Indian Lac Research Articles

CULTURABLE ENDOSYMBIOTIC BACTERIA FROM THE INDIAN LAC INSECT, KERRIA LACCA (KERR)

The Indian lac insect, Kerria lacca (Kerr) (Coccoidea: Tachardiidae) is a commercially important phytosuccivorous and sessile scale insect. Lac insects are cultured on suitable host plants in India and some Southeast Asian countries to produce lac. The lac insect harbours a number of endosymbionts. Isolation of culturable microbial endosymbionts and their identification through 16S rRNA has revealed sex and host-related differences of microbial species. Bacillus boroniphilus, Enterobacter cloacae and Staphylococcus sp. were found only in the lac insects reared on the plant host Cajanus cajan, whereas Bacillus firmus, Lysinibacillus xylanilyticus, Bacillus horneckiae and Bacillus velezensis were recorded only from Flemingia macrophylla. B. firmus and L. xylanilyticus were female-specific and B. horneckiae and B. velezensis were male-specific with Flemingia macrophylla as host; E. cloacae was female-specific and Bacillus boroniphilus and Staphylococcus sp. were male specific with C. cajan. Biochemical characteristics of the isolates, their genetic relationship with their taxonomic kin and their probable role, based on the information available about these endosymbionts in other hosts, have been studied.

Assessing the infestation-induced response on the plant host by the Indian lac insect

Lac, the only natural resin of animal origin is produced by the Indian lac insect- Kerria lacca. It is the by-product of the complex natural interaction between the lac insect and its host plant. Despite several studies on the perspectives of the chemistry of lac and its production, very little work has been carried out to understand the biology of lac and its associated plant taxa. The present work has been designed to understand the preliminary response if any, of the host plant against lac insect infestation. Structural studies and metabolic profiling such as the determination of total phenolics, flavonoids, antioxidants, FTIR, and GC-MS were carried out. The anatomical investigations revealed coagulation/deposition of metabolites in the infested sites. The infested bark showed higher phenolic, flavonoid content, and antioxidant activity in comparison to non-infested bark which was corroborated by Fourier-transform infrared spectroscopy (FTIR) and GC-MS biochemical analysis. This preliminary study will shed some light on understanding the lac plant host's physiological response and the putative mechanism used by the lac insect in overcoming the plant response.

Weather based forecasting model for emergence of Aprostocetus purpureus (Cameron) – a parasitoid of lac insect, Kerria lacca (Kerr)

Indian lac insect, Kerria lacca is a scale group beneficial insect which suffers by several natural enemies. The establishment of the relationship between the incidences of natural enemies on lac insects with weather variables is essential for formulating management strategies well in advance. The relationship between weather factors and the emergence of A. purpureus was studied from 2011–12 to 2020–21 on the rangeeni summer (baisakhi) lac crop. Correlation and regression analyses were done after pooling ten years data (2011-12 to 2020-21) during the critical lac growth period i.e. SMW 8 to SMW 20. The relative abundance of lac-associated fauna showed that three parasitoids ((Aprostocetus purpureus, Tachardiaephagus tachardiae, and Tyndarichus(=Parechthrodryinus) clavicornis)) and one predator (Eublemma amabilis) were abundant. Among them, A. purpureus recorded maximum percent infestation, which was 84% and 75% on ber and palas, respectively. Maximum number of A. purpureus was emerged during the sexual maturity period (8 to 20 SMW) of the summer lac crop. The incidence exhibited a significant negative correlation with maximum (Tmax) and minimum temperature (Tmin) and a significant positive correlation with morning and evening relative humidity (RH-II). Stepwise regression analysis showed Tmax and RH-II were the most important factors contributing to 68% variation in the incidence of A. purpureus on palas. The present study results indicated that environmental factors played a significant role in the incidence of parasitoids on lac insect.

DNA Barcoding, Morphological Description and Field Diagnostics of <i>Eublemma amabilis</i> (Lepidoptera: Erebidae)

Lac is an important commodity because of its versatile usage, biodegradability, and potentiality to sustain the livelihood of tribal community of India. Lepidopteran pest of Indian lac insect, Eublemma amabilis Moore [1884] (Lepidoptera: Erebidae) causes significant loss than any other lepidopteran pest thereof, both in wild and cultivated crop. Even though it’s a regular pest recorded for decades, its taxonomic studies are lacking. In the present study, we have redescribed E. amabilis added photographic illustrations of male and female genitalia and which is also supported by a novel mitochondrial cytochrome oxidase I DNA barcode.

The role of Wolbachia and the environment on sex determination of the Indian lac insect, Kerria lacca (Coccoidea: Tachardiidae)

• In Kerria lacca , presence of endosymbiont Wolbachia is developmental-stage dependent. • Administration of antibiotic enhances the sex ratio of the lac insect multifold. • The lac insect sex ratio is influenced by environmental factors. • Mode of metamorphosis together with Wolbachia dynamics explains sex ratio changes. The Indian lac insect, Kerria lacca (Kerr) exhibits a highly variable sex ratio, apparently influenced by various environmental factors, in the absence of a chromosomal sex determination mechanism. The endosymbiont Wolbachia, well known as sex ratio distorter, was reported in this species but its role in the sex determination of the host has hitherto remained unexplored. Distinct variation in the presence of Wolbachia was observed in relation to the developmental stages and the sexes of K. lacca as well as the host plant species; adult males of W + whereas adult females are W -. The post-settlement environmental factors such as host plant, season and genetic makeup affect the sex ratio in this insect, probably mediated through Wolbachia dynamics. Denser settlement of the crawlers led to comparatively higher Wolbachia presence with an elevated sex ratio. Suppression of Wolbachia through antibiotic administration resulted in a nearly three-fold increase in the sex ratio. Distinct transitions in the proportion of insects with heterochromatic genome (cytological males) have been observed during the first instar on different host plants, indicating that changes in the sex ratio occur during early development phase. Environmental factors appear to influence Wolbachia , which in turn brings about sex ratio changes, probably mediated through changes in juvenile hormone levels, as the male and female lac insects show distinct modes of metamorphosis.

Effect of different growing media on seed germination and growth of Indian lac tree (Schleichera oleosa) in nursery conditions

Effect of different growing media on seed germination and growth of Indian lac tree (Schleichera oleosa) in nursery conditions

Studying Saraha: Technical and Multi-Analytical Investigation of the Painting Materials and Techniques in an 18th Century Tibetan Thangka

Scientific analyses of the traditional materials and methods in thangka production are uncommon, as thangkas are sacred objects, the sampling of which is discouraged, in order to preserve their integrity. This study builds on this important ethical challenge and presents a three-stage methodology that systematically delves deeper into each layer of the composition, successfully enabling the investigation of different phases of production. In the first stage, visual examination of the painting, including observations under magnification, was used to assess its condition. In the next step, the infrared reflected (IRR) and short-wave infrared (SWIR) images revealed the underdrawing and instances of modifications as well as colour notations. Additionally, ultraviolet-induced visible luminescence (UVL), infrared-reflected false colour (IRRFC) and X-ray images provided important preliminary information on the colourants present, the nature of the underdrawing, and the painting technique. In the final stages, fibre optic reflectance (FORS), Fourier-transform infrared (FTIR) and Raman spectroscopies confirmed the identity of many of the pigments (cinnabar/vermillion, minium, iron oxide, malachite, azurite, indigo, Indian lac), the ground and the binder used for the blue and green paint layers. In addition, key details of practices and materials were revealed, that may indicate provenance or other information of scholarly importance. These will constitute a helpful comparison to existing and future studies of other thangkas.

RECORD OF NATURAL INFESTATION OF THE INDIAN LAC INSECT, Kerria lacca (KERR) (COCCOIDEA:TACHARDIIDAE) ON Prosopis cineraria (L.) DRUCE., Calliandra surinamensis BENTH. AND Ficus carica L. IN PRAYAGRAJ DISTRICT OF EASTERN UTTAR PRADESH

The Indian lac insect, Kerria lacca (Kerr) family Lacciferidae (Kerriidae) and order Hemiptera is economically important insect exploited commercially for lac, a resin refined into shellac and other products. Natural scarlet dye obtained after

Broodlac Quality Assessment and Forecasting of Crawler Emergence in Indian LAC Insect <i>Kerria lacca</i>

Assessment of quality and freshness of broodlac produced by the Indian lac insect Kerria lacca (Kerr) is an important parameter in its pricing. The present study provides an assessment of broodlac quality with 25, 50, 75 and 100% of lac encrustation harvested from summer (baisakhi), rainy (katki) on palas, winter (aghani) on ber, and summer season (jethwi) on kusum. The curve fitting model based on the rate of weight reduction can be used for prediction of freshness. Among the four lac crops, maximum rate of reduction was observed with summer kusmi (jethwi) and rangeeni (baisakhi) crops, and a good quality of broodlac was observed with katki crop on palas. Crawler emergence, stages of yellow spots and number of days for actual crawler emergence varied in different seasons. The present study differentiates the embryonic development into six stages with yellow spots through microscopic images. This reveals that the stage 1 coincides with earlier defined stage 2 and 3, and stage 2, 3, 4 and 5 are of stages 4, 5, 6 and 7 and 8, respectively.

RETRACTED ARTICLE: Surveillance of Suspicious Activities of Chinese Army Near Indian LAC

According to the agreement of border sharing between India and China, China was supposed to be posted till finger 8 position and India till finger 4 position and between the finger 8 and finger, 4 was considered no man’s land in which only patrolling is allowed. But in recent days, chinese has crossed finger 8 position and has set up its camp near finger 4 position means in no man’s land, therefore, violating the agreement and thus resulting in a critical situation between two countries. Also on 15th June 2020, a group of soldiers went into their camp for sorting this issue of border sharing but were brutally killed by Chinese soldiers in which 20 of our soldiers lost their lives. Therefore, taking into consideration the current situation, it is now very important to monitor and analyze the activities of the Chinese army so that India can prepare itself for any further problems. Surveillance is not about detecting objects in an image but to analyze suspicious activity in a scenario. Therefore, the proposed surveillance dashboard can be used to analyze the activities done by the Chinese army within a specific number of days when inputted satellite images. The stage of a manual analysis of these activities will be reduced. The system is based on Content-Based Image Retrieval using deep learning with real-time feature extraction property.

Essential amino acid profiling of the four lac hosts belonging to genus Flemingia: its implications on lac productivity.

The Indian lac insect (Kerria lacca), a hemipteran, phloem sap sucking sedentary insect is an important bioresource which thrives on tender twigs of more than 400 plant species belonging to various genera and families. The most common commercial host plants for lac cultivation are big trees hence cultivation was concentrated mainly to dense forests across the country till last decade. Recently, a new bushy host plant belonging to the genus Flemingia has been introduced so that lac can be cultivated on farmlands like other cash crops. The insect is sedentary and feeds on the phloem sap of the host plants, the only source of its nutrition. Interestingly, the biological attributes of the insect as well as the qualitative and quantitative production of lac is influenced by the host plant on which the insect feeds upon. The present study was thus aimed at deciphering the effect of phloem sap constituents obtained from four plant host taxa belonging to the same genus Flemingia viz. F. semialata, F. macrophylla, F. bracteata and F. chapar (essential amino acids only-EAAs) on lac productivity. Moreover, a newer method for phloem sap collection i.e. Dot-blot in addition to the facilitated exudation using EDTA was also investigated. Dot-blot method for phloem sap collection also came out to be a promising method for field studies; although slightly higher concentration of EAAs were obtained from EDTA method, thus the later was used for further analysis. Phloem sap of four plant host taxa belonging to the same genus Flemingia were qualitatively and quantitatively analysed for seven EAAs (Arginine, Glycine, Leucine, Methionine, Phenylalanine Tyrosine and Valine). Amino acid concentration regime and further analysis done using statistical tools (ANOVA and PCA) points out the EAA concentration in the phloem sap is in congruency with the lac production data obtained through previous studies as F. semialata > F. macrophylla > F. chapar > F. bracteata. The present study thus scientifically points out that F. semialata can be a promising plant for lac cultivation on the basis of higher EAA content as compared to the rest three.

Fibre optic reflectance spectroscopy and multispectral imaging for the non-invasive investigation of Asian colourants in Chinese textiles from Dunhuang (7th-10th century AD)

The archaeological complex of Dunhuang (northwestern Gansu, China) is considered a pearl on the Silk Road and the content of its caves revolutionised oriental studies. The British Museum hosts a significant number of textiles and textile fragments from the site. Although mostly catalogued and studied from the point of view of textile production, weaving techniques and iconography, they have never undergone dye analysis, which is here presented for the first time. Due to the value, preciousness and fragility of these textiles, sampling is not always an option. Therefore, a non-invasive approach combining multispectral imaging (MSI) and fibre optic reflectance spectroscopy (FORS) was tested. The final aims of this research were to explore the advantages and limitations of such an approach with application to Asian dyes and ultimately to provide new insights into the technological skills related to dyestuff production and use in ancient China.To achieve these aims, twenty-nine reference specimens of Asian dyes were sourced, including sappanwood (Biancaea sappan), sandalwood (Pterocarpus santalinus), different species of madder (Rubia spp.), rosewood (Dalbergia sp.), rhubarb (Rheum emodi), dragon's blood (Daemonorops sp.), Indian lac (Kerria lacca), gromwell (Lithospermum erythrorhyzon), safflower (Carthamus tinctorius), amur-cork tree (Phellodendron amurense), gamboge (Garcinia hanburyi), violet (Viola yedoensis), pagoda tree (Sophora japonica). Silk was dyed using these raw materials and the reference samples were analysed by colorimetry before and after ageing. These measurements highlighted the most light-sensitive dyes to be turmeric, safflower, sappanwood and sandalwood. The reference samples were also used to create a database of reflectance and apparent absorption spectra obtained by FORS, as well as to record the reflectance, luminescence and absorption properties of specific dyes by MSI. The combined information obtained from MSI and FORS brought to the fore the signature behaviour of certain dyes (protoberberine-based dyes, safflower, gromwell, tannins, madder, sappanwood, Indian lac and indigo), which enabled their unequivocal identification and the mapping of their distribution in thirty-one Dunhuang textiles from the British Museum's collection.The data presented in this work will be beneficial to other researchers and conservators investigating textiles from the Silk Road using a non-invasive approach and for the assessment of textiles and their suitability for display.

Detection of Wolbachia Phage (WO) in Indian Lac Insect [Kerria lacca (Kerr.)] and Its Implications.

Wolbachia, a maternally inherited bacterium induces reproductive alterations in its hosts such as feminization of males, male killing and parthenogenesis. It is the most diverse endosymbiont infecting more than 70% of the insects ranging from pests to pollinators. Kerria lacca-a hemipteran is a sedentary, oriental insect known to produce lac-the only resin of animal origin. The present study was conducted to screen the presence of Wolbachia and its associated phages in the two infrasubspecific forms (four insect lines) of K. lacca viz. kusmi and rengeeni differing from each other on the basis of host preference. Wolbachia and its associated phage were found to be prevalent in all the insect lines analyzed. We, hereby, report the presence of WO-phage (Wolbachia phage) for the first time in K. lacca. Further, phylogenetic data differentiated the kusmi and rengeeni infrasubspecific forms into two different groups on the basis of WO-phage sequences.

Isolation and identification of culturable bacteria from honeydew of Indian lac insect, Kerria lacca (Kerr) (Hemiptera: Tachardiidae)

Indian lac insect, Kerria lacca belongs to a distinct group of phytosuccivorous insects feeding on sugar-rich phloem sap and excreting excess sugar in the form of large quantities of honeydew, which serves as an excellent medium for microbial growth. This work has made a novel attempt to isolate and identify culturable bacterial species associated with lac insect honeydew, which can be further used to unravel their significance by identifying and characterizing semiochemicals or bacterial volatiles involved in signalling the natural enemies of lac insects and can be effectively used in biological pest control. To isolate microorganisms present in honeydew, droplets of honeydew were collected, diluted and spotted on suitable culture media. Further, 16S rRNA sequencing method was undertaken for the identification of isolated bacterial species. Following BLAST analysis, the nucleotide sequences were submitted to GenBank. Ten different bacterial species were isolated in which species of Enterobacter was the most abundant isolate identified. Others were Klebsiella variicola, Stenotrophomonas maltophilia, Pantoea cypripedii, Pseudomonas stutzeri, Pantoea dispersa, Bordetella trematum and Cedecea davisae.

Bacterial flora associated with Kerria lacca (Kerr)

The application of biochemical and molecular methods in this study to investigate the microbiota of Indian lac insect, K. lacca has provided the basis to explore the community of bacterial taxa. Eight different bacterial species were isolated and categorized as endosymbiont, gut bacterium or subsurface bacterium. Three of them viz. Paenibacillus barengoltzii, Pseudomonas fulva and Pantoea ananatis were exclusive to males, whereas Bacillus cereus, Solibacillus silvestris and Curtobacterium citreum were found in females only. Majorly, the associations were sex-specific and only Bacillus megaterium and Arthrobacter subterraneus were common to both the sexes. Bacillus megaterium, Arthrobacter subterraneus and Pantoea ananatis were found to be the most abundant bacterial species. This work has made an attempt to identify the microbes associated with lac insect and to characterize them using molecular and biochemical approaches. The location of these bacteria in insect body and their significance in lac insect life cycle has also been discussed.

Spectroscopic and mass spectrometric approach to define the Cyprus Orthodox icon tradition - The first known occurrence of Indian lac in Greece/Europe

The present work reports decisive new data regarding the characterization of the paint materials used by Cypriot icon painters in Orthodox icons from the 12th to the 18th century. Eight samples of red paint, collected from seven different icons originally placed in three different churches in Morphou and Nicosia (Cyprus) have been analysed to identify the painting technique.The identification of pigments and fillers was performed using micro-Raman spectroscopy and Fourier Transform Infrared spectroscopy (FT-IR). In order to determine the binding medium, Pyrolysis Gas Chromatography-Mass Spectrometry (Py/GC/MS) and Gas Chromatography-Mass Spectrometry (GC/MS) were used. The study of the organic colorants was performed by Liquid Chromatography-tandem Mass Spectrometry (LC-MS/MS) analysis. Both inorganic pigments and organic colorants were used to create the red hues in the icons. Cinnabar was identified in almost all the paint samples studied, although in some cases red lead was also found. European cochineal, kermes, redwood and Indian lac were identified in the red paints, in combination with inorganic pigments. Moreover, stratigraphic analyses allowed us to assess that animal glue and calcium sulphate were used for the preparation layers, while egg was used as binding medium in the red paint layers. The identification of Indian lac in a 12th century icon is the first known occurrence reported in the literature of this organic colorant in the European tradition.

A Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry Method for the Identification of Anthraquinones: the Case of Historical Lakes.

This study deals with the identification of anthraquinoid molecular markers in standard dyes, reference lakes, and paint model systems using a micro-invasive and nondestructive technique such as matrix-assisted laser desorption/ionization time-of-flight-mass spectrometry (MALDI-ToF-MS). Red anthraquinoid lakes, such as madder lake, carmine lake, and Indian lac, have been the most widely used for painting purposes since ancient times. From an analytical point of view, identifying lakes in paint samples is challenging and developing methods that maximize the information achievable minimizing the amount of sample needed is of paramount importance. The employed method was tested on less than 0.5 mg of reference samples and required a minimal sample preparation, entailing a hydrofluoric acid extraction. The method is fast and versatile because of the possibility to re-analyze the same sample (once it has been spotted on the steel plate), testing both positive and negative modes in a few minutes. The MALDI mass spectra collected in the two analysis modes were studied and compared with LDI and simulated mass spectra in order to highlight the peculiar behavior of the anthraquinones in the MALDI process. Both ionization modes were assessed for each species. The effect of the different paint binders on dye identification was also evaluated through the analyses of paint model systems. In the end, the method was successful in detecting madder lake in archeological samples from Greek wall paintings and on an Italian funerary clay vessel, demonstrating its capabilities to identify dyes in small amount of highly degraded samples. Graphical Abstract ᅟ.

The “Coptic” textiles of the “Museo Egizio” in Torino (Italy): a focus on dyes through a multi-technique approach

The Coptic textile collection of the Museo Egizio in Torino (Italy) has been the object of a broad project aimed at investigating the production techniques, at documenting the conservation state and at reconsidering the attributed age. The collection was also analysed by non-invasive and micro-invasive techniques with the aim of detecting the dyes that have been employed to obtain the colours, in order to complete the set of technological information available for each textile. The data collected in the present work have been compared with published results from other Coptic textile collections, with the aim of highlighting a possible link between the age of the textile and the dyes that were employed. Moreover, the combined use of spectroscopic and chromatographic techniques allowed us to compare the results for the non-invasive and the micro-invasive approaches and to go deeper into the dyeing technology by detecting unexpected combinations of dyes. In particular, the use of a double dyeing with madder and Indian lac dye was revealed in some Roman-Byzantine and Byzantine textiles from the collection.

Textile technology in Nepal in the 5th-7th centuries CE: the case of Samdzong

The first results of textile and dye analyses of cloth remains recovered in Samdzong, Upper Mustang, Nepal, are presented. The site consists of ten shaft tombs, dated between the 400-650 CE, cut into a high cliff face at an elevation of 4000 m asl. The dry climate and high altitude favoured the exceptional preservation of organic materials. One of the objects recovered from the elite Samdzong 5 tomb complex is composed of wool fabrics to which copper, glass and cloth beads are attached and probably constitutes the remains of a complex decorative headwear, which may have been attached to a gold/silver mask. SEM was used to identify the fibre sources of the textiles, which are all of animal nature. Two of the textiles are made of degummed silk. There is no evidence for local silk production suggesting that Samdzong was inserted into the long-distance trade network of the Silk Road. HPLC-DAD analysis permitted identification of a variety of organic dyes, including Indian lac, munjeet, turmeric and knotweed/indigo, while cinnabar was identified through micro Raman spectrometry. The results indicate that locally produced materials were used in combination with those likely imported from afar, including China and India.Statement of significance Textiles are not commonly found in most archaeological contexts, yet when preserved in organic state they can be analysed using modern scientific techniques such as Scanning Electron Microscopy and High Performance Liquid Chromatography to uncover information about their fibre and dye sources. This information can be used to reconstruct the economic and social aspects of textile production and exchange in the past. Identification of silk fibres and munjeet and Indian lac dyes in the textile finds from Samdzong, Nepal suggests that imported materials were used in combination with those locally produced.Data availability The authors confirm that all data underlying the findings are fully available without restriction. All relevant data are contained within the paper.

Lac dye as a potential anti-neoplastic agent.

Certain anthraquinone compounds are effectively used for treatment of cancer. The present study describes the inhibitory activity of lac dye, a mixture of polyhydroxy anthraquinone compounds (Laccaic acid A, B, C, D and E), produced by the Indian lac insect, Kerria lacca (Kerr). In vitro testing for anticancer activity of lac dye was done at four concentrations (10, 20, 40, 80 μg/ml) on ten human malignant cell lines including six human leukemia cell lines, U973, Raji, K562, Jurkat, MOLT 4 and HL60 using SRB assay. Both crimson and yellow lac dye inhibits proliferation of Human leukemia cell lines, Raji, U937, K562, HL60 and Jurkat, therefore can be considered as a potential anticancer agent for leukemia.