Botanical Types Research Articles

Influence of oak wood on the chemical and organoleptic profile of white wines

The use of oak wood in various forms for the production of white wines has become an increasingly popular practice among winemakers. As a result of extraction, condensation and other processes, their chemical composition and sensory profile were modified. Phenolic compounds and volatile aromatic substances passed from the wood to the wine improving the aroma and taste and enhancing its antioxidant activity. Their quantity has depended on the botanical type and origin of the wood and the method of its heat treatment. In the production of white wines several technologies have been applied - conducting the alcoholic fermentation and aging on lees in barrels, completing the fermentation in tanks and racking the wine to the barrels, starting the fermentation in tanks but in the middle of the process racking the fermenting juice to the barrels. Into the wine, of phenols mainly passed gallotannins and ellagitannins and of aromatic substances – eugenol, isoeugenol, vanillin, furan derivatives, lactones, etc. Further to the use of barrels, the application of alternative oak forms has become more and more popular, which in the case of white wines allowed avoiding the volatile components oxidation. The result was a quality product with better technological properties. The wines acquired hints of wood without losing their freshness and fruity character.

Effect of Drought and Pluvial Climates on the Production and Stability of Different Types of Peanut Cultivars in Guangdong, China

The production and breeding of peanuts was restricted by the frequently extreme climatic conditions in Guangdong province, China. To understand the influence of drought and pluvial climates on peanut traits and yield, a phenotypic investigation of seventy peanut cultivars was conducted from 2018 to 2022; comprehensive field meteorological data collection, and typical drought (2021) and pluvial (2022) climates were recorded. The results revealed that the cultivars achieved the highest single plant pod weight (SPPW) and single plant seed weight (SPSW) of 61.03 g and 45.84 g, respectively, in drought conditions, followed by the control, and finally the pluvial. The SPPW, SPSW and eight agronomy traits exhibited significant differences across the different climatic conditions. Correlation analysis revealed the yield traits and key yield-related traits were positively or negatively correlated with soil water content (SWC), total global radiation (TGR), total precipitation (TP) and total net radiation (TNR). The intermediate and Spanish type cultivars were more stable and productive than the other botanical types of cultivars, commercial varieties exhibited better performance than landraces, and seven cultivars were identified with good production potential, under drought and pluvial conditions. Our study showed that pluvial climate was detrimental to peanut yield, and the SPPW and SPSW were significantly influenced by climates with genotype differences.

ПАЛИНОЛОГИЧЕСКОЕ ИССЛЕДОВАНИЕ МЁДА В РЕСПУБЛИКЕ БУРЯТИЯ

In recent years, agricultural production has been given the strategic task of intensifying and strengthening the production of a quality food product. Many experts testify that indicators regulated by documents, although providing sufficient information about the quality of honey, they do not allow characterizing some properties, for example, its botanical origin. The botanical type of honey is substantiated by the nectar collection area. In practice, examples are given when natural honey is falsified by introducing various impurities into it. For example, they falsify with sugar, flour, starch, sugar syrup, starch and sugar syrup, artificial honey. Palynological analysis is the most objective method for assessing the botanical origin of honey, on which many of its value and quality characteristics depend. The goal is to study the palynological composition of honey obtained in the conditions of the Republic of Buryatia, identify falsification and confirm naturalness. The studies were carried out using a Carl Zeiss Primo Star microscope. The morphometric study was carried out on a Carl Zeiss Primo Star microscope at a magnification of about 10, volume 5 in the program Zeiss Efficien tNavigation (ZEN). Results: Microscopic examination revealed crystals of various shapes in the samples: needle-shaped, star-shaped, irregular and regular geometric shapes. In natural honey, crystals should only be needle-shaped and star-shaped. According to the results of the research, it was revealed that crystals were found in 40 % of the studied samples, namely, regular and irregular geometric shapes. This indicates that honey samples are falsified and made by bees using sugar syrup or sugar. Based on a palynological study, pollen grains were found in all 20 samples studied, corresponding to their botanical origin. Solitary pollen grains of other associated plants were also found. This method of analyzing honey allows you to detect falsification of a product with sugar or powdered sugar under a microscope, as well as fungal spores, pollen grains of plants.

Quantitative botanical characterization of Saragolla wheat landraces from Abruzzo and Puglia Regions of Italy

AbstractSince the 1600s, a pasta wheat locally known as Sargolla, Saragollio or Saragollo has been cultivated in central and southern regions of Italy, including Abruzzo and Puglia. To identify if distinctive Saragolla landraces are present in Abruzzo and with a view to registering them as Conservation Varieties in the Abruzzo Regional Register of Genetic Resources, a quantitative botanical characterization of 11 farm-saved seed samples from Abruzzo and a previously defined Saragolla landrace from Puglia was performed. All twelve samples were grown for two years at the CREA-CI field station in Foggia, Puglia (Italy), and were morphologically characterized according to the criteria used by the botanist Johan Percival in 1921. In the 12 samples, we identified 9 taxonomically different botanical types, the most frequent of which were the italicum (32%), found in every population from Abruzzo. The leucurum and the affine were the main components of the previously defined Saragolla from Puglia. Results of the multivariate analysis indicated that the samples collected from farms in Abruzzo are more similar to each other than to the landrace of Saragolla from Puglia and support the registration of a specific Saragolla landrace for the Abruzzo region. The diachronic comparison of the botanical diversity of the currently cultivated Saragolla, with that observed in Sicily and Algeria at the beginning of 1900, suggests that a high degree of genetic diversity still exists in the 11 Saragolla populations from the Abruzzo Region.

Optimizing Procedures for Antioxidant Phenolics Extraction from Skin and Kernel of Peanuts with Contrasting Levels of Drought Tolerance.

Peanut is an affordable legume known for its nutritional value and phenolic content. The kernel and skin of 14 peanut genotypes contrasting in drought tolerance had their phenolic profiles determined and reactive oxygen species (ROS) scavenging activity evaluated. Firstly, temperature and % EtOH to extract antioxidant phenolic compounds were optimized using response surface methodology (RSM). The optimized extraction conditions, 60 °C and 35% EtOH for kernels and 40 °C and 60% EtOH for skins, were further adopted, and phenolic compounds were identified and quantified using high-performance liquid chromatography coupled with electrospray ionization-quadrupole-time of flight-mass spectrometry (HPLC-ESI-QTOF-MS) and high-performance liquid chromatography with photodiode array detector (HPLC-PDA). As a result, phenolic acids and glycosidic/non-glycosidic flavonoids were found. Principal component analysis was conducted, and the pairwise score plot of the skin extracts based on individual phenolic compounds showed a trend of genotype clustering based not only on drought tolerance but also on botanical type of germplasm. Therefore, our results demonstrate the status quo for antioxidant phenolic compounds of peanut genotypes contrasting in drought tolerance grown under natural field conditions.

Optical properties of honey: FTIR spectroscopy and refractometry

115 samples of honey of various botanical types, geographical origin and harvest year (2019– 2021) were analysed using attenuated total reflection infrared spectroscopy of impaired total reflection, refractometry and biochemical analysis. Initial honey samples in liquid and crystallised states were investigated. Crystalline D-glucopyranose (glucose), D-fructopyranose (fructose), their 40% solutions and invert sugar were used as auxiliary substances. Biochemical analysis was used to determine the glucose content in honey samples. Based on the obtained data, a relationship between the results of biochemical analysis and refractometry (refractive index, the content of invert sugars, humidity) was established. We deduced equations that allow the content of glucose and fructose in honey to be evaluated by the refractive index. Studying honey by IR spectroscopy showed that all investigated samples, regardless of the botanical and geographical origins, can be classified into three groups dominated by: I – glucose, II – fructose and III – mixed, with a close content of two monosaccharides. This allowed the bands characteristic of α- and β-pyranose forms of glucose and fructose to be identified, as well as the nature of their changes depending on the ratio of both monosaccharides in honey as a result of their crystallisation to be assessed. It is noted that the ratio of monosaccharides determines not only the stability of the liquid crystal structure and crystallisation rate in honey but also their optical, biochemical and nutritional properties, which are important for the preferred use of honey in medical and pharmacopoeial practice, dietetics and cosmetology.

Nutritional Composition and Health Benefits of Various Botanical Types of Melon (Cucumis melo L.).

Characterizing the diverse melon cultivars for nutrition aids in crop improvement and promoting a healthy diet. Here, we used in vitro assays to characterize the nutritional qualities and health-beneficial effects of 30 melon (Cucumis melo L.) genotypes, including 10 improved cultivars, 16 landraces, and 4 wild types collected from different parts of India. Two landraces (Sidoota and Alper Green) had the highest (12.20 and 11.25) total soluble solids (TSS) contents. The Sidoota and Pappusa landraces had high reducing sugar contents (2.84 and 2.81 mg g−1 fresh weight [FW]). The highest polyphenols contents (22.0 mg g−1 FW) were observed in the landraces Mage Kaayi-2, Budamekaayi, and Small Melon. Reflecting on the primary and secondary metabolite contents, the Mekke Kaayi and Giriyala landraces exhibited high 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity (97.6 and 91% at 100 μg mL−1). Additionally, seven of the landraces showed significant nitric oxide (NO) induction activity (>80% inhibition at 200 μg mL−1), indicating their potential health benefits, and seven showed considerable angiotensin-converting enzyme (ACE) inhibition activity (highest in Kashi Madhu), indicating their potential usefulness in reducing hypertension. Genotypes with high health beneficial compounds identified in this study can be used for breeding improved melon cultivars to promote these fruits as well as a healthy diet.

Front Face Fluorescence Spectroscopy Combined with PLS‐DA Allows to Monitor Chemical Changes of Edible Vegetable Oils during Storage at 60 °C

AbstractIn this work, front‐face fluorescence spectroscopy coupled with chemometric tools are used for the monitoring of rapeseed, sunflower, extra virgin olive, and linseed oil. The oil samples are heated at 60 °C for up to 15 days. The fluorescence excitation spectra are scanned in the range of 270–420 nm and 250–310 nm after emission wavelength set at 450 and 330 nm, respectively. The primary and secondary lipid oxidation products are also determined on the same oil samples. By applying partial least square discriminant analysis (PLS‐DA) to each spectral collection obtained after emission set at 450 and 330 nm, clear discrimination of edible oil samples according to their botanical types, and sometimes aging, is obtained.Practical Applications: This study shows that oxidation of rapeseed, sunflower, extra virgin olive, and linseed oil monitored different trends due to their difference in composition. The presented results demonstrated the capability of the front‐face fluorescence spectroscopy to differentiate four vegetable oils contained in capped and uncapped flasks indicating the evolution of primary and secondary indicators of oxidation during the storage period. The technology can be exploited at the industrial level for monitoring in a non‐destructive manner the level of oxidation of edible oil.

A genomics resource for genetics, physiology, and breeding of West African sorghum.

Local landrace and breeding germplasm is a useful source of genetic diversity for regional and global crop improvement initiatives. Sorghum (Sorghum bicolor L. Moench) in western Africa (WA) has diversified across a mosaic of cultures and end uses and along steep precipitation and photoperiod gradients. To facilitate germplasm utilization, a West African sorghum association panel (WASAP) of 756 accessions from national breeding programs of Niger, Mali, Senegal, and Togo was assembled and characterized. Genotyping-by-sequencing (GBS) was used to generate 159,101 high-quality biallelic single nucleotide polymorphisms (SNPs), with 43% in intergenic regions and 13% in genic regions. High genetic diversity was observed within the WASAP (π = .00045), only slightly less than in a global diversity panel (GDP) (π = .00055). Linkage disequilibrium (LD) decayed to background level (r2 < .1) by ∼50 kb in the WASAP. Genome-wide diversity was structured both by botanical type and by populations within botanical type with eight ancestral populations identified. Most populations were distributed across multiple countries, suggesting several potential common gene pools across the national programs. Genome-wide association studies (GWAS) of days to flowering (DFLo) and plant height (PH) revealed eight and three significant quantitative trait loci (QTL), respectively, with major height QTL at canonical height loci Dw3 and SbHT7.1. Colocalization of two of eight major flowering time QTL with flowering genes previously described in U.S. germplasm (Ma6 and SbCN8) suggests that photoperiodic flowering in West African sorghum is conditioned by both known and novel genes. This genomic resource provides a foundation for genomics-enabled breeding of climate-resilient varieties in WA.

Geographical origin and botanical type honey authentication through elemental metabolomics via chemometrics

The trace and rare earth elements content of 93 honeys of different botanical type and origin have been studied through ICP-MS. Discriminant Analysis (DA) was successful for botanical type and geographical origin classification while Cluster Analysis (CA) was successful only for botanical type. Through Probabilistic Neural Network (PNN) analysis, 85.3% were correctly classified by the network according to their geographical origin and 73.3% according to their organic characterization. A Partial Least Squares (PLS) model was constructed, giving a prediction accuracy of more than 95%. Information obtained using Rare Earths (Y, La, Ce, Pr, Nd, Sm, Eu, Gd, Tb, Dy, Ho, Er, Tm, Yb, Lu) and trace elements (Li, Mg, Mn, Ni, Co, Cu, Sr, Ba, Pb) via chemometric evaluation facilitated classification of honey samples.

Fatty acid desaturase-2 (ahFAD2) mutant alleles in peanut (Arachis hypogaea L.) pre-breeding lines: an insight into the source, features, discourse, and selection of novel pre-breeding lines

High oleic peanuts and derived food products offer longer shelf life benefits to the food processing industry in addition to multiple health benefits to the consumers. The two mutant alleles, ahFAD2A and ahFAD2B control composition of oleic, linoleic and palmitic acid content in peanut. A total of 563 peanut pre-breeding lines were tested for the presence ahFAD2A and ahFAD2B mutant alleles using allele specific markers. The ahFAD2A mutant allele was present in 82 lines, while none of these lines had ahFAD2B mutant allele. Among botanical types, ahFAD2A mutant allele was more frequent in lines with Virginia growth habit than Spanish bunch although no correlation of ahFAD2A mutant allele with high oleic acid content and growth habit could be established. Oleic and linoleic acid content in 82 pre-breeding lines ranged from 39.70 to 62.70% and 17.76 to 31.95%, respectively, with maximum oleic to linoleic acid ratio of 4. Oleic acid was found to be negatively correlated with linoleic and palmitic acid. Further, pre-breeding lines with ahFAD2A mutant allele, high oleic content and high oleic to linoleic ratio were investigated and novel lines were identified for resistance to late leaf spot, short duration, higher pod yield and other yield related traits. These novel pre-breeding lines can be used as a potential donor in peanut improvement programme and to diversify the primary gene pool including initiating further research on induction of fresh ahFAD2B mutant allele.

Profiling of Nutraceuticals and Proximates in Peanut Genotypes Differing for Seed Coat Color and Seed Size.

A total of 60 genotypes of peanut comprising 46 genotypes selected from ICRISAT mini core collection and 14 elite cultivars with differing kernel color and size were used to profile the nutritional parameters such as proximates (moisture, fat, ash, crude protein, crude fiber, carbohydrate content) and nutraceuticals (total polyphenol content and total antioxidant activity). The genotypes showed varied kernel color ranging from white to purple. Kernel skin color was quantified using colorimetry, and the color parameters were expressed as CIELAB color parameters. In total, nine morphological traits, six yield related traits, eight nutritional traits and eleven color parameters were observed across 60 genotypes. The sixty genotypes were grouped into ten clusters based on the color strength. Among them, Cluster-III with dark red seeds had the maximum fat content and total polyphenol content (TPC). Cluster-VI with light pink colored seeds had high antioxidant activity (AOA) and Cluster-X with white colored seeds had highest moisture and crude protein content. Color strength (K/S) was found to be positively correlated with TPC. Another color parameter, redness/greenness (a*) was found to be positively correlated with AOA. However, seed size was positively correlated with the crude protein content, but not with any other nutritional traits under study. The population studies based on the genotypic data indicated two distinct groups pertaining to botanical types of peanut. The marker-trait association (MTA) using single marker analysis indicated 75 major MTAs for most of the nutritional traits except for moisture content. The markers associated with nutritional parameters and other important yield related traits can further be utilized for genomics-assisted breeding for nutrient-rich peanuts.

Botanical types and composition of grazing field in the captive breeding area for Timor deer (Cervus timorensis) at Wosu Village, Central Sulawesi

A research has been conducted to determine the botanical types and composition of the grazing field of a captive breeding area for Timor deer (Cervus timorensis) at Wosu village, Bungku Barat district, Morowali district, Central Sulawesi province from April 26 to May 16, 2018. Forage samples were taken with a destructive sampling method using a pair of 1 x 1m frame. The dry matter content of the forage samples was analyzed at the animal nutrition and feeding laboratory, Tadulako University. Data were analyzed, and results were presented descriptively. The results showed that the botanical composition of the was 83% kinds of grass and 17% weeds. The grasses were dominated by Imperata cylindrica, Cyperus rotundus, Cyperus kyllingia Endl., and Paspalum conjugatum.

Phenolic Composition Influences the Health-Promoting Potential of Bee-Pollen

Information on compositional, nutritional and functional properties of bee-pollen, as a health-promoting food, is essential for defining its quality. Concerning the nutritional importance of phenolic compounds, the aim of this study was to determine the phenolic profile and antioxidant activity of twenty-four bee-pollen samples collected from different regions of Serbia. High-performance thin-layer chromatographic (HPTLC) fingerprinting was used for profiling of bee-pollen samples according to the botanical type. HPTLC hyphenated with image analysis and a pattern recognition technique confirmed the grouping of samples caused by the specific phenolic composition of pollens of different botanical origin. Flavonoid glycosides in bee-pollen samples were identified by applying ultra-high-performance liquid chromatography (UHPLC) coupled with linear ion trap-Orbitrap mass spectrometry (LTQ Orbitrap MS). Eight out of twenty-seven flavonol glycosides were identified in bee-pollen samples for the first time. All analyzed bee-pollen samples showed a high number of phenolic compounds which may have therapeutic potential.

English

Groundnut farming has always been an essentially major agricultural venture, both commercially and in subsistence, to the inhabitants of the Northern Regions of Ghana. The current study sought to assess the relationship and genetic diversity, through a phylogenetic analysis, among twenty-five (25) Ghanaian groundnut genotypes, in reference to US groundnut mini-core collection, based on the Kompetitive allele-specific PCR (KASP) protocol (a high throughput genotyping platform for marker-assisted selection, studies [LightCycler 480II]) and axiom single nucleotide polymorphism (SNP)&nbsp; array version 2 (48,000 SNP features). It further sought to classify the groundnuts into sub-species based on botanical types, market types, general uses as well as genetic diversity to late leaf spot resistance. The Qiagen Dneasy DNA extraction Kit was used for DNA extraction. Results of the study indicate that majority of the Ghanaian groundnuts evaluated belonged to the Virginia (hypogaea) (large-seeded), Spanish/Valencia (vulgaris/fastigiata) and Mixed (Virginia x Valencia) types based on characteristics, including growth habit (erect/semi-erect/upright/bunch or trailing/spreading) seed or pod size/shape and disease resistance. Thus, significant genetic diversity in morphological characteristics, botanical and market varieties, growth habit and seed size were found among the groundnuts studied. Information generated from this study can be used to develop new groundnut breeding lines that combine these traits to produce higher yields. Marker-assisted selection procedures could help enhance this process based on the availability of QTLs and genes for the traits and markers developed in that regard. Key words: Axiom, genetic diversity, genotypes, groundnut, Kompetitive allele-specific PCR (KASP), mini-core collection, phylogenetic analysis, disease resistance, single nucleotide polymorphism (SNP). &nbsp;

Screening of Arachis ssp. against Aspergillus flavus with in vitro seed colonization, EST-SSRs and generic primers and efficacy of different fungicidal control measures

Aflatoxin produced in groundnut due to the infection by the fungus Aspergillus flavus is a major issue with respect to human health as well as export potential. Genetic resistance against this fungus is an important trait needed to counter the issue of Aflatoxin. In the present investigation, 50 Arachis subspecies genotypes including five different Arachis botanical types viz., spanish bunch, virginia bunch and valencia (cultivated) as well as two wild species viz., A. peruviana and A. aequatoriana were tested for their resistance to A. flavus. Compared to the reported resistant varietyICG1326 (J11), only three genotypes viz., ICG10933, ICG12625, TG26 showed resistance to fungal infection. Specific EST-SSR marker shown to be associated with resistance to A. flavus as well as generic molecular markers known to be associated with fungal resistance was tested in the selected genotypes. But, no strong association between markers and fungal resistance could be established. Also, among different fungicides tested for their ability to control A. flavus, carbendazim + thiram was found to be effective.

Establishment and evaluation of a peanut association panel and analysis of key nutritional traits.

Breeding programs aim to improve the yield and quality of peanut (Arachis hypogaea L.); using association mapping to identify genetic markers linked to these quantitative traits could facilitate selection efficiency. A peanut association panel was established consisting of 268 lines with extensive phenotypic and genetic variation, meeting the requirements for association analysis. These lines were grown over 3 years and the key agronomic traits, including protein and oil content were examined. Population structure (Q) analysis showed two subpopulations and clustering analysis was consistent with Q-based membership assignment and closely related to botanical type. Relative Kinship (K) indicated that most of the panel members have no or weak familial relatedness, with 52.78% of lines showing K = 0. Linkage disequilibrium (LD) analysis showed a high level of LD occurs in the panel. Model comparisons indicated false positives can be effectively controlled by taking Q and K into consideration and more false positives were generated by K than Q. A preliminary association analysis using a Q + K model found markers significantly associated with oil, protein, oleic acid, and linoleic acid, and identified a set of alleles with positive and negative effects. These results show that this panel is suitable for association analysis, providing a resource for marker-assisted selection for peanut improvement.

Peatland substrates in northern KwaZulu-Natal: a study of the forming environments, properties and an approach towards classification

Peatlands in South Africa are rare and fulfil important ecological functions but are threatened by degradation. Because of this peatlands have gained increasing attention from scientists during the last two decades. However, knowledge of the peatland substrates and the means to classify them is limited. This study was conducted in peatlands of the Maputaland Coastal Plain. The approach uses the German Soil Mapping directive ‘KA5’, which acknowledges the accumulation process of substrates and botanical peat types. The aims were to investigate which substrates occur, whether these have affinities to certain hydrogeomorphic wetland types (HGMTs), and to determine the physical and chemical properties. Seven peatlands in five different HGMTs were investigated along 19 transects with 141 soil profiles and 674 horizons. In total 15 different peatland substrates were encountered and characterised, amongst them raphia peat, ficus peat and peat-gyttja, which have not been described previously. Mean values were determined for organic matter content, bulk density, degree of decomposition, porosity, pH and carbon/nitrogen ratio. A substrate reference scheme was also developed and classifies the encountered substrates with respect to their occurrence in HGMTs.

Fourier-Transform Imaging of Cotton and Botanical and Field Trash Mixtures

Botanical and field cotton trash comingled with Upland cotton lint can greatly reduce the marketability and quality of cotton. Trash found comingled with cotton lint during harvesting, ginning, and processing is of interest to the textile community. In the current study attenuated total reflectance-Fourier transform infrared (ATR-FTIR) spectroscopic imaging was employed as an analytical technique to analyze cotton trash. Some benefits of this technique were its non-destructive nature and lack of required sample preparation. The technique used in this study, specifically ATR-FTIR spectroscopic chemical imaging, allows for three-dimensional spectral and spatial data to be obtained. In the current study, cotton in mixtures with botanical and field trash types have been identified spectrally and spatially using ATR-FTIR imaging. Botanical trash types (trash derived from the cotton plant) were evaluated and identified independently from cotton, even though both contained cellulose. The field trash types were easily identified from cotton due to their differences in chemical composition. This study can complement current cotton qualitative studies by adding spectral and spatial information to sample analysis.

Isotope Ratio Mass Spectrometry Coupled to Element Analyzer and Liquid Chromatography to Identify Commercial Honeys of Various Botanical Types

The aim of the present work was to investigate whether honeys of various botanical characteristics could be discriminated and identified on the basis of the isotopic data. For this purpose, 45 commercial honey samples of various botanical types were analyzed using isotope ratio mass spectrometry (IRMS) coupled to element analyzer (EA) and liquid chromatography (LC). δ13C values of honey; corresponding extracted protein and individual sugars (sucrose, glucose, and fructose) of honey; reducing sugar; and sucrose contents were determined and utilized to demonstrate the authenticity of commercial honeys of different botanical types. Results indicated that a very strong correlation (δ13Cprotein = 0.862δ13Choney + 0.800, R2 = 0.944) was observed between δ13C honey values and δ13C protein values of real honey samples. δ13Choney and δ13Cprotein should range from −27.2 to −23.5 and −26.9 to −23.6‰ for pure honeys of various botanical types investigated in this study. The use of isotopic compositions and some systematic differences as authenticity criterion permits the honey samples of various botanical types to be reliably detected.