Gut Content Analysis Research Articles

Diet of Andean Leaf-Eared Mice (Phyllotis) Living at Extreme Elevations on Atacama Volcanoes: Insights From Metagenomics, DNA Metabarcoding, and Stable Isotopes.

On the flanks of > 6000 m Andean volcanoes that tower over the Atacama Desert, leaf-eared mice (Phyllotis vaccarum) live at extreme elevations that surpass known vegetation limits. The diet of these mice in these barren, hyperarid environments has been the subject of much speculation. According to the arthropod fallout hypothesis, sustenance is provided by windblown insects that accumulate in snowdrifts ("aolian deposits"). Mice may also feed on saxicolous lichen or forms of cryptic vegetation that have yet to be discovered at such high elevations. We tested hypotheses about the diet of mice living at extreme elevations on Atacama volcanoes by combining metagenomic and DNA metabarcoding analyses of gut contents with stable isotope analyses of mouse tissues. Genomic analyses of contents of the gastrointestinal tract of a live-captured mouse from the summit of Volcán Llullaillaco (6739 m) revealed an opportunistic but purely herbivorous diet, including lichens. Although we found no evidence of animal DNA in gut contents of the summit mouse, stable isotope data indicate that mice from elevations at or near vegetation limits (~5100 m) include a larger fraction of animal prey in their diet than mice from lower elevations. Some plant species detected in the gut contents of the summit mouse are known to exist at lower elevations at the base of the volcano and in the surrounding Altiplano, suggesting that they may occur at higher elevations beneath the snowpack or in other cryptic microhabitats.

Common Food Sources for Macrobenthic Communities in Different Bottom Covers: Submerged Macrophytes and Benthic Cyanobacteria in a Japanese Temperate Lake.

Macrobenthic communities in a lake are affected by the type of bottom cover such as macrophytes or algae. In the southern basin of Lake Biwa, mats of the benthic cyanobacteria (BC) Microseira wollei widely cover the lake bottom and are interspersed with submerged macrophytes (SMs). Because different macrobenthos species appear to occur at those bottoms, we investigated the composition of the communities. Furthermore, as M. wollei is supposed to be inedible to consumer organisms owing to its hard sheath and toxins, the food possibility of the cyanobacteria and macrophytes for macrobenthos was analyzed. In this study, macrobenthos were collected, identified molecularly, counted in the BC and SM zones, and analyzed for carbon and nitrogen stable-isotopic compositions. In the BC zone, asellids and amphipods dominated the macrobenthic community, while chironomids dominated the SM zone. The stable-isotope analysis revealed that M. wollei was a minor food source for macrobenthos and other higher-level consumers, with some macrophytes, particulate organic matter and bottom sediment potentially being a major source. The dominance of crustacean macrobenthos in M. wollei mats suggested that they provided a refuge from predation for crustaceans, whereas SMs were not sufficiently abundant to achieve this. Although different macrobenthic communities in the BC and SM zones were likely supported by common food sources, with the exeption of M. wollei, the present study was conducted over a short period and lacked advanced methods for gut content analysis. Therefore, further monitoring and food web analysis are required.

The contribution of fish to the diet of loggerhead sea turtles in the western Mediterranean revisited

Early juvenile loggerhead turtles (Caretta caretta) rely on gelatinous zooplankton, whereas individuals larger than 40 cm curved carapace length are adapted to crush hard-shelled invertebrates. Nevertheless, fish were reported to be the staple food of loggerhead turtles in the western Mediterranean 30 years ago. Here, the temporal consistency of such a fish-based diet of loggerhead turtles is assessed through gut content analysis and stable isotope analysis of samples from the Mediterranean coast of Spain spanning three decades. The gut contents of 134 juvenile loggerhead turtles (curved carapace length range: 27–71 cm) from three different periods (1991, 1999–2008 and 2010–2017) were analyzed, as well as a subsample of the same turtles (n = 10 in each period) for both bulk and compound-specific stable isotope ratios (CSIA-AA). Gut content analysis revealed a decline in the frequency of occurrence and numerical abundance of fish and an increasing contribution of gastropods and bivalves throughout time, although pelagic tunicates were always the most frequently observed prey. The δ15Nbulk of turtle bone also dropped throughout the study period, but the values of the stable isotope ratio of N in phenylalanine (δ15NPhe) indicated that 52.5% of that variability was due to a baseline shift over time. Accordingly, the trophic position estimated from CSIA-AA did not follow the decreasing pattern of δ15Nbulk, but fluctuated throughout time. The overall evidence indicates that fish consumption by loggerhead turtles in the study region declined through time, but the trophic position of loggerhead turtles did not change simultaneously. This is probably because low trophic prey such pelagic tunicates and filter-feeding bivalves and suspension-feeding gastropods were the bulk of the diet during the whole study period and fish played a minor role, even when their frequency of occurrence peaked. Past levels of high fish consumption might be due to high levels of fishery discards, currently declining because of the recent reduction of the fishing fleet.

Functional Organisation, Predator-prey Interactions, and Gut Content Analysis for Selected Groups of Benthic Macroinvertebrates in High Elevation Ponds in St. Ann Parish, Jamaica

Functional Organisation, Predator-prey Interactions, and Gut Content Analysis for Selected Groups of Benthic Macroinvertebrates in High Elevation Ponds in St. Ann Parish, Jamaica

Functional Organisation, Predator-prey Interactions, and Gut Content Analysis for Selected Groups of Benthic Macroinvertebrates in High Elevation Ponds in St. Ann Parish, Jamaica

Functional Organisation, Predator-prey Interactions, and Gut Content Analysis for Selected Groups of Benthic Macroinvertebrates in High Elevation Ponds in St. Ann Parish, Jamaica

Hydrology and trophic flexibility structure alpine stream food webs in the Teton Range, Wyoming, USA

AbstractUnderstanding biotic interactions and how they vary across habitats is important for assessing the vulnerability of communities to climate change. Receding glaciers in high mountain areas can lead to the hydrologic homogenization of streams and reduce habitat heterogeneity, which are predicted to drive declines in regional diversity and imperil endemic species. However, little is known about food web structure in alpine stream habitats, particularly among streams fed by different hydrologic sources (e.g., glaciers or snowfields). We used gut content and stable isotope analyses to characterize food web structure of alpine macroinvertebrate communities in streams fed by glaciers, subterranean ice, and seasonal snowpack in the Teton Range, Wyoming, USA. Specifically, we sought to (1) assess community resource use among streams fed by different hydrologic sources, (2) explore how variability in resource use relates to feeding strategies, and (3) identify which environmental variables influenced resource use within communities. Average taxa diet differed among all hydrologic sources, and food webs in subterranean ice‐fed streams were largely supported by the gold alga Hydrurus. This finding bolsters a hypothesis that streams fed by subterranean ice may provide key habitat for cold‐water species under climate change by maintaining a longer growing season for this high‐quality food resource. While a range of environmental variables associated with hydrologic source (e.g., stream temperature) were related to diet composition, hydrologic source categories explained the most variation in diet composition models. Less variable diets within versus among streams suggest high trophic flexibility, which was further supported by high levels of omnivory. This inherent trophic flexibility may bolster alpine stream communities against future changes in resource availability as the mountain cryosphere fades. Ultimately, our results expand understanding of the habitat requirements for imperiled alpine taxa while empowering predictions of their vulnerability under climate change.

To pool or not to pool: Pooled metabarcoding does not affect estimates of prey diversity in spider gut content analysis

Abstract Gut content metabarcoding has provided important insights into the food web ecology of spiders, the most dominant terrestrial arthropod predators. In small invertebrates, like spiders, gut content analysis is often performed on whole body DNA extracts of individual predators, from which prey sequences are selectively enriched and sequenced. Since many spider species are generalist predators, large numbers of samples comprising individual spider specimens must be analysed to recover an exhaustive image of a spider species' prey spectrum, which is costly and time‐consuming. Pooled processing of bulk samples of multiple specimens has been suggested to reduce the necessary workload and cost while still recovering a representative estimate of the prey diversity. However, it is still unclear if pooling approaches lead to bias in recovering the prey spectrum and if the results are comparable with data from individually processed spiders. Here, we test the effects of metabarcoding pooled spider gut content on the recovered taxonomic diversity and composition of prey. Using a newly adapted primer pair, which efficiently enriches COI barcode sequences of diverse arthropod prey groups while suppressing spider amplification, we test if pooling leads to reduced taxonomic diversity or skewed estimates of prey composition. Our results show that pooling and individual processing recover highly correlated taxonomic diversity and composition of prey. The only exception are very rare prey items which were less well recovered by pooling. Our results support pooling as a cost‐effective and time‐efficient approach to recover the diet of generalist predators for population‐level studies of spider trophic interactions.

Niche partitioning of invasive Amur sleeper (Perccottus glenii) amongst native fish communities in three different freshwater ecosystems

The invasive fish, Amur sleeper, poses a significant and growing threat to Central European freshwater ecosystems. Despite its rapid spread, the ecological implications of its invasion have been poorly explored. Recent findings confirm its presence in various Estonian freshwater systems, raising concerns about its imminent expansion into larger lakes. To better understand its potential ecological impacts, we explored the isotopic niche of the Amur sleeper in comparison with native fish species co-existing in three Estonian freshwater ecosystems. We employed carbon and nitrogen stable isotope analyses alongside gut content analyses. Our findings show that the Amur sleeper’s diet in newly-invaded Estonian water bodies predominantly comprises benthic macroinvertebrates, although it may also include fish, confirming its role as a predator in the local food web. Notably, Amur sleeper populations exhibited clear isotopic niche partitioning in three invaded ecosystems. A logistic regression model, based on stomach content analyses, revealed an ontogenetic diet shift from benthivorous to piscivorous feeding habits from small to large specimens. Amur sleeper exhibits voracious, non-selective feeding habits, which can negatively impact native freshwater communities. The ability to occupy a distinct isotopic niche, with minimal overlap with native fish populations, may reduce interspecific competition, facilitating the spread and establishment of Amur sleeper in newly-invaded habitats. Managing the spread of this invasive species thus becomes even more critical to safeguard the integrity of native aquatic ecosystems.

Plastics at an Offshore Fish Farm on the South Coast of Madeira Island (Portugal): A Preliminary Evaluation of Their Origin, Type, and Impact on Farmed Fish

Plastic pollution is a global problem affecting all ecosystems, and it represents most of the marine litter. Offshore aquaculture is a sector particularly vulnerable to this issue. To investigate this concern, the present study employed videography to monitor macroplastics at an offshore fish farm on Madeira Island (Portugal) and analysis of fish gut content to evaluate macroplastic ingestion by farmed sea bream Sparus aurata. Our analysis revealed that the majority of identified plastic debris originated from domestic use (66.66%) and fisheries/aquaculture activities (24.99%). While the number of dead fish suitable for sampling was limited (1.05% of the total mortality), macroplastic debris ingestion was identified in 5.15% of the total mortalities and reported for the first time in species in offshore farming conditions. Fish ingested fragmented plastic sheets, with the amount positively correlated with fish weight (r = 0.621, p = 0.031, n = 12). Notably, the stretched length of these fragments exceeded 50% of the standard length of most fish. Inconsistencies were observed in the number of samples collected per cage and per week. To ensure robust results, these discrepancies should be rectified in future studies. Additionally, extending the sampling period to encompass all seasons would be beneficial for a more comprehensive understanding of seasonal variations in plastic occurrence.

Puerarin ameliorates colitis by direct suppression of macrophage M1 polarization in DSS mice

BackgroundUlcerative colitis (UC) is a type of inflammatory bowel disease primarily affecting the colon and rectum. The clinical symptoms of UC include persistent diarrhea, abdominal pain, and rectal bleeding, with chronic inflammation often limited to the mucosal layer of the colon. Macrophages play a significant role in the pathogenesis of UC in response to the presence of gut microbiota. Puerarin is an active compound derived from the root of pueraria lobata, a traditional Chinese herbal medicine, and exhibits potent anti-inflammatory properties in various diseases and disease models including UC-like colitis in mice. However, how the molecule achieves its therapeutic effect in colitis by re-polarizing macrophages remains poorly understood. PurposeUtilizing in vivo and in vitro experimental methods along with multi-omics analysis, we aimed to elucidate the potential mechanism by which puerarin targets macrophages to treat colitis. MethodsThe inflammation induced by DSS was assessed both locally in the gut and systemically, and the anti-inflammatory effect of puerarin was evaluated using molecular and histological assays such as H&E staining, qPCR, ELISA, Western blot, and immunofluorescence. Intestinal permeability parameters were measured by in vivo imaging, immunofluorescence, Western blot, qPCR, and PAS staining. The central role of macrophages in colitis was investigated through macrophage depletion/infusion using cytological methods. The direct effects of puerarin on the macrophages were examined by CCK8, flow cytometry, and qPCR in vitro. Additionally, 16S rRNA sequencing and metabolomics analysis of gut contents were conducted. Identification of key pathogenic flora was facilitated by a trans-omic approach and validated both in vitro and in vivo. ResultsPuerarin exerted a direct and robust suppression of M1-like polarization of macrophages in vitro, which was sufficient to confer therapeutic benefits in terms of colonic lesions and systemic inflammation in DSS mice. Puerarin also reduced the abundance of Akkermansia muciniphila in the gut, which was significantly upregulated in DSS mice in our experimental context. Further study demonstrated that puerarin effectively suppressed M1-like macrophage activation induced by Akkermansia muciniphila secreted protein Amuc_2172, thereby altering the pathology in the DSS model. ConclusionOur data suggest that the pathogenesis of DSS colitis is mediated by host cellular responses to toxic foreign molecules and the gut microbiota, and targeting specific cell populations, such as macrophages, with puerarin holds potential therapeutic value.

What's on the menu? A novel molecular gut content analysis to investigate the feeding behavior of phytophagous insects.

The relationship between phytophagous insects and plants is a central aspect of food webs and ecosystem functioning. The introduction of new species into an environment can have significant impacts on the food web of a native ecosystem. In many cases, there is a lack of knowledge on the biology and feeding behavior of invasive species prior their introduction and in the invaded regions. Gut content analyses of insects have provided valuable information on the host spectrum of insects. However, current approaches are time-consuming and costly. Here, we describe a new molecular gut content analysis (GCA) approach using the Oxford Nanopore (ONT) Flongle sequencing platform to characterize the plant DNA present in the gut of the highly polyphagous insect species Halyomorpha halys. We demonstrate that this technique efficiently amplifies and correctly identifies plant DNA in a mock community. We performed a feeding experiment to determine the sensitivity of this approach and to assess how long the plant DNA can be detected. All plants used in the feeding experiment were correctly identified and detected after 56 days. Surprisingly, we also detected various plant genera that were not included in the feeding experiment and thus were likely ingested months before the experiment. Our study suggests that the GCA using the ONT Flongle sequencing platform represents a rapid and cost-efficient diagnosis of the dietary preferences, host range, and the diversity of consumed plant species of pest insects with high precision.

New data on trophic associations of dung beetles (Coleoptera, Scarabaeidae, Scarabaeinae) and lemurs (Primates, Lemuroidea) in Madagascar revealed by metabarcoding.

Resource use and diet specialisation of Madagascan dung beetles have been little studied especially concerning the possible associations between specific dung beetle and lemur species. Pilot studies have demonstrated that amplicon sequencing is a promising tool for the lemur inventories. In the present contribution, we report the results of the gut content analysis of three endemic Madagascan dung beetles species: Helictopleurusclouei (Harold), Epilissusapotolamproides (Lebis) and Nanosdubitatus (Lebis). Amplicon metagenomics revealed trophic associations of these species with Eulemursanfordi (Archbold), Eu.fulvus (É.Geoffroy Saint-Hilaire) and Cheirogaleuscrossleyi (Grandidier), respectively. The reads of other mammal species, revealed by the analysis, including putative contaminations, are discussed.

Stable isotope analysis reveals fish juveniles as a temporal main resource consumed by invasive pumpkinseed (Lepomis gibbosus)

AbstractThe pumpkinseed Lepomis gibbosus (Linnaeus, 1758) is a small centrarchid fish species from North America that has invaded most European basins, with pronounced expected impacts. Analyses of pumpkinseed diet in its invasive range using gut content analysis (GCA) show macrozoobenthos as the most common dietary item, suggesting a competitive effect on native fish communities. Our study uses a combination of GCA and stable isotope analysis (SIA) to document pumpkinseed diet at two sites in its invaded range and relates pumpkinseed diet preferences with parasitic load. SIA revealed juvenile fish as a major prey item at one of the sites (63 and 50% of the diet of small and the large fish), while GCA showed pumpkinseed preying mostly on macroinvertebrates (chironomids, gastropods, Ephemeroptera, and Zygoptera larvae) at both sites. While infrequent infection by trophically transmitted parasitic nematodes could be related to low consumption of zooplankton, substantial infection by metacercariae of Posthodiplostomum centrarchi Hoffman1958 appears to reflect relatively high consumption rates of its intermediate host, physid snails. Using SIA allowed to demonstrate that pumpkinseed feeding in its non-native range can be based on piscivory, emphasising the need for combining multiple methods when studying the impact of non-native species on food webs.

Diet of Andean leaf-eared mice (Phyllotis) living at extreme elevations on Atacama volcanoes: insights from metagenomics, DNA metabarcoding, and stable isotopes.

On the flanks of >6000 m Andean volcanoes that tower over the Atacama Desert, leaf-eared mice (Phyllotis vaccarum) live at extreme elevations that surpass known vegetation limits. What the mice eat in these barren, hyperarid environments has been the subject of much speculation. According to the arthropod fallout hypothesis, sustenance is provided by windblown insects that accumulate in snowdrifts ('aolian deposits'). It is also possible that mice feed on saxicolous lichen or forms of cryptic vegetation that have yet to be discovered at such high elevations. We tested hypotheses about the diet of mice living at extreme elevations on Atacama volcanoes by combining metagenomic and DNA metabarcoding analyses of gut contents with stable-isotope analyses of mouse tissues. Genomic analyses of contents of the gastrointestinal tract of a live-captured mouse from the 6739 m summit of Volcán Llullaillaco revealed evidence for an opportunistic but purely herbivorous diet, including lichens. Although we found no evidence of animal DNA in gut contents of the summit mouse, stable isotope data indicate that mice native to elevations at or near vegetation limits (~5100 m) include a larger fraction of animal prey in their diet than mice from lower elevations. Some plant species detected in the gut contents of the summit mouse are known to exist at lower elevations at the base of the volcano and in the surrounding Altiplano, suggesting that such plants may occur at higher elevations beneath the snowpack or in other cryptic microhabitats.

Unveiling the diet of two generalist stink bugs, Halyomorpha halys and Pentatoma rufipes (Hemiptera: Pentatomidae), through metabarcoding of the ITS2 region from gut content.

The use of DNA metabarcoding has become an increasingly popular technique to infer feeding relationships in polyphagous herbivores and predators. Understanding host plant preference of native and invasive herbivore insects can be helpful in establishing effective integrated pest management (IPM) strategies. The invasive Halyomorpha halys and native Pentatoma rufipes are piercing-sucking stink bug pests that are known to cause economic damage in commercial fruit orchards. In this study, we performed molecular gut content analysis (MGCA) on field-collected specimens of these two herbivorous pentatomids using next-generation amplicon sequencing (NGAS) of the internal transcribed spacer 2 (ITS2) barcode region. Additionally, a laboratory experiment was set up where H. halys was switched from a mixed diet to a monotypic diet, allowing us to determine the detectability of the initial diet in a time series of ≤3 days after the diet switch. We detected 68 unique plant species from 54 genera in the diet of two stink bug species, with fewer genera found per sample and a smaller diet breadth for P. rufipes than for H. halys. Both stink bug species generally prefer deciduous trees over gymnosperms and herbaceous plants. Landscape type significantly impacted the observed genera in the diet of both stink bug species, whereas season only had a significant effect on the diet of H. halys. This study provides further insights into the dietary composition of two polyphagous pentatomid pests and illustrates that metabarcoding can deliver a relevant species-level resolution of host plant preference. © 2024 Society of Chemical Industry.

Resource overlap and infrequent predation on key pests show vulnerability in cotton biological control services

Promoting arthropod biodiversity to increase ecosystem services through ecological intensification is a challenge for agriculture. And recent evidence suggests that standard pesticide applications not only harm natural enemies but may also fail to deliver long-term pest control solutions. To fuel ecological intensification and build predictive frameworks for pest management incorporating estimates of pest abundance, natural enemy abundance and their associated interactions is essential. Within this framework, there is a need to shift the focus from a single pest and predator to consider the community of predatory arthropods that interact with communities of prey. We took a network-based approach to investigate community interactions of predatory arthropods that feed on key pests and alternative prey in cotton over a three-year period. We merged prey activity, generalist predator communities collected from cotton canopies, and reconstructed trophic interactions with DNA detection frequencies estimated from molecular gut content analysis. Overall, many predator diets overlap, resulting in similar foraging patterns on groups of cotton pests. Moreover, predation on key cotton pests, such as stink bugs and white flies, was low. Therefore, ecological intensification that increases specialized arthropod predators within the community should improve biological control service delivery.

Consistencies in the dietary and isotopic niche of spotted seatrout, Cynoscion nebulosus, across a salinity gradient within a coastal Louisiana estuary.

Estuaries are essential habitats for recreational and commercial fish that are shaped by both natural and anthropogenic processes. In Louisiana a combination of climate change and planned coastal restoration actions is predicted to increase freshwater introduction to coastal estuaries. As such there is a need to quantify the relationships between estuarine fish ecology and salinity to aid in predicting how species will respond to shifts in salinity. We investigated the relative abundance and dietary niches of adult (24.5 ± 5.4 cm standard length) spotted seatrout Cynoscion nebulosus across varying salinity regimes (oligohaline, mesohaline, and polyhaline) within Barataria Bay, Louisiana, using a combination of net sampling and gut content and stable isotopes analysis. We found that the relative abundance of C. nebulosus was lowest at the oligohaline site, translating to approximately five fewer fish captured for every single psu decrease in a site's average annual salinity. In contrast, we found that diets and, to a lesser extent, isotopic niches had a high degree of overlap across sites with differing salinity regimes. Fish and penaeid shrimp were the most common and important prey taxa recovered from guts at all sites. The small isotopic differences found among sites were likely due to spatial variation in hydrogeochemical baselines, and the observed isotopic overlap provides support for the idea that C. nebulosus move between adjacent salinity regimes and forage throughout Barataria Bay. Our results contribute to a greater understanding of the salinity preference and trophic ecology of C. nebulosus that can aid in predicting their responses to future salinity and habitat changes within Barataria Bay associated with predicted climate change and planned coastal restoration actions.

Untangling the complex food webs of tropical rainforest streams.

Food webs depict the tangled web of trophic interactions associated with the functioning of an ecosystem. Understanding the mechanisms providing stability to these food webs is therefore vital for conservation efforts and the management of natural systems. Here, we first characterised a tropical stream meta-food web and five individual food webs using a Bayesian Hierarchical approach unifying three sources of information (gut content analysis, literature compilation and stable isotope data). With data on population-level biomass and individually measured body mass, we applied a bioenergetic model and assessed food web stability using a Lotka-Volterra system of equations. We then assessed the resilience of the system to individual species extinctions using simulations and investigated the network patterns associated with systems with higher stability. The model resulted in a stable meta-food web with 307 links among the 61 components. At the regional scale, 70% of the total energy flow occurred through a set of 10 taxa with large variation in body masses. The remaining 30% of total energy flow relied on 48 different taxa, supporting a significant dependency on a diverse community. The meta-food web was stable against individual species extinctions, with a higher resilience in food webs harbouring omnivorous fish species able to connect multiple food web compartments via weak, non-specialised interactions. Moreover, these fish species contributed largely to the spatial variation among individual food webs, suggesting that these species could operate as mobile predators connecting different streams and stabilising variability at the regional scale. Our results outline two key mechanisms of food web stability operating in tropical streams: (i) the diversity of species and body masses buffering against random and size-dependent disturbances and (ii) high regional diversity and weak omnivorous interactions of predators buffering against local stochastic variation in species composition. These mechanisms rely on high local and regional biodiversity in tropical streams, which is known to be strongly affected by human impacts. Therefore, an urgent challenge is to understand how the ongoing systematic loss of diversity jeopardises the stability of stream food webs in human-impacted landscapes.

Food and Feeding Habits of Gerres filamentosus in the Cochin Backwaters

A total of 601 specimens of Gerres filamentosus were collected from the sampling sites and were subjected to gut content analysis. Guts were visually classified according to the fullness of the stomach. Occurrence, gravimetric and volumetric methods were followed to quantify the gut contents. Feeding intensity was assessed using methods such as the Gastro rosomatic index, mean index of feeding intensity, and index of fullness. Sex wise, month and length group-wise fluctuations in the feeding intensity were also studied. It can be seen that the percentage of actively feeding fish was observed to be high during pre-monsoon and monsoon months. Feeding intensity was at its peak during April and July 2019, and the main food was found to be prawn juveniles, polychaete worms and other detritus matter.

Diversified cropping strengthens herbivore regulation by providing seasonal resource continuity to predators

Abstract Agricultural practices shape arthropod communities in arable fields, consequently influencing their interactions and the resulting ecosystem services, in particular pest regulation. Predatory arthropods play a pivotal role by preying on herbivores, soil fauna, and on other predators. However, the intricate mechanisms through which agricultural practices shape the dietary preferences of predators, and regulate herbivore populations remain complex and inadequately understood. We assessed how fertilisation with organic fertiliser and extending crop rotations with perennial ley affected predation pressure across prey taxa. We mapped predator and prey trophic linkages with molecular analysis of carabid predator gut contents, and measured densities and taxonomic richness of predators, herbivores, and soil fauna in 19 cereal fields during three samplings across the growing season. We derived two food web structure metrics: prey vulnerability that is the average number of predators feeding on a selected prey, and predator trophic redundancy, that is dietary overlap. Prey vulnerability was compared among soil fauna, herbivores, and other predator species (that is interspecific intraguild predation) over the growing season, and across treatments. The mechanistic underpinnings of observed shifts in vulnerability of herbivorous prey at different crop stages were identified using information criteria to select among candidate variables related to the richness, density and interaction structure of the different guilds during both the current, and the previous crop stages. Agricultural diversification via organic fertilisation combined with perennial ley in the crop rotation decreased the vulnerability of both intraguild prey and soil fauna prey, and stabilised herbivore vulnerability. Mechanistically, the vulnerability of herbivorous prey at crop ripening emerges from the combination of predator richness and trophic redundancy during this sampling round, rather than from carryover effects from previous crop stages. Synthesis and applications: Our results suggest that locally provided resource continuity through diversified cropping practices bolster biological pest regulation, thus underline the importance of lesser disturbance in arable ecosystems for the provision of ecosystem services. Enhanced predator species richness together with availability of alternative prey through the season underpins this enhanced pest regulation.