Analysis of Fluorescent-Stained Lipid Droplets with 3D Reconstruction for Hepatic Steatosis Assessment.

Lipid droplets (LDs) are specialized organelles that mediate lipid storage and play a very important role in suppressing lipotoxicity and preventing dysfunction caused by free fatty acids (FAs). The liver, given its critical role in the body's fat metabolism, is persistently threatened by the intracellular accumulation of LDs in the form of both microvesicular and macrovesicular hepatic steatosis. The histologic characterization of LDs is typically based on lipid-soluble diazo dyes, such as Oil Red O (ORO) staining, but a number of disadvantages consistently hamper the use of this analysis with liver specimens. More recently, lipophilic fluorophores 493/503 have become popular for visualizing and locating LDs due to their rapid uptake and accumulation into the neutral lipid droplet core. Even though most applications are well-described in cell cultures, there is less evidence demonstrating the reliable use of lipophilic fluorophore probes as an LD imaging tool in tissue samples. Herein, we propose an optimized boron dipyrromethene (BODIPY) 493/503-based protocol for the evaluation of LDs in liver specimens from an animal model of high-fat diet (HFD)-induced hepatic steatosis. This protocol covers liver sample preparation, tissue sectioning, BODIPY 493/503 staining, image acquisition, and data analysis. We demonstrate an increased number, intensity, area ratio, and diameter of hepatic LDs upon HFD feeding. Using orthogonal projections and 3D reconstructions, it was possible to observe the full content of neutral lipids in the LD core, which appeared as nearly spherical droplets. Moreover, with the fluorophore BODIPY 493/503, we were able to distinguish microvesicles (1 µm < d ≤ 3 µm), intermediate vesicles (3 µm < d ≤ 9 µm), and macrovesicles (d > 9 µm), allowing the successful discrimination of microvesicular and macrovesicular steatosis. Overall, this BODIPY 493/503 fluorescence-based protocol is a reliable and simple tool for hepatic LD characterization and may represent a complementary approach to the classical histological protocols.

In vivo toxicity assessment of Remazol Gelb-GR (RG-GR) textile dye in zebrafish embryos/larvae (Danio rerio): Teratogenic effects, biochemical changes, immunohistochemical changes.

Dyes, which are very important for various industries, have very adverse effects on the aquatic environment and aquatic life. However, there are limited studies on the toxic properties of dyes on living things. This research elucidated the sublethal toxicity of acute exposure of the textile dye remazol gelb-GR (RG-GR) using zebrafish embryos and larvae for 96 h. The 96 h-LC50 for RG-GR in zebrafish embryos/larvae was determined to be 151.92 mg/L. Sublethal 96 hpf exposure was performed in RG-GR concentrations (0.5; 1.0; 10.0; 100.0 mg/L) to determine the development of toxicity in zebrafish embryos/larvae. RG-GR dye affected morphological development, and decreased heart rate, hatching, blood flow, and survival rates in zebrafish embryos/larvae. The immunopositivity of 8-hydroxy 2 deoxyguanosine (8-OHdG) in larvae exposed to RG-GR at high concentrations was found to be intense. Depending on the RG-GR dose increase, some biochemical parameters such as glutathione peroxidase (GSH) level, acetylcholinesterase (AChE) activity, catalase (CAT) activities, superoxide dismutase (SOD), and nuclear factor erythroid 2 (Nrf-2) levels were detected to be decreased in larvae, while malondialdehyde (MDA) content, nuclear factor kappa (NF-kB), tumor necrosis factor-α (TNF-α), DNA damage (8-OHdG level), interleukin-6 (IL-6) and apoptosis (Caspase-3) levels were found to be increased. The experimental results revealed that RG-GR dye has high acute toxicity on zebrafish embryo/larvae.

Optimization for minimizing the cost of ozonation of highly concentrated textile dyeing wastewater in a bubble column reactor.

Ozonation is one of the advanced oxidation methods that provide effective decolorization and detoxification of the dyeing wastewater without causing any sludge formation. Despite being a good alternative to biodegradation, ozonation suffers from a high operating cost. This study conducted the ozonation process at high initial dye concentrations and optimized the process parameters (such as initial ozone concentration, initial dye concentration, and pH) to minimize the operating cost in terms of the overall power consumption of the process. The ozonation of Reactive Blue dye was performed in a bubble column reactor at various process conditions. A central composite design (CCD)-based response surface method (RSM) statistical tool was used to optimize the process. An empirical correlation for the specific power consumption (defined as electricity consumed per unit mass of dye removed from a unit volume of dyeing wastewater) was developed and verified. It was found that the specific power consumption during ozonation can be lowered significantly (by ~25-30%) if the dyeing water was treated at high initial dye concentrations.

A techno-economic approach for eliminating dye pollutants from industrial effluent employing microalgae through microbial fuel cells: Barriers and perspectives.

Microbial fuel cells are biochemical factories which besides recycling wastewater are electricity generators, if their low power density can be scaled up. This also adds up to work on many factors responsible to increase the cost of running a microbial fuel cell. As a result, the first step is to use environment friendly dead organic algae biomass or even living algae cells in a microbial fuel cell, also referred to as microalgal microbial fuel cells. This can be a techno-economic aspect not only for treating textile wastewater but also an economical way of obtaining value added products and bioelectricity from microalgae. Besides treating wastewater, microalgae in its either form plays an essential role in treating dyes present in wastewater which essentially include azo dyes rich in synthetic ions and heavy metals. Microalgae require these metals as part of their metabolism and hence consume them throughout the integration process in a microbial fuel cell. In this review a detail plan is laid to discuss the treatment of industrial effluents (rich in toxic dyes) employing microbial fuel cells. Efforts have been made by researchers to treat dyes using microbial fuel cell alone or in combination with catalysts, nanomaterials and microalgae have also been included. This review therefore discusses impact of microbial fuel cells in treating wastewater rich in textile dyes its limitations and future aspects.

Emerging biotechnological potentials of DyP-type peroxidases in remediation of lignin wastes and phenolic pollutants: a global assessment (2007-2019).

Dye decolourizing peroxidase (DyP) is an emerging biocatalyst with enormous bioremediation and biotechnological potentials. This study examined the global trend of research related to DyP through a bibliometric analysis. The search term 'dye decolourizing peroxidase' or 'DyP-type peroxidase' was used to retrieve published articles between 2007 and 2019 from the Web of Science (WoS) and Scopus databases. A total of 62 articles were published within the period, with an annual growth rate of 17·6%. The highest research output was observed in 2015, which accounted for about 13% of the total output in 12 years. Germany published the highest number of articles (n = 10, 16·1%) with a total citation of 478. However, the lowest number of published articles among the top 10 countries was observed in India and Korea (n = 2, 3·2%). Research collaboration was low (collaboration index = 4·08). In addition to dye decolourizing peroxidase(s) and DyP-type peroxidase(s) (n = 33, 53·23%), the top authors keywords and research focus included lignin and lignin degradation (n = 10, 16·1 %). More so, peroxidase (n = 59, 95·2%), amino acid sequence (n = 27, 46·8%), lignin (n = 24, 38·7%) and metabolism (n = 23, 37·1%) were highly represented in keywords-plus. The most common conceptual framework from this study include characterization, lignin degradation and environmental proteomics. Apart from the inherent efficient dye-decolourizing properties, this study showed that DyP has emerging biotechnological potentials in lignin degradation and remediation of phenolic environmental pollutants, which at the moment are under explored globally.

Cost-effective in-situ remediation technologies for complete mineralization of dyes contaminated soils.

Dyes & dyes intermediates are one of the important and critically polluted industrial sectors, generating a large environmental liability. Therefore characterization of microbial community structure and diversity of contaminated soils helps to optimize the bioremediation strategies and performance. This study investigated the roles of environmental variables and contamination levels in shaping microbial community structure at an abandoned aged. In total, 20 bacterial and 4 fungal phyla were identified across soils with different physiochemical properties and concentration of the contaminants. Taxonomic analysis revealed the Proteobacteria, Firmicutes, Ascomycota and Basidiomycota represented the lineages and accounted for 49.5%-51.2%, 5.1%-15.3%, 2.5%-3.8% and 1.6%-2.1% of the total population, respectively. Microbial community (site 1) achieved decolorization rate of 972 ±â€¯1.21 mg dyes kgsoil-1d-1 after 15d and 692 ±â€¯1.4 mg kg-soil-1d-1 COD removal rate after 30 day of the experiment. Microbial community mineralized intermediates, 4-amino benzenesulfonate, 6-amino naphthalene sulfonate and benzene/naphthalene sulfonate in TCA cycle and stimulate the growth of selected soil bacteria, primary nutrient as well as plant growth of contaminated soil.

Optimization of Direct Blue-14 dye degradation by Bacillus fermus (Kx898362) an alkaliphilic plant endophyte and assessment of degraded metabolite toxicity.

Alkaliphilic bacteria possesses the ability to survive in the extreme conditions with high salt concentrations. The adaptability of alkaliphilic bacteria to extreme conditions has made them predominant degrader in the field of biodegradation. A moderately alkaliphilic endophyte was isolated from Centella asiatica with a potential to degrade a di-azo dye Direct Blue-14(DB-14). The isolate was identified as Bacillus fermus with 97% similarity strain Xmb064. On optimization, maximum of 92.76% biodegradation was attained with dye concentration at 68.78 ppm supplemented with 1 g of sucrose and 2.5% (v/v) of inoculum for 72 h incubation. Characterization of the biodegraded product carried out using UV-vis spectrophotometry, FT-IR and LC-MS confirmed the destabilization of di-azo bond followed with the degradation of DB-14. Cytogenotoxicity studies revealed the biodegraded products to be less toxic. The current study is the first report on the optimization, biotransformation and cytogenotoxicity of DB-14 by B. fermus strain Centella.

Influence of auxochrome group in disperse dyes bearing azo groups as chromophore center in the biotransformation and molecular docking prediction by reductase enzyme: Implications and assessment for environmental toxicity of xenobiotics.

Synthetic azo dyes have increasingly become a matter of great concern as a result of the genotoxic and mutagenic potential of the products derived from azo dye biotransformation. This work evaluates the manner in which reducing enzymes produced by Escherichia coli (E. coli) act on three disperse dyes bearing azo groups, namely Disperse Red 73 (DR 73), Disperse Red 78 (DR 78), and Disperse Red 167 (DR 167). UV-Vis spectrophotometry, high-performance liquid chromatography with diode array detector (HPLC-DAD), and liquid chromatography mass spectrometry (LC-MS/MS) were applied towards the identification of the main products. Seven days of incubation of the azo dyes with the tested enzymes yielded a completely bleached solution. 3-4-Aminophenyl-ethyl-amino-propanitrile was detected following the biotransformation of both DR 73 and DR 78. 4-Nitroaniline and 2-chloro-4-nitroaniline were detected upon the biotransformation of DR 73 and DR 78, respectively. The main products derived from the biotransformation of DR 167 were dimethyl 3,3'-3-acetamido-4-aminophenyl-azanedyl-dipropanoate and 2-chloro-4-nitroaniline. The results imply that DR 73 lost the CN- substituent during the biotransformation. Furthermore, theoretical calculations were also carried out aiming at evaluating the interaction and reactivity of these compounds with DNA. Taken together, the results indicate that DR 73, DR 78, and DR 167 pose health risks and serious threats to both human beings and the environment at large as their biotransformation produces harmful compounds such as amines, which have been widely condemned by the International Agency for Research on Cancer.

Photocatalytic mineralization and degradation kinetics of sulphamethoxazole and reactive red 194 over silver-zirconium co-doped titanium dioxide: Reaction mechanisms and phytotoxicity assessment.

The photodegradation and phytotoxicity of the pharmaceutical antibiotic, sulphamethoxazole (SMX) and the azo-dye reactive-red-194 (RR194) under visible-light irradiation of TiO2 nanoparticles modified by silver and zirconium was investigated. The results indicated that sulphamethoxazole and its toxic degradation by product, 3-amino-5-methylisoxazole and RR-194 could be degraded efficiently by the co-doped Zr/Ag-TiO2 catalyst. PL studies and ROS generation results suggested that the effective charge separation was carried out while irradiation of the modified TiO2 nanoparticles. Phytotoxicity tests demonstrated lower percentage of germination in P. vulgaris (40%), V. radiata (30%) and P. lunatus (30%) of the seeds treated with 50 ppm of SMX, compared to the seeds treated with the degradation products (100%). The results with 50 ppm of RR-194 also showed lower percentage of germination in P. vulgaris (40%), V. radiata (50%) and P. lunatus (30%) compared to the degradation products (100%). Furthermore, significant increase in root and shoot development was observed in the seeds treated with the degraded products when compared with SMX and RR-194. Overall, this study contributes to further understanding the photodegradation mechanisms, degradation products and environmental fate of SMX and RR-194 in water which helps in the evaluation and mitigation of the environmental risk of SMX and RR-194 for water reuse and crop irrigation.

Viability Assessment of Cryptosporidium parvum Oocysts by Vital Dyes: Dry Mounts Overestimate the Number of "Ghost" Oocysts.

Viability assessment of Cryptosporidium parvum oocysts is crucial for evaluation of the public health significance of this important zoonotic protozoon. Viability is commonly assessed in wet mounts after acid pretreatment and staining with fluorogenic vital dyes. However, in some studies, oocyst viability is evaluated in dry mounts after staining in suspension. Here, we evaluate the effect of acid pretreatment in nine replicate samples and compare the assessment of oocyst viability after evaluation in wet and dry mounts, respectively. Although acid pretreatment had no significant effect on the viability scores, data obtained by scoring oocysts in dry mounts resulted in ∼25% underestimation of the proportion of viable oocyst (82.5% ± 0.9% [wet mount +acid], 57.7% ± 2.3% [dry mount, ÷ acid], 76.0% ± 1.7% [wet mount, ÷ acid]), while the proportions of nonviable oocysts (DAPI+/PI+) were comparable for wet and dry mounts (9.7% ± 0.4% [wet mount +acid], 12.1 ± 1.5% [dry mount, ÷ acid], 15.5% ± 1.1% [wet mount, ÷ acid]).

Serine Protease Zymography: Low-Cost, Rapid, and Highly Sensitive RAMA Casein Zymography.

To detect serine protease activity by zymography, casein and CBB stain have been used as a substrate and a detection procedure, respectively. Casein zymography has been using substrate concentration at 1 mg/mL and employing conventional CBB stain. Although ordinary casein zymography provides reproducible results, it has several disadvantages including time-consuming and relative low sensitivity. Improved casein zymography, RAMA casein zymography, is rapid and highly sensitive. RAMA casein zymography completes the detection process within 1 h after incubation and increases the sensitivity at least by tenfold. In addition to serine protease, the method also detects metalloprotease 7 (MMP7, Matrilysin) with high sensitivity.

Assaying Cellular Viability Using the Neutral Red Uptake Assay.

The neutral red uptake assay is a cell viability assay that allows in vitro quantification of xenobiotic-induced cytotoxicity. The assay relies on the ability of living cells to incorporate and bind neutral red, a weak cationic dye, in lysosomes. As such, cytotoxicity is expressed as a concentration-dependent reduction of the uptake of neutral red after exposure to the xenobiotic under investigation. The neutral red uptake assay is mainly used for hazard assessment in in vitro toxicology applications. This method has also been introduced in regulatory recommendations as part of 3T3-NRU-phototoxicity-assay, which was regulatory accepted in all EU member states in 2000 and in the OECD member states in 2004 as a test guideline (TG 432). The present protocol describes the neutral red uptake assay using the human hepatoma cell line HepG2, which is often employed as an alternative in vitro model for human hepatocytes. As an example, the cytotoxicity of acetaminophen and acetyl salicylic acid is assessed.

Clinical application of a rapid cervical cancer screening method: Folate receptor-mediated staining of cervical neoplastic epithelia.

AIM: A newly developed cervical cancer screening method - folate receptor-mediated epithelium staining utilizes methylene blue internalized by folate receptor-mediated endocytosis to stain cervical intra-epithelial neoplasia. We test the clinical feasibility of this method in this study. METHODS: A total of 14 344 women who were at least 21 years old and had been sexually active, participated in the study. In gynecological clinics, participants underwent cervical cancer screening with folate receptor-mediated epithelium staining followed by cytology sampling. The color change of methylene blue in the cervical neoplastic epithelium can then be detected by the cotton swabs placed inside the cervix. A change of color to blue, bluish black or black is positive, whereas a change of color to green or no color change indicates negative result. Three hundred and twenty-three women who were positive with either or both tests had histopathologic diagnosis. RESULTS: The sensitivity, specificity, positive predictive value and negative predictive value of folate receptor-mediated epithelium staining for cervical intra-epithelial neoplasia grade 2 and worse was 85.7%, 76.4%, 61.3% and 92.5%, respectively. Folate receptor-mediated epithelium staining had moderate agreement with cytology thresholded at atypical squamous cells, unable to exclude a high grade intra-epithelial lesion and was more sensitive that the later (85.7% vs 74.5% for intra-epithelial neoplasia grade 2 and worse; 89.2% vs 75.4% for intra-epithelial neoplasia grade 3 and worse). CONCLUSION: Folate receptor-mediated epithelium staining is capable of detecting cervical precancerous and cancerous lesions rapidly and cost-effectively.

Remediation of textile azo dye acid red 114 by hairy roots of Ipomoea carnea Jacq. and assessment of degraded dye toxicity with human keratinocyte cell line.

Bioremediation has proven to be the most desirable and cost effective method to counter textile dye pollution. Hairy roots (HRs) of Ipomoea carnea J. were tested for decolourization of 25 textile azo dyes, out of which >90% decolourization was observed in 15 dyes. A diazo dye, Acid Red 114 was decolourized to >98% and hence, was chosen as the model dye. A significant increase in the activities of oxidoreductive enzymes was observed during decolourization of AR114. The phytodegradation of AR114 was confirmed by HPLC, UV-vis and FTIR spectroscopy. The possible metabolites were identified by GCMS as 4- aminobenzene sulfonic acid 2-methylaniline and 4- aminophenyl 4-ethyl benzene sulfonate and a probable pathway for the biodegradation of AR114 has been proposed. The nontoxic nature of the metabolites and toxicity of AR114 was confirmed by cytotoxicity tests on human keratinocyte cell line (HaCaT). When HaCaT cells were treated separately with 150 µg mL(-1) of AR114 and metabolites, MTT assay showed 50% and ≈100% viability respectively. Furthermore, flow cytometry data showed that, as compared to control, the cells in G2-M and death phase increased by 2.4 and 3.6 folds respectively on treatment with AR114 but remained unaltered in cells treated with metabolites.

Environmental assessment of the degradation potential of mushroom fruit bodies of Pleurotus ostreatus (Jacq.: Fr.) P. Kumm. towards synthetic azo dyes and contaminating effluents collected from textile industries in Karnataka, India.

Pleurotus ostreatus (Jacq.: Fr.) P. Kumm. is one of the edible mushrooms currently gaining attention as environmental restorer. The present study explores the potential of P. ostreatus (Jacq.: Fr.) P. Kumm. in degradation of textile dyes and effluents. The mushroom cultivation was carried out using paddy bed as substrate. The fully grown mushroom fruit bodies were used as a bioremediation agent against two industrially important azo dyes such as nylon blue and cotton yellow and few effluents collected from various textile industries in Karnataka, India. The ideal growth parameters such as temperature, pH, and dye concentrations for effective degradation were carried out. One of the main enzymes, laccase, responsible for biodegradation, was partially characterized. The degradation was found to be ideal at pH 3.0 and temperature at 26-28 °C. This study demonstrated a percentage degradation of 78.10, 90.81, 82.5, and 64.88 for dye samples such as nylon blue (50 ppm), cotton yellow (350 ppm), KSIC effluents, and Ramanagar effluents at 28 °C within 15th days respectively in comparison with other temperature conditions. Similarly, a percentage degradation of 35.99, 33.33, 76.13 and 25.8 for nylon blue (50 ppm), cotton yellow (350 ppm), Karnataka Silk Industries Corporation (KSIC) effluents and Ramnagar effluents were observed at pH 3.0 within 15 days, respectively (p < 0.05). Thus, the current study concluded that the utilization of P. ostreatus (Jacq.: Fr.) P. Kumm. at ideal environmental conditions is a cost-effective and eco-friendly approach for the degradation of various azo dyes and textile effluents which are harmful to the ecosystem.

Assessment of cervical passage of vital dyes in pregnant, nonpregnant, and mated rats and mice.

Risk assessment for indirect exposure to small molecule pharmaceuticals in semen to the conceptus has traditionally been handled by calculations based on assumptions that any embryo-fetal exposure would be secondary to maternal absorption and redistribution. This study was designed to assess the potential for transcervical passage of drugs from semen. Reproductive tracts of rodents were examined following vaginal dosing with vital dyes during the estrous cycle, mating, and pregnancy. Toluidine Blue was not observed beyond the cervix after vaginal administration in pregnant rats; additionally, it did not pass the cervix in rats during any phase of estrous. In order to address the effects of semen, rats were dosed at receptivity and mated. Vital dyes were not visually evident in the uterus despite vaginal and sperm plug staining. This study provides evidence that direct transcervical passage is not a substantial route of direct embryo-fetal exposure for small molecule drugs in semen.

A low-cost wheat bran medium for biodegradation of the benzidine-based carcinogenic dye Trypan Blue using a microbial consortium.

Environmental release of benzidine-based dyes is a matter of health concern. Here, a microbial consortium was enriched from textile dye contaminated soils and investigated for biodegradation of the carcinogenic benzidine-based dye Trypan Blue using wheat bran (WB) as growth medium. The PCR-DGGE analysis of enriched microbial consortium revealed the presence of 15 different bacteria. Decolorization studies suggested that the microbial consortium has high metabolic activity towards Trypan Blue as complete removal of 50 mg∙L-1 dye was observed within 24 h at 30 ± 0.2 °C and pH 7. Significant reduction in TOC (64%) and COD (88%) of dye decolorized broths confirmed mineralization. Induction in azoreductase (500%), NADH-DCIP reductase (264%) and laccase (275%) proved enzymatic decolorization of dye. HPLC analysis of dye decolorized products showed the formation of six metabolites while the FTIR spectrum indicated removal of diazo bonds at 1612.30 and 1581.34 cm-1. The proposed dye degradation pathway based on GC-MS and enzyme analysis suggested the formation of two low molecular weight intermediates. Phytotoxicity and acute toxicity studies revealed the less toxic nature of the dye degradation products. These results provide experimental evidence for the utilization of agricultural waste as a novel low-cost growth medium for biodegradation of benzidine-based dyes, and suggested the potential of the microbial consortium in detoxification.

A high throughput colorimetric assay of ß-1,3-D-glucans by Congo red dye.

Mushroom strains contain complex nutritional biomolecules with a wide spectrum of therapeutic and prophylactic properties. Among these compounds, ß-d-glucans play an important role in immuno-modulating and anti-tumor activities. The present work involves a novel colorimetric assay method for ß-1,3-d-glucans with a triple helix tertiary structure by using Congo red. The specific interaction that occurs between Congo red and ß-1,3-d-glucan was detected by bathochromic shift from 488 to 516 nm (>20 nm) in UV-Vis spectrophotometer. A micro- and high throughput method based on a 96-well microtiter plate was devised which presents several advantages over the published methods since it requires only 1.51 µg of polysaccharides in samples, greater sensitivity, speed, assay of many samples and very cheap. ß-D-Glucans of several mushrooms (i.e., Coriolus versicolor, Ganoderma lucidum, Pleurotus ostreatus, Ganoderma carnosum, Hericium erinaceus, Lentinula edodes, Inonotus obliquus, Auricularia auricular, Polyporus umbellatus, Cordyseps sinensis, Agaricus blazei, Poria cocos) were isolated by using a sequence of several extractions with cold and boiling water, acidic and alkaline conditions and quantified by this microtiter plate method. FTIR spectroscopy was used to study the structural features of ß-1,3-D-glucans in these mushroom samples as well as the specific interaction of these polysaccharides with Congo red. The effect of NaOH on triple helix conformation of ß-1,3-D-glucans was investigated in several mushroom species.

Rapid and sensitive detection of human astrovirus in water samples by loop-mediated isothermal amplification with hydroxynaphthol blue dye.

BACKGROUND: The aim of this paper was to develop a reverse transcription loop-mediated isothermal amplification (RT-LAMP) method for rapid, sensitive and inexpensive detection of astrovirus. RESULTS: The detection limit of LAMP using in vitro RNA transcripts was 3.6 × 10 copies·µL⁻¹, which is as sensitive as the presently used PCR assays. However, the LAMP products could be identified as different colors with the naked eye following staining with hydroxynaphthol blue dye (HNB). No cross-reactivity with other gastroenteric viruses (rotavirus and norovirus) was observed, indicating the relatively high specificity of LAMP. The RT-LAMP method with HNB was used to effectively detect astrovirus in reclaimed water samples. CONCLUSIONS: The LAMP technique described in this study is a cheap, sensitive, specific and rapid method for the detection of astrovirus. The RT-LAMP method can be simply applied for the specific detection of astrovirus and has the potential to be utilized in the field as a screening test.