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Several phytoseiid mite species are important natural enemies used in biological control strategies. In the present study, Cytb mtDNA sequences of various populations of two species, Phytoseiulus macropolis and P. persimilis, were compared to determine whether the specimens collected in Brazil could belong to P. persimilis as this latter species is reported in South America but not in Brazil. The Cytb marker was used because of its high evolution rate, assumed to capture intraspecific variation. No overlap between intra- and interspecific distances was observed but the distances were quite low for interspecific variation. This can be due to the particular biology of Phytoseiulus species and this shows the difficulty to apply a universal threshold in genetic distances to conclude about the existence of one or several species. Cytb mtDNA sequences were also considered to assess intraspecific variation. The DNA sequences of P. persimilis populations were very similar, probably because they all originated from the West Palearctic region or because of a prevalence of commercialized specimens in natura. For P. macropilis, higher genetic distances were observed and differentiation was noted according to geographic location and, to a smaller extent, pyrethroid resistance. To determine how DNA variation might impact the protein function (CytB fragment considered), the amino acid compositions of the populations studied were compared. No diagnostic mutation was observed between pyrethroid resistant and susceptible populations, whereas four mutations were identified between populations of P. macropilis separated by 1300 km (different climatic conditions). The impact of such mutations is discussed but knowledge is scarce, which makes it difficult to root testable hypotheses. The protein analysis clearly opens new perspectives in Phytoseiidae studies., (© 2021. The Author(s), under exclusive licence to Springer Nature Switzerland AG.)

Modelling studies on climate change predict continuous increases in atmospheric carbon dioxide concentration [CO2] and increase in temperature. This may alter carbon-based phytochemicals such phenolics and modify plant interactions with herbivorous. We investigated the effects of enhanced [CO2] and local climatic variation on young coffee plants, Coffea arabica L. cv Catuaí vermelho IAC-144 and Obatã vermelho IAC-1669-20, cultivated in the FACE (Free-Air Carbon Dioxide Enrichment) facility under two atmospheric [CO2] conditions. Coffee leaves were evaluated for total soluble phenolics (TSP), chlorogenic (5-CQA) and caffeic (CAF) acids, diversity and population size of mites, along two dry and two rainy seasons. Elevated atmospheric CO2 (e[CO2]) significantly decreased 5-CQA in cv. Catuaí but did not affect cv. Obatã. Species richness and population size of mites in coffee leaves were not affected by e[CO2] but were strongly related to the seasonal variability of coffee leaf phenolics. In general, high levels of phenolics were negatively correlated with population size while the mite species richness were negatively correlated with 5-CQA and TSP levels. Our findings show that [CO2] enhancement affects phenolics in coffee plants differentially by cultivars, however seasonality is the key determinant of phenolics composition, mite species richness and population size.

Neospora caninum is a protozoan that can cause reproductive problems in several animal species. Although N. caninum infection has been reported in swine, the pathogenesis and clinical signs are not fully known in this species. The objective of this work was to evaluate the effect of experimental infection with tachyzoites of the N. caninum strain Nc1 in swine matrices at different stages of gestation. For that purpose, 12 gilts, seronegative for N. caninum and T. gondii, were selected and allocated into four groups of three animals each. Animals in group A were not inoculated (control) and animals in groups B, C, and D were inoculated intravenously with of 2.9 × 10 7 tachyzoites, 30 days before conception, and at 45 and 90 days of gestation, respectively. Temperature, heart rate, blood, saliva, and vaginal mucus samples from the animals were collected periodically until the time of delivery for the investigation of IgG and IgM antibodies against N. caninum using IFAT and PCR to detect the parasite DNA. All gilts sero-converted from 5 and 7 DPI (days postinoculation) to IgM and IgG, respectively. Two gilts showed hypothermia on the 5th and 7th DPI, and five inoculated animals had leukocytosis on the 7th DPI. It was possible to detect DNA of N. caninum in samples of saliva (33/84), vaginal mucus (17/84), and blood (2/84). Based on serology (IgM) and PCR, three animals in group B showed evidence of reappearance of the infection during pregnancy. It is concluded that N. caninum can cause clinical signs in infected swine females, in addition to indicating saliva as a suitable diagnostic biological material for the detection of N. caninum DNA in this animal species.

Phytoseiid mites are efficient predators of mites and small pest insects. Understanding the dispersion and distribution pattern of phytoseiid mites is essential to promote the conservation of these natural enemies and support their use in biological control. Population genetic studies using molecular markers such as microsatellites have proved to be extremely informative to address questions about population structure and dispersion patterns of predatory mites. The objective of this work was to develop specific microsatellite markers for the predatory mite Phytoseiulus macropilis, aiming at improving field dispersion studies. For this purpose, the genomic DNA was extracted from the whole body of a pool of 260 adult females and used to build the genomic microsatellites-enriched library, using biotinylated probes (CT) 8 and (GT) 8 . In total 26 pairs of primers were synthesized and screened across 30 adult females of P. macropilis for characterization. Seven loci were polymorphic, revealing from two to six alleles per locus. Cross amplifications were successfully obtained in the species Phytoseiulus persimilis, Amblyseius swirskii and Proprioseiopsis sp. The molecular markers obtained are the first developed for P. macropilis-they are effective for the detection and quantification of genetic variation, and show high transferability, thus can be used in genetic and molecular studies of this and other species of the same genus and also of close genera.

The two-spotted spider mite, Tetranychus urticae Koch, is one of the most important pests on a wide range of crops worldwide. Studies on stability of resistance and possible fitness costs associated with etoxazole resistance were carried out in T. urticae to provide basic information necessary to define effective acaricide resistance management strategies for this pest. Selection for resistance to etoxazole was performed in a population of T. urticae collected from a commercial rose field, in Holambra County, in the State of São Paulo, Brazil. After five rounds of selection for resistance, the resistance ratio (R/S) at the LC50 reached 8739 fold value in comparison with a susceptible strain (Brazabrantes S Strain). The stability of etoxazole resistance was studied under laboratory conditions, using a population with initial frequency of 75 % of resistant mites. The frequencies of etoxazole resistance were evaluated monthly for a period of 13 months. The frequency of etoxazole resistance decreased from 75 to 37 % in this period. Comparison of biological traits between resistant and susceptible strains indicated the presence of fitness costs associated with etoxazole resistance. The resistant strain showed lower fecundity and a higher proportion of males in the population. The figures for net reproductive rate (R 0), intrinsic rate of natural increase (r m) and finite rate of increase (λ) were higher in the susceptible strain. The instability of etoxazole resistance is a favorable aspect for the acaricide resistance management in the spider mite.

Phytoseiulus macropilis Banks (Acari: Phytoseiidae) is an effective predator of Tetranychus urticae Koch (Acari: Tetranychidae). The objectives of this research were to study the stability of fenpropathrin resistance and the cross-resistance relationships with different pyrethroids, and also to evaluate the effect of synergists [piperonyl butoxide (PBO), diethyl maleate (DEM) and S,S,S-tributyl phosphorotrithioate (DEF)] on fenpropathrin resistant and susceptible strains of this predaceous mite. The stability of fenpropathrin resistance was studied under laboratory conditions, using P. macropilis populations with initial frequencies of 75 and 50% of resistant mites. The percentages of fenpropathrin resistant mites were evaluated monthly for a period of up to 12 months. A trend toward decreased resistance frequencies was observed only during the first 3-4 months. After this initial decrease, the fenpropathrin resistance was shown to be stable, maintaining constant resistance frequencies (around 30%) until the end of the evaluation period. Toxicity tests carried out using fenpropathrin resistant and susceptible strains of P. macropilis indicated strong positive cross-resistance between fenpropathrin and the pyrethroids bifenthrin and deltamethrin. Bioassays with the synergists DEM, DEF and PBO were also performed. The maximum synergism ratio (SR = LC50 without synergist/LC50 with synergist) detected for the three evaluated synergists (PBO, DEM, DEF) was 5.86 (for DEF), indicating low influence of enzyme detoxification processes in fenpropathrin resistance.

Brevipalpus phoenicis (Geijskes) is associated with the transmission of Citrus leprosis which is considered the main viral disease for the Brazilian citrus production. Mites of the families Stigmaeidae and Phytoseiidae coexist in various agricultural crops, often promoting the biological control of pest mites. The aim of this work was to study the interactions of Neoseiulus californicus (McGregor) (Phytoseiidae) and Agistemus brasiliensis Matioli, Ueckermann & Oliveira (Stigmaeidae), in the presence or absence of B. phoenicis. Two experiments were carried out. In the first, a N. californicus female was placed in each leaf disc arena, with eggs of B. phoenicis and A. brasiliensis as food sources. In the second, an A. brasiliensis female was placed in each arena, with eggs of B. phoenicis and N. californicus as food sources. Adults of both predators were able to consume both types of eggs available as food sources, but they fed on considerably higher proportions of B. phoenicis than on eggs of the predator. Eggs of A. brasiliensis were not a suitable food source for N. californicus, which produced only 0.1 egg per female per day when only eggs of that species were present in the experimental unit. The results suggest that eggs of N. californicus were a suitable food source for A. brasiliensis, which oviposited 1.12 eggs per day, when only eggs of N. californicus were provided to the stigmaeid mite. The possible interactions among N. californicus, A. brasiliensis and B. phoenicis in citrus orchards are discussed.

The twospotted spider mite (TSSM), Tetranychus urticae Koch, is one of the main pests on strawberry crops in Brazil. TSSM can be difficult to control due to acaricide resistance. The objective of this work was to compare the effect of conventional and integrated strawberry production (ISP) systems on mite abundance and acaricide resistance. The control of TSSM in ISP was based on the release of Neoseiulus californicus (McGregor) or application of a selective acaricide (propargite), when TSSM monitoring indicated the timing for the release of predaceous mites (1-3 mites per leaflet on 30% leaflets) or chemical intervention (>10 mites per leaflet). Only acaricides (abamectin, fenpyroximate) were applied in the conventional system. Integrated control of TSSM were sufficient to maintain a significantly lower pest infestation level, resulting in a sixfold reduction in the frequency of acaricide applications, and consequently, a lower selection pressure for acaricide resistance. Strategies for the management of TSSM in strawberry fields are described and discussed.

Among the ectoparasites of cattle, Rhipicephalus (Boophilus) microplus (Canestrini) (Acari: Ixodidae) remains a major cause of economic losses to livestock. The chemical control with acaricides is still the most efficient method available to control ticks. The aims of this study were to diagnose resistance to amitraz in 16 tick populations from the States of São Paulo (14) and Paraná (2), using the larval immersion technique (LIT), and evaluate the effect of synergists [piperonyl butoxide (PBO), diethyl maleate (DEM), triphenyl phosphate (TPP)] on amitraz resistant and susceptible strains of cattle tick. Most of the evaluated populations (68.7 %) showed to be resistant to amitraz, with resistance ratios ranging from 2.14 to 132. The results suggest that the test procedure by LIT is sensitive and adequate for detection and monitoring of amitraz resistance in cattle tick. No synergistic effect was observed for the synergists PBO, DEM and TPP, on the amitraz resistant (Poa) strain of cattle tick, indicating that increased detoxification metabolism was not involved in this resistance.

Family Health Support Centers (NASF) were created in Brazil to increase the case-resolution capacity of primary healthcare. Prior to their implementation in the West Side of the city of São Paulo, Brazil, a series of workshops were held for primary healthcare professionals to prepare a proposal for such centers. Hermeneutic analysis was used to study the transcribed material. The thematic categories were: role, constitution, and functioning of the NASF, relationship with family health teams, and interdisciplinarity. The participants' expected the NASF to be an empowering device for comprehensiveness of care, intervening in an existing culture of unnecessary referrals while fostering linkage with other levels of care. The participants also expected the NASF to contribute to the discussion on health professionals' training and stimulating reflection with policymakers on health indicators based exclusively on the number of consultations. These indicators fail to reflect the impact on the services' activities and the quality of care offered to the population in the coverage area.

Studies on artificial laboratory selections with milbemectin, stability of milbemectin resistance and possible cross-resistance with abamectin were carried out with Tetranychus urticae Koch to provide basic information for a milbemectin resistance management program. Selections for resistance and susceptibility to milbemectin were performed in a population of T. urticae, collected from a commercial chrysanthemum field in the State of São Paulo, Brazil. After six selections for resistance and five selections for susceptibility, susceptible (S) and resistant (R) strains of T. urticae to milbemectin were obtained. The resistance ratio (R/S) at the LC(50) reached 409-fold value. The stability of milbemectin resistance was also studied under laboratory conditions, using a population with initial frequency of 75% of resistant mites. The frequencies of milbemectin resistance were evaluated monthly for a period of 7 months. In order to observe possible correlation between milbemectin and abamectin resistance, the frequencies of abamectin resistance were also evaluated for that population, during the same period. The frequency of milbemectin resistance decreased from 75 to 14.5%, while the percentage of abamectin resistant mites decreased from 57 to 9.1%, in 7 months. The frequencies of milbemectin and abamectin resistance were also evaluated in 25 field populations of T. urticae, collected from several crops in the State of São Paulo. The frequencies of milbemectin resistance varied from 4.1 to 89.5%, and of abamectin, from 7.0 to 90.5%. A positive and significant correlation was observed between the frequencies of milbemectin and abamectin resistance, indicating positive cross-resistance between these acaricides. The results indicate that abamectin should be avoided for managing milbemectin resistance in T. urticae. This is the first report on milbemectin resistance in T. urticae in Brazil.

Wnt and Sonic Hedgehog (Shh) signals are known to pattern the somite into dermomyotomal, myotomal and sclerotomal cell fates. By employing explants of presomitic mesoderm cultured with constant levels of Wnt3a conditioned medium and increasing levels of Shh, we found that differing levels of Shh signaling elicit differing responses from somitic cells: the lowest level of Shh signaling allows dermomyotomal gene expression, intermediate levels induce loss of dermomyotomal markers and activation of myogenic differentiation, and higher levels induce loss of myotomal markers and activation of sclerotomal gene expression. In addition, we have found that in the presence of high levels of Wnt signaling, instead of inducing sclerotomal markers, Shh signals act to maintain the expression of dermomyotomal and myotomal markers. One of the sclerotomal genes induced by high levels of Shh signaling is Nkx3.2. Forced expression of Nkx3.2 blocks somitic expression of the dermomyotomal marker Pax3 both in vitro and in vivo. Conversely, forced expression of Pax3 in somites can block Shh-mediated induction of sclerotomal gene expression and chondrocyte differentiation in vitro. Thus we propose that varying levels of Shh signaling act in a morphogen-like manner to elicit differing responses from somitic cells, and that Pax3 and Nkx3.2 set up mutually repressing cell fates that promote either dermomyotome/myotome or sclerotome differentiation, respectively.

The objective of this work was to study the population dynamics of Brevipalpus phoenicis (Geijskes) and predaceous mites (Phytoseiidae and Stigmaeidae), as well as the interactions among these mite species, in a coffee plantation in the State of São Paulo, Brazil. Tydeids were also evaluated because of the high frequency of these mites on coffee plants. Samples of leaves, branches and fruits were taken fortnightly, from April 2001 to June 2003, from plants randomly chosen in the coffee plantation. B. phoenicis mites were found on leaves in higher number during the drier periods of the year. Among the predaceous mites, Euseius citrifolius Denmark & Muma and Euseius concordis (Chant) were the most frequent species on the surface of leaves. During the evaluations, 72,534 domatia were cut and opened to remove the mites, from 6,360 leaves examined. Zetzellia malvinae Matioli, Ueckermann & Oliveira was the most frequent predator species found inside domatia. Significant correlations were observed between population dynamics of, among others, E. concordis and B. phoenicis, Z. malvinae and B. phoenicis, and Z. malvinae and E. concordis. Significant correlations were also verified between the number of domatia and the population densities of B. phoenicis, E. concordis, Lorryia sp. and Z. malvinae. Interactions between predator-prey and predator-predator on coffee plants are discussed. The influence of the meteorological factors temperature and precipitation on the most frequent mite species is also discussed.

The purpose of this study was to evaluate the performance of Neoseiulus californicus (McGregor) for the control of Tetranychus urticae Koch in commercial strawberry fields, under greenhouse conditions, in association or not with the use of acaricides. The N. californicus strain used in this study was tolerant or resistant to several pesticides. Three experiments were carried out in the State of São Paulo, Brazil. For the first experiment, the initial infestation of T. urticae was 87.1 active stages per leaflet. Two applications of propargite were made on the first and the 14th day of the experiment. Approximately 2 h after each propargite application, N. californicus was released at a rate of 3.0 and 1.9 adult mites per plant, respectively, for each application. The population of T. urticae decreased from 87.1 to 2.8 mites per leaflet in the first three weeks. After this period, the population of T. urticae was maintained at low levels (

The infesting species and their infestation indices of fruit flies were determined for eleven guava genotypes (Psidium guajava L.). From March to April 2000, ten mature fruits of each genotype were harvested at weekly intervals from insecticide unsprayed trees located in the municipality of Monte Alegre do Sul, SP, Brazil. Fruits were brought to the laboratory, weighed and placed in individual plastic cups containing sand at the bottom to obtain the tephritid pupae. About 95% of guavas produced fruit fly puparia. Of the 682 Anastrepha females recovered, four species were identified: A. fraterculus (Wied.) (86.5%), A. obliqua (Macquart) (10.8%), A. bistrigata Bezzi (1.8%) and A. sororcula Zucchi (0.9%). Three species of parasitoids Braconidae (Opiinae) were recovered: Doryctobracon areolatus (Szépligeti), Doryctobracon brasiliensis (Szépligeti) and Utetes anastrephae (Viereck). The genotypes differ in level of infestation depend on the collecting time. The genotypes 'L2P4 Vermelha', 'Ruby Suppreme' and 'Webber Suppreme' showed the lowest susceptibility to tephritids in terms of puparia per fruit. The variability of infestation among the guava genotypes and the reasons for increasing fruit fly infestations along the time were discussed.

The purpose of this study was to evaluate the cytochrome P450-dependent monooxygenase activities in methidathion resistant and susceptible strains of Amblyseius womersleyi Schicha. Artificial laboratory selections for resistance and susceptibility to methidathion were performed in an organophosphate resistant strain of A. womersleyi (Kanaya strain). Selections for susceptibility were also performed in a susceptible strain of this predaceous mite (Ishigaki Strain). After the selection process, the LC(50) of methidathion for the selected strains of A. womersleyi were 816 mg/l (Kanaya R), 4.61 mg/l (Kanaya S) and 1.59 mg/l (Ishigaki S). The monooxygenase activities were determined biochemically by the O-deethylation of 7-ethoxycoumarin (7-EC). The monooxygenase activity in adult females of Kanaya R strain (51.1 pmol/30 min/mg protein) was 3.60- and 5.42-fold higher than the activity observed for Kanaya S and Ishigaki S strains, respectively. Significant correlation between monooxygenase activity and LC(50) (mg/l) of methidathion was observed analyzing 16 populations of A. womersleyi with different susceptibilities to methidathion. Monooxygenase activity was also evaluated in different life stages (egg, larva, protonymph, deutonymph and adult) of A. womersleyi. The lowest activity was observed for the larval stage, which presented the highest susceptibility to methidathion. Protonymph, deutonymph and adult presented the highest monooxygenase activities. These stages were the most tolerant to methidathion. Monooxygenase activities of the Kanaya R strain were higher than of the Kanaya S strain in all developmental stages. The present study can be helpful for the implementation of a program involving release of insecticide-resistant populations of A. womersleyi in the field. The monooxygenase activity determination is easier and quicker than the estimation of LC(50), requiring fewer mites.

Prior work has established that transient Shh signals from the notochord and floor plate confer a competence in somitic tissue for subsequent BMP signals to induce chondrogenesis. We have therefore proposed that Shh induces a factor(s) that renders somitic cells competent to chondrify in response to subsequent BMP signals. Recently, we have shown that forced expression of Nkx3.2, a transcriptional repressor induced by Shh, is able to confer chondrogenic competence in somites. In this work, we show that administration of Shh or forced Nkx3.2 expression induces the expression of the transcription factor Sox9 in the somitic tissue. Forced expression of Sox9 can, in turn, induce robust chondrogenesis in somitic mesoderm, provided that BMP signals are present. We have found that in the presence of BMP signals, Sox9 and Nkx3.2 induce each other's expression. Thus, Nkx3.2 may promote axial chondrogenesis by derepressing the expression of Sox9 in somitic mesoderm. Furthermore, forced expression of either Sox9 or Nkx3.2 not only activates expression of cartilage-specific genes in somitic mesoderm, but also promotes the proliferation and survival of the induced chondrocytes in the presence of BMP signals. However, unlike Nkx3.2, Sox9 is able to induce de novo cartilage formation in non-cartilage-forming tissues. Our findings suggest that Shh and BMP signals work in sequence to establish a positive regulatory loop between Sox9 and Nkx3.2, and that Sox9 can subsequently initiate the chondrocyte differentiation program in a variety of cellular environments.

The study was carried out to evaluate the effect of some insecticides and fungicides on the interaction between predacious mites of the families Phytoseiidae and Stigmaeidae on citrus. Euseius citrifolius Denmark and Muma, Euseius concordis (Chant) (Phytoseiidae), and Agistemus aff. bakeri (Stigmaeidae) were the most abundant species of predacious mites. No significant reduction of the population of stigmaeids was detected following the pesticide treatment. On the contrary, the population of those mites increased after the application of methidathion, petroleum oil, deltamethrin, cupric oxychloride and cuprous oxide, as compared to the population of those mites in the control plots. The chemicals promoting increased stigmaeid populations are some of those that promoted reduction of phytoseiid populations. A significant negative correlation was found between the numbers of phytoseiids and stigmaeids on citrus leaves immediately before treatments, and 33, 50, 83 and 105 days after treatment. A clear negative linear correlation between the number of predacious mites from both families was detected for benomyl, cupric oxychloride, cuprous oxide, thiophanate methyl, and control. Phytoseiid-stigmaeid interaction on citrus orchard is discussed.