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1.  The Influence of Natural and Anthropic Environmental Variables on the Structure and Spatial Distribution Along Longitudinal Gradient of Macroinvertebrate Communities in Southern Brazilian Streams 
Southern Brazilian rivers and streams have been intensively affected by human activities, especially agriculture and the release of untreated domestic sewage. However, data about the aquatic macroinvertebrates in these streams are scarce and limited to only certain groups. In addition, studies focusing on the structure and spatial distribution of these communities are lacking. This study analyzed the effects of natural and anthropic variables on the community structure of macroinvertebrates along a longitudinal gradient in three microbasins located in a region of landscape transition in the state of Rio Grande do Sul, Brazil. Sampling was conducted in the Vacacaí-Mirim River (August 2008) and in the Ibicuí-Mirim and Tororaipí rivers (August 2009) following an environmental gradient including 1st, 2nd, 3rd, and 4th order segments. Local natural factors that were analyzed include water temperature, pH, electrical conductivity, dissolved oxygen, substrate granulometry, and the presence of aquatic vegetation. Anthropic variables that were analyzed include including bank erosion, land use, urbanization, riparian deforestation, and fine sediments input. A total of 42 families and 129 taxa were found, with predominance of environmentally tolerant taxa. Geological context (landscape transition and large hydrographic basins) tended to influence natural environmental factors along the rivers' longitudinal gradients. However, changes in anthropic variables were not affected by these geological differences and therefore did not correlate with patterns of spatial distribution in macroinvertebrate communities. Only 1st order stream segments showed a community composition with high richness of taxa intolerant to anthropic disturbance. Richness as a whole tended to be higher in 3rd to 4th order set of segments, but this trend was a result of local anthropic environmental disturbances. Future inventories conducted in similar landscape transition regions of Brazil, for conservation purposes, must consider stream segments of different orders, microbasins, and major basins in order to obtain data that faithfully reflect the regional diversity. Additionally, it is necessary to consider environmental gradients of land use and anthropic impacts in order to suggest appropriate strategies for conserving the environmental integrity of streams.
doi:10.1673/031.014.13
PMCID: PMC4199531  PMID: 25373160
aquatic insects; landscape; multiple scales; Neotropical region; river order
2.  Impact of Temperature on Postdiapause and Diapause of the Asian Gypsy Moth, Lymantria dispar asiatica 
Lymantria dispar asiatica (Vnukovskij) (Lepidoptera: Lymantridae) is one of three gypsy moth subspecies found in East Asia. Understanding the diapause and postdiapause phases of its eggs is important in characterizing its life cycle. The effects of different constant temperatures for different lengths of times on field-collected, postdiapause eggs were tested during the first year. In the second year, the effects of the same treatments on laboratory-raised eggs in diapause were investigated. The effects of temperature on percent egg hatching, time to hatching, and hatching duration were determined. When field-collected eggs were held at 0 and 5°C, they terminated postdiapause within 11 days. The percent hatching tended to decline with an increased duration of exposure at temperatures greater than 5°C. Diapause terminated slowly (> 37 days) and with a high percentage of hatching for postdiapause eggs held at 10°C. There was a positive correlation between temperature and the speed of postdiapause development for field-collected eggs held at constant temperatures between 10 and 25°C. However, the number of days to the first hatch was significantly longer than for eggs treated with lower temperatures before being transferred to 25°C. Freshly oviposited eggs treated at a constant 0 or 5°C for 200 days were unable to develop into pharate larva. However, eggs treated at a constant 20 or 25°C for 200 days developed into pharate larva but did not hatch even after a subsequent chill. This result suggests why L. dispar asiatica is not found in tropical areas and helps us to predict the distribution of the gypsy moth in China.
doi:10.1673/031.014.05
PMCID: PMC4199355  PMID: 25373152
egg hatch; forest pest; gypsy moth distribution
3.  Effect of Honey Solution and Water Acquisition on Survival of Starved Solenopsis Mealybug, Phenacoccus solenopsis 
The current study examined the effects of honey solution and water access on feeding behavior and survival of starving solenopsis mealybugs, Phenacoccus solenopsis Tinsley (Hemiptera: Pseudococcidae). The electrical penetration graph technique and an artificial membrane system were used to check whether P. solenopsis could imbibe free water or other liquid, such as the honey solution used here, in its natural environment. The recorded electrical penetration graph waveforms revealed that P. solenopsis could continuously imbibe water-honey solution for several hours, which indicated that honey solution and water acquisition could possibly occur when P. solenopsis had access to such liquids in its natural environment. Waveforms of water-honey solution feeding alternated between two distinct feeding phases in a regular pattern, which was assumed to reflect inherent habits of feeding attempts. The effects of honey solution and water acquisition on survival of P. solenopsis was also examined. Comparison between P. solenopsis in different treatments (starved, water feeding, honey solution feeding, and cotton plant feeding) suggested that 1) P. solenopsis could accept but did not favor feeding on water or the honey solution, and 2) this feeding could prolong its survival, but had no effect on body size.
doi:10.1673/031.014.01
PMCID: PMC4199356  PMID: 25373148
artificial membrane; cotton mealybug; electrical penetration graph technique; invasive pest; starvation
4.  Evaluation of the Influence of the Antibiotic Ciprofloxacin in the Development of an Old World Screwworm Fly, Chrysomya putoria 
Chrysomya putoria (Wiedemann) (Diptera: Calliphoridae), an Old World screwworm fly, is a species with potential for maggot therapy practice and has been described in myiasis and forensic entomology studies. The objective of the present study was to assess the action of different ciprofloxacin concentrations on the growth and development of C. putoria. First instar maggots of the third generation were raised on 60 g of chicken gizzard homogenate in 65% agar diet and received ciprofloxacin chloridrate. Each concentration of the antibiotic tested (3.33 µg/mL, 6.66 µg/mL, and 13.33 µg/mL) and the control (no antibiotic) were replicated four times (40 maggots/ replication). The control received distilled water instead of the antibiotic. Maggots were kept in an acclimatized chamber at 30° C during the day and 28° C at night, with 70 ± 10% RH and a 14:10 L:D photoperiod. They were weighed in batches of five and stored in test tubes sealed with nylon fabric and elastic. Microsoft Excel and STAT were used for the analysis. The variation among the maggot weight means and the duration of the maggot stage, pupal stage, and time to total development (neo-larvae to adult) were analyzed by Student's t-test (α = 5%). The viabilities and the normality rates were compared using ANOVA, and the expected sex ratio frequency was tested by the chi-squared test (χ2). There was no significant difference among the four treatments regarding mean individual maggot weight, mean duration of the maggot inoculation until abandonment, the duration of the maggot and pupal stages, and the total duration of all stages. The sex ratios found in the four treatments did not differ from the expected. Only treatment 2 (6.66 µg/mL concentration of ciprofloxacin) differed significantly from the control in maggot and total viability. The antibiotic did not seem to alter C. putoria development in the post-embryonic period.
doi:10.1673/031.014.03
PMCID: PMC4199357  PMID: 25373150
blowflies; drug; maggots; pupae
5.  Multiplex PCR in Determination of Opiinae Parasitoids of Fruit Flies, Bactrocera sp., Infesting Star Fruit and Guava 
Malaysia is a tropical country that produces commercial fruits, including star fruits, Averrhoa carambola L. (Oxalidales: Oxalidaceae), and guavas, Psidium guajava L. (Myrtales: Myrtaceae). There is a high demand for these fruits, and they are planted for both local consumption and export purposes. Unfortunately, there has been a gradual reduction of these fruits, which has been shown to be related to fruit fly infestation, especially from the Bactrocera species. Most parasitic wasps (Hymenoptera: Braconidae: Opiinae) are known as parasitoids of fruit fly larvae. In this study, star fruits and guavas infested by fruit fry larvae were collected from the Malaysian Agricultural Research and Development Institute. The parasitized larvae were reared under laboratory conditions until the emergence of adult parasitoids. Multiplex PCR was performed to determine the braconid species using two mitochondrial DNA markers, namely cytochrome oxidase subunit I and cytochrome b. Two benefits of using multiplex PCR are the targeted bands can be amplified simultaneously using the same reaction and the identification process of the braconid species can be done accurately and rapidly. The species of fruit flies were confirmed using the COI marker. The results obtained from our study show that Diachasmimorpha longicaudata (Ashmead) (Hymenoptera: Braconidae), Fopius arisanus (Sonan), and Pysttalia incisi (Silvestri) were parasitoids associated with Bactrocera carambolae (Drew and Hancock) (Diptera: Tephritidae) infested star fruits. Fopius arisanus was also the parasitoid associated with Bactrocera papayae (Drew and Hancock) infested guavas. Maximum parsimony was been constructed in Opiinae species to compare tree resolution between these two genes in differentiating among closely related species. The confirmation of the relationship between braconids and fruit fly species is very important, recognized as preliminary data, and highly necessary in biological control programs.
doi:10.1673/031.014.07
PMCID: PMC4199358  PMID: 25373154
biological control programs; braconids; parasitic wasps
6.  Selection and Validation of Reference Genes for Functional Studies in the Calliphoridae Family 
The genera Cochliomyia and Chrysomya contain both obligate and saprophagous flies, which allows the comparison of different feeding habits between closely related species. Among the different strategies for comparing these habits is the use of qPCR to investigate the expression levels of candidate genes involved in feeding behavior. To ensure an accurate measure of the levels of gene expression, it is necessary to normalize the amount of the target gene with the amount of a reference gene having a stable expression across the compared species. Since there is no universal gene that can be used as a reference in functional studies, candidate genes for qPCR data normalization were selected and validated in three Calliphoridae (Diptera) species, Cochliomyia hominivorax Coquerel, Cochliomyia macellaria Fabricius, and Chrysomya albiceps Wiedemann. The expression stability of six genes (Actin, Gapdh, Rp49, Rps17, α-tubulin, and GstD1) was evaluated among species within the same life stage and between life stages within each species. The expression levels of Actin, Gapdh, and Rp49 were the most stable among the selected genes. These genes can be used as reliable reference genes for functional studies in Calliphoridae using similar experimental settings.
doi:10.1673/031.014.02
PMCID: PMC4199359  PMID: 25373149
BestKeeper; Cochliomyia; Chrysomya; gene expression; geNorm; NormFinder
7.  Evaluating Buprestid Preference and Sampling Efficiency of the Digger Wasp, Cerceris fumipennis, Using Morphometric Predictors 
In-ground colonies of the native digger wasp, Cerceris fumipennis Say (Hymenoptera: Crabronidae), were sampled over two years in four New York State counties to characterize prey range, primarily their preying on beetles in the metallic wood-boring family, Buprestidae. These records were also used to evaluate beetle sampling efficiency by comparing collected beetles to historic county records and to identify limitations of wasp-mediated sampling in study areas. Overall, 1,530 beetles representing three families and 44 beetle species were collected from C. fumipennis. Five of these species (Agrilus cuprescens (Ménétriés) (Coleoptera: Buprestidae), A. pensus Horn, Buprestis nutalli Kirby, Chrysobothris scabripennis Gory and Laporte, Dicerca pugionata (Germar)) were new prey records for C. fumipennis. The wasps exhibited a strong preference for larger beetle genera (e.g., Dicerca, Buprestis), which accounted for 68% of beetles caught. Agrilus and Chrysobothris were the next dominant genera, accounting for 16% and 11%, respectively. A 4–19 mm prey size range is proposed, as all beetles collected were within this range despite the availability of prey outside of this range. Cerceris fumipennis caught 43% of the 42 buprestids species present in museum records from the four census counties as well as an additional 23 buprestid species that were not represented in museum records. Of the 22 buprestid species identified in museum collections that were not caught by C. fumipennis in the census counties, only one was within the proposed size range and active during the C. fumipennis flight season (late June through August). Overall, sampling C. fumipennis colonies over two summers at five sites resulted in 32% of the recorded buprestid species in New York State being caught, indicating that monitoring colonies is an efficient and viable means of quantifying buprestid assemblages.
doi:10.1673/031.014.04
PMCID: PMC4199360  PMID: 25373151
biomonitoring; biosurveillance; Crabronidae; insect museum collections; nest provisioning; wasp foraging behavior
8.  Sterile Insect Technique and F1 Sterility in the European Grapevine Moth, Lobesia botrana 
Newly emerged adults of the European grapevine moth, Lobesia botrana (Denis and Schiffermuller) (Lepidoptera: Tortricidae), were irradiated with various doses of gamma radiation and crossed to unirradiated counterparts of the opposite sex. Fecundity was decreased when unirradiated females were mated with either 300- or 350-Gy-irradiated males. Adult males that were irradiated with 400 Gy and mated with unirradiated females retained a residual fertility of 2.7%. The radiation dose at which irradiated females were found to be 100% sterile when mated with unirradiated males was 150 Gy. The inherited effects in the F1 progeny of irradiated male parents were examined at 100, 150, and 200 Gy. Fecundity and fertility of the F1 progeny of males irradiated with 150 Gy and inbred or crossed with irradiated and unirradiated moths were also recorded. A significant reduction in fertility was observed when F1 males mated with either F1 or unirradiated females. According to sterility index, F1 females who mated with F1 males had greater sterility than when F1 females were crossed to 150-Gy-irradiated males. Based upon the results of this study, 150 Gy of gamma radiation would be the optimal dose to use in a sterile insect technique and F1 sterility program against L. botrana.
doi:10.1673/031.014.08
PMCID: PMC4199361  PMID: 25373155
gamma radiation; inherited sterility; pest management; radiation biology
9.  Wolbachia Infection and Lepidoptera of Conservation Concern 
Conservation of at-risk species requires multi-faceted and carefully-considered management approaches to be successful. For arthropods, the presence of endosymbiotic bacteria, such as Wolbachia (Rickettsiales: Rickettsiaceae), may complicate management plans and exacerbate the challenges faced by conservation managers. Wolbachia poses a substantial and underappreciated threat to the conservation of arthropods because infection may induce a number of phenotypic effects, most of which are considered deleterious to the host population. In this study, the prevalence of Wolbachia infection in lepidopteran species of conservation concern was examined. Using standard molecular techniques, 22 species of Lepidoptera were screened, of which 19 were infected with Wolbachia. This rate is comparable to that observed in insects as a whole. However, this is likely an underestimate because geographic sampling was not extensive and may not have included infected segments of the species' ranges. Wolbachia infections may be particularly problematic for conservation management plans that incorporate captive propagation or translocation. Inadvertent introduction of Wolbachia into uninfected populations or introduction of a new strain may put these populations at greater risk for extinction. Further sampling to investigate the geographic extent of Wolbachia infections within species of conservation concern and experiments designed to determine the nature of the infection phenotype(s) are necessary to manage the potential threat of infection.
doi:10.1673/031.014.06
PMCID: PMC4199377  PMID: 25373153
captive rearing; endangered species; endosymbiont; translocation
10.  The Dufour's Gland and the Cuticle in the Social Wasp Ropalidia marginata Contain the Same Hydrocarbons in Similar Proportions 
Queens in many social insects are known to maintain their status through chemicals (pheromones) and cuticular hydrocarbons and have been the focus of many investigations that have looked at the chemicals involved in queen signaling. In the primitively eusocial wasp Ropalidia marginata Lepeletier (Hymenoptera: Vespidae), the Dufour's gland has been shown to be involved in queen signaling, and Dufour's gland hydrocarbons have been found to be correlated with fertility. Hence, this study analyzed the cuticle of R. marginata along with the Dufour's gland in order to compare their hydrocarbon profiles. The results show that the Dufour's gland and cuticle contained the same set of hydrocarbons in similar proportions (for the majority of compounds). Patterns pertaining to fertility signaling present in cuticular hydrocarbons were also similar to those present in the Dufour's gland hydrocarbons. Furthermore, the haemolymph contained the same hydrocarbons as found in the Dufour's gland and cuticle in similar proportions, thereby providing an explanation as to why the hydrocarbon profiles of the Dufour's gland and cuticle are correlated.
doi:10.1673/031.014.09
PMCID: PMC4199378  PMID: 25373156
cuticular hydrocarbons; fertility signaling; haemolymph
11.  Comparative Isoenzyme Electrophoreses between the Brown-Spotted Locust, Cyrtacanthacris tatarica, and the Desert Locust, Schistocerca gregaria 
The desert locust, Schistocerca gregaria (Forskål) (Orthoptera: Acrididae), and the brownspotted locust, Cyrtacanthacris tatarica (Linné) (Orthoptera: Acrididae), were collected from Saudi Arabia to investigate their relationships. Native polyacrylamide gel electrophoreses of five arbitrarily chosen metabolic enzymes extracted from the leg muscles of the two locust taxa were conducted. These enzymes were acid phosphatase (Acph), alcohol dehydrogenase (Adh), β esterase (β est), malic enzyme (Mal) and malate dehydrogenase (Mdh). Twenty presumptive gene loci and 26 polymorphic alleles were recorded. Acph did not discriminate between the two locust species, while the other four isoenzymes discriminated between them. Most of the alleles were monomeric, but Mal and Mdh exhibited dimeric alleles in the samples of C. tatarica. β est fractions were more expressed in C. tatarica, and the three enzymes β est, Mal, and Mdh discriminated clearly between the two species. The similarity coefficient that was calculated according to the number of sharing alleles between the two locusts was found to be 0.69. The isoenzyme variation presented herein seemed to reflect either their physiological adaptation or the taxonomic consequences between the two taxa. Collecting more isoenzymes for more samples could have taxonomic value.
doi:10.1673/031.014.20
PMCID: PMC4199529  PMID: 25373167
gene locust; guardian locust; Saudi Arabia
12.  Effects of Roads on Castanopsis carlesii Seedlings and Their Leaf Herbivory in a Subtropical Forest in China 
The effects of a forest road on Castanopsis carlesii (Hemsley) Hayata (Fagales: Fagaceae) seedlings and their leaf herbivory were investigated in a subtropical forest at Jiulianshan National Nature Reserve, Jiangxi, China. A total of 1124 seedlings, 33949 leaves, 468 leaf mines, and 205 leaf galls were found. Generally, individual numbers, tree heights, and leaf numbers of C. carlesii seedlings became lower with increasing distances from the road. These results might indicate that old seedlings were fewer and survival rate of seedlings was lower in forest interiors. Leaf miners preferred the seedlings close to the forest road, while leaf gallers preferred the seedlings about 2 m from the road. Species diversity of leaf miners was higher in the forest interior area, while species diversity of leaf gallers was higher near the road. However, both leaf miners and leaf gallers decreased in general from the road to the interior forest. There were interspecific differences in the effects of roads on leaf miner species and leaf galler species. The effects of the road on seedlings and insects could be explained by varying microhabitat conditions and different ecological strategies.
doi:10.1673/031.014.17
PMCID: PMC4199530  PMID: 25373164
leaf miners; leaf gallers; road ecology; species diversity
13.  Morphological Variation on Isolated Populations of Praocis (Praocis) spinolai 
In this study, the morphological variations of four geographically isolated populations of Praocis (Praocis) spinolai Gay & Solier (Coleoptera: Tenebrionidae) in the transitional coastal desert, Chile, were studied. The study was conducted in the coastal area of Punta de Choros and Los Choros-Archipelago, which includes three islands: Choros, Damas, and Gaviota. 113 specimens of the species P. (P.) spinolai belonging to the four locations sampled were collected analyzed with geometric morphometrics techniques to explore the pattern of shape variation on the different isolated environments. The principal component analysis revealed a well-defined pattern of variation between the populations analyzed. Moreover, differences between populations emerged also from the canonical variation analysis and were confirmed by the Procrustes ANOVA. All analyses performed confirmed the existence of a pattern of variation, due to the isolation of the populations and to environmental effects. The islands are subject to more arid pressures than the continent, where there is a more stable environment and the presence of coastal wetlands and the coastal range of mountains act together and enable fog condensation. This study indicates the existence of a clear pattern of variation, which indicates an evolutionary trend among the population examined.
doi:10.1673/031.014.11
PMCID: PMC4199532  PMID: 25373158
coastal desert; epigean tenenebrionids; geometric morphometrics; Pingüino de Humboldt National Reserve
14.  Toxicological and Biochemical Characterizations of AChE in Phosalone-Susceptible and Resistant Populations of the Common Pistachio Psyllid, Agonoscena pistaciae 
The toxicological and biochemical characteristics of acetylcholinesterases (AChE) in nine populations of the common pistachio psyllid, Agonoscena pistaciae Burckhardt and Lauterer (Hemiptera: Psyllidae), were investigated in Kerman Province, Iran. Nine A. pistaciae populations were collected from pistachio orchards, Pistacia vera L. (Sapindales: Anacardiaceae), located in Rafsanjan, Anar, Bam, Kerman, Shahrbabak, Herat, Sirjan, Pariz, and Paghaleh regions of Kerman province. The previous bioassay results showed these populations were susceptible or resistant to phosalone, and the Rafsanjan population was most resistant, with a resistance ratio of 11.3. The specific activity of AChE in the Rafsanjan population was significantly higher than in the susceptible population (Bam). The affinity (KM) and hydrolyzing efficiency (Vmax) of AChE on acetylthiocholine iodide, butyrylthiocholine iodide, and propionylthiocholine odide as artificial substrates were clearly lower in the Bam population than that in the Rafsanjan population. These results indicated that the AChE of the Rafsanjan population had lower affinity to these substrates than that of the susceptible population. The higher Vmax value in the Rafsanjan population compared to the susceptible population suggests a possible over expression of AChE in the Rafsanjan population. The in vitro inhibitory effect of several organophosphates and carbamates on AChE of the Rafsanjan and Bam populations was determined. Based on I50, the results showed that the ratios of AChE insensitivity of the resistant to susceptible populations were 23 and 21.7-fold to monocrotophos and phosphamidon, respectively. Whereas, the insensitivity ratios for Rafsanjan population were 0.86, 0.8, 0.78, 0.46, and 0.43 for carbaryl, eserine, propoxur, m-tolyl methyl carbamate, and carbofuran, respectively, suggesting negatively correlated sensitivity to organophosphate-insensitive AChE. Therefore, AChE from the Rafsanjan population showed negatively correlated sensitivity, being insensitive to phosphamidon and monocrotophos and sensitive to N-methyl carbamates.
doi:10.1673/031.014.18
PMCID: PMC4199533  PMID: 25373165
AChE insensitivity; enzyme inhibition; kinetic parameters; organophosphate resistance
15.  Keys to the Species of Mydaeinae (Diptera: Muscidae) from China, with the Description of Four New Species 
Four new species of Mydaeinae, Mydaea franzosternita n. sp., Myospila apicaliciliola n. sp., Myospila maoershanensis n. sp., and Myospila subflavipennis n. sp., are described and illustrated here for the first time. A key to the genus of Mydaeinae from China and keys to species of genera from Mydaeinae are provided.
doi:10.1673/031.014.22
PMCID: PMC4199534  PMID: 25373169
Chinese species; classification; Mydaea; Myospila
16.  The Reproductive Performance of the Mupli Beetle, Luprops tristis, in Relation to Leaf Age of the Para Rubber Tree, Hevea brasiliensis 
An analysis of host plant leaf age preferences and phenology studies led to the predictions that tender rubber plant leaves are essential for the completion of the life cycle of the Mupli beetle, Luprops tristis Fabricius (Coleoptera: Tenebrionidae) and that low tender leaf availability during the post-dormancy stage will limit the beetle population. Analyses of the effects of feeding the beetles leaves of various ages, nitrogen (N) content, and moisture content on fecundity and the duration of post-dormancy survival were carried out. The results showed that tender leaf availability during the post-dormancy phase of L. tristis is a critical factor that determines the survival of L. tristis adults and the subsequent generation. The control of powdery mildew (Odium hevea) disease-mediated premature leaf fall in rubber plantations may regulate the beetle population. A peak in fecundity during the early phase of post-dormancy is proposed as an adaptive mechanism of L. tristis to synchronize egg production and feeding with tender leaf availability in rubber plantations. Variations in nutrient levels and moisture content between deciduous rubber tree leaves of different ages are attributed to the leaf nutrient resorption mechanism of senescing leaves. These results established that tender leaves with high N and moisture levels are essential for post-dormancy survival and that N influences fecundity. The results of the experiments could aid decision making regarding the population management and control of L. tristis in rubber plantations.
doi:10.1673/031.014.12
PMCID: PMC4199535  PMID: 25373159
fecundity; leaf age performance; leaf nutrient resorption; leaf substrate quality; survival
17.  Polydomy in the ant Ectatomma opaciventre 
Tropical ants commonly exhibit a hyper-dispersed pattern of spatial distribution of nests. In polydomous species, nests may be satellites, that is, secondary structures of the main nest, where the queen is found. In order to evaluate whether the ant Ectatomma opaciventre Roger (Formicidae: Ectatomminae) uses the strategy of building polydomous nests, the spatial distribution pattern of 33 nests in a 1,800 m2 degraded area located in Rio Claro, SP, Brazil, were investigated using the nearest neighbor method. To complement the results of this investigation, the cuticular chemical profile of eight colonies was analyzed using Fourier transform infrared photoacoustic spectroscopy (FTIR-PAS). The nests of E. opaciventre presented a hyper-dispersed or regular distribution, which is the most common in ants. The analysis of the cuticular hydrocarbons apparently confirmed the hypothesis that this species is polydomous, since the chemical profiles of all studied colonies with nests at different sites were very similar to the chemical signature of the single found queen and were also different from those of colonies used as control.
doi:10.1673/031.014.21
PMCID: PMC4199536  PMID: 25373168
colony organization; cuticular hydrocarbons; nestmate recognition; satellite nests; spatial distribution
18.  A Comparison of the Life-History Traits between Diapause and Direct Development iNdividuals in the Cotton Bollworm, Helicoverpa armigera 
In order to understand the differences of life-history traits between diapause and direct development individuals in the cotton bollworm, Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae), the development time, body size, growth rate, and adult longevity were investigated between the two populations, which were induced under 12:12 L:D and 16:8 L:D photoperiods, respectively, at 20, 22, and 25°C. The results indicated that the larval development time, pupal weight, adult weight, and growth rate were significantly different between diapause and direct developing individuals. The diapause developing individuals had a significantly higher pupal and adult weight and a longer larval time compared with direct developing individuals. However, the growth rate in diapause developing individuals was lower than that in the direct developing individuals. Analysis by GLM showed that larval time, pupal and adult weight, and growth rate were significantly influenced by both temperature and developmental pathway. The pupal and adult weights were greater in males than females in both developmental pathways, exhibiting sexual size dimorphism. The dimorphism in adult weight was more pronounced than in pupal weight because female pupae lost more weight at metamorphosis compared to male pupae. Protogyny was observed in both developmental pathways. However, the protogyny phenomenon was more pronounced at lower temperatures in direct developing individuals, whereas it was more pronounced in diapause developing individuals when they experienced higher temperatures in their larval stage and partial pupal period. The adult longevity of diapause developing individuals was significantly longer than that of direct developing individuals. The results reveal that the lifehistory strategy was different between diapause and direct developing individuals.
doi:10.1673/031.014.19
PMCID: PMC4199537  PMID: 25373166
developmental pathways; protogyny; theremal reaction
19.  Survey of Mymarommatidae and Their Occurrence in Agricultural Systems in Brazil 
Mymarommatidae surveys were carried out through the use of yellow pan traps in crops of green dwarf coconut, Cocos nucifera L. (Arecales: Arecaceae), papaya, Carica papaya L. (Brassicales: Caricaceae), citrus, Citrus spp. L. (Sapindales: Rutaceae), and guava, Psidium guajava L. (Myrtales: Myrtaceae), in the northern Espirito Santo State, Brazil. 146 specimens of mymarommatids were collected, of which 71, 55, 16, and 4 exemplars were obtained in the area cultivated with guava, papaya, citrus, and coconut, respectively. The mean numbers of mymarommatids collected in the period from April to June 2011 were significantly higher than those obtained in the other nine months. Two genera, Mymaromma and Mymaromella, were identified The most abundant genus was Mymaromma, comprising 93.8% of the total collection; however, the genus Mymaromella was encountered in all crops. This is the first record of the presence of mymarommatids in these agricultural systems.
doi:10.1673/031.014.15
PMCID: PMC4199538  PMID: 25373162
Hymenoptera; parasitoid; yellow pan traps
20.  P Element Activity and Molecular Structure in Drosophila melanogaster Populations from Firtina Valley, Turkey 
In order to study P element dynamics in natural populations of Drosophila melanogaster, 88 isofemale lines were examined from the Firtina Valley, Turkey. The P-M gonadal dysgenesis characteristics and the molecular patterns of P and KP elements were analyzed. Gonadal dysgenesis tests showed a slight variation both for P activity and P susceptibility, however the results showed a predominant M' phenotype for this region. The P and KP element were also characterized by polymerase chain reaction. The molecular analyses showed that all the populations examined had the entire 1.15 kb KP element. The molecular patterns of KP elements were the same for the populations studied. No clear relationship was found between phenotype and genomic P element composition. The correlations between the level of gonadal dysgenesis percentage (as an index for P activity and P susceptibility) and several geoclimatic factors were tested, and no general effects of altitude, temperature, rainfall, or humidity were found. The theoretical P' strain, which is very rare in natural populations, was also recorded for this region.
doi:10.1673/031.014.16
PMCID: PMC4199539  PMID: 25373163
geoclimatic variables; transposable elements
21.  Serum-Free Culture of the Suspension Cell Line QB-Tn9-4s of the Cabbage Looper, Trichoplusia ni, is Highly Productive for Virus Replication and Recombinant Protein Expression 
Serum-free cultures of insect cells play an important role in the fields of protein engineering, medicine, and biology. In this paper, the suspension cell line QB-Tn9-4s of Trichoplusia ni (Hübner) (Lepidoptera: Noctuidae) was successfully adapted to serum-free Sf-900 III medium and passaged for 52 generations. The adapted QB-Tn9-4s cells grew faster. Their population doubling time shortened from 27.4 hr in serum-containing medium to 24.1 hr, and their maximal density increased by 1.83-fold, reaching 3.50 × 106 cells/mL in serum-free culture in T-flasks. The cells readily adapted to spinner culture, with maximum cell density of 4.40 × 106 cells/mL in a spinner flask. Although the infection rate of Autographa californica multiple nucleopolyhedrovirus and production of occlusion bodies (OBs) of the adapted QB-Tn9-4s cells were 91.0% and 85.4 OBs/cell, respectively, similar to those of QB-Tn9-4s cells cultured in serum-containing medium and control BTI-Tn5B1-4 cells, their budded virus titer was 4.97 × 107 TCID50/mL, significantly higher than those of the latter two. In addition, the expression levels of β-galactosidase at six days post-infection and secreted alkaline phosphatase at seven days postinfection in the adapted QB-Tn9-4s cells reached 2.98 ± 0.15×104 IU/mL and 3.34 ± 0.13 IU/mL, respectively, significantly higher than those of QB-Tn9-4s and control BTI-Tn5B1-4 cultured in serum-containing media. The above findings establish a foundation for industrial production of virus and recombinant proteins in QB-Tn9-4s serum-free culture.
doi:10.1673/031.014.24
PMCID: PMC4199540  PMID: 25373171
insect cell lines; population doubling time; virus production
22.  The Fern-Feeder Aphids (Hemiptera: Aphididae) from China: A Generic Account, Descriptions of One New Genus, One New Species, One New Subspecies, and Keys 
Fern-feeder aphids (Hemiptera: Aphididae) in China are represented by 13 species in 10 genera, including a new genus, Vietaphis gen. nov., a new species, Vietaphis aliquantus sp. nov., from Guizhou and Tibet on Plagiogyria japonicum, and a new subspecies, Amphorophora scabripes galba ssp. nov., from Guizhou on Pentarhizidium intermedium. Two genera, Amphorophora Buckton and Idiopterus Davis, and four species or subspecies, Amphorophora ampullate bengalensis Hille Ris Lambers and Basu, Idiopterus nephrelepidis Davis, Micromyzodium polypodii Takahashi, and Myzus filicis Basu, are reported for the first time in China. Apterae and alatae of Myzus filicis are redescribed herein, and with host plant notes. The fern-feeder aphid genus Neomacromyzus Lee is considered a junior synonym of Idiopterus. Furthermore, Neomacromyzus cyrtomicola Lee is transferred to the genus Idiopterus, as Idiopterus cyrtomicola (Lee), comb. nov., which is herein considered a junior synonym of Idiopterus nephrelepidis Davis. Keys to Chinese fern-feeder species are provided. Morphological figures and biometrical data of Vietaphis aliquantus sp. nov., Amphorophora scabripes galba ssp. nov., and Myzus filicis are presented.
doi:10.1673/031.014.23
PMCID: PMC4199544  PMID: 25373170
23.  Spermatozoon Ultrastructure of Hangingflies, Bittacus strigosus and Bittacus stigmaterus 
In the present study, spermatozoon ultrastructure was documented in two species of hangingflies, Bittacus strigosus Hagen (Mecoptera: Bittacidae) and B. stigmaterus Say. Structures considered important to phylogenetic assessment that were observed in B. strigosus and B. stigmaterus included a short bilayered acrosome, elongated nucleus, tube-like glycocalyx, centriole adjunct material, accessory bodies, two mitochondrial derivatives, extra axonemal rods, globular units, and 9+2 arrangement of microtubules in the axoneme. Comparisons were made to Bittacus planus Cheng, which was previously examined by electron microscopy (Xie and Hua 2010). Similarities among the ultrastructural characteristics of the three Bittacus species support the monophyly of this genus. Displacement of a mitochondrial derivative by an accessory body was documented for the first time. This paper includes clarifications on differences between accessory bodies and extra axonemal rods, which are issues important to phylogenetic placement.
doi:10.1673/031.014.10
PMCID: PMC4204384  PMID: 25373157
electron microscopy; insect phylogeny; sperm
24.  Quantification of Supercolonial Traits in the Yellow Crazy Ant, Anoplolepis gracilipes 
Supercoloniality is a social structure displayed by many invasive ant species, but there has been surprisingly little research quantifying the extent to which individual species display traits underlying such social organisation. This study quantifies three traits for the yellow crazy ant, Anoplolepis gracilipes Smith (Hymenoptera: Formicidae): little or no aggression between workers from different nests; the exchange of workers among nests; and resource exchange among nests, as well as supercolony structure arising from patterns of distribution and density of detections. Supercolonies displayed a structural continuum from being small (< 10 ha) and “aggregated” with great continuity among detections through to being large (> 10,000 ha) and “diffuse” with little continuity among detections. Smaller supercolonies had greater ant densities than larger supercolonies. In laboratory trials, no aggression was observed between workers from different nests sourced from different supercolonies, and paired nests merged within 24 hours. Workers lacked nest fidelity by rapidly populating artificial nests containing alien queens. The daily worker turnover rate per nest was estimated to be below 20%. Resources were readily moved among nests, with a resource being detected up to 13 m away from a source within 24 hours, and as far as 32 m after four days. The rate and distance of resource movement increased with increasing worker and nest density. This research has demonstrated that A. gracilipes displays supercoloniality equivalent to that of the well-studied Argentine ant Linepithema humile. Quantification of these traits is required for other supercolonial species to improve our understanding of this social strategy, especially for invasive ants to aid in understanding factors that promote invasion success and to improve management.
doi:10.1673/031.014.25
PMCID: PMC4204385  PMID: 25373172
colony; invasion; nest fidelity; polygyny; supercolony; resource flow
25.  Identification and Analysis of the Pigment Composition and Sources in the Colored Cocoon of the Silkworm, Bombyx mori, by HPLC-DAD 
This study used the larval tissues and colored cocoons of silkworms, Bombyx mori L. (Lepidoptera: Bombycidae), that were fed leaves of cultivated mulberry, Husang 32, as experimental material. The pigment composition and content in colored cocoons and tissues of the 5th instar larvae and the mulberry leaves were rapidly detected using organic solvent extraction and reverse phase high-performance liquid chromatography with diode array detection. It was found that the mulberry leaf mainly contained four types of pigment: lutein (30.86%), β-carotene (26.3%), chlorophyll a (24.62%), and chlorophyll b (18.21%). The silk glands, blood, and cocoon shells of six yellow-red cocoons were used as the experimental materials. The results showed that there were generally two kinds of carotenoids (lutein and β-carotene) in the silk gland and cocoon shell, a little violaxanthin was detected in silk gland, and the pigment found in the blood was mainly lutein in all varieties of silkworm tested. Chlorophyll a and b had not been digested and utilized in the yellow-red series of silkworm. The method used to detect visible pigments reported here could be used to breed new colors of cocoons and to develop and utilize the pigments found in mulberry.
doi:10.1673/031.014.31
PMCID: PMC4206223  PMID: 25373178
β-carotene; chlorophyll; lutein; mulberry leaf

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