Global distribution and host range of the endoparasitic mite genus Locustacarus (Acari: Podapolipidae) with description of a new species from Iran parasitizing grasshoppers (Orthoptera: Acrididae).

Mites (Acari) represent important parasites for a diverse range of hosts. Within the Acari, the Prostigmata represent a diverse suborder of the order Trombidiformes with about 20 000 species, including parasitic forms on both vertebrates and invertebrates. Within the Prostigmata, the genus Locustacarus (Heterostigmata: Podapolipidae) is particularly known as an intratracheal parasite of bumblebees and grasshoppers. In a survey on prostigmatic mites (Acari: Trombidiformes: Prostigmata) associated with insects in the eastern parts of Iran, one new species of the endoparasitic Locustacarus was collected associated to the grasshopper Aiolopus thalassinus (Fabricius) (Orthoptera: Acrididae). This new species, Locustacarus aiolopi Rahmatzaei & Hajiqanbar n. sp., is described here and compared morphologically with other species of the genus. This new species represents the first record of a grasshopper-associated Locustacarus species in the Palearctic. We further provide a comprehensive review of the global distribution of this genus as well as its host range.

Malpighamoeba infection compromises fluid secretion and P-glycoprotein detoxification in Malpighian tubules.

Malpighian tubules, analogous to vertebrate nephrons, play a key role in insect osmoregulation and detoxification. Tubules can become infected with a protozoan, Malpighamoeba, which damages their epithelial cells, potentially compromising their function. Here we used a modified Ramsay assay to quantify the impact of Malpighamoeba infection on fluid secretion and P-glycoprotein-dependent detoxification by desert locust Malpighian tubules. Infected tubules have a greater surface area and a higher fluid secretion rate than uninfected tubules. Infection also impairs P-glycoprotein-dependent detoxification by reducing the net rhodamine extrusion per surface area. However, due to the increased surface area and fluid secretion rate, infected tubules have similar total net extrusion per tubule to uninfected tubules. Increased fluid secretion rate of infected tubules likely exposes locusts to greater water stress and increased energy costs. Coupled with reduced efficiency of P-glycoprotein detoxification per surface area, Malpighamoeba infection is likely to reduce insect survival in natural environments.

Molecular Identification of Oxyspirura Petrowi Intermediate Hosts by Nested PCR Using Internal Transcribed Spacer 1 (ITS1).

Recently, the heteroxenous eyeworm, Oxyspirura petrowi, has gained attention due to its prevalence in the declining game bird, Northern bobwhite (Colinus virginianus), but the intermediate hosts of many nematodes remain unknown. However, identifying the intermediate host of O. petrowi with traditional techniques would be difficult and time-consuming, especially considering there are more than 80 potential orthopteran hosts just in Texas. To screen a large number of samples quickly and effectively, primers for nested PCR (nPCR) were developed using the internal transcribed spacer 1 (ITS1) region. Then the nPCR was used to identify which of the 35 species collected from the Order Orthoptera were potential intermediate hosts of O. petrowi. With this technique, 18 potential intermediate hosts were identified. Later, we collected live specimens of species that tested positive to confirm the presence of larvae, but larvae were not found in the live specimens, nor in the extra tissue of the species that had tested positive for O. petrowi DNA. Despite this, this study demonstrated that nPCR is more sensitive than traditional techniques and can be a valuable tool in determining the intermediate hosts of parasites.

Morphology and Phylogenetic Position of Two New Gregarine Species (Apicomplexa: Eugregarinorida) Parasitizing the Lubber Grasshopper Taeniopoda centurio (Drury, 1770) (Insecta: Orthoptera: Romaleidae) in Mexico.

Eugregarines are understudied apicomplexan parasites of invertebrates inhabiting marine, freshwater, and terrestrial environments. Most currently known terrestrial eugregarines have been described parasitizing the gut from less than 1% of total insect diversity, with a high likelihood that the remaining insect species are infected. Eugregarine diversity in orthopterans (grasshoppers, locusts, katydids, and crickets) is still little known. We carried out a survey of the eugregarines parasitizing the Mexican lubber grasshopper, Taeniopoda centurio, an endemic species to the northwest of Mexico. We described two new eugregarine species from the gut of the host: Amoebogregarina taeniopoda n. sp. and Quadruspinospora mexicana n. sp. Both species are morphologically dissimilar in their life-cycle stages. Our SSU rDNA phylogenetic analysis showed that both species are phylogenetically distant to each other, even though they parasitize the same host. Amoebogregarina taeniopoda n. sp. clustered within the clade Gregarinoidea, being closely related to Amoebogregarina nigra from the grasshopper Melanoplus differentialis. Quadruspinospora mexicana n. sp. clustered within the clade Actinocephaloidea and grouped with Prismatospora evansi, a parasite from dragonfly naiads. Amoebogregarina taeniopoda n. sp. and Q. mexicana n. sp. represent the first record of eugregarines found to infect a species of the family Romaleidae.

A parasitological evaluation of edible insects and their role in the transmission of parasitic diseases to humans and animals.

From 1 January 2018 came into force Regulation (EU) 2015/2238 of the European Parliament and of the Council of 25 November 2015, introducing the concept of "novel foods", including insects and their parts. One of the most commonly used species of insects are: mealworms (Tenebrio molitor), house crickets (Acheta domesticus), cockroaches (Blattodea) and migratory locusts (Locusta migrans). In this context, the unfathomable issue is the role of edible insects in transmitting parasitic diseases that can cause significant losses in their breeding and may pose a threat to humans and animals. The aim of this study was to identify and evaluate the developmental forms of parasites colonizing edible insects in household farms and pet stores in Central Europe and to determine the potential risk of parasitic infections for humans and animals. The experimental material comprised samples of live insects (imagines) from 300 household farms and pet stores, including 75 mealworm farms, 75 house cricket farms, 75 Madagascar hissing cockroach farms and 75 migrating locust farms. Parasites were detected in 244 (81.33%) out of 300 (100%) examined insect farms. In 206 (68.67%) of the cases, the identified parasites were pathogenic for insects only; in 106 (35.33%) cases, parasites were potentially parasitic for animals; and in 91 (30.33%) cases, parasites were potentially pathogenic for humans. Edible insects are an underestimated reservoir of human and animal parasites. Our research indicates the important role of these insects in the epidemiology of parasites pathogenic to vertebrates. Conducted parasitological examination suggests that edible insects may be the most important parasite vector for domestic insectivorous animals. According to our studies the future research should focus on the need for constant monitoring of studied insect farms for pathogens, thus increasing food and feed safety.

Gene expression changes elicited by a parasitic B chromosome in the grasshopper Eyprepocnemis plorans are consistent with its phenotypic effects.

Parasitism evokes adaptive physiological changes in the host, many of which take place through gene expression changes. This response can be more or less local, depending on the organ or tissue affected by the parasite, or else systemic when the parasite affects the entire host body. The most extreme of the latter cases is intragenomic parasitism, where the parasite is present in all host nuclei as any other genomic element. Here, we show the molecular crosstalk between a parasitic chromosome (also named B chromosome) and the host genome, manifested through gene expression changes. The transcriptome analysis of 0B and 1B females of the grasshopper Eyprepocnemis plorans, validated by a microarray experiment performed on four B-lacking and five B-carrying females, revealed changes in gene expression for 188 unigenes being consistent in both experiments. Once discarded B-derived transcripts, there were 46 differentially expressed genes (30 up- and 16 downregulated) related with the adaptation of the host genome to the presence of the parasitic chromosome. Interestingly, the functions of these genes could explain some of the most important effects of B chromosomes, such as nucleotypic effects derived from the additional DNA they represent, chemical defense and detoxification, protein modification and response to stress, ovary function, and regulation of gene expression. Collectively, these changes uncover an intimate host-parasite interaction between A and B chromosomes during crucial steps of gene expression and protein function.

Molecular detection of Oxyspirura larvae in arthropod intermediate hosts.

To determine potential intermediate hosts of Oxyspirura petrowi, a common nematode eyeworm of wild gallinaceous birds, various arthropod species including red harvester ants, beetles, wood cockroaches, crickets, grasshoppers, katydids, and desert termites were screened for the presence of O. petrowi using specific polymerase chain reaction (PCR) primers targeting the internal transcribed spacer 2 region (ITS2) of the eyeworm ribosomal deoxyribonucleic acid (rDNA). This is the first study to investigate the intermediate hosts of O. petrowi utilizing molecular techniques. We determined 38% (13/34) of the cockroaches, 27% (3/11) of the crickets, and 23% (68/289) of the grasshoppers which were positive for O. petrowi. Identifying potential intermediate hosts of O. petrowi is essential to better understanding the epizoology of the eyeworm's transmission mechanics and to controlling infections in wild gallinaceous birds.

Condition-dependence and sexual ornamentation: Effects of immune challenges on a highly sexually dimorphic grasshopper.

Sexual ornaments contribute substantially to phenotypic diversity and it is particularly relevant to understand their evolution. Ornaments can assume the function of signals-of-quality that the choosy sex uses to evaluate potential mating partners. Often there are no obvious direct benefits and investment into mate choice is primarily rewarded by beneficial alleles that are inherited to the offspring. Inter-sexual communication via sexual ornaments requires honesty of the sexual signal, yet the question of what maintains honesty remains only partially solved. One solution is that honesty is maintained by trait expression being dependent on individual condition, since condition-dependent trait expression offers an effectively inexhaustible source of genetic variability. Here we test in the highly sexually dimorphic club-legged grasshopper Gomphocerus sibiricus if putative sexual ornaments, in particular the striking front-leg clubs, are more strongly affected by a lipopolysaccharide (LPS) immune challenge than putatively not sexually selected traits. Our results show overall little condition-dependent expression of morphological and song traits, with sexually selected traits exhibiting effects comparable to nonsexually selected traits (with the possible exception of stridulatory file length and syllable-to-pause ratio in advertisement songs). Interestingly, field observations of individuals of lethally parasitized individuals suggest that a very strong environmental challenge can specifically affect the expression of the front-leg clubs. The presence of 1% of males in natural populations with missing or heavily deformed clubs plus 5% with minor club deformations furthermore indicate that there are risks associated with club development during final ecdysis and this might act as a filter against deleterious alleles.

Adaptive significance of precocious pupation in the bean blister beetle, Epicauta gorhami (Coleoptera: Meloidae), a hypermetamorphic insect.

Larvae of the bean blister beetle, Epicauta gorhami Marseul (Coleoptera: Meloidae), feed on grasshopper eggs in soil and undergo hypermetamorphosis. They normally enter diapause as a pseudopupa at the fifth instar, a form characteristic of hypermetamorphosis for meloid beetles. However, fourth-instar larvae exposed to long days and high temperature avoid pseudopupal diapause and pupate directly from the fourth instar. Fourth-instar larvae also tend to pupate precociously with a smaller body size if they are deprived of food. In these larvae, the critical day-length controlling induction of pseudopupal diapause becomes shorter than that for fully-fed larvae. In this study, we examined how the reaction norm of food-deprived E. gorhami larvae functions in nature by rearing insects from the egg stage outdoors in different seasons with manipulation of the food supply. The results indicated that most fully-fed larvae entered pseudopupal diapause, whereas food-deprived larvae tended to pupate precociously without entering diapause, especially early in the season. The resulting smaller adults reproduced early in the autumn and their progeny attained the pseudopupal stage before winter, indicating that the reaction norm may have an adaptive role in controlling seasonal development in the face of food shortages, producing a bivoltine life cycle.

Nematodes (Mermithidae) parasitizing grasshoppers (Orthoptera: Acrididae) in the Pampean region, Argentina / Nematóides (Mermithidae) parasitando gafanhotos (Orthoptera: Acrididae) nos Pampas região, a Argentina

Abstract This work provides the results of a survey of entomonematodes parasites of grasshoppers in grasslands of the Pampean Region, Argentina. Nymphs of Staurorhectus longicornis Giglio-Tos, Laplatacris dispar Rhen, 1939, Dichroplus elongatus Giglio-Tos, 1894 and Metaleptea brevicornis (L.) (Orthoptera: Acrididae) were collected. Mermithidae was the only family registered with seven species: Agamermis decaudata Cobb, Steiner and Christie, 1923, Amphimermis bonaerensis Miralles and Camino, 1983, Amphimermis dichroplusi Camino and Lange, 1997, Amphimermis ronderosi Camino and Lange, 1997, Hexamermis coclhearius Stock and Camino, 1992, Hexamermis ovistriata Stock and Camino, 1992, and Longimermis acridophila Camino and Stock, 1989. The values of parasitism ranged between 1-12%, and intensity not overcome the number of 5.0 nematodes per larva. The nematodes observed showed specificity, not registering the same species of parasite in more than one host species. The Pampean region constituted an area with high diversity of mermithids where new species could be consider as bioregulator agents of this troublesome insect pests in agricultural areas of Argentina.

Resumo Este trabalho apresenta os resultados de uma pesquisa entomonematode parasitando gafanhotos em pastagens da região pampeana, Argentina. Ninfas de Staurorhectus longicornis Giglio-Tos, Laplatacris dispar Rhen de 1939, Dichroplus elongatus Giglio-Tos, 1894 e Metaleptea brevicornis (L.) (Orthoptera: Acrididae) foram coletados. Mermithidae era a única família registrado com sete espécies: Agamermis decaudata Cobb, Steiner and Christie, 1923, Amphimermis bonaerensis Miralles and Camino, 1983, Amphimermis dichroplusi Camino and Lange, 1997, Amphimermis ronderosi Camino and Lange, 1997, Hexamermis coclhearius Stock and Camino, 1992, Hexamermis ovistriata Stock and Camino, 1992, e Longimermis acridophila Camino and imagem, de 1989. Os valores de parasitismo variou entre 1-12%, e intensidade não superar o número de 5,0 nematóides por larva. Os nemátodos observados demonstraram especificidade, não registar as mesmas espécies de parasita em mais do que uma espécie de hospedeiro. A região pampeana constituída uma área com alta diversidade de mermithids onde novas espécies poderiam ser consideradas como agentes bio-reguladores deste incômodos insetos pragas em áreas agrícolas da Argentina.

Nematodes (Mermithidae) parasitizing grasshoppers (Orthoptera: Acrididae) in the Pampean region, Argentina.

This work provides the results of a survey of entomonematodes parasites of grasshoppers in grasslands of the Pampean Region, Argentina. Nymphs of Staurorhectus longicornis Giglio-Tos, Laplatacris dispar Rhen, 1939, Dichroplus elongatus Giglio-Tos, 1894 and Metaleptea brevicornis (L.) (Orthoptera: Acrididae) were collected. Mermithidae was the only family registered with seven species: Agamermis decaudata Cobb, Steiner and Christie, 1923, Amphimermis bonaerensis Miralles and Camino, 1983, Amphimermis dichroplusi Camino and Lange, 1997, Amphimermis ronderosi Camino and Lange, 1997, Hexamermis coclhearius Stock and Camino, 1992, Hexamermis ovistriata Stock and Camino, 1992, and Longimermis acridophila Camino and Stock, 1989. The values of parasitism ranged between 1-12%, and intensity not overcome the number of 5.0 nematodes per larva. The nematodes observed showed specificity, not registering the same species of parasite in more than one host species. The Pampean region constituted an area with high diversity of mermithids where new species could be consider as bioregulator agents of this troublesome insect pests in agricultural areas of Argentina.

MaMk1, a FUS3/KSS1-type mitogen-activated protein kinase gene, is required for appressorium formation, and insect cuticle penetration of the entomopathogenic fungus Metarhizium acridum.

Entomopathogenic fungi have great potential for development as insecticides. However, large-scale use of mycoinsecticides is partially limited by poor efficiency. In many fungal pathogens, the yeast and fungal extracellular signal-regulated kinase (YERK1) subfamily is crucial to the fungal pathogenicity. In this study, a Fus3/Kss1-type mitogen-activated protein kinase (MAPK) gene MaMk1 (GenBank accession No. EFY93607) was identified in Metarhizium acridum, which encodes a member of the YERK1 subfamily. Targeted gene disruption was used to analyze the function of MaMk1 in fungal growth, conidial yield and virulence. Growth assays showed that MaMk1 disruption did not affect fungal growth and conidial yield on potato dextrose agar (PDA) plates. Bioassays by topical inoculation showed that a MaMk1-disruption mutant entirely lost its pathogenicity for the locusts, likely because of failure to penetrate the insect cuticle, which might have been caused by inability to form appressoria during infection. However, bioassays by injection showed no significant difference in virulence among the wild type (WT), ΔMaMk1 mutant and complementary transformant. ΔMaMk1 mutant failed to penetrate the cuticle outwards and sporulate on the locust cadaver. These results suggest that MaMk1 is required for penetration of the insect cuticle both into the hemocele and outside from the hemocele, but is dispensable for fungal growth in insect hemolymph. Gene expression pattern analysis showed that MaMk1 disruption downregulated expression of Mad1 and Mpl1, but did not reduce expression of Pr1 in M. acridum.

Prevalence and Molecular Identification of Nematode and Dipteran Parasites in an Australian Alpine Grasshopper (Kosciuscola tristis).

In alpine Australia, Orthoptera are abundant, dominant herbivores, important prey species, and hosts for parasites and parasitoids. Despite the central role of orthopterans in alpine ecosystems, the impact of parasites on orthopteran populations is under-explored. In this study we describe the relationship between parasite prevalence and host sex, body size and year of collection. We accessed an existing, preserved collection of 640 Kosciuscola tristis collected from across its range between 2007 and 2011. Upon dissection we collected juvenile parasites and used molecular tools to identify them to three families (Nematoda; Mermithidae, and Arthropoda: Diptera: Tachinidae and Sarcophagidae). The prevalence of nematodes ranged from 3.5% to 25.0% and dipterans from 2.4% to 20.0%. Contrary to predictions, we found no associations between parasite prevalence and grasshopper sex or size. Although there was an association between prevalence of both nematodes and dipterans with year of collection, this is likely driven by a small sample size in the first year. Our results provide a foundation for future studies into parasite prevalence within the alpine environment and the abiotic factors that might influence these associations.

Variable effects of dipteran parasitoids and management treatment on grasshopper fecundity in a tallgrass prairie.

Grasshoppers host a number of parasitoids, but little is known about their impact on grasshopper life history attributes or how those impacts may vary with land use. Here, we report on a three-year survey of nine grasshopper species in a tallgrass prairie managed with fire and bison grazing treatments. We measured parasitoid prevalence and the impact of parasitoid infection on grasshopper fecundity to determine if grasshopper-parasitoid interactions varied with management treatment. Adult female grasshoppers were collected every three weeks from eight watersheds managed with different prescribed burning and grazing treatments. Grasshopper fecundity with and without parasitoids was estimated through dissections of reproductive tracts. Dipteran parasitoids from two families (Nemestrinidae and Tachinidae) were observed infecting grasshoppers. We found significant effects of grazing treatment, but not burn interval, on grasshopper-parasitoid interactions. Parasitoids were three times more abundant in watersheds with bison grazing than in ungrazed watersheds, and the relative abundance of nemestrinid and tachinid flies varied with grazing treatment. Parasitoid prevalence varied among grasshopper species from <0.01% infected (Mermiria bivittata) to 17% infected (Hypochlora alba). Parasitoid infection reduced individual grasshopper fecundity, with stronger effects on current reproduction than on past reproduction. Furthermore, current fecundity in parasitized grasshoppers was lower in grazed watersheds compared to ungrazed watersheds. Nemestrinid parasitoids generally had stronger impacts on grasshopper fecundity than tachinid parasitoids, the effects of which were more variable.

World distribution and host range of Podapolipoides spp. (Acari: Heterostigmatina: Podapolipidae), with the description of a new species.

The ectoparasitic mite Podapolipoides anacridii n. sp. (Heterostigmatina: Podapolipidae), collected from the basal hindwings of Anacridium sp. (Orthoptera: Acrididae) in central Iran, is described. Podapolipoides Regenfuss, 1968 is defined, and the distribution, host-specificity and host range of its constituent species are discussed.

Acanthamoeba produces disseminated infection in locusts and traverses the locust blood-brain barrier to invade the central nervous system.

BACKGROUND: Many aspects of Acanthamoeba granulomatous encephalitis remain poorly understood, including host susceptibility and chronic colonization which represent important features of the spectrum of host-pathogen interactions. Previous studies have suggested locusts as a tractable model in which to study Acanthamoeba pathogenesis. Here we determined the mode of parasite invasion of the central nervous system (CNS). RESULTS: Using Acanthamoeba isolates belonging to the T1 and T4 genotypes, the findings revealed that amoebae induced sickness behaviour in locusts, as evidenced by reduced faecal output and weight loss and, eventually, leading to 100% mortality. Significant degenerative changes of various tissues were observed by histological sectioning. Both isolates produced disseminated infection, with viable amoebae being recovered from various tissues. Histological examination of the CNS showed that Acanthamoeba invaded the locust CNS, and this is associated with disruption of the perineurium cell/glial cell complex, which constitutes the locust blood-brain barrier. CONCLUSIONS: This is the first study to demonstrate that Acanthamoeba invades locust brain by modulating the integrity of the insect's blood-brain barrier, a finding that is consistent with the human infection. These observations support the idea that locusts provide a tractable model to study Acanthamoeba encephalitis in vivo. In this way the locust model may generate potentially useful leads that can be tested subsequently in mammalian systems, thus replacing the use of vertebrates at an early stage, and reducing the numbers of mammals required overall.

Novel in vitro and in vivo models to study central nervous system infections due to Acanthamoeba spp.

Acanthamoeba granulomatous encephalitis is a serious human infection with fatal consequences. The most distressing aspect of Acanthamoeba granulomatous encephalitis is the limited improvement in mortality. The underlying neurobiology is at present not well understood and treatment options are often of limited efficacy. There is therefore a real need to obtain more knowledge regarding the pathogenesis and pathophysiology of Acanthamoeba granulomatous encephalitis and to develop new chemotherapeutic approaches. However, the difficulties in using mammalian models to study this infection have hindered our search for therapeutic interventions. Recent availability of the blood-brain barrier, in vitro and use of locust as an in vivo model will undoubtedly allow us to investigate disease pathogenesis, mechanisms of parasite traversal across the blood-brain barrier and new drug therapies. It is argued that the models described here can offer several advantages in terms of speed, cost, technical convenience, and ethical acceptance. Furthermore, they are extremely valuable tools to discriminate molecules participating from both sides of the host-parasite interaction and will generate potentially useful leads in the identification of new potential drugs, as well as testing drug toxicity.

Novel model for the in vivo study of central nervous system infection due to Acanthamoeba spp. (T4 genotype).

In this study it was shown for what is believed to be the first time that the African migratory locust can be used as a model for the study of Acanthamoeba pathogenesis. Mature adult locusts were injected intra-abdominally with 10 mul suspension of 10(6) Acanthamoeba (a clinical isolate of the T4 genotype) in culture medium, or with the same volume of sterile culture medium. Locusts injected with Acanthamoeba showed significant weight loss and reduced production of faeces compared with control locusts. Furthermore, injection of amoebae killed all of the locusts within 17 days at room temperature, although the speed of kill was temperature and dose dependent. When samples of faecal pellets and various tissues of infected locusts were cultured on non-nutrient agar plates containing bacterial lawns, live amoebae were recovered from haemolymph, flight muscle and fat body samples, but not from faeces. When brains dissected from locusts were incubated with an anti-amoebic drug (100 muM chlorhexidine) to kill extracellular amoebae, and then washed, homogenized and cultured on bacteria-seeded non-nutrient agar plates, only lysates from amoebae-infected locusts were positive for Acanthamoeba. This strongly suggests that amoebae invade the locust brain and, indeed, trophozoites of Acanthamoeba could be identified within the brain in histological sections of brains from infected locusts, but not from uninfected locusts. These findings support the view that locusts can be used as a model for the study of Acanthamoeba pathogenesis in vivo.

A new Encephalitozoon species (Microsporidia) isolated from the lubber grasshopper, Romalea microptera (Beauvois) (Orthoptera: Romaleidae).

We describe a new microsporidian species, Encephalitozoon romaleae n. sp., isolated from an invertebrate host, the grasshopper Romalea microptera, collected near Weeks Island, Louisiana, and Jacksonville, Florida. This microsporidian is characterized by specificity to the gastric caecae and midgut tissues of the host and a life cycle that is nearly identical to that of Encephalitozoon hellem and Encephalitozoon cuniculi. Mature spores are larger (3.97 x 1.95 microm) than those of other Encephalitozoon species. Polar filament coils number 7 to 8 in a single row. Analysis of the small subunit (SSU) rDNA shows that E. romaleae fits well into the Encephalitozoon group and is a sister taxon to E. hellem. This is the first Encephalitozoon species that has been shown to complete its life cycle in an invertebrate host.