Yanina Mariottini

Morphology and taxonomy of the microsporidium Liebermannia covasacrae n. sp. from the grasshopper Covasacris pallidinota (Orthoptera, Acrididae)

During a survey for grasshopper pathogens in Argentina in 2005-2006, individual Covasacris pallidinota from halophylous grasslands in Laprida, Buenos Aires province were found to be infected with a microsporidium. Infection was restricted to the salivary gland epithelial cells. The microsporidium produced ovocylindrical spores averaging 2.6+/-0.28 x 1.4+/-0.12 microm (range 2.2-3.4 x 1.1-1.7 microm), which resembled in size and shape the spores of Liebermannia patagonica and L. dichroplusae, two recently described species that also parasitize Argentine grasshoppers. The life cycle of the microsporidium included the formation of polynucleate, diplokaryotic, moniliform, merogonial plasmodia wrapped in flattened cisterns of the host endoplasmic reticulum (ER). Plasmodia divided to produce diplokaryotic cells. The latter underwent elongation, dissociation of diplokarya counterparts, vacuolization, dismantling of the host ER envelope, and deposition of electron-dense material outside the plasma membrane. The resultant binucleate sporogonial plasmodia divided into two uninucleate sporoblasts, which eventually transformed into spores. Uninucleate spores contained a lamellar polaroplast, embraced by an elongated polar sac, anchoring disc, 3-5 polar filament coils, and a cluster of anastomizing tubules (sporoblast trans-Golgi, posterosome) at the posterior end. Sequence similarity of the SSU rDNA of the newly discovered microsporidium (Genbank accession no. EU709818) to L. patagonica and L. dichroplusae was 99% and 97%, respectively, suggesting that the three species belong to one genus. All three species fell into one clade in SSU rDNA-based phylogenetic trees produced by neighbor joining, maximum parsimony, and maximum likelihood analyses with 100% statistical support. We assign the name Liebermannia covasacrae to this microsporidium. It can be easily differentiated from both congeners by host species, tissue tropism, type of sporogony, and several features of morphology. Comparison of the three Liebermannia spp. demonstrates that the nuclear phase (dikaryotic versus monokaryotic spores) and type of sporogony (polysporous versus disporous) may vary in closely related species.

Status of the alien pathogen Paranosema locustae (Microsporidia) in grasshoppers (Orthoptera: Acridoidea) of the Argentine Pampas

Abstract After experimental introductions from North America in 1978–1982, the biocontrol agent Paranosema locustae became established in grasshopper communities of the western Pampas region of Argentina. The use and establishment of P. locustae in Argentina constitute both a case of neoclassical or new association biological control (use of an alien species against native pests) and a case of pathogen pollution (anthropogenic introduction–establishment of an infectious disease in populations of native species). Since P. locustae is a multihost pathogen among grasshoppers, its presence in the western Pampas represents an additional factor disrupting grasshopper communities according to the differential susceptibility of each species and possibly threatening some species. Microscopic examination of 504 grasshopper samples (the mean number of individuals per sample = 185) belonging to 43 species from 93 localities throughout the Pampas revealed an establishment area of approximate 90,000 km2 from about 35° North to 38° South and from 61° East to 65° West. Field infections by P. locustae have now been detected in 21 grasshopper species in the western Pampas. Susceptible species with geographic distributions mostly restricted to the establishment area and with numerically small populations, like the melanopline Scotussa daguerrei, are predicted to be the ones facing higher risks of negative impacts.

Postembryonic development and food consumption of Dichroplus elongatus Giglio-Tos and Dichroplus maculipennis (Blanchard) (Orhtoptera: Acrididae: Melanoplinae) under laboratory conditions.

Dichroplus maculipennis (Blanchard) and D. elongatus Giglio-Tos are two of the most important melanoplines in Argentina, both ecologically and economically. The postembryonic development and forage loss (consumption of Bromus brevis Ness + fallen material) caused by older nymphs (instars IV, V, VI) and adults of both species were studied under controlled conditions (30ºC, 14L:10D, 40% RH). Five nymphal instars were recorded in D. elongatus, and six in D. maculipennis. Total nymphal development was similar in both species (D. elongatus: 32 ± 0.70 days; D. maculipennis: 34.5 ± 0.37 days). Daily consumption increased from nymphal instars to pre-reproductive adult stage. In both species, pre-reproductive females had higher consumption rates than other stages considered (D. elongatus: 30.6 ± 0.56 mg dry weight/day; D. maculipennis: 48.7 ± 0.74 mg dry weight/day). In the reproductive stage, consumption decreased significantly in both sexes. When feeding, D. maculipennis let some plant material to drop, increasing total loss. The percentage of fallen material was greater in reproductive adults, representing 3.9% and 2.9% of the total daily loss for males and females, respectively. Females and males of D. maculipennis were heavier than those of D. elongatus (P < 0.05), and daily consumption was significantly higher (P < 0.05). Regardless sex and reproductive status, adults of D. maculipennis consumed 29.1 ± 0.64 mg dry weight/day on average, while one of D. elongatus 20.0 ± 0.3 mg dry weight/ day.

The biology and some population parameters of the grasshopper, Ronderosia bergi, under laboratory conditions.

Abstract Some biological and population parameters of Ronderosia bergi (Stål) (Orthoptera: Acrididae: Melanoplinae) were estimated by monitoring five cohorts of the first generation (F1) of individuals born in captivity from grasshoppers collected in the South of Misiones province, northeastern Argentina, and held under controlled conditions (30° C, 14:10 L:D, 40% RH). The mean embryonic development time was 40.6 ± 1.7 days. Five nymphal instars were recorded. Total duration of nymphal development was 30.8 ± 0.54 days. The mean lifespan of cohorts was 22.6 ± 0.7 weeks. The number of egg-pods per female was 7.6 ± 1.44, and the amount of eggs per egg-pod was 16.45 ± 0.85. Mean fecundity was 125 ± 5.83 eggs per female with an oviposition rate of 1.55 ± 0.57 eggs/female/day. Survivorship curves showed that mortality was concentrated in the final weeks of adulthood, and the life expectancy curve decreased accordingly. The population parameters estimated gave the following values: the net rate of reproduction (R0) was 46.75 ± 11.2, generation time (T) was 18.87 ± 1.67 weeks, duplication time (D) was 3.31 ± 0.34, the intrinsic rate of population growth (rm) was 0.21 ± 0.021 and the finite rate of population increase (&lgr;) was 1.24 ± 0.026. The reproductive values (Vx) indicated that the largest contribution of females to the subsequent generation was between weeks 15 and 25.

Longevity and fecundity of Dichroplus maculipennis (Orthoptera, Acrididae) at non-outbreaking and outbreaking situations

Dichroplus maculipennis is one of the most characteristic and damaging grasshopper species of Argentina, mainly in areas of the Pampas and Patagonia regions. We estimated and compared the longevity and fecundity of adult female D. maculipennis under controlled conditions (30oC, 14L:10D, 40% RH) from individuals collected as last instar nymphs (VI) in the field and with a known recent history of low and high density conditions. Densities of D. maculipennis at the collecting sites were 0.95 individuals per m2 in 2006 and 46 ind/m2 in 2009, representing non-outbreaking and outbreaking situations, respectively. Adult female longevity in 2006 (67.96 ± 3.2 days) was significantly higher (p 0.05). The fecundity curves showed that the highest values were at weeks 11 and 13 for the 2006 females, and at week 6 for those of 2009. Since the daily oviposition rate at low and high densities was not significantly different, the diminished fecundity rate at high density is attributable to their reduced longevity.

Survival and fecundity of Dichroplus maculipennis and Ronderosia bergi (Orthoptera: Acrididae: Melanoplinae) following infection by Beauveria bassiana (Ascomycota: Hypocreales) under laboratory conditions

Abstract This study examined the effects of strain Beauveria bassiana (LPSC 1067) on nymphal development time, fecundity and adult survival in Dichroplus maculipennis and Ronderosia bergi under laboratory conditions. It was observed that infection with 1×103 conidia/ml altered nymphal development time, fecundity and adult survival in both species. Mortality of D. maculipennis during third through the last instar (sixth) was significantly higher among treated nymphs (66±3.8%) than in controls (15±1.7%). Similarly, mortality in R. bergi during third through the last instar (fifth) was higher in treated nymphs (71±2.8%) than in controls (19±1.5%). Nymphal development times of both infected D. maculipennis and R. bergi were longer than controls. On the other hand, among survivors of both the species, control adults lived longer than infected adults. Finally, control grasshoppers of both species were much more successful reproductively than infected grasshoppers.

Beauveria bassiana (Ascomycota: Hypocreales) Introduced as an Endophyte in Corn Plants and Its Effects on Consumption, Reproductive Capacity, and Food Preference of Dichroplus maculipennis (Orthoptera: Acrididae: Melanoplinae)

In this study, the effects of strain Beauveria bassiana (LPSC 1067) as an endophyte in corn plants on consumption, fecundity, and food preference of Dichroplus maculipennis were examined. We observed that the daily consumption by grasshoppers fed with control plants was almost twice that of those that were fed treated plants. Significant differences in fecundity of grasshoppers that were fed with treated plants compared with those that only fed on control plants were also observed. The number of eggs laid per female fed with control plants was 27.2, while the number of eggs laid per female that were fed during 15 d with treated plants was 17.7. Similar results were observed when the number of embryonated eggs was evaluated. The highest number of embryonated eggs were recorded in those females that only fed on control plants (96%) while fewer embryonated eggs were recorded in grasshoppers fed for 15 d with treated plants only (25%). In relation to food preference the average consumption rate for D. maculipennis females on control corn plants was 303.8 ± 24.5 mg while it was only 25 ± 2.1 mg on plants treated with B. bassiana as an endophyte. In summary, we observed that B. bassiana as a corn plant endophyte negatively affected the daily consumption rate, fecundity and food preference of D. maculipennis.

Morphometric Differentiation in a Field Population of Dichroplus maculipennis (Orthoptera: Acrididae: Melanoplinae) under Outbreak and Non-Outbreak Situations

Abstract Dichroplus maculipennis is one of the most damaging grasshopper species of Argentina. Individuals of this species at high density are historically known to show aggregation behavior and dispersal flights, attributes that might suggest that it does exhibit to some extent phase polyphenism in relation to population density. Phase polyphenism is a complex phenomenon and the amplitude of phase change is usually species-dependent. Morphological differences between gregarious and solitarious locusts can be measured and analyzed in order to characterize the phase status. The objective of this study was to evaluate morphometric differences between individuals of a D. maculipennis field population in the southern Pampas region of Argentina during non-outbreak and outbreak conditions including the magnitude of sexual size dimorphism related to variation in density. Collected individuals in the outbreak condition totaled 804 (422 females, 382 males) and those in non-outbreak condition were 325 (141 females, 184 males). Six morphometric characters were measured and two ratios (F/C and E/F) usually used to discriminate between solitarious and gregarious individuals in true locust species were calculated. Results show that size traits of D. maculipennis change over time at the population level, and that these changes correlate with outbreak vs non-outbreak populations. Females and males of D. maculipennis in outbreak conditions are significantly larger than in non-outbreak conditions. Furthermore, significant change over time was recorded in values of the two ratios calculated. D. maculipennis showed female biased sexual size dimorphism in both outbreak and non-outbreak conditions. There is a smaller difference in body size among females and males in outbreak conditions than in females and males in non-outbreak conditions. These results may be an indication of the presence of density-dependent phenotypic plasticity in this species, but additional experiments are required in order to establish a causal relationship between population density and changes in size traits.

Infectivity, viability and effects of Paranosema locustae (Microsporidia) on juveniles of Dichroplus maculipennis (Orthoptera: Acrididae: Melanoplinae) under laboratory conditions

Infectivity and effects on host of a long-term stored aqueous suspension of Paranosema locustae on juveniles of Dichroplus maculipennis, a pest grasshopper in parts of the Pampas and Patagonia, were evaluated. Infections developed in 90–97.8% of treated individuals. Mortality increased with time, reaching highest values at 30–40 days post-inoculation (79.5–100%). Infected nymphs showed significantly slower development.