Cochineal Research Articles

Characterizing the Interaction of Carmine with Human Serum Albumin Using Biophysical and Molecular Docking Approaches

Carmine (CRM) is regarded as a safe coloring agent derived from the cochineal insect and is used to impart red color in a variety of products ranging from food, beverages, and cosmetics to pharmaceutical drugs. However, various toxicological implications are associated with the use of CRM, viz. anaphylaxis, utricaria, rhinitis, asthma and other cytotoxic and genotoxic ramifications. Therefore, it is imperative to evaluate the toxicity of CRM at the molecular level. A number of biophysical and biochemical methods have been used to examine the impact of CRM on human serum albumin (HSA). The formation of a CRM–HSA complex was revealed by intrinsic fluorescence analysis. The resulting circular dichroism spectra showed that CRM binding reduces the amount of alpha helices present in HSA. The synchronous fluorescence spectroscopy revealed significant microenvironment changes around the tyrosine (Tyr) and tryptophan (Trp) residues in HSA upon CRM binding. The binding of CRM takes place at or close to site I of HSA while also having transient interactions at the subdomain IIA–IIB interfaces determined by the drug displacement and molecular docking studies. Our present research described in the paper, we believe, is significant to understand the CRM-induced toxicity in people.

Exploring the effects of cochineal and indigotin extracts on the properties of plasma electrolytic oxidation coating

The aim of this study is to investigate the effectiveness of chromophoric organic compounds in modifying the surface of AZ31 Mg alloy using the plasma electrolytic oxidation (PEO) process. In particular, the study examines the deposition of cochineal extract and indigotin extract on PEO coatings and evaluates their impact on the morphology, structure, and properties of the resulting coatings. The PEO coatings were analyzed by scanning electron microscopy (SEM), energy dispersive spectroscopy (EDS), and X-ray diffraction (XRD), while the anti-corrosive properties were characterized by potentiodynamic polarization (POL) measurements. The results indicate that the deposition of chromophores of cochineal extract on PEO coatings leads to a smoother surface morphology, while indigotin deposition results in a partially sealed surface with large cracks. The combined solution of both the organic extracts results in a higher carbon content but does not effectively seal the surface, likely due to the formation of larger molecules that cannot fully penetrate the pores of the PEO coating. Overall, this study demonstrates the potential of chromophoric organic compounds for surface modification of Mg alloys and highlights the importance of careful selection and characterization of the post-treatment compounds.

The successful biological control of Cylindropuntia fulgida var. mamillata (A. Schott ex Engelm.) Backeb. (Cactaceae) (coral or boxing glove cactus) at two field sites in Queensland, Australia

Cylindropuntia fulgida var. mamillata (A. Schott ex Engelm.) Backeb. (Cactaceae) (coral or boxing glove cactus) is one of eight invasive species of Cylindropuntia in Australia that have been targeted for biological control. The ‘cholla’ lineage of Dactylopius tomentosus (Lamarck) (Hemiptera: Dactylopiidae) was first released in South Africa in 2008. This lineage is the second of a series of host-specific cochineal insects to be released to control Cylindropuntia species in Australia and was selected to target C. fulgida var. mamillata. Approval to field release this lineage was granted in November 2015 and the cochineal was first released at Leander Station, near Longreach, Queensland in March 2016 and at Booligar Station, near Hebel, Queensland in April 2016. An intensive monitoring program was undertaken at both sites to evaluate the establishment, rate and direction of spread and impact of the D. tomentosus ‘cholla’ lineage on C. fulgida var. mamillata. The cochineal established very well at Leander Station but needed to be re-released at Booligar Station following heavy rain and flooding soon after the first release. Once established, populations of the cochineal at both sites built up very quickly and spread with the prevailing winds, infesting 100% of plants at each of the monitoring sites within 14 months of release. The impact on plants was substantial, with 95% of plants dying at Leander Station within 19 months of the cochineal being released. At Booligar Station, about 98% of plants had died within 18 months of the second cochineal release. Of the plants that were still alive, all had been severely damaged by the cochineal and were showing signs of poor health. These results and observations at other release sites, both in Australia and South Africa, suggest D. tomentosus ‘cholla’ lineage should be an effective agent for the control of C. fulgida var. mamillata.

High performance liquid chromatography as a micro-destructive technique for the identification of anthraquinone red dyestuffs in cultural heritage objects

Abstract This article describes two of the most common natural sources of red anthraquinones that have been used since antiquity for textile dyeing and as lake pigments, namely cochineal insect and the roots of Rubia and Relbunium plants. Carminic acid is the main component of carmine, the red colorant obtained from cochineal. Alizarin and purpurin are found in the extracts of Rubia roots, but Relbunium roots contain only purpurin. These red anthraquinones have been identified in a variety of cultural heritage objects. High performance liquid chromatography with diode array detection (HPLC-DAD) is described as the most suitable analytical technique for the identification of these dyestuffs, particularly in micro-samples from textiles or pigment layers in paintings and polychrome sculptures. Finally, application of HPLC-DAD analysis to the identification of lake pigments in micro-samples of mural paintings of two eighteenth century Andean churches in Peru is described.

De novo biosynthesis of carminic acid in Saccharomyces cerevisiae

Carminic acid is a natural red dye extracted from the insect Dactylopius coccus. Due to its ideal dying effect and high safety, it is widely used in food and cosmetics industries. Previous study showed that introduction of polyketide synthase (OKS) from Aloe arborescens, cyclase (ZhuI) and aromatase (ZhuJ) from Streptomyces sp. R1128, and C-glucosyltransferase (UGT2) from D. coccus into Aspergillus nidulans could achieve trace amounts of de novo production. These four genes were introduced into Saccharomyces cerevisiae, but carminic acid was not detected. Analysis of the genome of A. nidulans revealed that 4′-phosphopantetheinyl transferase (NpgA) and monooxygenase (AptC) are essential for de novo biosynthesis of carminic acid in S. cerevisiae. Additionally, endogenous hydroxylase (Cat5) from S. cerevisiae was found to be responsible for hydroxylation of flavokermesic acid to kermesic acid. Therefore, all enzymes and their functions in the biosynthesis of carminic acid were explored and reconstructed in S. cerevisiae. Through systematic pathway engineering, including regulating enzyme expression, enhancing precursor supply, and modifying the β-oxidation pathway, the carminic acid titer in a 5 L bioreactor reached 7580.9 μg/L, the highest yet reported for a microorganism. Heterologous reconstruction of the carminic acid biosynthetic pathway in S. cerevisiae has great potential for de novo biosynthesis of anthraquinone dye.

Physicochemical and sensory characteristics of mortadella manufactured with <em>Caiman yacare (Caiman yacare)</em> meat and natural food colorants

This study evaluated the effects of the natural food colorants from annatto (Bixa orellana) (0.3%, 0.6%, and 0.9%) and cochineal carmine (Dactylopius coccus) (0.05%, 0.10% and 0.15%) on the physicochemical and sensory characteristics of mortadella made with Caiman yacare (Caiman yacare) meat. All mortadella formulations were prepared in accordance with Brazilian law. There were significant differences (P < 0.05) between treatments in pH, color parameters C* and h*, and texture profile. The scores determined in the sensory evaluation also showed differences in color, flavor, texture, and overall impression of mortadella formulations with different concentrations of both natural food colorants (P < 0.05). The mortadella containing 0.15% cochineal carmine received the highest scores for color (7.91), flavor (7.49), texture (7.43) and overall impression (7.58), thus presenting the best physicochemical and sensory characteristics among all formulations studied.

Challenges Inherent in Controlling Prickly Pear Species; a Global Review of the Properties of Opuntia stricta, Opuntia ficus-indica and Opuntia monacantha.

Opuntia species (prickly pear) were deliberately introduced to many countries around the world for fruit, cochineal dye production, living fencing or as ornamentals. They are now some of the world's most significant weeds, particularly in regions with warm and or dry climates, as they pose threats to economic and environmental assets. In addition, they can cause considerable health issues for humans and animals. Opuntia spp. have prolific reproduction abilities, being able to reproduce both vegetatively and by seed. They have generalist pollination and dispersal requirements, which promotes their establishment and spread. Opuntia stricta, O. monacantha and O. ficus-indica are the most globally widespread of the Opuntia spp. In many countries, biological control agents, particularly the cactus moth (Cactoblastis cactorum) and various cochineal insects from the Dactylopius genus, have successfully reduced land-scape scale populations. On a smaller scale, controlling these weeds by either injecting or spraying the cladodes with herbicides can provide effective control. Care must be taken during herbicide treatments as any untreated areas will regenerate. While biological control is the most cost and time effective control method for landscape-scale infestations, further research into the combined efficacy of herbicides, fire, grubbing and pre-burial techniques would be beneficial for land managers to control small-scale and establishing populations. It would also be useful to have greater knowledge of the potential seedbank longevity and seed ecology of these species so that integrated management strategies can be developed to not only deal with initial populations but also the subsequent seedling regrowth.

Current status of the cactus scale Diaspis echinocacti Bouché (Hemiptera: Diaspididae) on Opuntia ficus‐indica and its first morphological and microscopic description in Morocco

AbstractA total of 460 sites in 28 provinces located in the northern part of Morocco, where the prickly pear cactus Opuntia ficus‐indica is a natural crop, were surveyed from autumn 2020 to winter 2022. In 2020 and the first half of 2021 the cochineal scale Dactylopius opuntiae (Cockerell, 1896) (Hemiptera: Dactylopiidae) was the only scale recorded on pads of O. ficus‐indicata. However, high infestation of pads and fruits of O. ficus‐indica with Diaspis echinocacti Bouché (Hemiptera: Diaspididae) was subsequently recorded in three sites. D. echinocacti is described based on the morphology of the male and female adults, and microscopic examination of 60 slide‐mounted adult females. The presence of D. echinocacti on prickly pear cactus in these three sites in Morocco represents a serious potential threat for cactus in the country. Two predators, Rhyzobius lophanthae Blaisdell (1892) (Coleoptera: Coccinellidae) and Cybocephalus sp. (Coleoptera: Nitidulidae), and two hymenopteran parasitoids, the ectoparasitoid Aphytis sp. (Hymenoptera: Encyrtidae) and an endoparasitoid Encyrtidae, were recorded to be associated with cactus scale. Further work is needed to identify all these predators/parasitoids to species level and to evaluate their potential for biological control.

Histologic changes in liver tissue of sheep fed spineless cactus resistant to Dactylopius opuntiae

This study aimed to assess the effects of diets containing spineless cactus genotypes resistant to carmine cochineal insect (Dactylopius opuntiae) on the liver condition of sheep. Thirty-six non-castrated Santa Inês male sheep (six months of age and average initial body weight of 22.0 ± 2.9 kg) were assigned to a completely randomized design, with three treatments and 12 replicates. The animals were fed a diet with Tifton hay as exclusive roughage (control) and two more diets in which the hay was partially replaced by ‘Miúda’ or ‘Orelha de Elefante Mexicana’ (OEM.) spineless cactus. The animals were randomly slaughtered at 86 days after 16-h solid fast. The use of spineless cactus, regardless of genotype, increased the liver weight and caused inflammatory processes and necrosis on the liver parenchyma. Spineless cactus in diets for sheep confined for 86 days, despite causing liver tissue damage, such as inflammation and death process of hepatocytes, does not compromise weight gain.

New progress in the extraction and identification of Mexican cochineal (Dactylopius coccus) in fresh dyed wool and dyed wool with artificially accelerated aging and color description

AbstractThe main purpose of this study was to determine if a method developed in the laboratory by HPLC‐ESI‐QToF was able to identify the characteristic compounds of the Mexican cochineal (Dactylopius coccus) in freshly dyed wool fiber and following an artificially accelerated aging process. Wool fibers were dyed with various formulations of popular interest (red‐orange, Mexican red, cochineal red, and purple). These formulations are currently used by indigenous communities in the south of Mexico. Some dyed wools were aged in an UV chamber, in order to simulate the deterioration that a fiber dyed in a previous time could have. Notable results were the identification of the main component, carminic acid, and the respective biomarkers of the Mexican cochineal (dcII, dcIV, and dcVII) with a small amount of sample and in a short time, in three of the four formulations before and after aging, thus providing a method that can probably be used for conservation purposes of cultural heritage. It was concluded that when alum is used as a mordant and as a carbonate pH fixer, the fixation of cochineal on the wool is not very strong. This is one of the few works where Mexican cochineal has been studied, simulating the deterioration due to the passage of time in wool samples of different colors with the same natural dye. As a way of describing the color more objectively, the color was measured with the CIE L*A*B* scale in the four formulations of interest, in recently dyed and artificially aged wool.

Resistance of Opuntia ficus-indica cv ‘Rojo Pelon’ to Dactylopius coccus (Hemiptera: Dactylopiidae) under greenhouse condition

Opuntia ficus-indica (L.) Mill.), is called "beles" in the Tigray region of Ethiopia, where it is used for multiple purposes, such as food, fodder, and fences; however, in recent years the introduction of the cochineal (Dactylopius coccus Costa) to this region has caused it to become a plague that has affected thousands of hectares, causing an economic and social problem. Six cultivars: three from Opuntia ficus-indica (‘Atlixco’, ‘Chicomostoc’, and ‘Rojo Pelón’) and three others O. cochenillifera (‘Nopalea’ and ‘Bioplástico’) and O. robusta (‘Robusta’) were tested for resistance to D. coccus in completely randomized design (CRD) experiment with six replications under greenhouse conditions, for two seasons (cycles). Matured cladodes were infested by attaching a paper bag with ten ovipositing female D. coccus. The number of insects at different stages and yields were recorded, log transformed (insect count data), and subjected to statistical analysis. The number of nymphs (stages I and II) was significantly different in both cycles (P=0.0000). The insects died at the nymph I stage at ‘Rojo Pelón’ and ‘Robusta’, in contrast, they completed their life cycle at ‘Atlixco’, ‘Chicomostoc’, and ‘Nopalea’ cultivars. Insects at ‘Bioplástico’ cultivar remained in the nymph I stage the whole experimental period; the molting was hampered. Although some crawlers started settling (nymph I) at ‘Rojo Pelón’, they couldn’t survive and developed, and this confirms that this O. ficus-indica cultivar is resistant to D. coccus.

Safety and efficacy of a feed additive consisting of carmine for cats and dogs (Mars Petcare GMbH).

Following a request from the European Commission, EFSA was asked to deliver a scientific opinion on the re‐evaluation of the safety and efficacy of carmine (sensory additive, functional group: colourants – substances that add or restore colour in feeds) for cats and dogs. The additive consists of aluminium lakes of carminic acid (carmines), which are complexes of aluminium and carminic acid. Carminic acid, produced from the female insect Dactylopius coccus Costa, is the colouring principle and is present in the final additive at 50%. The additive does not pose a risk concerning genotoxicity and is considered safe for dogs and cats at 264 and 220 mg Carmine/kg feed, respectively (which correspond to 132 and 110 mg carminic acid/kg feed, respectively). Due to the nature of the additive, the FEEDAP Panel concluded that allergic reactions may occur in the target species following the ingestion of feeds containing the additive. The FEEDAP Panel concluded that the exposure of users by inhalation is very likely, and that carmine is a respiratory and skin sensitiser; however, due to the lack of data, a conclusion cannot be reached on the potential skin and eye irritation of the additive. The additive is considered to be efficacious in feeds for dogs and cats under the proposed conditions of use.

Dactylopius coccusCosta pest ofbeles, and ways for sustainable pest management and development in Tigray, Ethiopia

ISHS X International Congress on Cactus Pear and Cochineal: Cactus - the New Green Revolution in Drylands Dactylopius coccus Costa pest of beles, and ways for sustainable pest management and development in Tigray, Ethiopia

CULTIVO in vitro DE CÉLULAS EMBRIONARIAS DE COCHINILLA (Dactylopius coccus Costa) A DIFERENTES pH

Con el objetivo de determinar el pH óptimo al que cultivos celulares de cochinilla (Dactylopius coccus Costa) registran la máxima producción de ácido carmínico in vitro, se evaluaron varios niveles de pH (4.5, 5.0, 5.5, 6.0 y 6.5) y de fracciones de centrifugado (0, 2, 5 y 10 min) de células maceradas de embriones de cochinilla establecidos en medio nutritivo de Schneider. El contenido de ácido carmínico presente en cada muestra fue determinado mediante cromatografía líquida de alta resolución. La extracción del pigmento de las muestras se realizó con una solución de HCl 2M. Los límites de detección del pigmento fueron de 1.0 mg L-1 a 120.0 mg L-1. No se observó un efecto significativo del pH en el contenido de ácido carmínico ni en el número de líneas celulares, aunque el pH intermedio (5.5) tendió a dar mejores resultados.

Hiding in plain sight: Cryptic enemies are found on cochineal (Hemiptera: Dactylopiidae), a scale insect of economic and cultural significance.

Cochineal is the common name for cactus‐feeding scale insects in the Dactylopiidae. These ruby‐red insects include the domesticated dye insect Dactylopius coccus. Dactylopius coccus and congeners have been introduced around the world, some accidentally, to become pests of prickly pear cactus species (Opuntia), and some intentionally, for dye production or biological control of pest Opuntia. In the northern Sonoran Desert (Tucson, AZ, USA), we studied the enemy complex of D. opuntiae and D. confusus on Opuntia and characterized two cryptic enemies, a coccinellid beetle predator and a parasitoid wasp. (1) Hyperaspis sp. The coccinellid predator Hyperaspis trifurcata shares a niche with a similar, typically all‐black beetle. Morphological data, crossing tests, and phylogenetic results showed the black beetle to be a distinct, undescribed species in the genus Hyperaspis, with a rare spotted phenotype that is similar in appearance to H. trifurcata. Crossing tests among black and spotted forms showed the spotted morph is inherited as a single‐locus dominant allele. (2) Formicencyrtus thoreauini. Rearing of this ant‐like parasitoid wasp (Encyrtidae) in pure culture of D. opuntiae showed it to be a semi‐gregarious primary parasitoid of cochineal. To our knowledge, this is the first confirmed instance of a cochineal parasitoid. Observations of development show early instar larvae keep their posterior end within the egg chorion, attached to an aeroscopic plate with a connection to the cochineal body wall. Late instar larvae are eventually surrounded by a membrane, likely of larval origin. Wasps then pupate in a dry air‐filled chamber within the desiccated scale remains before chewing out as an adult. Both Hyperaspis sp. and F. thoreauini may have restricted distributions. Hyperaspis sp. does not appear to be a member of the cochineal community in Mexico or Texas, and scant records suggest F. thoreauini may also be restricted to the Southwestern USA.

Insecticidal activity of some plant essential oils against the Opuntia cochineal scale insect, Dactylopius opuntiae Cockerell (Hemiptera: Dactylopiidae)

Insecticidal activity of some plant essential oils against the Opuntia cochineal scale insect, Dactylopius opuntiae Cockerell (Hemiptera: Dactylopiidae)

Genotoxicity test of eight natural color additives in the Korean market.

BackgroundVarious natural color additives are preferred by many consumers over synthetic color additives because they are perceived to be safer. However, most do not have sufficient toxicity data for safety assurance. Color ingredients in particular have some structures suspected of being toxic. Eight natural color additives, gardenia red, blue, and yellow; lac color; cochineal extract; beet red; Curcuma longa Linne extract (Curcuma extract); and Monascus red, currently permitted for use in Korea, were selected and subjected to genotoxicity tests. Acceptable daily intake values have not been allocated to these color additives (except for cochineal extract) due to the lack of toxicity data. We used genotoxicity testing—the bacterial reverse mutation test (Ames test), in vitro mammalian chromosomal aberration test, and in vivo alkaline comet test—for minimum safety assurance.ResultsGardenia red and blue, cochineal extract, lac color, and beet red did not induce mutagenicity or chromosomal abnormalities. Gardenia yellow was mutagenic in the Ames test, but was not positive in the in vitro chromosomal aberration test or in vivo alkaline comet assay. Curcuma extract and Monascus red induced cytotoxicity in the Ames test at high concentrations in Salmonella typhimurium TA1537 and TA100, without showing mutagenicity. On cytotoxicity testing, Curcuma extract and Monascus red showed cytotoxicity at concentrations higher than 313 μg/ml in Chinese hamster ovary CHO-K1 cells and showed equivocal results in chromosomal aberration assay of the same cells. Curcuma extract and Monascus red produced significant increases in DNA damage at a dose of 2000 mg/kg b.w./day, and induced dose-dependent increases in % DNA in the tail and tail moment on in vivo comet assay.ConclusionsSix out of eight food colorants did not cause genotoxicity and cytotoxicity. However, Monascus red and Curcuma extract showed definite cytotoxicity and probable genotoxicity.

Electrochemical evaluation of mucilage and cochineal pigments as a hybrid film coating on aluminum surfaces

This work deals with the electrochemical evaluation of a hybrid coating based on mucilage and prickly pear cochineal (Dactylopius coccus) for corrosion protective applications. The Opuntia streptacantha mucilage was extracted by grinding prickly pear cochineal, and three coatings containing mucilague (CM), cochineal-mucilage (CMC), and cochineal-mucilage without Tween 80 (CMC-T) were formulated. The aluminum working electrodes (WE) were coated by immersion and then left to dry for 72 h at room temperature. The formed coats were assessed by electrochemical impedance spectroscopy, electrochemical noise, and potentiodynamic polarization curves after 24, 72, and 168 h of immersion in a 3.5 wt% NaCl solution. These electrochemical measurements were performed in triplicate to check reproducibility. The Tween 80 plasticizer-free cochineal-mucilage-based hybrid coating reveals more excellent protection against corrosion than uncoated aluminum. Optical micrographs were used to set apart the conditions of the hybrid coating after its valuation, which show the protection of the metallic surface on which it was not coated. Results showed that the hybrid coating has suitable properties as a barrier against corrosion due to its ability to block the aggressive species diffusion by trapping them in the coating structure, which prevents their contact with the metal surface. This property is attributable to better mucilage and cochineal film homogeneity. Also, it acts as a corrosion inhibitor due to its semi-permeable behavior where only water molecules flow through its pores. This hybrid coating showed an excellent corrosion-resistant behavior to be used to protect aluminum.

A report on the extinction of a population of Coleocephalocereus pluricostatus Buining & Brederoo due to ecological imbalance

Coleocephalocereus pluricostatus is an endemic cactus from eastern Brazil that occurs on naked gneiss/granite inselbergs in the Atlantic Forest biome. Here we report the extinction of a population of C. pluricostatus and the rapid decline of other populations of the species in eastern Minas Gerais, Brazil due to cochineal insects, diseases and mining operations. We offer some hypotheses to explain this dramatic decline and suggestions for monitoring the populations.

Natural Dyeing of Modified Cotton Fabric with Cochineal Dye.

Natural dyes are not harmful to the environment owing to their biodegradability. For dye application to textiles, salts are necessary as mordant or electrolytes and make an environmental impact. In this paper, the influence of cationization during mercerization to the dyeing of cotton fabric with natural dye from Dactylopius coccus was researched. For this purpose, bleached cotton fabric as well as fabric cationized with Rewin OS was pre-mordanted using iron(II) sulfate heptahydrate (FeSO4·7H2O) and dyed with natural cochineal dye with and without electrolyte addition. For the characterization of surface changes after cationization, an electrokinetic analysis on SurPASS was performed and compared to pre-mordanting. For determination of dye exhaustion, the analysis of dye solution was performed on a UV/VIS spectrophotometer Cary 50 Solascreen. Spectrophotometric analysis was performed using a Datacolor 850 spectrophotometer, measuring remission ”until tolerance” and the whiteness degree, color parameters, color depth (K/S), and colorfastness of dyed fabric were calculated. Levelness was determined by visual assessment. Cationized cotton fabrics showed better absorption and colorfastness. Pre-mordanting and cationization showed synergism. The electrolytes improved the process of dye absorption. However, when natural dyeing was performed on cotton fabric cationized during mercerization, similar chromacity, uniform color, and colorfastness were achieved with and without electrolyte, resulting in pure purple hue of cochineal. For achieving a violet hue, pre-mordanting with Fe-salt was needed. Therefore, salt can be reduced or even unnecessary, which makes this process of natural dyeing more environmentally friendly.