Overview of a classical biological control project directed against the red palm mite in Florida

Information is provided on the steps involved in classical biological control programs, with an outline of the steps achieved in the classical biological control of the red palm mite, Raoiella indica (Acari: Tenuipalpidae), in Florida. An overview is provided of the results and an accompanying paper by Bowman and Hoy (2012) describes the molecular analyses conducted to characterize phytoseiid predators of the red palm mite collected from Mauritius. The Mauritius phytoseiids were identified as Amblyseius largoensis, using morphological criteria, and compared to four populations of A. largoensis from Florida. The difficulties encountered in the systematics, rearing, and evaluation of the Mauritius phytoseiids are described. A method was tested for obtaining DNA from single mites without destroying the body so that voucher specimens can be maintained. Ultimately, the project was terminated due to constraints expected in obtaining permission to make releases.     

Toxicity of acaricides to Raoiella indica and their selectivity for its predator, Amblyseius largoensis (Acari: Tenuipalpidae: Phytoseiidae)

Raoiella indica Hirst (Acari: Tenuipalpidae) is considered a pest of coconut palm in Asia and the Middle East. This mite was recently introduced in the Americas, where it spread to several countries and expanded its range of hosts, causing heavy losses to coconut and banana production. The phytoseiid mite Amblyseius largoensis (Muma) is one of the predators most often encountered in coconut palms. Because the current prospects for the control of R. indica in the New World indicate the use of acaricides and the management of their natural enemies, the objective of this study was to evaluate the toxicity of selected acaricides to R. indica and the selectivity (i.e., toxicity to the predator relative to toxicity to the prey) for A. largoensis. Assays were performed by the immersion of banana leaf discs in acaricide solutions, followed by the placing of adult females of the pest or predator on the discs. Mortality of the mites was evaluated after 24 h, and the data obtained were subjected to probit analysis. Abamectin, fenpyroximate, milbemectin and spirodiclofen were the products most toxic to R. indica adults, whereas fenpyroximate and spirodiclofen were the most selective for A. largoensis.     

Prey-stage preferences and functional and numerical responses of Amblyseius largoensis (Acari: Phytoseiidae) to Raoiella indica (Acari: Tenuipalpidae)

Raoiella indica Hirst (Acari: Tenuipalpidae) is a phytophagous mite that recently invaded the Western Hemisphere. This mite is a multivoltine and gregarious species that can reach very high population densities and cause significant damage to various palm species (Arecaceae). The predatory mite Amblyseius largoensis (Muma) (Acari: Phytoseiidae) has been found associated with R. indica in Florida. This study evaluated A. largoensis for potential to control R. indica by (1) determining predator preferences among developmental stages of R. indica, and (2) estimating predator functional and numerical responses to varying densities of its most preferred prey-stage. Under no-choice conditions A. largoensis consumed significantly more eggs than other stages of R. indica. In choice tests A. largoensis showed a significant preference for R. indica eggs over all other prey stages. Amblyseius largoensis displayed a type II functional response showing an increase in number of prey killed with an increase in prey population density. Consumption of prey stabilized at approximately 45 eggs/day, the level at which oviposition by the predator was maximized (2.36?±?0.11 eggs/day; mean?±?SEM). Results of this study suggest that A. largoensis can play a role in controlling R. indica populations, particularly when prey densities are low.     

First Report of <Emphasis Type="Italic">Raoiella indica</Emphasis> (Hirst) (Acari: Tenuipalpide) in Southern Brazil

The red palm mite (RPM), Raoiella indica (Hirst) (Acari: Tenuipalpidae), was found for the first time in the Paraná State, in southern Brazil. The first observations occurred in September 2015, on strawberry (Fragaria × ananassa Duch) leaves, which is not considered a typical host plant of RPM. It is probable that its occurrence on this plant was serendipitous. Visual surveys for RPM were carried out on four typical host plants (banana, coconut, foxtail palm, and real palm), in five cities of the Paraná State (Bela Vista do Paraíso, Londrina, Maringá, Marialva, and Sarandi). RPM was found on each of the four typical host plants, in each of the five cities. Our survey extends RPM occurrence to the southern region of Brazil and indicates that the pest could be widespread in the country.     

Effect of coconut palm proximities and Musa spp. germplasm resistance to colonization by Raoiella indica (Acari: Tenuipalpidae)

Although coconut (Cocos nucifera L.) is the predominant host for Raoiella indica Hirst (Acari: Tenuipalpidae), false spider mite infestations do occur on bananas and plantains (Musa spp. Colla). Since its introduction, the banana and plantain industries have been negatively impacted to different degrees by R. indica infestation throughout the Caribbean. Genetic resistance in the host and the proximity of natural sources of mite infestation has been suggested as two of the main factors affecting R. indica densities in Musa spp. plantations. Greenhouse experiments were established to try to determine what effect coconut palm proximities and planting densities had on R. indica populations infesting Musa spp. plants. Trials were carried out using potted Musa spp. and coconut palms plants at two different ratios. In addition, fourteen Musa spp. hybrid accessions were evaluated for their susceptibility/resistance to colonization by R. indica populations. Differences were observed for mite population buildup for both the density and germplasm accession evaluations. These results have potential implications on how this important pest can be managed on essential agricultural commodities such as bananas and plantains.     

Unique advantages of using low temperature scanning electron microscopy to observe bacteria

Summary Electron microscopy (EM) has greatly helped to elucidate our understanding of bacterial structure and function. However, several recent studies have cautioned investigators about artifacts that result from the use of conventional EM preparation procedures. To avoid these problems, the use of low temperature scanning electron microscopy (LTSEM) was evaluated for examining frozen, fully hydrated specimens. Spinach leaves (Spinacia oleracea L. cv. New Jersey), which were naturally infected or inoculated with bacteria, were used as the experimental material. 1 cm segments of the infected leaves were plunge frozen in liquid nitrogen, transferred to a cryochamber for sputter coating and then moved onto a cryostage in an SEM. After observation, some of the frozen, hydrated leaf segments were transferred onto agar medium to determine whether preparation for LTSEM was nondestructive to the bacteria. The other tissue segments were chemically fixed by freeze-substitution. The results indicated that after cryopreparation and observation in the LTSEM: (i) viable bacteria, which were recovered from the leaf sample, could be cultured on agar medium for subsequent study, and (ii) the frozen samples could be freeze substituted and embedded so that transmission electron microscopic (TEM) observations could be carried out on the same specimen. In conclusion, frozen, hydrated leaf tissue infected with bacteria can be observed using LTSEM and then can be either processed for TEM observation to obtain further structural details or recovered to culture the pathogenic bacteria for supplementary studies.Abbreviations EPS extracellular polysaccharide - EM electron microscopy - LTSEM low temperature scanning electron microscopy - SEM scanning electron microscopy - TEM transmission electron microscopy - TSA tryptic soy agar - TSB tryptic soy broth Dedicated to Professor Eldon H. Newcomb in recognition of his contributions to cell biology     

Comparison of two populations of the pantropical predator Amblyseius largoensis (Acari: Phytoseiidae) for biological control of Raoiella indica (Acari: Tenuipalpidae)

The red palm mite, Raoiella indica Hirst (Acari: Tenuipalpidae), was recently introduced in the Americas. It spread quickly throughout coconut palm growing areas, expanding considerably its host range. The invasion of this species has caused high economic impact in several countries. In Brazil, extensive areas are expected to be affected. For logistical reasons and other concerns, chemical control does not seem desirable for the control of this pest in most Latin American countries. Biological control of R. indica by introducing exotic natural enemies seems to be an important control measure to be considered. Surveys in many countries have shown that Amblyseius largoensis (Muma) (Acari: Phytoseiidae) is a very common predator on coconut palms. This study compared the biology of a population of A. largoensis found for a long time in association with R. indica in La Reunion Island (Indian Ocean) with a population from Roraima State (northern Brazil), where R. indica was first found about two and a half years ago. No significant differences were observed between populations in relation to the duration of different immature stages or total survivorship. However, the oviposition period, prey consumption and net reproductive rate were significantly higher for the La Reunion population, warranting further investigation to determine whether that population should be released in Roraima to control the pest.     

Variability in response of four populations of Amblyseius largoensis (Acari: Phytoseiidae) to Raoiella indica (Acari: Tenuipalpidae) and Tetranychus gloveri (Acari: Tetranychidae) eggs and larvae

Raoiella indica (Acari: Tenuipalpidae) is a phytophagous mite that recently invaded the Neotropical region. A predatory mite Amblyseius largoensis (Acari: Phytoseiidae) has been found associated with R. indica in Florida. This study evaluated A. largoensis by determining its likelihood of consuming eggs and larvae of R. indica and Tetranychus gloveri (Acari: Tetranychidae) under no-choice and choice conditions. To detect variations in the response of A. largoensis to R. indica, four populations of predators were examined: (1) predators reared exclusively on R. indica in the laboratory for 2 years, (2) predators reared on T. gloveri in the laboratory for 2 months but reared on R. indica for 2 years previously, (3) predators collected from a field infested with R. indica, and (4) predators collected from a field that had never been infested with R. indica. Results of this study suggest that A. largoensis is likely to accept and consume high numbers of R. indica eggs regardless of their previous feeding experience. In contrast, all populations consumed relatively fewer R. indica larvae than the other prey tested. Predators previously exposed to R. indica were more likely to consume R. indica larvae. By contrast, predators not previously exposed to R. indica showed the lowest likelihood of choosing to feed on this prey item. Plasticity in the response of A. largoensis to R. indica larvae could be associated to selection, learning, or a combination of both. The possible implications of the observed differences in terms of biological control of R. indica are discussed.     

Study of lichens with different state of hydration by the combination of low temperature scanning electron and confocal laser scanning microscopies.

The use of techniques such as low temperature scanning electron microscopy (LTSEM) and confocal laser scanning microscopy (CLSM) allows the study of lichen thalli in different states of hydration and also near the natural state. The spatial organization of desiccated thalli, with reduced, very compact algal layers, is different from that of hydrated ones. Sometimes, the observation with transmission electron microscopy (TEM) of photobiont pyrenoids from desiccated thalli reports pyrenoids with a central part of a weak stained matrix lacking pyrenoglobuli, named "empty zones". "Empty zones" are not distinguishable with LTSEM and do not present immunolabelling with rubisco antibody in TEM. These zones could be originated by an expansion process during rehydration produced in chemical fixation.     

Low temperature scanning electron microscopy of the superficial envelope of canine chondrocytes in culture

Low temperature scanning electron microscopy (LTSEM) has been employed to examine the surface morphology of chondrocyte cultures on ceramic granules. Well-established cultures on porous hydroxyapatite consist of ceramic cores overlaid and interspersed with a cellular matrix of collagen and proteoglycan (Cheung, 1985); of especial interest is the superficial layer of cells. These cells are believed, on the basis of immuno-light microscopy (Gardner et al., 1987), to be coated by an hydrated porous envelope of collagen/proteoglycan which is likely to obscure cell outlines. This relationship is confirmed by enzymic digestion of the superficial material. Post-digestion LTSEM examination of the fully hydrated preparations establishes the existence of arrays of rounded structures identified as superficial cells.     

Development and reproduction of <Emphasis Type="Italic">Amblyseius largoensis</Emphasis> (Acari: Phytoseiidae) feeding on pollen, <Emphasis Type="Italic">Raoiella indica</Emphasis> (Acari: Tenuipalpidae), and other microarthropods inhabiting coconuts in Florida,USA

The red palm mite, Raoiella indica (Acari: Tenuipalpidae), is an important pest of palms (Arecaceae) and other species within the Zingiberaceae, Musaceae and Strelitziaceae families. Raoiella indica was discovered in the USA (Palm Beach and Broward counties, Florida) late in 2007, and it subsequently spread to other Florida counties. The predatory mite Amblyseius largoensis (Acari: Phytoseiidae) has been found associated with R. indica in Florida. In order to verify whether A. largoensis can develop and reproduce when feeding exclusively on R. indica, the biology of this predator was evaluated on various food sources, including R. indica. Five diets [R. indica, Tetranychus gloveri¸ Aonidiella orientalis, Nipaecocus nipae, oak (Quercus virginiana) pollen] and a no-food control were tested to determine the predators’ development, survivorship, oviposition rate, sex ratio and longevity at 26.5 ± 1°C, 70 ± 5% RH and a 12:12 L:D photophase. Amblyseius largoensis was able to complete its life cycle and reproduce when fed exclusively on R. indica. The development of immature stages of A. largoensis was faster and fecundity and survivorship were higher when fed on R. indica or T. gloveri compared to the other food sources. The intrinsic rate of natural increase of A. largoensis was significantly higher when fed on R. indica than on other diets. These results suggest that, despite earlier assessments, A. largoensis can play a role in controlling R. indica.     

Geographic distribution and host plants of Raoiella indica and associated mite species in northern Venezuela

The red palm mite (RPM), Raoiella indica Hirst (Acari: Tenuipalpidae), is an invasive pest in the New World, where it is currently considered a serious threat to coconut and banana crops. It was first reported from northern Venezuela in 2007. To determine its current distribution in this country, surveys were carried out from October 2008 to April 2010 on coconut (Cocos nucifera L.), banana (Musa spp.), ornamental plants and weeds in northern Venezuela. Higher population levels of RPM were registered on commercial coconut farms in Falcón and Sucre states but also on other plant species naturally growing along the coastal line in Anzoategui, Aragua, Carabobo, Monagas and Nueva Esparta states. Out of 34 botanical species evaluated, all RPM stages were observed only on eight arecaceous, one musaceous and one streliziaceous species, indicating that the pest developed and reproduced only on these plants. Mite specimens found on weeds were considered spurious events, as immature stages of the pest were never found on these. Amblyseius largoensis (Muma) (Acari: Phytoseiidae) was the most frequent predatory mite associated with RPM in all sampling sites. The results indicate that RPM has spread to extensive areas of northern Venezuela since its initial detection in Güiria, Sucre state. Considering the report of this pest mite in northern Brazil in the late 2009, additional samplings in southern Venezuela should be carried out, to evaluate the possible presence of RPM also in that region.     

Reproductive compatibility and genetic and morphometric variability among populations of the predatory mite,Amblyseius largoensis (Acari: Phytoseiidae), from Indian Ocean Islands and the Americas

The red palm mite (RPM), Raoiella indica Hirst (Acari: Tenuipalpidae), is an invasive phytophagous mite that was recently introduced into The Americas. The predatory mite Amblyseius largoensis Muma (Acari: Phytoseiidae) has been the only natural enemy consistently found in association with RPM. This study aimed to determine if A. largoensis populations from the Indian Ocean Islands (La Réunion and Mauritius) and the Americas (Brazil, Trinidad and Tobago and the USA) consist a taxonomic unit or a group of cryptic species. First, the morphological variability among the A. largoensis populations from these areas was evaluated through morphometric analyses of 36 morphological traits. Then, their genetic variability and phylogenetic relationships were assessed based on two target DNA fragments: the nuclear Internal Transcribed Spacer and the mithochondrial 12S rRNA. Finally, reproductive compatibility of the populations from La Réunion and Roraima, Brazil was evaluated. Morphometric differences between the A. largoensis specimens from La Réunion Island and the Americas were observed, most of them on the length of the setae. Molecular analysis indicated that the A. largoensis populations from the Indian Ocean Islands and the Americas belong to the same taxonomic entity, although to two well defined genetic groups. Crossings involving the A. largoensis populations from La Réunion Island and Roraima, Brazil revealed complete reproductive compatibility between these populations. Information on the morphometric and genetic variability among studied A. largoensis populations can be further exploited in future studies to follow colonization of Indian Ocean Islands populations in the Americas, in the case of field releases.     

Potential geographical distribution of the red palm mite in South America

Among pests that have recently been introduced into the Americas, the red palm mite, Raoiella indica Hirst (Prostigmata: Tenuipalpidae), is the most invasive. This mite has spread rapidly to several Caribbean countries, United States of America, Mexico, Venezuela, Colombia and Brazil. The potential dispersion of R. indica to other regions of South America could seriously impact the cultivation of coconuts, bananas, exotic and native palms and tropical flowers such as the Heliconiaceae. To facilitate the development of efficacious R. indica management techniques such as the adoption of phytosanitary measures to prevent or delay the dispersion of this pest, the objective of this paper was to estimate the potential geographical distribution of R. indica in South America using a maximum entropy model. The R. indica occurrence data used in this model were obtained from extant literature, online databases and field sampling data. The model predicted potential suitable areas for R. indica in northern Colombia, central and northern Venezuela, Guyana, Suriname, east French Guiana and many parts of Brazil, including Roraima, the eastern Amazonas, northern Pará, Amapá and the coastal zones, from Pará to north of Rio de Janeiro. These results indicate the potential for significant R. indica related economic and social impacts in all of these countries, particularly in Brazil, because the suitable habitat regions overlap with agricultural areas for R. indica host plants such as coconuts and bananas.     

Mite diversity on plants of different families found in the Brazilian Atlantic Forest

This work reports the occurrence of mites predominantly predatory, phytophagous and with varied feeding habits on plants of the Atlantic Forest vegetation type of the State of S?o Paulo. The objective was to estimate the possible role of the Atlantic Forest vegetation as reservoir of these groups of mites which are also found on plants of agricultural importance. Samples were taken from 187 plant species belonging to 73 families in three vegetation types of the Atlantic Forest from February of 2001 to October of 2002. A total of 2,887 mites belonging to 163 morpho-species of 16 families were collected. Mite diversity was high, especially of predatory mites; these corresponded to 1,562 specimens of 92 morpho-species. Within this group, Phytoseiidae comprised 71% of the specimens and 62% of the morpho-species. Phytophagous mites comprised 836 specimens of 36 morpho-species. Within this group, Tenuipalpidae comprised the larger proportion of specimens (61%) whereas Tetranychidae corresponded to the larger proportion of morpho-species (64%). Mites with varied feeding habits corresponded to 491 specimens of 36 morpho-species. In this group, the larger proportion of specimens (52%) consisted of Ascidae and the larger proportion of morpho-species (42%) consisted of Tydeinae (family Tydeidae). High abundance and high morpho-species richness of mites of predominantly predatory, phytophagous and variable feeding habits were observed on 17, five and nine plant species, respectively. The results obtained suggest the importance of plants of the studied vegetation as reservoirs of predatory mites.     

Phylogenetic investigation of the genus Raoiella (Prostigmata: Tenuipalpidae): diversity, distribution, and world invasions

The genus Raoiella is best known because of the red palm mite, R. indica, a major pest of palms spreading aggressively throughout the Americas. Not much was known about the biology, geographic origins, or evolutionary history of the genus when R. indica emerged as a major invasive pest. This paper attempts to address some of the basic historical questions regarding the palm mite as well as the genus. Molecular characters from COI and 28S regions were used to produce a phylogenetic hypothesis for the genus in an effort to understand its geographic origin. It also uses COI barcode data to delimit several potentially new species discovered by the authors in Australia. Results show a basal split between R. indica and all other Raoiella species, which indicates Africa or the Middle East as the most probable origin of the genus. Additionally, COI data suggests that at least eight new species are represented among the 20 Australian populations included in this study.     

Can the red palm mite threaten the Amazon vegetation?

The red palm mite Raoiella indica Hirst (Tenuipalpidae) was first reported in the New World in 2004, dispersing quickly and widely while adopting new plant species as hosts. Since then, it has caused severe damage in this region, especially to coconut (Cocos nucifera L.). It was first found in Brazil in 2009, in the northern Amazonian state of Roraima. In the present study, native and introduced plants were sampled between March 2010 and February 2011 in sites of the 15 Roraima municipalities, to estimate its distribution and the associated mite fauna. In addition, monthly samples were taken from a coconut plantation in Mucajaí throughout the same period, for an initial appraisal of the levels R. indica could reach. It was found in 10 municipalities, on 19 plant species of four families. Six species are reported for the first time as hosts. Among the associated predators, 89.1% were Phytoseiidae, most commonly Amblyseius largoensis (Muma), Iphiseiodes zuluagai Denmark & Muma and Euseius concordis (Chant). The highest densities of R. indica, 1.5 and 0.35 mites/cm² of leaflet (approx total of 331 and 77 mites/leaflet), were reached respectively in March 2010 and February 2011. The highest density of phytoseiids on coconut (0.009 mites/cm² or about 2 mites/leaflet) was reached in November 2010. The average densities of R. indica recorded for Roraima were comparable to those reported for countries in which the mite is reportedly economically damaging. The dispersal of R. indica through the Amazon forest may result in damage to cultivated and native palms, and plants of other families, if the projected increase in both the frequency and the severity of drought events occurs. Parts of the Amazon have undergone periods of low rainfall, a condition that appears to favour the biology of this mite. Its eventual arrival to northeastern Brazil may result in heavy economic and ecological losses.     

巴基斯坦为害杧果树的短须螨属一新种(蜱螨亚纲,细须螨科)

在对巴基斯坦杧果种植区苏加堡的杧果树螨类区系研究中,发现短须螨属(蜱螨亚纲：细须螨科)1新种：礼短须螨。文中记录并描述了该新种。新种的背饰特点是：前背中侧近分颈沟处具网状纹、外侧具纵向条纹;具6对末体背侧毛。雄螨 未采到。正模♀,1994年1月1日采自巴基斯坦,苏加堡杧果园的落叶中;副模1,1994年11月3日采自马尔坦南3km处的杧果树叶,曼苏采。模式标本保存于农业大学农业昆虫系螨类研究室,费沙拉堡,巴基斯坦。新种的背饰独特,略与短须螨属中具7对末体背侧毛的种相像,但具6对末体背侧毛的种皆无此种背饰。     

A comparison of conventional SEM techniques, low temperature SEM and the electroscan wet scanning electron microscope to study the structure of a biofilm of Streptococcus crista CR3

Biofilms of Streptococcus crista CR3 were generated on hydroxyapatite (HA) discs for 20 h in a continuous flow system with brain heart infusion broth dripped over the disc at a rate of 6 ml h^-1. This study compares the conventional scanning electron microscope (SEM) preparation techniques, of critical point drying and freeze-drying, with low temperature SEM (LTSEM) and Electroscan generated images of hydrated biofilms, which preserve the integrity of hydrated polymers.
Critical point drying and freeze-drying caused almost complete disappearance of the matrix of extracellular polymeric substances (EPS). Critical point drying, however, showed evenly spaced single or paired cocci remaining on the HA disc whereas freeze-drying caused the biofilm to detach from the HA leaving only patchy clumps of cells visible. By comparison LTSEM preserved the EPS better than critical point drying and freeze-drying, but holes were seen in the top and side of the biofilm and the EPS did show some shrinkage artefacts. An untreated wet biofilm viewed in the Electroscan showed an intact, hydrated, smooth matrix of EPS with cell shapes only visible indistinctly in a canopy of moist EPS. No holes were visible and no shrinkage artefacts were evident. Therefore, Electroscan imaging of the biofilm was the only method that preserved the integrity of the matrix with no apparent shrinkage artefacts.     

The relationship between cranial morphology,bite performance,diet and habitat in a radiation of dwarf chameleon (Bradypodion)

Many animals show unique morphological and behavioural adaptations to specific habitats. In particular, variation in cranial morphology is known to influence feeding performance, which in turn influences dietary habits and, ultimately, fitness. Dietary separation is an important means of partitioning ecological niches and avoiding inter‐ and intraspecific competition. Consequently, differences in dietary resources may help explain phenotypic divergence in closely‐related species occupying different habitats, as well as sexual dimorphism. We test this hypothesis on five phenotypic forms of a recent radiation of dwarf chameleons (Bradypodion) that vary extensively in habitat use and cranial morphology. By examining stomach contents, the dietary composition of each phenotypic form is compared to investigate potential differences in feeding strategies. Overall, chameleons in the present study exhibit considerable dietary overlap (at both inter‐ and intraspecific levels), indicating that diet is not a major driver of variation in cranial morphology within this radiation. However, the stomachs of closed‐canopy females were found to contain more prey items than male stomachs, possibly indicating that females require a greater caloric intake than their male counterparts.