Allosteric strand displacement isothermal amplification for the visual detection of Toxoplasma gondii in 30 minutes.

Early detection of Toxoplasma gondii (T. gondii) is critical due to a lack of effective treatment for toxoplasmosis.This study established a simple, cost-effective, and rapid colorimetric detection method for T. gondii. The entire testing process, from sample collection to results, takes only 0.5 h. These characteristics fulfill the demands of researchers seeking rapid target detection with minimal equipment reliance. For genomic extraction, this study evaluated the ability of two filter papers to capture genomes. A rapid genomic extraction device combined with the two filter papers was designed to simplify the genomic extraction process, which was completed within 10 min and increased the detection sensitivity tenfold. The method utilized a simplified primer design for isothermal amplification, namely allosteric strand displacement (ASD), and employed an underutilized commercial color indicator, Bromothymol Blue (BTB), for signal output. Compared with other reported indicators, BTB exhibited a more pronounced color change, shifting from blue to yellow in positive samples, facilitating easier visual differentiation. The reaction was completed in 20 min with a limit of detection (LOD) as low as 0.014 T. gondii per microliter.

Development of real time-pH sensitive intelligent indicators for monitoring chicken breast freshness/spoilage using real packaging practices.

Real-real time CO2-sensitive freshness indicators, phenol red (PR) and bromothymol blue (BTB) dyes, in three-layer system using cellulose based binder was developed to determine the freshness/spoilage of chicken breast. The developed indicators were used to monitor chicken meat spoilage packaged in polyamide/polyethylene (PA/PE) pouches under air and 100% nitrogen (N2) at 4 °C for 10 days. Changes in the ΔE and ΔRGB values of the indicators, CO2/O2 gas composition of packs, and chemical (TVBN, pH, trimethylamine), microbial, and sensory quality parameters of chicken breast meat were analyzed. The visual color change in the PR-based indicator was insufficient for the consumer to detect the spoilage with the naked eye in both simulation and food trial. However, three stage color (dark blue-turquoise-green) change was occurred in BTB-based indicators, and the color transition in the spoilage level of CO2 (10-15% (v/v)) is supported by the physicochemical, microbiological and sensorial properties of the chicken breast. The shelf life of chicken breast under air was limited to 4 days, while the shelf life under 100% N2 was 6 days which are supported by the visual color change of BTB indicator. The BTB-based indicators were found promising on a real packaging conditions and could be adapted to industrial scale for monitoring real-time freshness/spoilage of poultry, ensuring food safety.

Novel pH-responsive indicator films based on bromothymol blue-anchored chitin for shrimp freshness monitoring.

The cellulose nanofibers (CNFs) based pH-sensitive indicator films were developed by mixing guar gum (GG) with bromothymol blue-anchored chitin (BTB-Chitin) as an indicator to monitor shrimp freshness. The BTB-Chitin was prepared by grafting hydroxypropyltriethylamine groups (HPTA) to chitin first, then anchoring bromothymol blue (BTB) to prepare intelligent pH response BTB-Chitin. The 0.08 BTB-Chitin films had a good tensile strength of 11.76 MPa and the water contact angle values were 125°, which displayed obvious color response to pH buffers and acid base volatile gas. Besides, the homogeneous and flexible composite films showed good color stability and reversibility. The released amount of BTB was very low from the BTB-Chitin films in heptane and corn oil. The composite films had been degraded completely in 15 days in soil. The pH and volatile base nitrogen were measured to determine the degree decay of shrimp (Litopenaeus vannamei), and the prepared pH-sensitive films changed from yellow (fresh) to cyan (spoiled) with the freshness of shrimp decreased, indicating the BTB-Chitin films could detect the shrimp freshness in real-time and high visibility.

Near-IR-Regulated Composite Hydrogel with Real-Time Infection Monitoring and a Combined Antibacterial Effect for Efficient Wound Management.

Chronic wounds induced by bacterial infection have seriously affected the health of people in the world. So, it is meaningful to develop a novel strategy with real-time infection monitoring and excellent antibacterial performance for enhancing wound management. Herein, we constructed a composite hydrogel by loading the pH indicator bromothymol blue (BTB) and gold nanocages containing 2,2'-azobis[2-(2-imidazolin-2-yl)propane] dihydrochloride (Au NCs@AIPH) into a polyacrylamide-co-poly(acrylic anhydride-modified oxidized sodium alginate) (PAM-co-PAOSA) hydrogel. In vitro and in vivo experimental results demonstrated that the composite hydrogel could effectively detect bacteria and diagnose the infection status of a wound in real time by showing visible color changes. In addition, the composite hydrogel containing Au NCs@AIPH possessed an excellent photothermal effect under near-IR (NIR) laser irradiation. The photothermal effect further activated AIPH to generate toxic free radicals to form combined antibacterial therapy for accelerating wound healing. Moreover, the composite hydrogel showed great biocompatibility. Therefore, the multifunctional hydrogel provided a novel wound management strategy for bacterial infection diagnosis and combined therapy in an infected wound.

An antibacterial and intelligent cellulose-based label self-assembled via electrovalent bonds for a multi-range sensing of food freshness.

Intelligent labels provide customers with food freshness information. However, the existing label response is limited and can only detect a single kind of food. Here, an intelligent cellulose-based label with highly antibacterial activity for a multi-range sensing freshness was developed to overcome the limitation. Cellulose fibers were modified using oxalic acid to graft -COO- followed by binding chitosan quaternary ammonium salt (CQAS), the remaining charges of which attached methylene red and bromothymol blue to form response fibers and to further self-assemble into the intelligent label. CQAS electrostatically gathered the dispersed fibers, resulting in an increase in TS and EB of 282 % and 16.2 %, respectively. After that, the rest positive charges fixed the anionic dyes to broaden pH response range of 3-9 effectively. More significantly, the intelligent label exhibited highly antimicrobial activity, killing 100 % of staphylococcus aureus. The rapid acid-base response revealed the potential for practical application in which the label color from green to orange represented the milk or spinach from fresh to close to spoiled, and from green to yellow, and to light green indicated the pork fresh, acceptable, and close to spoiled. This study paves a way for the preparation of intelligent labels in large-scale and promote the commercial application to improve food safety.

A collagen-based theranostic wound dressing with visual, long-lasting infection detection capability.

Continuous wound monitoring is one strategy to minimise infection severity and inform prompt variations in therapeutic care following infection diagnosis. However, integration of this functionality in therapeutic wound dressings is still challenging. We hypothesised that a theranostic dressing could be realised by integrating a collagen-based wound contact layer with previously demonstrated wound healing capability, and a halochromic dye, i.e. bromothymol blue (BTB), undergoing colour change following infection-associated pH changes (pH: 5-6 ➔ >7). Two different BTB integration strategies, i.e. electrospinning and drop-casting, were pursued to introduce long-lasting visual infection detection capability through retention of BTB within the dressing. Both systems had an average BTB loading efficiency of 99 wt% and displayed a colour change within 1 min of contact with simulated wound fluid. Drop-cast samples retained up to 85 wt% of BTB after 96 h in a near-infected wound environment, in contrast to the fibre-bearing prototypes, which released over 80 wt% of BTB over the same time period. An increase in collagen denaturation temperature (DSC) and red shifts (ATR-FTIR) suggest the formation of secondary interactions between the collagen-based hydrogel and the BTB, which are attributed to count for the long-lasting dye confinement and durable dressing colour change. Given the high L929 fibroblast viability in drop-cast sample extracts (92 %, 7 days), the presented multiscale design is simple, cell- and regulatory-friendly, and compliant with industrial scale-up. This design, therefore, offers a new platform for the development of theranostic dressings enabling accelerated wound healing and prompt infection diagnosis.

Recyclable waste Dry-cell batteries derived carbon dots (CDs) for detection of Two-fold metal ions and degradation of BTB dye.

In modern era, electronic wastes are one of the major threats around us, most of them are reused with less efficiency instead of re-usage, and conversion into valuable products is highly recommended. In this work, we report an innovative approach for the synthesis of highly photoluminescent CDs from waste dry-cell batteries through one-step hydrothermal treatment for the detection and degradation of environmental pollutants. The as-prepared CDs were studied by X-ray photoelectron spectroscopy (XPS), HR-TEM studies, X-ray diffractometer (XRD), Raman spectrometer, FTIR spectroscopy, UV-visible spectrophotometer, and spectrofluorometric measurements. The calculated quantum yield for synthesized CDs was around 13 %. The CDs have uniform particle size distribution, strong photoluminescent behavior, and possess high stability against various environmental conditions. Also, CDs display the selective and sensitive detection of Cr6+ and Co2+ and ions with a detection limit of around 0.11 µM and 0.10 µM respectively. The possible mechanism of CDs was also examined. Moreover, the photocatalytic activity of CDs with Bromothymol Blue (BTB) dye was studied. The degradation efficiency of BTB dye can be achieved at around 84 % over 180 min under the irradiation of direct sunlight in presence of H2O2. To date, it's the first time we have recycled waste dry-cell batteries into CDs as an effective probe for the detection and decomposition of environmental pollution. Furthermore, this work provides not only an easier route to make good quality and improved photoluminescent CDs from waste material like used batteries and also paves way for the reconversion of global treating waste. Finally, the outstanding detection capability with multiple properties of as-prepared CDs provides various environmental applications like the detection of pollutants and carcinogenic polluted water treatment.

Silk fibroin-based colorimetric microneedle patch for rapid detection of spoilage in packaged salmon samples.

Spoiled salmon can cause foodborne diseases and severely affects human health. Herein, we report a pH-responsive colorimetric microneedle (MN) patch fabricated from bromothymol blue (BTB) and silk fibroin meth acryloyl (SilMA) (BTB/SilMA@MN patch) for sensing salmon spoilage. The needle tips of MN could penetrate food cling film and insert into fish to extract tissue fluids directly and transport the extracted fluids to the backing layer for color displaying. The color change of BTB/SilMA@MN patches depended on the pH variation resulting from the increase of total volatile basic nitrogen in salmon during storage. The color of MN patches changed from yellow to yellowish green and to final green, indicating salmon changed from fresh to medium fresh and then to putrefied, respectively. Salmon spoilage can be rapidly determined via naked eye recognition and also analyzed on a smartphone in a nondestructive way, allowing consumers to estimate food quality easily and reliably.

Development of a calcium hydroxide-dye kimchi ripening indicator and its application in kimchi packaging.

Intelligent packaging that provides real-time information on food quality is useful for consumers. We developed a kimchi ripening indicator that can determine the ripeness of kimchi inside packaging and evaluated its applicability and limitations. The indicator was made using calcium hydroxide, which captures CO2, and four pH-sensitive dyes (cresol red, bromothymol blue, bromocresol purple, and methyl red). Fourier-transform infrared spectra of the prepared powders showed shapes similar to that of calcium hydroxide, and the dyes were evenly distributed on the calcium hydroxide surfaces. When the developed indicators were evaluated for kimchi packaging application, the indicator made from synthesized calcium hydroxide and bromothymol blue was the most reliable and clearly reflected useful kimchi ripening information. The indicator developed in this study is judged to be practically usable at temperatures of 4-15 °C. However, its usefulness is limited in that the seller cannot change the packaging capacity or kimchi capacity.

High Spatial Resolution Chemical Imaging of Implant-Associated Infections with X-ray Excited Luminescence Chemical Imaging Through Tissue.

Microbial infections associated with implantable medical devices are a major concern in fracture fixation failure. Early diagnosis of such infection will allow successful eradication with antibiotics without an extra cost for a second surgery. Herein, we describe XELCI as a technique with high X-ray resolution, implant specificity, and chemical sensitivity to noninvasively image chemical concentrations near the surface of implanted medical devices. The devices are coated with chemically reporting surfaces. This chemically responsive surface consists of two layers coated on an implantable medical device; a pH-sensitive layer (bromothymol blue or bromocresol green incorporated hydrogel) which is coated over a red-light emitting scintillator (Gd2O2S: Eu) layer for monitoring. A focused X-ray beam irradiates a spot on the implant, and the red light generated by the scintillator (with 620 nm and 700 nm peaks) is transmitted through the sensing layer which alters the spectral ratio depending on the pH. An image is generated by scanning the X-ray beam across the implant and measuring the spectral ratio of light passing through the tissue point-by-point. We used this imaging technique for monitoring implant-associated infections previously on the bone surface of the femur with a modified implantable plate sensor. Now we are studying pH changes that occur from tibial intramedullary rod infections. Two different types of intramedullary rod designs are used in pre-pilot rabbit studies, and we learned that the XELCI technique could be used to monitor any chemical changes that occur not only on the bone surface but also inside the bone. Thus, this enables noninvasive, high spatial resolution, low background local pH imaging to study implant-associated infection biochemistry.

Preparation of pH-indicator films based on soy protein isolate/bromothymol blue and methyl red for monitoring fresh-cut apple freshness.

Intelligent pH-indicator films based on soy protein isolate (SPI) were prepared using pH-sensitive dyes (bromothymol blue and methyl red). The addition of mixed indicators imparts pH-indicator films with an appreciable microstructure, acceptable water resistance, and favorable optical properties. The incorporation of the mixed indicators did not lead to significant improvement in the mechanical properties of films due to weak ionic cross-linking by hydrogen bonding between the SPI macromolecules and low-molecular-weight indicators. Fourier-transform infrared spectroscopy indicated hydrogen bond-mediated intermolecular interactions, and scanning electron microscopy showed that BB/MR were well dispersed in the SPI film. The indicator addition hindered the sorption and passage of water molecules. The water vapor permeability, moisture sorption, moisture content, and total soluble matter were 4.32 to 6.12 ×10-12  g·cm/cm2 ·s·Pa, 36.70% to 73.33%, 25.28% to 44.11%, and 8.21% to 25.56%, respectively. Also, the addition of indicators reduced UV light transmittance with minimal effect on the transparency of the film. The presence of indicators enhanced the pH sensitivity, seen as a visible color reaction at different pHs (total color difference, ΔE > 5). When the pH-indicator film containing 8 ml/100 ml final film emulsions was used to monitor the fresh-cut apple freshness, a green color for fresh status was observed, which turned blue after 60 h. Collectively, our findings suggested that indicator-containing SPI films have the potential for monitoring the freshness of fruits. PRACTICAL APPLICATION: pH-indicator films can help consumers to identify the freshness of packaged food by a change in the color of the packaging material, which is easily visible to the unaided eye without the need for opening the packaging. This protects consumers' interests.

Colorimetric ammonia (NH3) sensor based on an alginate-methylcellulose blend hydrogel and the potential opportunity for the development of a minced pork spoilage indicator.

Hydrogels based on alginate and methylcellulose were developed as a colorimetric indicator for monitoring minced pork spoilage. The hydrogel was fabricated by an external gelation method using Ca2+ as the crosslinking agent. The pH-sensitive dye bromothymol blue was incorporated into the hydrogel to act as an indicator. The hydrogel's swelling index increased with an increasing ratio of methylcellulose, suggesting that the water uptake capacity is tunable by the polymer composition. The hydrogel's compression strength is directly proportional to the alginate content. The hydrogel indicator demonstrated a color change from orange to yellow (day 6) upon detecting total volatile basic nitrogen (TVB-N) built up in the package during minced pork storage at 4 °C, and the results showed a positive correlation between the color change, TVB-N and pH change of minced pork. This result demonstrated the potential application of the hydrogel as a spoilage indicator in intelligent packaging.

Carbon isotope ratio of organic acids in sake and wine by solid-phase extraction combined with LC/IRMS.

We developed an analytical procedure for determining the δ13C values of organic acids in sake and wine using solid-phase extraction combined with liquid chromatography/isotope ratio mass spectrometry (LC/IRMS). First, the solid-phase extraction (SPE) procedure was performed and various tests were conducted to extract organic acids from alcoholic beverages using the simulated sake sample. Under the optimal SPE procedure, high recovery rates (96-118%) and good accuracies (≤ 0.7‰) were thus achieved for the simulated sake and wine samples. Next, we determined the δ13C of organic acid (tartaric acid, malic acid, lactic acid, succinic acid) in 9 sake and 11 wine samples. Finally, the δ13C values of lactic acid in nine sake samples suggested that lactic acid had been added during the brewing process. The high correlation between the δ13C values of tartaric acid and malic acid in 11 wine samples was consistent with their common source, grapes. This analytical method may help to identify when organic acids have been added to sake and wine and to elucidate the process of organic acid production therein. Graphical abstract.

Development of a Dye-Based Device to Assess Poultry Meat Spoilage. Part I: Building and Testing the Sensitive Array.

The rationale behind the material and dye selection and the investigation of the properties of a solid-phase sensor array designed for following chicken meat spoilage is presented, having in mind that the final target must be the naked eye identification of the degradation steps. The device is obtained by fixing five acid-base indicators, m-cresol purple (1), o-cresol red (2), bromothymol blue (3), thymol blue (4), and chlorophenol red (5), and a sensing molecule specific for thiols, 5,5'-dithiobis(2-nitrodibenzoic acid), called Ellman's reagent, (6) on a commercial cellulose-based support. The dimensions of the sensor and the amount of dye sorbed on the solid are carefully studied. The preparation protocol to get reproducible sensing materials is established, based on the kinetic study and the color change investigation. The material stability and the capacity of changing color, according to the acid-base properties of the dyes, are tested. The sources of uncertainty, coming from the technique employed for signal data acquisition and treatment and from the intrinsic variability of the spots based on the commercial support, are established. The highest variability does not come from photo acquisition by a mobile phone, the effect of the illumination equipment, the partial least-squares (PLS) model employed to assess the amount of dye sorbed into the solid but from the variability of different spots and was found equal to 10%. The uncertainty is adequate for final employment since it is referred to as replicates under different conditions that are definitively judged almost always identical by naked eye evaluation, which is our last target for assessing a change of the colors associated with spoilage.

Development of a Dye-Based Device to Assess the Poultry Meat Spoilage. Part II: Array on Act.

This work presents a colorimetric dye-based array for naked-eye detection of chicken meat spoilage. The array is obtained by fixing five acid-base indicators, m-cresol purple (1), o-cresol red (2), bromothymol blue (3), thymol blue (4), and chlorophenol red (5), and a sensing molecule specific for thiols, 5,5'-dithiobis(2-nitrodibenzoic acid), called Ellman's reagent (6), on a cellulose-based support. The dyes, being permanently charged, are fixed on the support via ion-exchange. The entire degradation process of beast poultry meat, at ambient temperature and in a domestic fridge, is followed by the change of the color of the array, placed in the headspace over the meat samples. The device is set after selection of the most suitable starting form, which could be the acidic or the basic color of indicators, being the proper dye concentration and the dimension of the spots already established. Basing on sensors colors, we identified three levels of the degradation process of chicken meat, named SAFE, WARNING, and HAZARD. By instrumental analysis, we demonstrated that sensors response was correlated to volatile organic compounds (VOCs) composition in the headspace and, thus, to meat spoilage progress. We demonstrated that biogenic amines (BAs), commonly considered a critical spoilage marker, are indeed produced into the samples but never present in the headspace, even in traces, during the investigated time-lapse. The VOC evolution nevertheless allows one to assign the sample as WARNING and further HAZARD. Some indicators turned out to be more informative than others, and the best candidates for a future industrial application resulted in a bromothymol blue (3)-, chlorophenol red (5)-, and Ellman's reagent (6)-based array.

Intelligent Hybrid Hydrogels for Rapid In Situ Detection and Photothermal Therapy of Bacterial Infection.

Diseases induced by bacterial infections increasingly threaten the health of people all over the world; thus, it is urgent and significant to early diagnose and effectively eliminate infections to save people's lives. To this end, we synthesized an intelligent hydrogel that integrated in situ visualized diagnosis and photothermal therapy of bacterial infections. By simply and subtly incorporating pH-sensitive bromothymol blue (BTB) and near-infrared (NIR)-absorbing conjugated polymer (termed as PTDBD) into thermosensitive chitosan (CS)-based hydrogel, the synthesized BTB/PTDBD/CS hydrogel can diagnose the acidic microenvironment of Staphylococcus aureus (S. aureus) biofilm and infected wounds by showing visualized color change. After rapid diagnosis, the hydrogel can immediately treat the infection site by local hyperthermia under irradiation of NIR laser (808 nm) and even the stubborn biofilm that is difficult to eradicate. Since the dominating antibacterial mechanism is hyperthermia, the hybrid hydrogel shows broad-spectrum antibacterial activity against Gram-positive, Gram-negative, and drug-resistant bacteria. In addition, it has low cytotoxicity to normal cells and no effect on the main organs of mice. It paves a brand new avenue to develop smart and facile diagnosis and a treatment platform for bacterial infections.

Simultaneous quantification of peroxidase and ascorbic acid in biosamples with an automatic system based on a Fe(iii)/methylthymol blue-carbon dot simulative enzyme.

Peroxidase (POD) and ascorbic acid (AsA) usually coexist in organisms to synergistically protect them from reactive oxygen damage, and their contents undergo dynamic changes under different physiological conditions. What's more, the response of POD-catalytic activity in spectrophotometry has to be corrected using the content of concomitant AsA because we found that there is an extinction reaction between AsA and chromogenic products obtained from POD catalysis. With these implications, by skilfully using the chromogenic and the extinction phenomena in the guaiacol/POD/H2O2 reaction, an automatic analysis system for simultaneous quantification of POD (73-440 U L-1) and AsA (4-60 mg L-1) was successfully established based on flow injection analysis (FIA). Furthermore, under acidic conditions (0.5 mol L-1 of HCl), hydrothermal synthesis (250 °C for 1 h) was used for synthesizing new carbon dots (sPOD-CDs) of methylthymol blue (0.08 g L-1)/FeCl3 (0.8 g L-1), which is a simulative enzyme for POD, and it was first used for catalyzing the guaiacol/H2O2 reaction within the FIA system to replace natural HRP in the extinction reaction. This sPOD-CD solution has no background absorption and its concentration shows excellent correlation with simulative POD-activity. Finally, after optimization, this FIA system was utilized to testify that the reducibility of AsA is due to ascorbate ions and to determine POD and AsA in some plant samples. The standard addition recovery experiment showed that there was no interference from the matrix in real samples (recoveries: 95%-105%), and the obtained POD and AsA results were also consistent with the reference experiments (relative deviation ≤ 2.80%, t-test ≥ 0.07). The proposed FIA system is characterized by high sample-throughput (40 samples per h), better repeatability (relative standard deviation ≤ 1.4%), etc.

Automated analytical microsystem for the spectrophotometric monitoring of titratable acidity in white, rosé and red wines.

The design, construction and evaluation of a low-cost cyclic olefin copolymer (COC)-based continuous flow microanalyzer with optical detection to determine the titratable acidity content of wine is here presented. The analysis method is based on the monitoring of the blue coloration decrease of a buffered bromothymol blue (BTB) solution in the presence of the acidic compounds of wine. The microanalyzer monolithically integrates the required microfluidic motifs as well as an optical flow cell where the measurements are performed by using a miniaturized and versatile photometric detection system. Fluid management is totally automated by the use of computer-controlled microvalves, permitting the automatic calibration of the system as well as the automatic sampling, including in-line dilution and analysis. The reduced size of the whole system along with its high simplicity and automation make it suitable for its application to the on-line monitoring of titratable acidity during wine-making processes. With the optimal conditions, a linear range up to 0.50 g L-1 tartaric acid, a quantification limit (LOQ) of 0.01 g L-1 and a detection limit (LOD) of 0.004 g L-1 were obtained, covering the most common acidity content of musts and wines. A sampling rate up to 26 h-1 could be achieved, consuming less than 3 mL of inexpensive reagents per analysis and requiring no pretreatment of the sample. The microsystem has been successfully applied to the quantification of the titratable acidity content of several wine samples, being the results in excellent agreement with the ones obtained by the reference method.

A semen cassia gum-based film with visual-olfactory function for indicating the freshness change of animal protein-rich food.

Bromothymol blue (BTB) was fixed on the cationic cellulose fibers (CCFs) to prepare pH sensitive fibers (pH-SFs). The pH-SFs as intelligent indicator were added into the semen cassiae gum (SCG) as a weakly acidic matrix to prepare a visual-olfactory film. The 13C NMR results show that the CCFs were successfully obtained by introducing hydroxypropyltriethylamine groups which showed strong affinity to BTB molecules. Rheology results demonstrated that all of the film-forming solutions were shear-thinned fluids with non-Newtonian behavior. Scanning electronic microscopy showed that the addition of pH-SFs makes the film surface rougher and rougher. The addition of pH-SFs < 3% improved the tensile strength of the film. The visual-olfactory film was sensitive to ammonia with a highly visible color change from pale yellow to blue-green. The SCG-3SFs film pre-treated in NaOH solution changed from blue to pale yellow while the raw milk was close to spoilage. The light yellow SCG-3SFs film changed to blue-green as the freshwater shrimp changed from fresh to spoilage. The results indicate that the visual-olfactory film can be used for perceiving the freshness of milk and freshwater shrimp.