Host-range and cold temperature tolerance of Floracarus perrepae Knihinicki & Boczek (Acari: Eriophyidae), a potential biological-control agent of Lygodium microphyllum (Pteridophyta: Lygodiaceae)

Abstract The leaf galling eriophyid mite Floracarus perrepae is a widespread and damaging herbivore of Old World climbing fern, Lygodium microphyllum , across its native range in tropical and subtropical Asia and Australia; and was therefore selected as a candidate biological-control agent for the invasive fern in Florida, USA. The host testing of F. perrepae focused on Lygodium species from North America and the Neotropics, along with threatened or endangered ferns from Florida. Sporeling ferns were used for the initial no-choice screening and F. perrepae showed normal development on the Florida genotype of L. microphyllum with 33.0 ± 4.6 mites per marginal leaf roll gall, modest development on the North American native Lygodium palmatum with 29.0 ± 9.3 mites per roll, and minor development on six other fern species (4.0–0.3). Leaf rolls were induced only on Lygodium species and full rolls were common only on L. microphyllum . These same six non-target plant species were tested again as more mature plants in both no-choice and choice tests and F. perrepae developed only on L. microphyllum . Lethal minimum temperature and cold stress tests were also conducted on F. perrepae . These tests revealed that it would not likely establish north of USDA hardiness zone 8a (−9.5 to −12.2°C), which means it would not overlap with L. palmatum from temperate North America. Our studies conclude that F. perrepae is specific to L. microphyllum , poses little or no risk to native or cultivated ferns in North and South America, and therefore should be considered for release in Florida.     

Matching the origin of an invasive weed for selection of a herbivore haplotype for a biological control programme

The Florida Everglades have been invaded by an exotic weed fern, Lygodium microphyllum. Across its native distribution in the Old World tropics from Africa to Australasia it was found to have multiple location-specific haplotypes. Within this distribution, the climbing fern is attacked by a phytophagous mite, Floracarus perrepae, also with multiple haplotypes. The genetic relationship between mite and fern haplotypes was matched by an overarching geographical relationship between the two. Further, mites that occur in the same location as a particular fern haplotype were better able to utilize the fern than mites from more distant locations. From a biological control context, we are able to show that the weed fern in the Everglades most likely originated in northern Queensland, Australia/Papua New Guinea and that the mite from northern Queensland offers the greatest prospect for control.     

Limited field establishment of a weed biocontrol agent, Floracarus perrepae (Acariformes: Eriophyidae), against Old World climbing fern in Florida--a possible role of mite resistant plant genotypes

The leaflet galling mite Floracarus perrepae Knihinicki & Boczek was released on Lygodium microphyllum (Cav.) in 63 plots in Florida from 2008 to 2009. Mites transferred onto field plants in 34 plots, but failed to establish populations in the majority of plots. Leaflet galls were observed in only six plots, and in only two plots did mite populations persist for >12 mo. Rates of mite transfer onto field plants were similar for methods using direct transfer of galls versus approaches using passive transfer of mites from infested plants. Often leaflets on some L. microphyllum plants were heavily galled by F. perrepae, whereas leaflets on intertwined stems of other L. microphyllum plants were ungalled but exhibited a characteristic browning and scorching of the leaflet tips. Living mites were consistently present on the undersurface of scorched leaflet tips on ungalled plants, suggesting that this damage might be caused by mite feeding on L. microphyllum genotypes that did not support induction of leaflet galls. Plant nutritional status did not account for differences in galling response, because there were no differences in leaflet nitrogen between galled and ungalled stems. We review those factors known to affect the colonization of biological control agents, and discuss how they may have contributed to the lower than expected rate of F. perrepae establishment.     

Herpobasidium filicinum (Eocronartiaceae,Platygloeales) occurs on Dennstaedtia wilfordii (Dennstaedtiaceae) in Japan

A fungus parasitic on a fern, Dennstaedtia wilfordii (Dennstaedtiaceae), was found at the foothill of Mt. Fuji, Yamanashi Prefecture, Japan. Its hyphae spread within host mesophyll cells and through intercellular spaces, forming coiled haustoria in the epidermal and mesophyll cells. The hyphae emerged either through stomata or by disrupting epidermal cell junctions. The hyphae spreading over the abaxial leaf surface generated one-septate, thin-walled basidia. All the morphological features observed were characteristic of the genus Herpobasidium. The species identification of the fungus as H. filicinum by morphology was supported by molecular phylogenetic analyses of the D1/D2 region of the large subunit rRNA gene.     

The reproductive biology of the invasive ferns Lygodium microphyllum and L. japonicum (Schizaeaceae): implications for invasive potential

The effect of culture system and population source on sexual expression and sporophyte production was examined for two invasive fern species in Florida, USA, Lygodium microphyllum and L. japonicum (Schizaeaceae). Both species are currently spreading through Florida. Long-distance dispersal of ferns is thought to rely on successful intragametophytic selfing. Given the rate of spread observed in both Lygodium species, we hypothesized that both species are capable of intragametophytic selfing. To test this hypothesis, gametophytes of both species were grown in vitro as isolates, pairs, and groups. Both species were capable of intragametophytic selfing; 78% of L. microphyllum isolates produced sporophytes and over 90% of the L. japonicum isolates produced sporophytes. Lygodium microphyllum also displayed the ability to reproduce via intergametophytic crossing, facilitated by an antheridiogen pheromone. Sporophyte production was rapid across mating systems for both species, an advantage in Florida's wet and dry seasonal cycles. The high intragametophytic selfing rate achieved by both species has likely facilitated their ability to colonize and spread through Florida. The mixed mating system observed in L. microphyllum appears to give this species the ability to invade distant habitats and then adapt to local conditions.     

Oviposition choice and larval performance of Neomusotima conspurcatalis on leaflet types of the invasive fern,Lygodium microphyllum

Lygodium microphyllum (Cavanilles) R Brown (Lygodiaceae), Old World climbing fern, invades wildlands in central and southern Florida, USA, and causes considerable habitat disturbance. Efforts to develop an effective biological control strategy have focused on several folivorous insects and a leaf‐galling mite from southeast Asia and Australia. Neomusotima conspurcatalis Warren (Lepidoptera: Crambidae) is a foliage‐feeding moth that specializes on Lygodium species. First released in 2008, N. conspurcatalis successfully established in Florida and is dispersing throughout the L. microphyllum‐invaded range. We investigated whether N. conspurcatalis display any preference (oviposition or larval feeding) between two leaflet morphologies, i.e., fertile spore‐producing vs. sterile leaflets. Furthermore, we examined whether oviposition and no‐choice diets on either fern leaflet type conferred any possible selective advantages (e.g., weight, emergence success) to the offspring. Finally, we analyzed leaf nitrogen and carbon content and leaf volatiles to gauge differences between the leaflet types. In choice and no‐choice scenarios, larvae consume approximately the same amount of leaf area and display no preference between fertile and sterile leaflets. However, females deposited significantly more eggs on fertile than on sterile leaflets. Oviposition choice had no effect on larval development times, pupal weights, or successful eclosure. Nitrogen and carbon content did not differ between fertile and sterile samples, but sterile leaflets contain more 1‐octen‐3‐ol, which may act as an herbivory deterrent. The propagule attack imposed by N. conspurcatalis feeding on fertile leaflets is unlikely to provide adequate control, but could be beneficial as part of a suite of biological control agents and other control methods.     

Stylostome formation in trombiculid mites (Acariformes: Trombiculidae)

Stylostomes of the trombiculid mite larvae Neotrombicula pomeranzevi (Schluger), Hirsutiella zachvatkini (Schluger), Miyatrombicula esoensis (Sasa and Ogata) and Euschoengastia rotundata (Schluger) (Acariformes: Trombiculidae), formed in the host skin during feeding of the parasites on their natural hosts (voles) were studied histologically and histochemically. A stylostome is a variously shaped tube formed of solidified mite saliva that extends from the mouthparts of the parasite through the epidermis into the dermis of the host, and allows the mite to obtain its liquid food. The first step of stylostome formation is deposition of an eosinophilic cone, to which the larva’s chelicerae are glued. Organization of the stylostome depends on the mite species, and its walls may show weakly expressed longitudinal or transverse stratification. Histochemically, the stylostome is composed of complex glycoprotein with varying tinctorial properties through the width or the length of the stylostome’s walls. Beneath the distal end of the stylostome, irrespectively of its localization either in the epidermis or in the dermis of the host, a feeding cavity is formed as a result of the action of the hydrolytic components of the mite’s saliva forced through the stylostome into the wound. An inflammatory dermal reaction of moderate intensity is evolved during larval feeding and stylostome formation. It is manifested by the infiltration of the foci with neutrophiles, lymphocytes and macrophages and by dilation of capillaries of the terminal vessel bed and filling them by erythrocytes and other blood elements. Around the stylostome, necrosis of the epidermal cells occurs, leucocytes come to the damaged area and fuse with the necrotic epidermal cells, leading to the formation of the large scabs on the surface of the host’s skin. In the case of E. rotundata, single capsules having a terminal opening and containing feeding larva are formed on the abdomen of the hosts. The walls of the capsules are composed of the mite’s saliva flowing upon the surface of the host’s skin. At the bottom of the capsule, a stylostome perforating the epidermis is also present.     

Parasite in peril? A new species of mite in the genus Ophiomegistus Banks (Parasitiformes: Paramegistidae) on an endangered host,the pygmy bluetongue lizard Tiliqua adelaidensis (Peters) (Squamata: Scincidae)

Host‐parasite relationships are generally understudied in wild populations but have a potential to influence host population dynamics and the broader ecosystem, which becomes particularly important when the host is endangered. Herein we describe a new species of parasitic mite from the genus Ophiomegistus (Parasitiformes: Mesostigmata: Paramegistidae) of an endangered South Australian skink; the pygmy bluetongue lizard (Tiliqua adelaidensis). Adult mites were observed on lizard hosts in three different host populations, among which prevalence varied. No temporal trend in prevalence was evident over two spring‐summer seasons of monitoring. We hypothesise that the reliance on burrows as refuges by T. adelaidensis may be essential for the completion of the mite life cycle and also for horizontal transmission. The conservation implications of not only its effect on the host, but also its potential status as an endangered species itself, are considered.     

Apospory in leaf culture of staghorn fern (Platycerium bifurcatum)

The induction, origin, morphology, and ploidy of aposporous gametophytes produced on juvenile leaves of the fern Platycerium bifurcatum (Cav.) C. Chr. were studied. Leaf explants were grown on modified Murashige and Skoog medium with 0%, 0.01%, 0.1%, 1%, or 2% sucrose. A low sucrose concentration (0.01%) and wounding of the adaxial side of the leaf significantly increased the induction of aposporous gametophytes (90% of leaves produced gametophytes). Regeneration began as a proliferation of mainly epidermal cells on both sides of the leaf; subsequent development was similar to that shown by gametophytes originating from spores. Flow cytometric analysis of sporophytes and aposporous gametophytes revealed that both forms had the same ploidy level. On the basis of these findings, we propose a set of conditions which regularly and reproducibly induces apospory on most of the leaf explants of the fern P. bifurcatum.     

The fine structure of muscle attachments in an acarid mite, Caloglyphus mycophagus (Megnin) (Acarina)

The fine structure of the myo-cuticular junction in an acarid mite, Caloglyphus mycophagus, is described. The muscle fibres are attached to the cuticle via flattened, much invaginated, epidermal cells. Unlike the situation described for other arthropods, the stress across these epidermal cells does not appear to be transmitted by microtubules but rather by desmosome-like structures which form intraepidermal cell bridges where invaginations from the outer and inner surfaces of the epidermal cells lie close together. The muscles are attached to the inner surface of this complex desmosome and the outer surface is linked to the cuticle by extracellular fibrils.     

Laboratory host range testing of Neomusotima conspurcatalis (Lepidoptera: Crambidae) – a potential biological control agent of the invasive weed,Old World climbing fern,Lygodium microphyllum (Lygodiaceae)

Old World climbing fern, Lygodium microphyllum, is a serious invasive weed in south Florida. Development of biological control is vital for sustainable management of L. microphyllum. Neomusotima conspurcatalis was discovered in Hong Kong in 1997 and was subsequently found causing feeding damage on L. microphyllum in much of its native range in Asia. Quarantine testing of N. conspurcatalis used 37 non-Lygodium fern species representative of New World genera of cultivated ferns and fern allies, one gymnosperm, three crop species, six Lygodium species, and the primary host L. microphyllum. No significant oviposition or feeding was observed on any of the 41 non-Lygodium species evaluated. Oviposition and feeding occurred on all Lygodium species, but amounts were low and usually significantly less than observed on L. microphyllum. The exception was L. japonicum, which was preferred as an oviposition host. Neomusotima conspurcatalis was only able to complete development on L. japonicum and L. palmatum, but survival on these species was only half that occurring on L. microphyllum. Neomusotima conspurcatalis is a Lygodium specialist. Lygodium japonicum is an invasive weed in the United States. Lygodium palmatum is restricted to areas of the United States where freezing winter temperatures would be lethal to N. conspurcatalis. It was concluded that N. conspurcatalis would pose no threat to native or cultivated plants in North America or the Caribbean and should be considered for biocontrol of L. microphyllum. A release petition was submitted in 2005. An USDA-APHIS release permit for N. conspurcatalis was issued in 2007.     

CYTOKININ- AND GIBBERELLIC ACID-INDUCED EFFECTS ON THE DETERMINATION AND MORPHOGENESIS OF LEAF PRIMORDIA IN OPUNTIA POLYACANTHA (CACTACEAE)

Cytokinins and gibberellins are able to strongly influence the development of “leaf” primordia in the cactus Opuntia polyacantha. Under the influence of cytokinin, the primordia produced by cultured axillary bud apical meristems develop as normal, photosynthetic leaves, being composed of regular epidermal cells, guard cells, mesophyll and mucilage cells as well as vascular tissue. Under the influence of gibberellic acid (GA), the primordia develop as cactus spines, composed of thick-walled epidermal and fiber cells. Guard cells, vascular tissue and parenchyma do not occur. Thus GA is able to redirect leaf morphogenesis in O. polyacantha far more completely than has been reported for other plants. The mitotic activity of the primordia that will develop into spines is significantly higher (at the 5 % level) than the mitotic activity of the primordia that will develop into leaves. This is interpreted to indicate that the primordia are either leaf primordia or spine primordia from a very early age, and possibly are never uncommitted or undetermined primordia, as has been suggested for fern leaf primordia.     

Micro-morphological alterations in young rosette leaves of Dipsacus laciniatus L. (Dipsacaceae) caused by infestation of the eriophyid mite Leipothrix dipsacivagus Petanovic et Rector (Acari: Eriophyoidea) under laboratory conditions

The present study describes micro-morphological and histological changes to rosette leaves of the native Eurasian plant species Dipsacus laciniatus (Dipsacaceae) provoked by infestation of the eriophyid mite Leipothrix dipsacivagus Petanovic et Rector. Conspicuous injuries to the leaf tissue were induced by mites feeding on leaves of D. laciniatus rosettes that were propagated from seed under laboratory conditions. Anatomical injuries extended into epidermal cells on the upper and lower leaf surface as well as to the mesophyll layer of infested D. laciniatus leaves. Statistical analysis (by ANOVA and MANOVA) showed significant differences between control and infested plants, particularly in total leaf thickness. The most striking change observed was the decrease in thickness of infested leaves, beginning from the 8th week, associated with the progressive replacement of epidermal cells with an acellular layer. Measures of mite density on test leaves indicated that mites vacated leaves as russeting symptoms intensified. They also appeared to vacate all leaves, whether symptomatic or not, after populations peaked 10–12 weeks after infestation. Comparisons were made between these studies and those on naturally infested, field-collected D. laciniatus plants, as well as with similar studies of other mite-plant interactions.     

Host inflammatory response governs fitness in an avian ectoparasite, the northern fowl mite (Ornithonyssus sylviarum)

Vertebrate immune responses to ectoparasites influence pathogen transmission and host fitness costs. Few studies have characterized natural immune responses to ectoparasites and resultant fitness effects on the ectoparasite. These are critical gaps in understanding vertebrate-ectoparasite interaction, disease ecology and host-parasite co-adaptation. This study focused on an ectoparasite of birds—the northern fowl mite (NFM) (Ornithonyssus sylviarum). Based on prior evidence that chickens develop resistance to NFM, these experiments tested two hypotheses: (i) skin inflammation blocks mite access to blood, impairing development, reproduction and survival; and (ii) host immunogenetic variation influences the inflammatory response and subsequent effects on the ectoparasite. On infested hosts, histology of skin inflammation revealed increased epidermal cell number and size, immigration of leukocytes and deposition of serous exudates on the skin surface. Survival of adult mites and their offspring decreased as the area of skin inflammation increased during an infestation. Inflammation increased the distance to blood vessels beyond the length of mite mouthparts (100-160 μm) and prevented protonymphs and adults from reaching a blood source. Consequently, protonymphs could not complete development, evidenced by a significant inverse relationship between inflammation and protonymph feeding success, as well as an increasing protonymph/adult ratio. Adult females were unable to feed and reproduce, indicated by an inverse relationship between inflammation and egg production, and decreasing female/juvenile ratio. These combined impacts of host inflammation reversed NFM population growth. Intensity of inflammation was influenced by the genotype of the major histocompatibility complex (MHC), supporting previous research that linked these immunological loci with NFM resistance. Overall, these data provide a model for a mechanism of avian resistance to an ectoparasitic arthropod and the fitness costs to the parasite of that host defense.     

Behavioural responses of Varroa destructor (Acari: Varroidae) to extracts of larvae, cocoons and brood food of worker and drone honey bees, Apis mellifera (Hymenoptera: Apidae)

Abstract. Varroa destructor is a parasitic mite of the honey bee species Apis cerana Fabr . and A. mellifera L. Mature females reproduce on the immature stages of their hosts, producing more viable female offspring on drone hosts than on worker hosts. Thus, immature drones are more likely to be infested with mites than immature workers. To investigate the hypothesis that differences in host chemistries underlie the biased distribution of mites between worker and drone brood, the arrestment responses of mites to solvent extracts of a number of stimuli normally encountered by a mite during its life cycle were measured. Mites were arrested by cuticular extracts of worker and drone larvae obtained at 0, 24 and 48 h prior to the time when cell capping is completed. Mites were also arrested by extracts of worker and drone, brood food and cocoons, and by a blend of synthetic fatty acid esters previously shown to be active in the host acquisition process. In a wind tunnel bioassay, mites were attracted to odours from living fifth-instar worker and drone larvae, but not to volatiles from cocoons, brood food or a blend of fatty acid esters. The sex of the host was not an important factor affecting the behavioural responses of the mites in any assay. We conclude that host kairomones play a role in the host acquisition process, but we found no evidence to support the hypothesis that mites use these substances to differentiate between worker and drone brood.     

Anatomical Injury Induced by the Eriophyid Mite Aceria anthocoptes on the Leaves of Cirsium arvense

Anatomical injury of the leaves of the invasive species, Cirsium arvense (L.) Scop., caused by the eriophyid mite Aceria anthocoptes (Nal.), which is the only eriophyid mite that has been recorded on C. arvense worldwide, is described. The injury induced by the mite feeding on the leaves of C. arvense results in visible russeting and bronzing of the leaves. Other conspicuous deformations are folding and distortion of the leaf blade and curling of leaf edge, as well as gradual drying of leaves. The anatomical injury of the mature leaves of field-collected plants was limited to the epidermis of the lower leaf surface. However, on young leaves of experimentally infested plants, rust mite injuries extend to epidermal cells on both leaf surfaces and to those of deeper mesophyll layers. On these leaves, lesions on the lower leaf surface even affected the phloem of the vascular bundles. Leaf damage induced by A. anthocoptes is discussed with regard to the mite’s potential as a biological control agent of C. arvense.     

Modelling Fungal (Neozygites cf. Floridana) Epizootics in Local Populations of Cassava Green Mites (Mononychellus Tanajoa)

The fungus, Neozygitis cf. floridana is parasitic on the cassava green mite, Mononychellus tanajoa (Bondar) (Acari: Tetranychidae) in South America and may be considered for classical biological control of cassava green mites in Africa, where cassava is an important subsistence crop, cassava green mites are an imported pest and specific natural enemies are lacking. Spider mites generally have a viscous structure of local populations, a trait that would normally hamper the spread of a fungus that is transmitted by the contact of susceptible hosts with the halo of capilliconidia surrounding an infectious host. However, if infected mites search and settle to produce capilliconidia on sites where they are surrounded by susceptible mites before becoming infectious, then the conditions for maximal transmission in a viscous host population are met. Because the ratio between spider mites and the leaf area they occupy is constant, parasite-induced host searching behaviour leads to a constant per capita transmission rate. Hence, the transmission rate only depends on the number of infectious hosts. These assumptions on parasite-induced host search and constant host density lead to a simple, analytically tractable model that can be used to estimate the maximal capacity of the fungus to decimate local populations of the cassava green mite. By estimating the parameters of this model (host density, per capita transmission rate and duration of infected and infectious state) it was shown that the fungal pathogen can reduce the population growth of M. tanajoa, but cannot drive local mite populations to extinction. Only when the initial ratio of infectious to susceptible mites exceeds unity or the effective growth rate of the mite population is sufficiently reduced by other factors than the fungus (e.g. lower food quality of the host plant, dislodgement and death by rain and wind and predation), will the fungal pathogen be capable of decimating the cassava green mite population. Under realistic field conditions, where all of these growth-reducing factors are likely to operate, there may well be room for effective control by the parasitic fungus. This revised version was published online in November 2006 with corrections to the Cover Date.     

Environmental factors affecting the life-tables ofTetranychus urticae (Acari: Tetranychidae) III. Host-plant nutrition

First, the literature of the last two decades on nutritional effects of host plants on spider mites is briefly reviewed. Second, experiments are described that subjected micro-propagated apple trees to four different levels of each macronutrient N, P and K. Spider mites (Tetranychus urticae Koch) feeding on leaf disks of these plants were checked for their developmental time, egg production and longevity. Plant analysis revealed that the concentration of N, P and K corresponded to the respective treatments. The content of phenolic compounds in the leaves increased with N and P deficiency. In the N experiment, spider-mite preimaginal developmental rate and oviposition rate were both positively correlated with leaf N. Often, fecundity was positively correlated with N and carbohydrate content of the leaves, and negatively with the phenolic content. Longevity of the two-spotted spider mite was not significantly affected by any treatment. The K experiments yielded only minor differences in plant contents as well as in spider-mite biology.From these mite data, file-tables were constructed and statistically analyzed by the Jackknife technique. The life-table analysis showed a gradual decline in the intrinsic rate of natural increase (r _m) with N and P deficiency. With all experiments pooled,r _m was clearly correlated to leaf N and particularly to the content of phenolic compounds in the leaves. Nitrogen shortage had the most distinct influence on mite population growth: in a range of 1.5–3.0% leaf N,r _m increased by a factor of 4, the number of multiplications per generation (R ₀) by 11, and the doubling time of the population was prolonged 4-fold on severely N deficient leaves.     

Developmental Morphology of Platycerium coronarium (Koenig) Desv

The growth habit and certain developmental aspects of Platyceriumcoronarium (stag's horn fern) are described. Starting from theprimordial stage, the nest and pendulous leaves develop to maturityin 90 and 80 days respectively. The fertile lobe, which is partof the pendulous leaf, reaches its maximum size in 40 days.The morphogenesis of the nest leaf is more variable, and itmatures and deteriorates earlier than the pendulous leaf. Theacrostichoid sorus formation is completed in 3 weeks from inceptionand spore dispersal takes place when the fertile lobe is about100 days old. The area of the fertile lobe and number of sporesproduced were determined. On Knop's agar medium the gametophytesdevelop in 2 months and 85 per cent of them are unisexual (bothmale and female) and 15 per cent bisexual. Less than 1 per centof the gametophytes give rise to sporophytes. The juvenile leavesare simple, displaying open dichotomous venation; the firstnest and pendulous leaves are produced 24 months after the dateof spore germination. Platycerium coronarium, stag's horn fern, leaf development, morphogenesis, spore production