite successfully used as biocontrol agents against some lepidopterans. Likewise, a number of fungi are important natural enemies of these pests. An interesting approach to increase control efficacy could be the combination of a given nucleopolyhedrovirus (NPV) and a fungus, since they possess distinct modes of action. As a first step towards this goal, we assessed the interaction between NPV (either AgMNPV-79 or SfMNPV-6nd) and the entomopathogenic fungus Metarhizium rileyi (either CG1153 or CG381), using Anticarsia gemmatalis and Spodoptera frugiperda as hosts. In sequential applications of these pathogens, per os inoculation of an NPV (leaf discs with 2.5 × 104 occlusion bodies) either two days before or two days post-spraying of its counterpart fungal strain (5 × 103 conidia.cm-2 sprays) usually resulted in an antagonistic effect. When both pathogens were simultaneously applied at different combined dosages, usually an additive effect was seen. Interestingly, a number of dead larvae showing signs of co-infections (partially with soft integument and partially mummified) were recorded. However, mixes with lower dosages of both pathogens did not cause significantly higher insect mortalities compared to low dosages of the fungus applied alone. The advantages and disadvantages of the simultaneous applications of NPV and M. rileyi aiming at the management of either A. gemmatalis or S. frugiperda were discussed.

Current status and perspectives of fungal entomopathogens used for microbial control of arthropod pests in Brazil.

Entomopathogenic fungi play a central role in Brazil's biopesticide market. Approximately 50% of registered microbial biopesticides comprise mycoinsecticides and/or mycoacaricides consisting of hypocrealean fungi, with most based on Metarhizium anisopliae sensu stricto (s. str.) and Beauveria bassiana s. str. These fungi are mainly used to control spittlebugs in sugarcane fields and whiteflies in row crops, respectively, with annual applications surpassing three million hectares. Research also emphasizes the potential of fungal entomopathogens to manage arthropod vectors of human diseases. Most registered fungal formulations comprise wettable powders or technical (non-formulated) products, with relatively few new developments in formulation technology. Despite the large area treated with mycoinsecticides (i.e., approx. 2 million ha of sugarcane treated with M. anisopliae and 1.5 million ha of soybean treated with B. bassiana), their market share remains small compared with the chemical insecticide market. Nevertheless, several major agricultural companies are investing in fungus-based products with the aim at achieving more sustainable IPM programs for major pests in both organic and conventional crops. Government and private research groups are pursuing innovative technologies for mass production, formulation, product stability and quality control, which will support cost-effective commercial mycoinsecticides. Here, we summarize the status of mycoinsecticides currently available in Brazil and discuss future prospects.

Metarhizium humberi sp. nov. (Hypocreales: Clavicipitaceae), a new member of the PARB clade in the Metarhizium anisopliae complex from Latin America.

A new species, Metarhizium humberi, from the M. anisopliae complex and sister lineage of the M. anisopliae s.str. in the PARB clade, including M. pingshaense, M. anisopliae, M. robertsii and M. brunneum, is described based on phylogenetic analyses [translation elongation factor 1-alpha (5'TEF and 3'TEF), RNA polymerase II largest subunit (RPB1a), RNA polymerase II second largest subunit (RPB2a) and ß-tubulin (BTUB)]. Metarhizium humberi was first collected in 2001 in the Central Brazilian state of Goiás, later found to be a common fungus in soils in Brazil, and since then has also been isolated from coleopteran, hemipteran and lepidopteran insects in Brazil and Mexico. This new species, named in honor of Richard A. Humber, a well-known insect pathologist and taxonomist of entomopathogenic fungi, is characterized by a high insecticidal activity against different developmental stages of arthropod pests with importance in agriculture and vectors of diseases to human and animals.

Metarhizium alvesii sp. nov.: A new member of the Metarhizium anisopliae species complex.

A strain within the Metarhizium anisopliae species complex was isolated in 2009 from a soil sample in a banana plantation in the municipality of Quixeré, Northeastern region of Brazil. Previous studies showed that this insect-pathogenic strain does not fit with any current taxon within the M. anisopliae species complex, as determined by both genomic and by mass spectrometric analyses. In the present study, CG1123 (=ARSEF 13308) is shown to be morphologically indistinguishable from most species in this cosmopolitan species complex, whereas multilocus phylogeny confirmed its uniqueness and supports its recognition as a new species, Metarhizium alvesii, in honor of Sérgio Batista Alves, one of the founders of insect pathology in Brazil.

Evaluation of Pochonia chlamydosporia and Purpureocillium lilacinum for Suppression of Meloidogyne enterolobii on Tomato and Banana.

Meloidogyne enterolobii is one of the most important root-knot nematode in tropical regions, due to its ability to overcome resistance mechanisms of a number of host plants. The lack of new and safe active ingredients against this nematode has restricted control alternatives for growers. Egg-parasitic fungi have been considered as potential candidates for the development of bionematicides. In tissue culture plates, Pochonia chlamydosporia (var. catenulata and chlamydosporia) and Purpureocillium lilacinum strains were screened for their ability to infect eggs of the root-knot nematode M. enterolobii on water-agar surfaces. Reduction in the hatching of J2 varied from 13% to 84%, depending on strain. The more efficacious strains reduced hatchability of J2 by 57% to 84% when compared to untreated eggs, but average reductions were only 37% to 55% when the same strains were applied to egg masses. Combinations of fungal isolates (one of each species) did not increase the control efficacy in vitro. In experiments in which 10,000 nematode eggs were inoculated per plant, reductions in the number of eggs after 12 months were seen in three of four treatments in banana plants, reaching 34% for P. chlamydosporia var. catenulata. No significant reductions were seen in tomato plants after 3 mon. In another experiment with tomato plants using either P. chlamydosporia var. catenulata or P. lilacinum, the number of eggs was reduced by 34% and 44%, respectively, when initial infestation level was low (500 nematode eggs per plant), but tested strains were not effective under a moderate infestation level (5,000 eggs per plant). Under all infestation levels tested in this work, gall and egg mass indexes (MI) did not differ from the untreated controls, bringing concerns related to the practical adoption of this control strategy by farmers. In our opinion, if the fungi P. chlamydosporia and P. lilacinum are to be used as biocontrol tools toward M. entorolobii, they should focus on agricultural settings with low soil infestation levels and within an IPM approach.

Conidial vigor vs. viability as predictors of virulence of entomopathogenic fungi.

We tested the hypothesis that debilitated conidia exhibiting slow-germination (requiring>16h to germinate) are less virulent than vigorous conidia exhibiting fast germination (requiring⩽16h to germinate). Preparations of Beauveria bassiana s.l. strain CG 1027 with variable ratios of vigorous to debilitated conidia were assayed against third-instar larvae of Spodoptera frugiperda. As the proportion of debilitated conidia in test preparations increased, LC50 expressed in terms of total viable conidia increased, while LC50 expressed solely in terms of vigorous conidia remained constant, indicating that vigorous conidia were responsible for nearly all mortality observed in the assays. Larvae treated with conidia from low-quality batches (with high proportions of debilitated conidia) survived consistently longer than those treated with comparable doses of conidia from high-quality batches. These results confirm our previous hypotheses that inclusion of debilitated conidia in viability assessments can lead to overestimation of the quality (potency) of mycoinsecticide preparations and support our recommendation for use of short incubation periods for assessing viability whenever viability is relied upon as an indicator of product quality.

The fungistatic and fungicidal effects of volatiles from metathoracic glands of soybean-attacking stink bugs (Heteroptera: Pentatomidae) on the entomopathogen Beauveria bassiana.

This study was initially designed to evaluate the differential susceptibility of three soybean-attacking pentatomids to the entomopathogenic fungus Beauveria bassiana in standardized bioassays. Euschistus heros (Eh) was shown to be significantly less susceptible than Chinavia ubica (Cu), whereas Dichelops melacanthus (Dm) adults were highly susceptible to fungal infections. A deeper look at the mechanisms involved in the possible role of volatiles from metathoracic glands on fungal infections was undertaken, and gland extracts from Nezara viridula (Nv), a species known for its resilience to fungal infections, were also included in the assays. Atmospheres with volatiles from pentatomids with very low-susceptibility to B. bassiana infections (Eh and Nv) had a significant effect on speed of germination as shown in counts performed up to 22h post-inoculation, by which time 0.1 (control), 0.6 (Dm), 17.9 (Cu), 32.6 (Eh), and 43.4% (Nv) of conidia had not germinated. The fungistatic (inhibitory) and fungicidal (lethal) effects of Eh and Nv volatile-rich atmospheres were subsequently quantified in Petri dishes with either PDA or PDA medium amended with carbendazim, which allowed germination rates to be determined at 18 and 48h post-inoculation, respectively. As opposed to control, Eh volatile-rich atmosphere had a clear fungistatic effect, since germination rate was only 27.4% within 18h, but reached 99.4% at 48h post-inoculation. For Nv volatile-rich atmospheres, only 15.1% of conidia germinated within 18h, and by 48h post-inoculation, approx. 18% of conidia were unviable (neither germ tubes nor intumescence), whereas in the control treatment rates were >99% at both reading times. Therefore, the gaseous phase of defensive secretions from fungus-resilient pentatomids possess a strong inhibitory effect and may display a less pronounced lethal effect on fungal germination, as was the case for Nv.

MALDI-TOF mass spectrometry applied to identifying species of insect-pathogenic fungi from the Metarhizium anisopliae complex.

Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) has proven to be a powerful tool for taxonomic resolution of microorganisms. In this proof-of-concept study, we assessed the effectiveness of this technique to track the current gene sequence-based phylogenetic classification of species in the Metarhizium anisopliae complex. Initially the phylogenetic analysis of 5' strains by sequencing of the 59' end of the TEF-1α gene region revealed seven species within M. anisopliae sensu lato and two varieties outside this complex. Because initial studies on MS profiles from different cell types showed that mycelial fragments or conidia produced on nutrient-poor medium may yield too much background noise, all subsequent spectrometric analyses were performed with acidhydrolyzed conidia from 10-12 d old PDA cultures. The initial MALDI-TOF reference library included protein spectral profiles from nine taxonomically distinct, molecularly identified isolates sharing high genetic homology with the ex-type or ex-epitype isolates of these taxa in Metarhizium. A second reference library added one isolate each for M. anisopliae sensu stricto and M. robertsii. The second, larger reference library (including 11 taxa) allowed nearly perfect MALDI-TOF matching of DNA-based species identification for the 40 remaining isolates molecularly recognized as M. anisopliae sensu stricto (n = 19), M. robertsii (n = 6), M. majus (n = 3), M. lepidiotae (n = 1), M. acridum (n = 3), M. flavoviride var. pemphigi (n = 1), plus seven unidentified strains (six of them phylogenetically close to M. anisopliae sensu stricto and one outside the Metarhizium pingshaense-anisopliae-robertsii-brunneum clade). Due to the increasing frequency of phylogenetically (genomically) based taxonomic revisions of fungi, this approach is especially useful for culture collections, because once the protein profiles of Metarhizium isolates are obtained taxonomic updating of MALDI-TOF library data is easily accomplished by comparing stored profiles with those of newly proposed taxa.

Generation of reactive oxygen species by leukocytes of Prochilodus lineatus.

Prochilodus lineatus (curimbatá), from the Procholodontidae family, is a Brazilian freshwater fish, which is important commercially, nutritionally and ecologically. It is encountered in the Rio da Prata Bay in Southern South America. Studies on the immune system of this fish are scarce, but the physiological mechanisms of the species are analogous to those of other vertebrates. Thus, this work discusses the present study, which correlates P. lineatus leukocytes and the generation of reactive oxygen species after modulatory stimuli. Leukocytes were characterized by light and electron transmission microscopy and investigated by the generation of H2O2 and O2 (-), using phenol red, flow-cytometry and electron transmission histochemistry. The study determined that monocytes and neutrophils are the main cells responsible for generating O2 after stimulation with phorbol myristate acetate. Superoxide dismutase successfully inhibited the generation of reactive oxygen species in neutrophils and monocytes, but stimulated generation when in association with phorbol myristate acetate. Fish leukocyte samples from P. lineatus showed cross-reactivity with antibodies directed against human NADPH-oxidase antibody subunits (p47(phox) and p67(phox)). Thus, catalase enhanced the presence of p47(phox). Neutrophil mitochondria were shown to be generators of H2O2 (charged by cerium precipitate), being enlarged and changing their format. The present study contributes to a better understanding of the respiratory burst pathways in this species and suggests mitochondria as the organelle responsible for generation of reactive oxygen species.

Influence of some parameters on the germination assessment of mycopesticides.

The substantial negative impact of some parameters on the germination of low-quality conidia (high proportion of slow-germinating propagules) was demonstrated, whereas for high-quality batches their effect was small or even absent. Germination was increased as the initial hydration status of conidia immediately prior to suspension preparation was increased, being ca. 33% and 80% for dehydrated Metarhizium anisopliae propagules (water activity ≤0.314) from low- or high-quality batches after an 18 h incubation period, respectively, and 63% and 95% for hydrated propagules (water activity = 0.933). Germination of low-quality propagules also increased as the time dry conidia were kept in aqueous suspension prior to inoculation onto culture media (15 min, 3 or 24 h) or the incubation time at 25°C before counts (18, 48 or 72 h) was increased. Depending on treatment conditions, average germination of low-quality conidia varied from 53% to 98%. On the other hand, germination for high-quality conidia was always ≥94%. Regarding the relative humidity (RH) of the incubation atmosphere, the average germination rates for low-quality conidia on Potato Dextrose Agar (PDA) in Petri plates was 49%, while germination of these conidia on PDA blocks kept under lower RH inside plastic boxes was ≤23%. Use of lactophenol-staining and/or use of coverslips had a negative effect when germination assessment was performed for low-quality conidia, resulting in distorted counts or increased standard deviations compared to high-quality conidial batches. The occurrence of dislodged conidia (ungerminated conidia outside the inoculation zone due to hydraulic pressure exercised by addition of stains and/or coverslips added to the substrate by the time germination is assessed) was common place, whereas dislodged conidia were not seen in treatments with high-quality batches. This work underscores the importance of a number of parameters that anyone working with low-quality fungi needs to be cognizant of in their research.

On-farm Production of Microbial Entomopathogens for use in Agriculture: Brazil as a Case Study.

In Brazil, the production of beneficial microorganisms by growers exclusively for their own use is a practice known as "on-farm production". Regarding on-farm bioinsecticides, they were initially deployed for pests of perennial and semi-perennial crops in the 1970s but, since 2013, their use has extended to pests of annual crops such as maize, cotton, and soybean. Millions of hectares are currently being treated with these on-farm preparations. Local production reduces costs, meets local needs, and reduces inputs of environmentally damaging chemical pesticides, facilitating establishment of more sustainable agroecosystems. Critics argue that without implementation of stringent quality control measures there is the risk that the on-farm preparations: (1) are contaminated with microbes which may include human pathogens or (2) contain very little active ingredient, impacting on field efficacy. The on-farm fermentation of bacterial insecticides predominates, especially that of Bacillus thuringiensis targeting lepidopteran pests. However, there has been a rapid growth in the past 5 years in the production of entomopathogenic fungi, mostly for the control of sap-sucking insects such as whitefly (Bemisia tabaci (Gennadius)) and the corn leafhopper (Dalbulus maidis (DeLong and Wolcott)). In contrast, on-farm production of insect viruses has seen limited growth. Most of the ca. 5 million rural producers in Brazil own small or medium size properties and, although the vast majority still do not practice on-farm production of biopesticides, the topic has aroused interest among them. Many growers who adopt this practice usually use non-sterile containers as fermenters, resulting in poor-quality preparations, and cases of failure have been reported. On the other hand, some informal reports suggest on-farm preparations may be efficacious even when contaminated, what could be explained, at least partially, by the insecticidal secondary metabolites secreted by the pool of microorganisms in the liquid culture media. Indeed, there is insufficient information on efficacy and mode of action of these microbial biopesticides. It is usually the large farms, some with > 20,000 ha of continuous cultivated lands, that produce biopesticides with low levels of contamination, as many of them possess advanced production facilities and have access to specialized knowledge and trained staff. Uptake of on-farm biopesticides is expected to continue but the rate of adoption will depend on factors such as the selection of safe, virulent microbial strains and implementation of sound quality control measures (compliance with emerging Brazilian regulations and international standards). The challenges and opportunities of on-farm bioinsecticides are presented and discussed.

Prevalence of tuberculosis, COVID-19, chronic conditions and vulnerabilities among migrants and refugees: an electronic survey.

OBJECTIVE: to analyze the prevalence of tuberculosis, coronavirus, chronic conditions and vulnerabilities among migrants and refugees in Brazil. METHOD: this is a cross-sectional study of the electronic survey type conducted with international migrants during the COVID-19 pandemic. Descriptive statistics was applied for the analysis, with calculation of position and dispersion measures. Regarding the categorical variables, relative and absolute frequencies were estimated. RESULTS: the study participants were 553 migrants and refugees, verifying 3.07%, 7.2% and 27.3% prevalence of tuberculosis, COVID-19 and chronic conditions, respectively. Among the vulnerabilities, 32% reported unemployment, 37.6% moved to Brazil as a result of the social situation in their countries and 33.6% were living as refugees or sheltered people. CONCLUSION: tuberculosis, chronic diseases and COVID-19 presented higher prevalence values in migrants and refugees than in the general population. As this is a population group that still has significant difficulty accessing health services and social protection systems, based on diverse evidence, the study will subsidize public policies, Nursing care and the incorporation of new routines in the service.

Ultrasound evaluation of the fetal nasal bone: what is the most appropriate first-trimester cut-off point for aneuploidy screening?

PURPOSE: To establish fetal nasal bone length cut-off points for first trimester aneuploidy screening based on a normal curve of a Brazilian population. METHODS: The following tests were proposed: presence or absence of the nasal bone (NB); 2.5 and 5.0 NB percentiles relative to the normal curve; and 0.70, 0.75 and 0.80 multiples of the median (MoM) values defined in the receiver operating characteristic (ROC) curve. Nasal Bone tests were based on positive and negative likelihood ratio value detection rates (LR); the confidence interval was 95% in all tests. Cases in which ultrasonographic images of the NB were absent were not taken into account when evaluating the 2.5 and 5.0 percentiles and the 0.70, 0.75 and 0.80 MoM. RESULTS: The sample consisted of 571 fetuses (10-14 weeks). After exclusions (11) and loss of follow-up (53), the study sample was reduced to 507 patients. There were 23 Down syndrome patients among 41 aneuploid fetuses. The sensitivity of the qualitative NB test (absent vs. present) was 34.1%, and the specificity was 99.1% (+LR 37.89, -LR 0.66). An image of the nasal bone was absent in 52.2% of fetuses with the Down syndrome (+LR 58.00, -LR 0.48). CONCLUSIONS: The best tool for aneuploidy screening was the qualitative NB test (absent vs. present). Ultrasonography of the NB is a component of aneuploidy screening, and should not be used alone.