Non-destructive Sampling Methods Research Articles

Loss of carbon stock in the forests of Uttarakhand due to unprecedented seasonal forest fires

Unprecedented seasonal forest fires pose a significant threat to the carbon stocks of diverse ecosystems, particularly in regions like Uttarakhand, west Himalaya. Understanding the impact of varying fire frequencies on different forest types is crucial for effective conservation and management strategies. This study aims to assess the loss of carbon stock in three distinct forest types—Sal, Pine, and Mixed across an elevation gradient in Uttarakhand, facing unprecedented seasonal forest fires. By investigating pre- and post-fire conditions, analyzing biomass dynamics, and mapping fire frequencies, the research aims to provide insights into the complex interplay of fire regimes and forest resilience. The investigation covers vegetation analysis, biomass assessment, and fire frequency mapping. Biomass and carbon stock calculations were carried out using a non-destructive sampling method. Fire frequency maps were generated using Landsat satellite imagery spanning a decade, integrating MODIS hotspot data for classification. The study reveals distinct patterns in biomass changes across Sal, Pine, and Mixed forests in response to varying fire frequencies. Sal forests exhibit resilience to low-intensity fires, while Pine forests show higher sensitivity. Carbon stock contributions of dominant species varied significantly, with Sal and Chir-Pine forests emerging as crucial contributors. High fire frequencies lead to substantial carbon stock reduction in all forest types. The findings emphasize the sensitivity of aboveground biomass to fire frequency, with significant carbon stock loss observed in higher fire frequency classes. The study underscores the importance of nuanced conservation strategies tailored to distinct forest types and species characteristics. This research provides valuable insights for policymakers, forest managers, and conservationists in formulating targeted conservation and management approaches.

Estimated Tree Carbon Reserves and Saplings in the Nagari Sumpur Kudus Social Forestry Area, Sijunjung Regency

The forest within The Social Forestry of Nagari Sumpur Kudus, Sijunjung Regency has the ability to store carbon reserves. The forest area located in the highlands shows a lot of biodiversity, influenced by the ability of the forest to maintain carbon stocks and diverse plant species in the area. The purpose of this study is to determine the total biomass and total carbon stocks stored in the Social Forestry Area of Nagari Sumpur Kudus, Sijunjung Regency. This research was conducted from May to October 2023. Taking at tree and sapling strata uses a nondestructive sampling method. From the results, it is obtained that the three strata are as many as 14 families and 14 species. The sapling strata is obtained from as many as 17 families and 37 species. The total above-ground biomass is l78,97 ton/ha consisting of 177,83 ton/ha tree biomass and 1,13 ton/ha sapling biomass. The total above-ground carbon stock is 86,11 ton/ha consisting of tree carbon stock 85,58 ton/ha and sapling carbon stock 0,53 ton/ha. The Social Forestry Area of Nagari Sumpur Kudus, Sijunjung Regency has the potential to store carbon stocks which is included in the medium category and need to be maintained.

The development of non-destructive sampling methods of parchment skins for genetic species identification.

Parchment, the skins of animals prepared for use as writing surfaces, offers a valuable source of genetic information. Many have clearly defined provenance, allowing for the genetic findings to be evaluated in temporal and spatial context. While these documents can yield evidence of the animal sources, the DNA contained within these aged skins is often damaged and fragmented. Previously, genetic studies targeting parchment have used destructive sampling techniques and so the development and validation of non-destructive sampling methods would expand opportunities and facilitate testing of more precious documents, especially those with historical significance. Here we present genetic data obtained by non-destructive sampling of eight parchments spanning the 15th century to the modern day. We define a workflow for enriching the mitochondrial genome (mtGenome), generating next-generation sequencing reads to permit species identification, and providing interpretation guidance. Using sample replication, comparisons to destructively sampled controls, and by establishing authentication criteria, we were able to confidently assign full/near full mtGenome sequences to 56.3% of non-destructively sampled parchments, each with greater than 90% of the mtGenome reference covered. Six of eight parchments passed all four established thresholds with at least one non-destructive sample, highlighting promise for future studies.

Buccal swabs for long-term DNA storage in conservation genetics of fish: One-and-a-half-year analysis timeframe

Conservation genetic research is essential for the management and recovery of endangered taxa. However, the invasive collection of biological material for DNA analysis is controversial. From an ethical perspective, non-destructive sampling methods leave the aquatic specimen alive and less invasive procedures minimize stress on the animals. DNA can be obtained from fish using minimally invasive techniques such as buccal swabs. Here we evaluated the performance of buccal swabs for long-term storage of DNA obtained from brown trout (Salmo trutta). The buccal swabs were stored at room temperature and cut into pieces, one part of which was used for extraction of an aliquot and the others were stored as a “biobank” of biological material. The elapsed time from sampling to molecular analysis was one and half year. The amplification of three different DNA targets was tested to assess the effectiveness of the extraction: mitochondrial DNA (the D-LOOP region), nuclear DNA (the LDH gene) and microsatellite DNA at multiple loci. The results showed high quantification (mean value: 281.84±72.4 ng/µL), indicating that DNA could be effectively extracted from the buccal swabs. Our study results suggest that buccal swabs for long-term storage of DNA at room temperature are promising for use in field conservation studies.

Numerical Simulation and Design of a Shaftless Hollow Pump for Plankton Sampling

It is important to marine ecology research that plankton samples are collected without damage, especially for time series samples. Usually, most fixed-point plankton samplers are made using a pump with paddle blades in order to increase the flow rate. But it can easily injure soft plankton. In this paper, a shaftless hollow sampling pump is designed, which can provide a highly efficient driving component for the plankton sampler. The numerical model of the sampling pump is established, and the flow rate of the sampling pump at different rotational speeds is simulated by the computational fluid dynamics method. In order to obtain a higher flow rate, the influence of internal and external cavity size, blade angle, and blade number on the flow rate of the sampling pump with a constant rotational speed of the blade was simulated and discussed. The results show that the flow rate at the internal cavity is positively correlated with the inlet and outlet pressure differences of the internal cavity, and the greater the negative pressure at the outlet of the internal cavity, the greater the flow rate. When the internal and external cavity sizes are h = 14 mm, d = 52 mm, blade angle θ = 45°, and number of blades s = 5, the flow rate of the sampling pump internal cavity reaches the maximum. Finally, the feasibility of the shaftless hollow sampling pump is verified by experiments. The shaftless hollow sampling pump can realize non-destructive sampling of plankton. This paper presents a theoretical design foundation for a new non-destructive siphon sampling method for marine plankton, which is of great significance for marine plankton sampling and subsequent research.

Nature Based Solutions: Carbon Sequestration by Standing Trees in Urban Parks and Gardens of Katni City in Madhya Pradesh, India

Aims: This study aims at estimating contribution of urban Parks and gardens by carbon sequestration and suggests suitable strategies that can be helpful in reducing climate change impacts in urban centres. The challenges of climate change can be efficiently overcome by the storage of carbon in terrestrial carbon sinks viz. plants, plant products and soils for longer periods. Selection of appropriate trees can help mitigate climate impact.
 Study Design: Non-destructive sampling method of biomass estimation was used to measure to GBH of individual trees.
 Place and Duration of Study: The study was conducted in Katni city, Madhya Pradesh in 2020.
 Methodology: The parks and gardens in Katni were categorized into three groups based on their sizes: (A) large parks/gardens, (B) medium parks/gardens, and (C) small parks/gardens. The height of the trees was estimated using a relascope, while the girth classes of the trees were measured at different heights, including the basal area at Diameter at Breast Height (DBH) and at a 2-meter height.
 Results: 26 parks were systematically selected from a total of 62, encompassing small, medium, and large-sized areas, representing 42% of the city's green cover. The sampled parks sequestered 340 tons of carbon, projecting an annual sequestration of 414 tons by all parks combined. Among the planted trees in these spaces were (Hyophorbe lagenicaulis), Fishtailed palm (Carota urens) Ashok (Polyalthia longifolia), Neem (Azardirachta indica), Amaltas (Cassia fistula), Champa (Magnolia champaca), Saptaparni (Alstonia scholaris), etc. Furthermore, the study identified ten suitable tree species—Bargad (Ficus benghalensis), Gulmohar (Delonix regia), Karanj (Millettia pinnata), Kassod (Cassia siamea), Mahaneem (Ailanthus excelsa), Neem (Azadirachta indica), Umar (Ficus racemosa), Peepal (Ficus religiosa), Peltaforum (Peltophorum pterocarpum) and Shisham (Dalbergia sissoo) ideal for planting in parks for climate mitigation. The study advocates for widespread planting of these identified indigenous species in urban areas to enhance environmental quality and contribute significantly to climate improvement efforts.
 Conclusion: This study provides valuable insights into the nature of carbon sequestration in urban green spaces, stressing the need for selection of tree species based on multiple criteria discussed in the paper. Holistic approaches in environmental management and conservation will help mitigate climate impact.

An original nondestructive sampling method to study the effect of gravity on the deposition of micron-sized large particles in exhaust gas recirculation (EGR) cooler fouling

Fouling is one of the barriers to developing more efficient and near-zero emission internal combustion engines. The micron-sized particulate matter is one of the roots of this fouling phenomenon in exhaust gas recirculation (EGR) coolers. This fouling is inadequately evaluated quantitatively, and its deposition mechanism is unknown. To investigate the effect of gravity on the deposition of micron-sized particles, an original nondestructive sampling fouling method and experiment apparatus have been developed to effectively obtain the upper and lower bottom fouling in the cooler in the direction of airflow, and the area proportion, count, and diameter of large particles in the fouling using image processing software. It was found that (i) the area proportion of large particles in the lower bottom fouling was almost 2.5 times higher than the upper bottom fouling; (ii) the count of large particles in the lower bottom fouling was higher, and the maximum diameter was larger, up to 639 μm; (iii) the mass of the lower bottom fouling was 1.5 times higher than the upper bottom fouling; (iv) gravity can significantly promote the deposition of micron-sized particles and should be considered in the design and arrangement of the EGR cooler to prevent fouling.

Establishing Allometric Equation based on 3D Scanning Technique - Non-destructive Sampling Method

Establishing Allometric Equation based on 3D Scanning Technique - Non-destructive Sampling Method

Classification of Plenodomus lingam and Plenodomus biglobosus in Co-Occurring Samples Using Reflectance Spectroscopy

Under natural conditions, mixed infections are often observed when two or more species of plant pathogens are present on one host. Thus, the detection and characterization of co-occurring pest species is a challenge of great importance. In this study, we focused on the development of a spectral unmixing method for the discrimination of two fungi species, Plenodomus lingam and Plenodomus biglobosus, the pathogens of oilseed rape. Over 24 days, spectral reflectance measurements from Petri dishes inoculated with fungi were conducted. Four experimental combinations were used: the first two were pure fungal samples, while the other two were co-occurring fungal samples. The results of the study show the possibility of distinguishing, based on spectral characteristics, between P. lingam and P. biglobosus not only in pure but also in co-occurring samples. We observed the changes in the reflectance of electromagnetic radiation from the tested fungi over time and a strong correlation between the reflectance and changes in the areas of the mycelia on the Petri dishes. Moreover, the wavelengths most useful for spectral classification of the tested fungal mycelia were selected. Finally, a spectral unmixing model was proposed, which enables the estimation of the areas of two pathogens in co-occurring samples based on the spectral characteristics of the entire plate with an error smaller than 0.2. To our knowledge, the present study is the first report examining the use of reflectance spectroscopy methods for classifying pathogens on the same Petri dish. The study results indicate the feasibility of reflectance spectroscopy as a nondestructive sampling method for plant pathogen detection and classification.

Estimation of Aboveground Carbon Stock in The Pertamina Hulu Rokan (PHR) Forest in Pinggir District, Bengkalis Regency, Riau Province

Extreme climate change affects human life. One of the causes of climate change is the increase of CO2 gas in the air. Forests can absorb CO2 from the air and store it in the form of tree biomass through the process of photosynthesis. One of the forests that can absorb and store carbon is Pertamina Hulu Rokan (PHR) forest, therefore it is necessary to conduct research on estimating the carbon content of PHR Forest area. The methods to be used in this research are non-destructive sampling and destructive sampling methods. The method with harvesting is only for seedlings, undergrowth and litter, while for saplings, poles, trees and necromass with the method without harvesting. Aboveground carbon storage in PHR forest was found to be 95.37 tonnes/ha. The largest carbon stocks were from trees (68.96 tonnes/ha), poles (13.06 tonnes/ha), necromas (5.70 tonnes/ha), litter (3.72 tonnes/ha), saplings (3.54 tonnes/ha) and the smallest carbon stocks were seedlings and understorey (0.39 tonnes/ha).

Evaluating Non-destructive Analysis and Sampling Methods for Identifying Plant and Animal Imprints in Late Pre-contact Wattle and Daub Rubble from the Yazoo Basin, Mississippi

ABSTRACT Past analyses of burned wattle and daub rubble from Native American house excavations have shown that plant and animal-related imprints sometimes occur in daub fragments. This paper evaluates the use of non-destructive analysis and sampling methods from a region-wide assemblage of burned daub recovered from a group of Pre-Contact Mississippian Period village and settlement sites in the Yazoo Basin of northwestern Mississippi, U.S.A. The quality and occurrence of imprints from this assemblage will be assessed, and how they may be considered in both future paleoenvironmental research and curation practices.

Vegetation Structure, Biomass, and Carbon Stock of Urban Forest of Bongohulawa National Wirakarya Campground in Gorontalo Regency - Gorontalo Province

An urban forest is one type of Urban Green Open Space (RTHKP) that reflects the character of nature and/or local culture with ecological, historical, and panoramic values that are unique to the level of technology application. The urban forest of the National Wirakarya Campsite is quite attractive, has a lovely view, is close to the road, and is easily accessible by the residents. This study aims to determine the structure of vegetation, biomass, and carbon values stored in the urban forest of the National Wirakarya Campsite, Bongohulawa, Gorontalo Regency, Gorontalo Province. The method used is a survey method with a quantitative descriptive research design. The sampling method used is a purposive sampling model based on land cover in the urban forests. For the measurement of the vegetation structure, the sampling of vegetation in the observation plots is carried out by using the number of sample plots that are placed regularly by calculating the Relative Density (RD), Relative Frequency (RF), and Relative Dominance (RDo), then the data obtained are tabulated to provide IVI. To measure the carbon stocks on the surface (stems), a non-destructive sampling method is used with an Allometric equation based on the plant species. The results showed, there were 13 families, 20 species, and 824 individuals. The vegetation structure in the urban forest of Bongohulawa National Wirakarya Campsite (PWN) has an IVI above 15 percent on average. The sawlog level is dominated by the Gmelina arborea (beechwood) species with an IVI of 98.36 percent, the pole level is dominated by the Swietenia mahagagoni (mahogany) with an IVI of 165.37 percent, the sapling level was dominated by Swietenia mahagoni with an IVI of 160.99 percent and the seedling level was dominated by Lantana camara (tembelekan) with an IVI of 32.25 percent. The content of biomass stored in the standing trees is 1,190.45 tons/ha, with the largest amount of biomass is at the sawlog level of 1,135.43 tons/year or 95.43 percent, the total biomass at the pole level is 45.10 tons/year or 3.79 percent and the amount of long-term biomass is 9.32 tons/year or 0.78 percent. The carbon stock stored (C-Stock) is 559.37 tons/ha and can absorb CO2 (CO2eq) of 2,052.88 tons/ha and provides converted O2 of 1,498.60 tons/ha.

Molecular identification of selected parrot eggs using a non-destructive sampling method

Molecular identification of selected parrot eggs using a non-destructive sampling method

Growth Performances of Cocoa (<em>Theobroma Cacao</em>) Varieties under Mature Oil Palm with Palm Oil Mill Effluent as Growing Media

Oil palm (Elaeis guineensis) is a leading edible oil-producing plant established in tropical countries which cause many negative impacts. Poor productivity of monoculture plantations and land and aquatic pollution caused by haphazard release of Palm Oil Mill Effluent (POME) are critical concerns. As a remedy, oil palm can be intercropped with cocoa by using POME as a potting media. In this study, we evaluated the growth of two different cocoa varieties with POME as the potting media under 20 years old oil palm shade. Two cocoa varieties (SCA6 x ICS6, and NA32 x ICS1) and three potting media (topsoil only, 40% topsoil with 60% POME, and 60% topsoil with 40% POME) were factorially combined in a randomized complete block design with three replicates under 40% shade, Faculty of Agriculture, University of Ruhuna. Growth parameters of the cocoa seedlings such as plant height, number of leaves, chlorophyll value, stem girth, root and shoot dry weights, root length, number of lateral roots per plant were measured using destructive and non-destructive sampling methods. The potting media consist of 40% topsoil with 60% POME showed significantly the highest plant height, number of leaves, chlorophyll value, stem girth, fresh and dry weight of shoot and roots and length of roots. Hence, potting media with 40% topsoil with 60% POME was recognized as the best potting media for the growth and development of cocoa seedlings of both varieties. Growth of cocoa seedling was well compatible under oil palm shade at full light condition with the use of POME as the potting media.

Economic value of carbon in mangrove ecosystem of Cawan Island, Indragiri Regency, Riau, Indonesia

This study aimed to calculate the economic value of carbon in the mangrove ecosystem of Cawan Island Indragiri Regency, Riau, Indonesia. Mangrove data retrieval was carried out at three stations using the 10 x 10 m quadratic transect method. A non-destructive sampling method used to estimate tree biomass, by measuring all diameters at breast height (DBH=1.3 m) of mangroves, then calculating using an allometric model. The types of mangroves found on Cawan Island are Rhizophora mucronata, Rhizophora apiculata, Xylocarpus granatum, Sonneratia alba, Sonneratia ovata, Lumnitzera racemosa, Bruguiera gymnorrhiza and Nypa fruticans. Rhizophora apiculata has the highest biomass of 2,207.48 tons.ha-1 with a carbon potential of 1,015.43 tons-C.ha-1 and carbon absorption of 3,723.24 tons-CO2.ha-1, in general, Cawan Island mangroves have a biomass value of 4,838.40 tons.ha-1 with a carbon potential of 2,199.54 tons-CO2.ha-1 and carbon absorption of 8,064.98 tons-CO2.ha-1. The total value of carbon sequestration produced by mangroves on Cawan Island is IDR.598,566,978,912.year-1.

Vegetation Analysis and Carbon Stock Estimation of Mangrove Island on Mangrove Ecotourism in Gebang Village, Lampung

Mangrove forests have an important role in reducing the effects of global warming. This study aims to analyze vegetation and estimate carbon stock in mangrove stands in the Mangrove Island of Petengoran Mangrove Ecotourism Area, Gebang Village, Lampung. The forest vegetation analysis method used is the plotted path method. Paths and plots were taken systematically with tree observation plots made measuring 20 m x 20 m. Carbon estimation was carried out by non-destructive sampling method using the allometric formula for Rhizophora apiculata log BBA= -1.315 + 2.614 log D. The results showed that the mangrove island zone was dominated by Rhizophora apiculata vegetation with a density of 325 ind/Ha. The research location has a silty clay loam soil texture with a temperature range of 25°C-32°C which is very suitable for the vegetation of R. apiculata. The estimated carbon stored in the vegetation was obtained from 46% of the standing biomass of R. apiculata, which was 86,283 tons C/Ha.KEYWORDSBiomass, Carbon, Mangroves, Petengoran, Vegetation

16S rRNA gene-based assessment of common broiler chicken sampling methods: Evaluating intra-flock sample size, cecal pair similarity, and cloacal swab similarity to other alimentary tract locations.

16S rRNA gene sequencing for characterization of microbiomes has become more common in poultry research and can be used to both answer specific research questions and help inform experimental design choices. The objective of this study was to use 16S rRNA gene sequencing to examine common sampling practices in broiler chicken studies such as: the required number of birds selected from a flock to adequately capture microbiome diversity, the differences between cecal pairs within the same bird, and whether cloacal swabs are representative of other alimentary tract (AT) locations. To do this, nine market age broilers were euthanized and immediately sampled in ten AT locations: crop, gizzard, proventriculus, duodenum, jejunum, ileum, cecal samples from each pouch, colon, and cloacal swab. DNA was extracted and subjected to 16S rRNA gene amplification and sequencing. Each location within the broiler AT hosts distinct microbial communities. When each sampling location was considered, it was found that sampling after 2.8 birds (range 2-4) resulted in less than 10% new amplicon sequencing variants (ASV) being added while sampling after 7.6 birds (range 6-10) increases new observed ASVs by less than 1%. Additionally, when cecal pairs from the same bird were evaluated, it was found that cecal pair mates are an adequate replication if interested in the total cecal microbiome but may be less useful if a rare lineage is of interest. Furthermore, when compared to other AT locations, the cecal microbiome was enriched in Firmicutes and Bacteroides while several lineages, most notably Lactobacillus, were under-represented. Finally, when cloacal swabs were compared to other AT locations, community similarity exhibited a direct distance relationship, i.e., the more aborad samples were the more similar they were to the swab. These findings indicate that while cloacal swabs can approximate overall changes in microbiome composition, they are not adequate for inferring changes to specific taxa in other parts of the AT tract-even those that are highly abundant within the microbial community. These data provide new insights guiding appropriate sample size selection within flocks and add to the consensus data regarding cecal pair similarity and destructive versus non-destructive sampling methods.

The environmental services of Pangarengan mangrove forest in Cirebon, Indonesia: conserving biodiversity and storing carbon

Abstract. Purwanto RH, Mulyana B, Sari PI, Hidayatullah MF, Marpaung AA, Putra ISR, Putra AD. 2021. The environmental services of Pangarengan mangrove forest in Cirebon, Indonesia: conserving biodiversity and storing carbon. Biodiversitas 22: 5609-5616. Pangarengan mangrove forest in Cirebon, Indonesia, plays a vital role in providing ecosystem services, such as conserving biodiversity and storing carbon. The area of the Pangarengan mangrove forest is relatively small, but it is home to diverse flora and fauna. Also, it can mitigate climate change by storing carbon. Unfortunately, no specific records exist on biodiversity and carbon stock in the Pangarengan mangrove forest. This study aimed to record the diversity of flora and fauna of the Pangarengan mangrove using a survey approach and to estimate its carbon stocks using a non-destructive sampling method, except for seedlings, on 78 sampling plots, distributed randomly along the banks of the Cipaluh River. The results revealed that the flora consisted of 24 species, and the fauna was composed of 12 species. The mangrove species in the Pangarengan mangrove forest was dominated by Rhizophora mucronata. The carbon stocks varied among species or the life stages of plants (seedlings, saplings, poles, and trees). The carbon stock of Avicennia marina trees was the highest (110.810 MgC/ha), and the lowest was the seedlings of the Avicennia alba (0.005 MgC/ha). Furthermore, the interpretation of the images showed that the Pangarengan mangrove forest area is 21.5 ha. The mangrove forest is located in the coastal area of the Java Sea, 2.4 km from the residential area of Pangarengan village. It is crucial to preserve the Pangarengan mangrove forest because it functions as a source of biodiversity and carbon storage.

A non-destructive sampling method for food authentication using gas chromatography coupled to mass spectrometry or ion mobility spectrometry

The study of volatile compounds obtained by gas chromatography (GC) coupled to mass spectrometry (MS) or ion mobility spectrometry (IMS) may be very useful to protect food quality, especially when using a non-destructive sampling method. In this work, the authentication of the highly appreciated dry-cured Iberian ham by those techniques was studied and compared. The results obtained show the suitability of a non-destructive sampling method coupled to headspace sampling (HS)-GC-IMS or HS-GC–MS to determine volatile markers in the feeding Iberian pig regime. Although both methods were suitable to differentiate the ham categories, HS-GC-IMS was more sensitive detecting a higher number of compounds than HS-GC–MS, which provided accurate qualitative results. The results of principal component analysis showed that ethanol, 2-propanol and 3-methylbutanol, identified by HS-GC-IMS, and 3-methylbutanal and heptane, identified by HS-GC–MS, could be considered potential markers to identify ham from different feeding regimes.

In Situ Non-Destructive Temporal Measurements of the Rhizosphere Microbiome ‘Hot-Spots’ Using Metaproteomics

Rhizosphere arguably embodies the most diverse microbial ecosystem on the planet, yet it is largely a functional ‘black box’ of belowground plant-microbiome interactions. The rhizosphere is the primary site of entry for subsurface injection of fixed carbon (C) into soil with impacts on local to global scale C biogeochemistry and ultimately Earth’s climate. While spatial organization of rhizosphere is central to its function, small scale and steep microbial and geochemical gradients within this dynamic region make it easily disrupted by sampling. The significant challenge presented by sampling blocks elucidation of discreet functions, drivers, and interactions within rhizosphere ecosystems. Here, we describe a non-destructive sampling method linked to metaproteomic analysis in order to measure temporal shifts in the microbial composition and function of rhizosphere. A robust, non-destructive method of sampling microbial hotspots within rhizosphere provides an unperturbed window into the elusive functional interactome of this system over time and space.