Resistance to hexythiazox in Brevipalpus phoenicis (Acari: Tenuipalpidae) from Brazilian citrus

The objective of this study was to collect baseline information for implementing an acaricide resistance management program of Brevipalpus phoenicis (Geijskes) to hexythiazox in Brazilian citrus groves. The egg susceptibility of B. phoenicis to hexythiazox was measured by a direct contact bioassay. The estimated LC50 for the S strain was 0.89 mg hexythiazox L(-1) of water (95% FL 0.75-1.03). After hexythiazox selection of a field-collected population associated with intense hexythiazox use, a resistance ratio greater than 10,000-fold was detected. Results from a survey revealed a great variability in the frequency of resistance in populations of B. phoenicis collected from citrus groves located in the State of S?o Paulo. No relationship was observed between the intensity of hexythiazox use and the frequency of resistance. Studies on dynamics of resistance showed that the resistance of B. phoenicis to hexythiazox is stable under laboratory conditions. Therefore, there is an urgent need to implement resistance management of B. phoenicis to hexythiazox in order to prolong its effective use in Brazilian citrus groves.     

Resistance of Brevipalpus phoenicis (Geijskes) (Acari: Tenuipalpidae) to acaricides that inhibit cellular respiration in citrus: cross-resistance and fitness cost

Acaricides that inhibit cellular respiration play an important role in the control of Brevipalpus phoenicis (Geijskes) in citrus groves in Brazil. Studies were conducted to evaluate: (a) the variability in the susceptibility among B. phoenicis populations collected from citrus groves to cyhexatin, azocyclotin, propargite and sulphur; (b) cross-resistance relationships between propargite and azocyclotin, cyhexatin, dinocap, pyridaben and sulphur; and (c) the fitness cost associated with propargite resistance in B. phoenicis under laboratory conditions. A residual-type contact bioassay was used to characterize the susceptibility. The susceptibility was estimated with diagnostic concentrations based LC(95) of each acaricide. The cross-resistance was evaluated by characterizing the concentration-mortality responses of susceptible (S) and propargite-resistant (Propargite-R) strains. The fitness cost was evaluated by measuring the biological parameters of S and Propargite-R strains on citrus fruits at 25 +/- 1 degrees C and fotophase of 14h. Significant differences in the susceptibility of B. phoenicis were detected at diagnostic concentration of cyhexatin (survivorship from 16.3% to 80.5%), azocyclotin (from 3.0% to 15.0%), propargite (from 1.0% a 71.6%) and sulphur (from 9.0% to 82.6%). A low intensity of cross-resistance was detected between propargite and the acaricides azocyclotin (1.8-fold), cyhexatin (4.6-fold), dinocap (3.5-fold) and pyridaben (3.5-fold). On the other hand, the intensity of cross-resistance to sulphur (> 111-fold) was very high. There was no fitness cost associated with B. phoenicis resistance to propargite, based on biological parameters evaluated. Therefore, the use of these acaricides should also be done very carefully in resistance management of B. phoenicis to acaricides.     

Coffee Ringspot Virus Vectored by Brevipalpus phoenicis (Acari: Tenuipalpidae) in Coffee

Coffee ringspot is characterized by conspicuous ringspot symptoms on leaves, berries, and less frequently on twigs. It is caused by coffee ringspot virus (CoRSV), a short, bacilliform virus (40 nm × 100–110 nm). The virus is not seed borne and is transmitted by Brevipalpus phoenicis (Geijskes). Transovarial transmission within the mite does not occur. CoRSV has been mechanically transmitted to Chenopodium amaranticolor Coste and Reynaud, C. quinoa Wildenow, Beta vulgaris L., and Alternanthera tenella Colla resulting in local lesions. Systemic infection within both C. amaranticolor and C. quinoa occurs. Virions are found in the nucleus or cytoplasm of infected cells, commonly associated with membranes. Occasionally, membrane bounded particles are found within the cisternae of the endoplasmic reticulum. A characteristic electron lucent, nuclear inclusion is commonly found in many infected cells. These cytopathic effects place CoRSV among the nuclear type of Brevipalpus-borne viruses. The disease has been reported in several Brazilian states (São Paulo, Paraná, Minas Gerais, and Federal District) and recently found in Costa Rica. A similar disease is known in the Philippines, but no information exists about its relationship to CoRSV. Coffee ringspot had no economical significance until recently when a large scale infection was reported in Minas Gerais that resulted in yield loss.     

Effect of culture media on virulence of Hirsutella thompsonii (Fischer) (Deuteromycetes) to control Brevipalpus phoenicis (Geijskes) (Acari: Tenuipalpidae)

The virulence of Hirsutella thompsonii (Fischer) to Brevipalpus phoenicis (Geijskes) was evaluated in laboratory, grown on complete and solid culture media (MC-S); complete and liquid culture media (MC-L); rice (APC) and powdered rice (APC-SM). Adults were confined to arenas prepared with citrus leaves in acrylic dishes containing water-agar. Conidial suspensions were prepared at different concentration (3.2 x 10(5) to 1 x 10(7) spores/ml) and applied on mites to establish the table curve-response on fourth day. For field evaluation, adults were maintained in arenas prepared with fruits which were placed in plants. In this test, four treatments were tried: H. thompsonii cultured on rice (APC) at two concentrations (20 kg/ha and 10 kg/ha), H. thompsonii produced by liquid fermentation (MC-L) (5 L/ha) and control (sterile water). Adult survival, number of eggs and nymphs per fruit were observed 10 and 20 days after the fungus application. The lowest LC25 value calculated was from pathogen produced in MC-S (1.9 x 10(5) conidia/ml).The LC25 values calculated to APC and APC-SM did not differ statistically. The LC25 values to MC-L and MC-S were 1.9 x 10(6) infective cells/ml and 2.2 x 10(5) conidia/ml. In the field, concentration and time to death differed between treatments and control. The applications resulted in reduction of adult survival and number of eggs.     

Biology of Agistemus brasiliensis Matioli, Ueckermann & Oliveira (Acari: Stigmaeidae) and its predation potential on Brevipalpus phoenicis (Geijskes) (Acari: Tenuipalpidae)

The present work aimed to study the biology of Agistemus brasiliensis Matioli, Ueckermann & Oliveira at the following temperatures, namely 15, 20, 25, 30 and 35 degrees C, fed with Brevipalpus phoenicis (Geijskes) and Typha pollen, in laboratory conditions. Life tables were calculated to evaluate the biological parameters. The optimal development of A. brasiliensis took place at 29 degrees C. The values of T (time of generation - days), R0 and r m at 30 degrees C were, 13.95, 16.25 and 0.20, respectively. The prey consumption of A. brasiliensis was studied at the densities of 5, 10, 20, 40 and 60 leprosis mite females per cage (3 cm in diameter) on citrus fruits at 29 degrees C. The maximum prey, namely 7.6 B. phoenicis females per day, were consumed at a density of 20 leprosis mites. At densities of above 40 leprosis mites per cage, A. brasiliensis oviposits 4.7 eggs per day, in comparison to 2.5 eggs per day at 20 mites per cage.     

Citrus Leprosis Virus Vectored by Brevipalpus phoenicis (Acari: Tenuipalpidae) on Citrus in Brazil

Citrus leprosis is caused by Citrus leprosis virus (CiLV) that is transmitted by mites in the genus Brevipalpus (Acari: Tenuipalpidae). This disease directly reduces production and the life span of the citrus plant. The main symptoms of the disease include lesions on fruits, leaves, and twigs or small branches, causing premature fruit drop, defoliation, and death of the twigs or branches leading to serious tree decline. Leprosis is a highly destructive disease of citrus, wherever it occurs. The Brazilian citrus industry spends over 100 million US dollars annually on acaricides to control the vector, Brevipalpus phoenicis (Geijskes). This review contains information about the history of the etiology of citrus leprosis, its geographical distribution, host range, the role of the mite vectors, viral morphology and relationships with the infected cell, and transmissibility of the virus by the mite. In addition, data on the mite-virus-plant relationship, disease damage, and strategies for controlling disease spread are presented.     

Mitochondrial DNA and RAPD polymorphisms in the haploid mite Brevipalpus phoenicis (Acari: Tenuipalpidae)

Brevipalpus phoenicis (Geijskes) (Acari: Tenuipalpidae) is recognized as the vector of citrus leprosis virus that is a significant problem in several South American countries. Citrus leprosis has been reported from Florida in the past but no longer occurs on citrus in North America. The disease was recently reported in Central America, suggesting that B. phoenicis constitutes a potential threat to the citrus industries of North America and the Caribbean. Besides B. phoenicis, B. obovatus Donnadieu, and B. californicus (Banks) have been incriminated as vectors of citrus leprosis virus and each species has hundreds of host plants. In this study, Brevipalpus mite specimens were collected from different plants, especially citrus, in the States of Florida (USA) and São Paulo (Brazil), and reared on citrus fruit under standard laboratory conditions. Mites were taken from these colonies for DNA extraction and for morphological species identification. One hundred and two Random Amplified Polymorphic DNA (RAPD) markers were scored along with amplification and sequencing of a mitochondrial cytochrome oxidase subunit I gene fragment (374 bp). Variability among the colonies was detected with consistent congruence between both molecular data sets. The mites from the Florida and Brazilian colonies were morphologically identified as belonging to B. phoenicis, and comprise a monophyletic group. These colonies could be further diagnosed and subdivided geographically by mitochondrial DNA analysis.     

Influence of soil applied nitrogen (NPKS 25: 5: 5:5) on Brevipalpus phoenicis Geijskes (Acari: Tenuipalpidae) mite incidence and damage symptoms on tea

Application of nitrogenous fertilisers (NPKS 25: 5: 5:5) for yield of tea and to induce tolerance to the red crevice ( Brevipalpus phoenicis Geijskes) mite was found to be rates between 150 and 200 Kg N ha^-1 yr^-1 rates above 400 Kg N ha^-1 encouraged the build up of mites on tea. Damage symptoms resulting from mite infestation declined with increase in application of nitrogenous fertilisers.
The levels of nitrogen in the leaf tissues increased with soil applied nitrogen and also was related to the mite numbers found on the leaves suggesting a link. The symptoms of the discoloured necrotic leaf base that accompanied mite infestation was significantly ( P ≤ 0.05) and linearly correlated (r = 0.60) to mite numbers in the control plot. The levels of applied nitrogen were negatively correlated to % leaf necrosis suggesting a tolerance leading to reduction in necrotic symptoms with fertiliser applications.     

Effects of radiation (Cobalt-60) on the elimination of Brevipalpus phoenicis (Acari: Tenuipalpidae) Cardinium endosymbiont

Brevipalpus phoenicis (Geijskes) (Acari: Tenuipalpidae) is a polyphagous mite with worldwide distribution and it is also a vector of several plant viruses. In citrus, B. phoenicis transmits Citrus leprosis virus (CiLV), the causal agent of leprosis, a disease that costs millions of dollars per year for its prevention and control. Brevipalpus phoenicis mites reproduce through thelytokous parthenogenesis, producing haploid females. This characteristic is attributable to the presence of an endosymbiont bacterium of the genus Cardinium; however, very little is known about the biological and ecological implications of the presence of this endosymbiont in Brevipalpus mites. In order to investigate the role of Cardinium in the transmission of CiLV to citrus plants, our goal was to eliminate the bacterium from the mite. We assessed the effectiveness of different doses of radiation from a Cobalt-60 source to cure B. phoenicis populations from Cardinium sp. The efficiency of irradiation on the elimination of the endosymbiont was determined by counting the number of females and males obtained in the F(1) generation after irradiation and confirming the presence of the endosymbiont by PCR. Both radiation treatments influenced the oviposition period and the number of eggs laid by irradiated females. Also, irradiation eliminated the Cardinium endosymbiont and increased the number of males in progeny of the exposed populations. Although macroscopic morphological abnormalities were not observed among the treated mites, the mortality was higher compared to the non-irradiated control group.     

Passion Fruit Green Spot Virus Vectored by Brevipalpus phoenicis (Acari: Tenuipalpidae) on Passion Fruit in Brazil

Passion fruit green spot disease was first identified in 1997 after a severe outbreak at Vera Cruz County, state of São Paulo, Brazil. Mature yellow fruits of Passiflora edulis Simms f. flavicarpa Degener showed characteristic green spots, 2–5 mm in diameter and patches of green tissues were present on senescent leaves. The devastating effect to passion flower is caused by necrotic lesions that encircle the stems and kill the plant. In severe cases, entire orchards of a few hectares in size have been completely destroyed. The disease was always preceded by heavy infestations of Brevipalpus phoenicis (Geijskes) (Acari: Tenuipalpidae). Transmission electron microscopy of affected tissues (fruits, leaves, and stems) consistently revealed the presence of short, bacilliform particles (50–70 nm × 100–120 nm) in the cisternae of the endoplasmic reticulum, as well as the presence of a dense viroplasm in the cytoplasm. This cytopathic effect has been found in several other Brevipalpus-transmitted or associated viruses and is classified as a cytoplasmic type of disease. Experimental reproduction of the leaf and stem symptoms was achieved by transferring B. phoenicis collected from affected field passion flower plants onto healthy plants. The evidence supports a viral etiology for the disease and the agent was named passion fruit green spot virus. Its relationship with other B. phoenicis related viruses continues to be studied. The disease was also found in the Brazilian states of Bahia, Sergipe, Rondonia, Minas Gerais, Rio de Janeiro, and in the Federal District. Use of one or more of the following acaricides (hexythiazox, fenbutatin-oxide, propargite, quinomethionate, or dicofol) has significantly reduced the incidence of the disease.     

Morphological Observations on Brevipalpus phoenicis (Acari: Tenuipalpidae) Including Comparisons with B. californicus and B. obovatus

The genus Brevipalpus has over 300 species worldwide. The three most important agricultural pest species in the genus, Brevipalpus californicus (Banks), B. obovatus Donnadieu, and B. phoenicis (Geijskes), have been consistently confused and misidentified for more than 50 years. The present study provides a discussion of the characters and character states used to separate these mites. Low-temperature scanning electron microscopy and traditional light microscopy techniques were used to illustrate the subtle morphological differences between these three species. Morphology of the dorsal propodosoma, opisthosoma, and leg chaetotaxy of all three species was examined and compared. The number of dorsal setae, the number of solenidia (omega) on tarsus II, and dorsal cuticular patterns were the most important characters in the identification of Brevipalpus species. B. phoenicis is similar to B. californicus in having two omega on tarsus leg II and different from B. obovatus which has only one omega on tarsus leg II and similar to B. obovatus in having only one pair of F setae (f3), but differing from B. californicus which has two pairs of F setae (f2-3). The dorsal opisthosomal and propodisomal cuticular patterns frequently used to distinguish between these three species are useful but one must be aware that age, feeding, and mounting techniques can affect the appearance of these characters.     

Predation of Galendromus helveolus (Acari: Phytoseiidae) on Brevipalpus californicus (Acari: Tenuipalpidae)

The false spider mite, Brevipalpus californicus (Banks), is a major citrus pest in Texas. This phytophagous mite causes damage to fruit, leaves and stems of citrus, and is also a vector of citrus leprosis virus. Galendromus helveolus (Chant) is one of the most prevalent predacious mite species found on Texas citrus. The predation potential of G. helveolus on different stages of B. californicus was evaluated in the laboratory. An individual immature G. helveolus mite consumed an average of 30.7 eggs, 53.6 larvae, or 22.7 nymphs of B. californicus before developing to an adult. An individual adult female G. helveolus consumed an average of 164.8 eggs, 369.6 larvae, or 80.9 nymphs of B. californicus. Both immature and adult G. helveolus never fed on adult stage of B. californicus and never completed development. The development times of the immature stages of G. helveolus were 4.5, 4.1 and 4.6 days when fed on eggs, larvae and nymphs of B. californicus, respectively. When G. helveolus fed on the larval stage of B. californicus, the adults had the longest longevity (18.5 days) and the highest fecundity (14.0 eggs/female).     

Population dynamics of phytophagous and predaceous mites on coffee in Brazil, with emphasis on Brevipalpus phoenicis (Acari: Tenuipalpidae)

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.     

Reproductive performance of the mite Brevipalpus phoenicis (Geijskes, 1939) on citrus and coffee, using life table parameters.

The flat-mite Brevipalpus phoenicis (Geijskes, 1939) (Acari: Tenuipalpidae) is considered important in citrus (Citrus spp.) and coffee plants (Coffea spp.) in Brazil, and is known as the leprosis and coffee ring spot mite, as being a vector of the Citrus Leprosis Rhabdovirus - CitLV and Coffee Ring Spot Virus - CoRSV. The objective of this work is to find out about the reproductive success of B. phoenicis on citric fruits and coffee leaves by fertility life table parameters and its biology. The experiments were carried out in laboratory conditions at 25 +/- 2 degrees C, 70 +/- 10% of relative humidity and 14 h of photophase. The lengths of embryonic and post-embryonic periods were different due to the host where the mite was reared. B. phoenicis showed better development and higher survival and fecundity in citric fruits than coffee leaves. The intrinsic rate of the population increase (r(m)) was 0.128 and 0.090 - females/female/day on citric fruits and coffee leaves, respectively. The citric fruits were more appropriate for the development of B. phoenicis than coffee leaves.     

Cryptic diversity in Brevipalpus mites (Tenuipalpidae)

Defining the taxonomic identity of organisms is a prerequisite for their study, and in the case of economically important species, misidentification may lead to the application of inappropriate prevention and control strategies. Flat mites of the Brevipalpus genus include several crop pests and the systematics of these mites represents a challenge for acarologists. Many of the most economically important Brevipalpus species have repeatedly been inaccurately identified. Such problematic classification has been attributed to the likely occurrence of cryptic species in the genus. In this study, we used an integrative approach that combined molecular analyses, including sequence‐based species delimitation, with detailed morphological identification using traits that have recently showed to be taxonomically informative. Sequences of mitochondrial cytochrome c oxidase subunit I (COI) were obtained from individuals collected from host plants belonging to 14 genera and 13 families across 29 locations in the Americas (Brazil, Chile, USA). The phylogenetic analyses included previously published Brevipalpus sequences from GenBank, and the final data set was classified into 44 haplotypes. Six putative species were recognised by COI‐based species delimitation analysis, and morphological evidence supported each of these species. The integrative approach revealed the occurrence of cryptic species in the Brevipalpus genus and contributed to the clarification of previously noted incongruences. The results presented here allow for the evaluation of taxonomic characteristics in a phylogenetic context and indicate new characters for the differentiation of Brevipalpus species. In addition, Brevipalpus incognitus n. sp. Ferragut & Navia, a cryptic species detected in this study, is described based on morphological and molecular traits. Implications of the advances in Brevipalpus systematics presented herein with respect to pest management are briefly discussed.     

Brevipalpus mites (Acari: Tenuipalpidae): vectors of invasive, non-systemic cytoplasmic and nuclear viruses in plants

Multi-directional interactions occur among plant hosts, Brevipalpus mites and the plant viruses they transmit. Such interactions should be considered when evaluating the severity of a disease such as citrus leprosis. The current understanding of Brevipalpus-transmitted viruses relies on the capability of the vector to transmit the disease, the persistence of the virus in the host plant and the ability of the disease to spread. Previously, we discussed the Citrus leprosis virus (CiLV) and its importance and spread over the past decade into new areas of South and Central America, most recently into southern Mexico and Belize. Here, we address key questions to better understand the biology of the mite vector, fitness costs, and the peculiarities of Brevipalpus mite reproduction, virus survival, transmissibility and spread, and the expansion of the host plant range of Brevipalpus species vectoring the disease.     

Evaluation of the predatory mite Amblyseius hainanensis (Acari: Phytoseiidae) and artificial rainfall for the management of Brevipalpus obovatus (Acari: Tenuipalpidae)

Brevipalpus obovatus Donnadieu is an important pest mite on tea plants in South China. In the current study, predatory mites of B. obovatus in the tea gardens of Guangzhou were extensively surveyed. In total, 13 species of predatory mites (four families with seven genera) were recorded. The population proportion of Amblyseius hainanensis Wu et Qian was the highest (68.6?%), followed by that of Anystis baccarum (L.) (8.4?%) and A. theae Wu (6.3?%). The effects of starvation time, habitat size and pest population density on the predatory efficiency of the most dominant species, A. hainanensis, feeding on B. obovatus were assessed. In addition, the effectiveness of artificial rainfall in reducing B. obovatus populations was evaluated. After starvation for 48?h, the predatory efficiency of A. hainanensis was significantly higher than those that had been starved for 24 or 72?h when 30-50 B. obovatus eggs were made available. The predation of A. hainanensis on B. obovatus also increased with increasing prey density. The number of prey attacked by A. hainanensis in a 3.2?cm(2) habitat was significantly higher than in a 6.3?cm(2) habitat. The average predation of A. hainanensis was 31.7 eggs per day when offered 100 B. obovatus eggs on a tea leaf. This decreased to 17.8 eggs per day when four A. hainanensis shared 100 B. obovatus eggs. B. obovatus populations can be reduced by artificial rainfall, with the reduction affected by rainfall intensity. With an intensity of 40?mm in 15?min, 90.2?% mortality of B. obovatus occurred; lower mortalities were recorded (13.3 and 29.8?%) when the intensity was 2 or 4?mm in 15?min. Combination of the predatory mite A. hainanensis and artificial rainfall for the integrated pest management of B. obovatus is discussed.