The authors would like to thank the U.S. Department of Agriculture Forest Service for funding this project through USFS Agreement 13-CS-11090800-022. Support for ECZ was provided by NSF-DBI-1263050. ECZ assisted in the development of the research concept, collected all field data, conducted laboratory analysis, and produced the original manuscript. MWE assisted in the development of the research concept and study design, assisted in directing field data collection and laboratory analysis, and heavily edited the manuscript. KPS assisted with study design, directed the field and laboratory work, assisted with data analysis, and reviewed the manuscript.

1. Placement of a trap on the tree
<ol>
	<li>Measure the diameter of a tree at breast height. At breast height in each cardinal direction, for an area the size of the pre-manufactured sticky trap (glue board), use a bark shaver to remove bark until an area the size for the sticky trap is smooth enough to staple the sticky trap onto the tree so that there is no space for arthropods to crawl under the trap. Label the back of the trap using a dark colored permanent marker with the date, trap number, location and other pertinent information.
	<ol>
		<li>To trap arthropods, either (a) capture both flying and crawling arthropods, by opening and removing the sides and cover of the sticky trap by cutting the cardboard along the edge of the sticky material, (b) or exclude flying arthropods from landing directly on the trap, by opening the trap as directed on the box.</li>
	</ol>
	</li>
	<li>Place one trap on each previously shaved location so that the openings are oriented vertically (one opening facing up, the other opening facing down) to maximize capture of arthropods crawling up and down the tree boles. For traps with the tops removed to capture both flying and crawling arthropods, orient traps so the end that was the opening prior to the removal of the cardboard cover is oriented vertically, to maintain trapping consistency.</li>
	<li>Staple traps to the tree by placing one staple at each corner and one staple in the center bottom and center top of the trap. Start stapling in the bottom right corner, then the bottom center, the top right corner, the top right center, the bottom left corner, and finally the top left corner. Be careful to ensure the entire bottom and top of the traps are flush against the tree to minimize arthropods crawling under the trap.</li>
	<li>Leave traps in place for desired amount of time. Be certain all traps are left in place the same amount of time. 
	NOTE: In areas where arthropods are extremely abundant, for example during moth outbreaks, traps may become saturated within hour or days. Under these circumstances, traps will need to be regularly replaced prior to be saturated to maintain constant capture probability.</li>
</ol>
2. Removing the trap from the tree
<ol>
	<li>After the desired amount of time trapping, cover the entire trap, except for the staples, with polymeric cellulose film (e.g., cellophane). 
	NOTE: Placing the film on the traps prior to removal will reduce the likelihood of disturbing the trapped arthropods.</li>
	<li>Remove each trap by taking a large flat screwdriver and prying each staple partially from the tree, adequate to facilitate the grasping of the staples using needle nose pliers. Take large needle nose pliers or a similar grasping tool and pull the staples from the tree.</li>
	<li>Place the traps in a rigid box of some type for transportation to a laboratory for analysis. If traps are to be stored for more than 12 h, store traps in a freezer to preserve content.</li>
</ol>
3. Laboratory analysis
<ol>
	<li>Using a dissecting scope, examine content of a trap recording the number of individuals to desired taxonomic level.</li>
	<li>Use sorted arthropods to estimate richness (total number of taxonomic groups), diversity indices, or abundance (total arthropods). If estimated biomass is a desired result, measure length and width of arthropods to the nearest mm and use published length/width, biomass regressions to estimate biomass32,33,34.</li>
	<li>Subtract the total width of the 4 traps from the diameter at breast height for each tree to estimate trapping effort (proportion of tree covered by the traps) for each tree.</li>
	<li>Because samples from multiple traps on the same tree are not independent, either sum samples from the same tree or include individual tree as a random variable in all analysis to avoid pseudo-replication.</li>
</ol>

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The authors have nothing to disclose.

Although alternative techniques such as suction or sweep nets have been used, most previously published attempts at quantifying arthropods on tree boles used some version of either quantifying arthropods by visually inspecting tree boles in the field, using chemical pesticides to kill arthropods in a specified area then quantifying the recovered arthropods, or placing funnel traps or a sticky substance directly onto the tree19,23,25</...

Terrestrial arthropods play an important role in our environment. In addition to being of scientific interest in their own right, arthropods can be both detrimental and beneficial to other trophic levels (i.e., crops, horticultural plants, native vegetation, and food for insectivorous organisms1,2,3,4). Thus, understanding the factors that influence arthropod community development and abundance is critical to farmers5, pest control managers6, foresters4, plant biologists7, entomologists8, and wildlife and conservation ecologists that study community dynamics and manage insectivorous organisms9. Arthropod communities vary in species composition and abundance both temporally and spatially across a variety of ecological landscapes including plant communities, plant species, and across various regions of individual plants. For example, studies have demonstrated significant differences in arthropod community metrics between the roots, bole and stems, and foliage, within the same individual tree10,11. These findings are not surprising considering that different parts of the same plant, e.g., leaves versus barks of a tree, provide different resources for which arthropods have adapted to exploit. Thus, each part of the plant can support a different arthropod community. Because foliage dwelling arthropods can have such a large socioeconomic and environmental impact, substantial effort has been expended to measure community metrics using both qualitative and quantitative approaches12. Alternatively, much less effort has been expended to develop approaches of quantifying corticolous (bark-dwelling) arthropod communities.
Like foliage-dwelling arthropod communities, corticolous arthropod communities can be important from both a socioeconomic and environmental viewpoint. Some forest diseases that are caused or facilitated by corticolous arthropods can be detrimental to economically viable timber harvest4. Additionally, corticolous arthropods can be an important component of the food chain in forest communities13,14. For example, forest dwelling arthropods are the primary food source for many insectivorous bark gleaning song birds15,16. Thus, understanding the factors that influence communities of corticolous arthropods is of interest to foresters and both basic and applied ecologists.
Understanding factors that influence arthropod community composition and abundance often requires the capture of individuals. Capture techniques can generally be categorized into qualitative techniques that only detect presence of a species for estimates of species range, richness, and diversity17, or semi-quantitative and quantitative techniques that allow for an index or estimate of abundance and density of individuals within a taxonomic group18,19. Semi-quantitative and quantitative techniques allow researchers to estimate or at least consistently sample a specified sample area and estimate probability of detection or assume detection probability is non-directional and adequate as to not obscure the researcher&#39;s ability to detect spatial or temporal variation in abundance. Semi-quantitative and quantitative techniques for quantifying corticolous arthropods include suction or vacuum sampling of a specific area20,21,22, systematic counting of visible arthropods18,23, sticky traps24, various funnel or pot-type traps8,25, and entrance or emergent holes26,27.
A number of spatial and temporal factors are thought to lead to variation in corticolous arthropod communities11,14,28,29. For example, texture of tree bark is thought to influence the community structure of tree-dwelling arthropods14. Because of the more diverse surface area of the trunks of trees with more furrowed bark, trees with more furrowed bark are thought to support a greater diversity and abundance of arthropods14.
With this article we report a new semi-quantitative approach of enumerating corticolous arthropods that could be used to describe and test hypotheses regarding variation in corticolous arthropod communities across time and space with adequate precision to detect differences among tree species. Using sticky traps attached to the trunks of trees, we compared the abundance, total length (a surrogate for body mass), richness, and diversity of the arthropod community on the bole of white oak (Quercus alba), pignut hickory (Carya glabra), sugar maple (Acer saccharum), American beech (Fagus grandifolia), and tulip poplar (Liriodendron tulipifera) trees, trees that vary in bark texture.
This study was conducted in the Ozark and Shawnee Hills ecological sections of the Shawnee National Forest (SNF) in southwestern Illinois. During July 2015, we identified 18 (9 dominated by oak/hickory and 9 dominated by beech/maple) sites with the USFS stand cover map for the SNF (allveg2008.shp) in ArcGIS 10.1.1. In the xeric sites, the dominant species were pignut hickory and white oak and in mesic sites, the dominant species were American beech, sugar maple, and tulip poplar. To compare bole arthropod community among tree species, at each data collection site, we identified the three of the five (white oak, pignut hickory, sugar maple, American beech and tulip poplar) focal species trees &#62;17 cm diameter at breast height (d.b.h.) closest to the center of a 10 m radial circle. If fewer than three appropriate trees were present, the circle was expanded and the closest tree fitting the criteria was selected. For each tree chosen, we installed four sticky traps at breast height, one facing in each cardinal direction: north, south, east and west.
We collected arthropod data from the boles of 54 individual trees (12 pignut hickories, 15 white oaks, 8 American beeches, 12 sugar maples, and 7 tulip poplars) among the 18 sites. We grouped arthropods according to a simplified guild classification by diagnostic morphological characteristics indicative of closely related orders from current phylogenetic records, similar to that of &#34;operational taxonomic units&#34;30,31 (Appendix A). Based on this classification, we captured representatives of 26 guilds in our traps that were each in place for 9 days (Appendix A). Because our study focused on trophic interactions between tree species, corticolous arthropods, and bark-gleaning birds, we removed all arthropods smaller than 3 mm from analysis because their importance as a food resource is minimal for bark-gleaning birds. We used a mixed model that included either arthropod length (surrogate to body mass), abundance, Shannon diversity and, richness as the dependent variable, tree species and effort (proportion of tree covered with traps) as fixed variables, and site as a random variable. Because all traps from a single tree were combined as one sample, individual trees were not included as a random variable.

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quantifying corticolous arthropods using sticky traps

Terrestrial arthropods play an important role in our environment. Quantifying arthropods in a way that allows for a precise index or estimate of density requires a method with high detection probability and a consistent sampling area. We used manufactured sticky traps to compare abundance, total length (a surrogate for biomass), richness, and Shannon diversity of corticolous arthropods among the boles of 5 tree species. Efficacy of this method was adequate to detect variation in corticolous arthropods among tree species and provide a standard error of the mean that was &#60;20% of the mean for all estimates with sample sizes from 7 to 15 individual trees of each species. Our results indicate, even with these moderate sample sizes, the level of precision of arthropod community metrics produced with this approach is adequate to address most ecological questions regarding temporal and spatial variation in corticolous arthropods. Results from this method differ from other quantitative approaches such as chemical knockdown, visual inspection, and funnel traps in that they provide an indication of corticolous arthropod activity over a relatively long-term, better including temporary bole residents, flying arthropods that temporarily land on the tree bole and crawling arthropods that use the tree bole as a travel route from the ground to higher forest foliage. Furthermore, we believe that commercially manufactured sticky traps provide more precise estimates and are logistically simpler than the previously described method of directly applying a sticky material to tree bark or applying a sticky material to tape or other type of backing and applying that to the tree bark.

Based on the mixed model results, the model that included tree species best explained variation in total arthropod length, abundance, and diversity, neither of independent variables explained substantial variation in richness, although the models that included tree species trapping effort were competitive with the null model (Table 1). In addition, proportion of the tree trapped appears to have no influence on abundance, total length, and Shannon diversity, with only mini...

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Quantifying Corticolous Arthropods Using Sticky Traps

We describe a semi-quantitative approach of measuring characteristics of corticolous (bark-dwelling) arthropod communities. We placed commercially manufactured sticky traps on tree boles to estimate abundance, total length (a surrogate to biomass), richness, and Shannon diversity for comparison among tree species.

Terrestrial arthropods play an important role in our environment. Quantifying arthropods in a way that allows for a precise index or estimate of density ...

quantifying-corticolous-arthropods-using-sticky-traps

Research

JoVE Journal

Environment

5.4K Views.  Southern Illinois University. Dit protocol kan worden gebruikt om de korte en lange termijn gemeenschapsstructuur van corticolous geleedpotigen te schatten. Deze techniek is handig voor het vastleggen van zowel vliegende als kruipende geleedpotigen en vergemakkelijkt eenvoudige laboratoriumanalyse. Om te beginnen met deze procedure meet de diameter van een boom op borsthoogte.Gebruik een schors scheerapparaat om te beginnen met het verwijderen van de schors op deze hoogte in elke kardinale richting in een gebied te...

Video: Quantifying Corticolous Arthropods Using Sticky Traps

Administering and Detecting Protein Marks on Arthropods for Dispersal Research

Monitoring arthropod movement is often required to better understand associated population dynamics, dispersal patterns, host plant preferences, and other ecological interactions. Arthropods are usually tracked in nature by tagging them with a unique mark and then re-collecting them over time and space to determine their dispersal capabilities. In addition to actual physical tags, such as colored dust or paint, various types of proteins have proven very effective for marking arthropods for ecological research. Proteins can be administered internally and/or externally. The proteins can then be detected on recaptured arthropods with a protein-specific enzyme-linked immunosorbent assay (ELISA). Here we describe protocols for externally and internally tagging arthropods with protein. Two simple experimental examples are demonstrated: (1) an internal protein mark introduced to an insect by providing a protein-enriched diet and (2) an external protein mark topically applied to an insect using a medical nebulizer. We then relate a step-by-step guide of the sandwich and indirect ELISA methods used to detect protein marks on the insects. In this demonstration, various aspects of the acquisition and detection of protein markers on arthropods for mark-release-recapture, mark-capture, and self-mark-capture types of research are discussed, along with the various ways that the immunomarking procedure has been adapted to suit a wide variety of research objectives.

Proteins are often used to mark arthropods for dispersal research. Methods for administering internal and external protein marks on arthropods are demonstrated. Protein mark detection techniques, either through indirect or sandwich enzyme-linked immunosorbent assays (ELISAs), are also illustrated.

Monitoring arthropod movement is often required to better understand associated population dynamics, dispersal patterns, host plant preferences, and other ...

Individual Culturing of Tigriopus Copepods and Quantitative Analysis of Their Mate-guarding Behavior

Copepods of the genus Tigriopus, which are common zooplankton in rocky tide pools, show precopulatory mate-guarding behavior where a male clasps a potential mate to form a pair. While this phenomenon has attracted interest of researchers, methods for its analysis have not been well described. Here we describe procedures for: 1) individual culturing and staging of Tigriopus juveniles and adults, and 2) video-based analysis of their mate-guarding behavior. The culturing method enables experimental control of paring experience of animals as well as the ability to track their development before behavioral tests. The analysis method allows quantitative evaluation of several aspects of the mate-guarding behavior, including capturing attempts by males and swimming trajectory of mate-guarding pairs. Although these methods were originally established for ethological studies on Tigriopus, with proper modifications they can also be applied to studies of other zooplankton in different research fields, such as physiology, toxicology, and ecological genetics.

Individual Culturing of Tigriopus Copepods and Quantitative Analysis of Their Mate-guarding Behavior

Mate-guarding behavior plays an important role in reproduction of intertidal copepods of the genus Tigriopus. However, methods for studying this behavior have not been well described. Here we describe methods for: 1) individual culture of virgin Tigriopus animals, and 2) quantitative analysis of their mate-guarding behavior.

Copepods of the genus Tigriopus, which are common zooplankton in rocky tide pools, show precopulatory mate-guarding behavior where a male clasps a ...

Key Elements of Photo Attraction Bioassay for Insect Studies or Monitoring Programs

Optimized visual attractants will increase insect trapping efficiency by using the target insect&#39;s innate behaviors (positive photo-taxis) as a means to lure the insect into a population control or monitoring trap. Light emitting diodes (LEDs) have created customizable lighting options with specific wavelengths (colors), intensities, and bandwidths, all of which can be customized to the target insects. Photo-attraction behavioral bioassays can use LEDs to optimize the attractive color(s) for an insect species down to specific life history stages or behaviors (mating, feeding, or seeking shelter). Researchers must then confirm the bioassay results in the field and understand the limited attractive distance of the visual attractants.
The cloverleaf bioassay arena is a flexible method to assess photo attraction while also assessing a range of natural insect behaviors such as escape and feeding responses. The arena can be used for terrestrial or aerial insect experiments, as well as diurnal, and nocturnal insects. Data collection techniques with the arena are videotaping, counting contact with the lights, or physically collecting the insects as they are attracted towards the lights. The assay accounts for insects that make no-choice and the arenas can be single (noncompetitive) color or multiple (competitive) colors. The cloverleaf design causes insects with strong thigmotaxis to return to the center of the arena where they can view all the options in a competitive LED tests. The cloverleaf arena presented here has been used with mosquitoes, bed bugs, Hessian fly, house flies, biting midges, red flour beetles, and psocids. Bioassays are used to develop accurate and effective insect traps to guide the development and optimization of insect traps used to monitor pest population fluctuations for disease vector risk assessments, the introduction of invasive species, and/or be used for population suppression.

Photo-attraction bioassay arenas are used to determine the optimal light color(s) to maximize insect attraction; however bioassays and methods are specific to target insect behaviors and habitats. Customizable equipment and modifications are explained for nocturnal or diurnal and terrestrial or aerial insects.

Optimized visual attractants will increase insect trapping efficiency by using the target insect&#39;s innate behaviors (positive photo-taxis) as a means ...

Behavior

A Method for Quantifying Foliage-Dwelling Arthropods

Terrestrial arthropods play an important role in our environment. Quantifying arthropods in a way that allows for a precise index or estimates of density requires a method with high detection probability and a known sampling area. While most described methods provide a qualitative or semi-quantitative estimate adequate for describing species presence, richness, and diversity, few provide an adequately consistent detection probability and known or consistent sampling areas to provide an index or estimate with adequate precision to detect differences in abundance across environmental, spatial, or temporal variables. We describe how to quantify leaf-dwelling arthropods by sealing the leaves and end of branches in a bag, clipping and freezing the bagged material, and rinsing the previously frozen material in water to separate arthropods from the substrate and quantify them. As we demonstrate, this method can be used at a landscape scale to quantify leaf-dwelling arthropods with adequate precision to test for and describe how spatial, temporal, environmental, and ecological variables influence arthropod richness and abundance. This method allowed us to detect differences in density, richness, and diversity of leaf-dwelling arthropods among 5 genera of trees commonly found in southeastern deciduous forests.

We describe how to quantify leaf dwelling arthropods by sealing the leaves and end of branches in a bag, clipping and freezing the bagged material, and rinsing the previously frozen material in water to separate arthropods from the substrate for quantification.

Terrestrial arthropods play an important role in our environment. Quantifying arthropods in a way that allows for a precise index or estimates of density ...

A Visual Guide for Studying Behavioral Defenses to Pathogen Attacks in Leaf-Cutting Ants

The complex lifestyle, evolutionary history of advanced cooperation, and disease defenses of leaf-cutting ants are well studied. Although numerous studies have described the behaviors connected with disease defense, and the associated use of chemicals and antimicrobials, no common visual reference has been made. The main aim of this study was to record short clips of behaviors involved in disease defense, both prophylactically and directly targeted towards an antagonist of the colony following infection. To do so we used an infection experiment, with sub-colonies of the leaf-cutting ant species Acromyrmex echinatior, and the most significant known pathogenic threat to the ants&#39; fungal crop (Leucoagaricus gongylophorus), a specialized pathogenic fungus in the genus Escovopsis. We filmed and compared infected and uninfected colonies, at both early and more advanced stages of infection. We quantified key defensive behaviors across treatments and show that the behavioral response to pathogen attack likely varies between different castes of worker ants, and between early and late detection of a threat. Based on these recordings we have made a library of behavioral clips, accompanied by definitions of the main individual defensive behaviors. We anticipate that such a guide can provide a common frame of reference for other researchers working in this field, to recognize and study these behaviors, and also provide greater scope for comparing different studies to ultimately help better understand the role these behaviors play in disease defense.

We present a visual guide to disease defense behaviors in leaf-cutting ants, with individual clips and accompanying definitions, illustrated in an experimental infection scenario. Our main aim is to help other researchers recognize key defensive behaviors and to provide a common understanding for future research in this field.

The complex lifestyle, evolutionary history of advanced cooperation, and disease defenses of leaf-cutting ants are well studied. Although numerous studies ...

Low-Cost Automated Flight Intercept Trap for the Temporal Sub-Sampling of Flying Insects Attracted to Artificial Light at Night

Sampling methods are selected depending on the targeted species or the spatial and temporal requirements of the study. However, most methods for passive sampling of flying insects have a poor temporal resolution because it is time-consuming, costly and/or logistically difficult to perform. Effective sampling of flying insects attracted to artificial light at night (ALAN) requires sampling at user-defined time points (nighttime only) across well-replicated sites resulting in major time and labor-intensive survey effort or expensive automated technologies. Described here is a low-cost automated intercept trap that requires no specialist equipment or skills to construct and operate, making it a viable option for studies that require temporal sub-sampling across multiple sites. The trap can be used to address a wide range of other ecological questions that require a greater temporal and spatial scale than is feasible with previous trap technology.

To study the impacts of artificial light at night (ALAN) on nocturnal flying insects, sampling needs to be confined to nighttime. The protocol describes a low-cost automated flight intercept trap that allows researchers to sample at user-defined periods with increased replication.

Sampling methods are selected depending on the targeted species or the spatial and temporal requirements of the study. However, most methods for passive ...

Vertical T-maze Choice Assay for Arthropod Response to Odorants

Given the economic importance of insects and arachnids as pests of agricultural crops, urban environments or as vectors of plant and human diseases, various technologies are being developed as control tools. A subset of these tools focuses on modifying the behavior of arthropods by attraction or repulsion. Therefore, arthropods are often the focus of behavioral investigations. Various tools have been developed to measure arthropod behavior, including wind tunnels, flight mills, servospheres, and various types of olfactometers. The purpose of these tools is to measure insect or arachnid response to visual or more often olfactory cues. The vertical T-maze oflactometer described here measures choices performed by insects in response to attractants or repellents. It is a high throughput assay device that takes advantage of the positive phototaxis (attraction to light) and negative geotaxis (tendency to walk or fly upward) exhibited by many arthropods. The olfactometer consists of a 30 cm glass tube that is divided in half with a Teflon strip forming a T-maze. Each half serves as an arm of the olfactometer enabling the test subjects to make a choice between two potential odor fields in assays involving attractants. In assays involving repellents, lack of normal response to known attractants can also be measured as a third variable.

A vertical, T-maze olfactometer is described for assaying the behavioral response of arthropods. The olfactometer allows the experimenter to measure choices performed by test subjects when subjected to two potential odor fields. Both attraction to and repulsion from odorants can be measured with this device.

Given the economic importance of insects and arachnids as pests of agricultural crops, urban environments or as vectors of plant and human diseases, ...

Neuroscience

Linking Predation Risk, Herbivore Physiological Stress and Microbial Decomposition of Plant Litter

The quantity and quality of detritus entering the soil determines the rate of decomposition by microbial communities as well as recycle rates of nitrogen (N) and carbon (C) sequestration1,2. Plant litter comprises the majority of detritus3, and so it is assumed that decomposition is only marginally influenced by biomass inputs from animals such as herbivores and carnivores4,5. However, carnivores may influence microbial decomposition of plant litter via a chain of interactions in which predation risk alters the physiology of their herbivore prey that in turn alters soil microbial functioning when the herbivore carcasses are decomposed6. A physiological stress response by herbivores to the risk of predation can change the C:N elemental composition of herbivore biomass7,8,9 because stress from predation risk increases herbivore basal energy demands that in nutrient-limited systems forces herbivores to shift their consumption from N-rich resources to support growth and reproduction to C-rich carbohydrate resources to support heightened metabolism6. Herbivores have limited ability to store excess nutrients, so stressed herbivores excrete N as they increase carbohydrate-C consumption7. Ultimately, prey stressed by predation risk increase their body C:N ratio7,10, making them poorer quality resources for the soil microbial pool likely due to lower availability of labile N for microbial enzyme production6. Thus, decomposition of carcasses of stressed herbivores has a priming effect on the functioning of microbial communities that decreases subsequent ability to of microbes to decompose plant litter6,10,11.
We present the methodology to evaluate linkages between predation risk and litter decomposition by soil microbes. We describe how to: induce stress in herbivores from predation risk; measure those stress responses, and measure the consequences on microbial decomposition. We use insights from a model grassland ecosystem comprising the hunting spider predator (Pisuarina mira), a dominant grasshopper herbivore (Melanoplus femurrubrum),and a variety of grass and forb plants9.

We present methods to evaluate how predation risk can alter the chemical quality of herbivore prey by inducing dietary changes to meet demands of heightened stress, and how the decomposition of carcasses from these stressed herbivores slows subsequent plant litter decomposition by soil microbes.

The quantity and quality of detritus entering the soil  determines the rate of decomposition by microbial communities as well as  recycle rates of ...

Biology

Laboratory Protocol for Genetic Gut Content Analyses of Aquatic Macroinvertebrates Using Group-specific rDNA Primers

Analyzing food webs is essential for a better understanding of ecosystems. For example, food web interactions can undergo severe changes caused by the invasion of non-indigenous species. However, an exact identification of field predator-prey interactions is difficult in many cases. These analyses are often based on a visual evaluation of gut content or the analysis of stable isotope ratios (&#948;15N and &#948;13C). Such methods require comprehensive knowledge about, respectively, morphologic diversity or isotopic signature from individual prey organisms, leading to obstacles in the exact identification of prey organisms. Visual gut content analyses especially underestimate soft bodied prey organisms, because maceration, ingestion and digestion of prey organisms make identification of specific species difficult. Hence, polymerase chain reaction (PCR) based strategies, for example the use of group-specific primer sets, provide a powerful tool for the investigation of food web interactions. Here, we describe detailed protocols to investigate the gut contents of macroinvertebrate consumers from the field using group-specific primer sets for nuclear ribosomal deoxyribonucleic acid (rDNA). DNA can be extracted either from whole specimens (in the case of small taxa) or out of gut contents of specimens collected in the field. Presence and functional efficiency of the DNA templates need to be confirmed directly from the tested individual using universal primer sets targeting the respective subunit of DNA. We also demonstrate that consumed prey can be determined further down to species level via PCR with unmodified group-specific primers combined with subsequent single strand conformation polymorphism (SSCP) analyses using polyacrylamide gels. Furthermore, we show that the use of different fluorescent dyes as labels enables parallel screening for DNA fragments of different prey groups from multiple gut content samples via automated fragment analysis.

Most common gut content analyses of macroinvertebrates are visual. Requiring intense knowledge about morphological diversity of prey organisms, they miss soft bodied prey and, due to strong comminution of prey, are nearly impossible for some organisms, including amphipods. We provide detailed, novel genetic approaches for macroinvertebrate prey identification in the diet of amphipods.

Analyzing food webs is essential for a better understanding of ecosystems. For example, food web interactions can undergo severe changes caused by the ...

Maintaining Laboratory Cultures of Gryllus bimaculatus, a Versatile Orthopteran Model for Insect Agriculture and Invertebrate Physiology

Gryllus bimaculatus (De Geer)&#160;is a large-bodied cricket distributed throughout Africa and Southern Eurasia where it is often wild-harvested as human food. Outside its native range, culturing G. bimaculatus is feasible due to its dietary plasticity, rapid reproductive cycle, lack of diapause requirement, tolerance for high-density rearing, and robustness against pathogens. Thus, G. bimaculatus can be&#160;a versatile model for studies of insect physiology, behavior, embryology, or genetics.
Cultural parameters, such as stocking density, within-cage refugia, photoperiod, temperature, relative humidity, and diet, all impact cricket growth, behavior, and gene expression and should be standardized. In the burgeoning literature on farming insects for human consumption, these crickets are frequently employed to evaluate candidate feed admixtures derived from crop residues, food-processing byproducts, and other low-cost waste streams.
To support ongoing experiments evaluating G. bimaculatus growth performance and nutritional quality in response to variable feed substrates, a comprehensive set of standard protocols for breeding, upkeep, handling, measurement, and euthanasia in the laboratory was developed and is presented here. An industry-standard cricket feed has proven nutritionally adequate and functionally appropriate for the long-term maintenance of cricket breeding stocks, as well as for use as an experimental control feed. Rearing these crickets at a density of 0.005 crickets/cm3 in screen-topped 29.3 L polyethylene cages at an average temperature of 27 &#176;C on a 12 light (L)/12 dark (D) photoperiod, with moistened coconut coir serving both as hydration source and oviposition medium has successfully sustained healthy crickets over a 2-year span. Following these methods, crickets in a controlled experiment yielded an average mass of 0.724 g 0.190 g at harvest, with 89% survivorship and 68.2% sexual maturation between stocking (22 days) and harvest (65 days).

Maintaining Laboratory Cultures of Gryllus bimaculatus, a Versatile Orthopteran Model for Insect Agriculture and Invertebrate Physiology

This paper outlines basic methods to standardize important factors such as density, feed availability, hydration source, and environmental controls for the long-term rearing of laboratory cultures of the edible cricket, Gryllus bimaculatus.

Gryllus bimaculatus (De Geer)&#160;is a large-bodied cricket distributed throughout Africa and Southern Eurasia where it is often wild-harvested as human ...

Quantifying Corticolous Arthropods Using Sticky Traps

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