Detection of Pesticides in Foods Using Chemiluminescence

Detection of Pesticides in Foods Using Chemiluminescence Sensitive And Selective Detection of Pesticides in Foods Using Chemiluminescence Zhou Yan Introduction Pesticides are widely used in agriculture to protect seeds and crops before and after harvesting. They have contributed significant economic benefits to society. At the same time, widespread use of pesticides has created serious problems regarding their effect on the environment and human health.[1]. Even low levels of contaminants can cause adverse effects on humans, animals, plants, and ecosystems. The application of pesticides has increased appreciably during the past few years, resulting in a potential risk for the human health.[2] .During recent decades, the number of publications on pesticide residue analysis has increased significantly, and important advances have been made in the development of methods for a wide variety of sample types[3]. N-methylcarbamates (NMCs) are commonly employed as pesticides, the structures show in figure 1. Their main characteristics of NMCs are low stability, high toxicity, and non-accumulation in organic tissues. Some of the NMCs, including carbaryl, carbofuran and methiocarb, are considered in the final list of compounds to be considered for periodic re-evaluations by the Joint FAO/WHO Meeting on Pesticide Residues (JMPR)[4]. Fig.1 Stucture of the selected N-methylcarbamates Analysis of pesticides in food, environmental, forensic and clinical samples is a difficult task because of the low concentration values and matrix complexity of the target compounds. Gas chromatography (GC) has been widely used for pesticide analysis in food, clinical and environmental samples due to its high selectivity and sensitivity [5]. However, some analytes are not suitable for GC separation because of their low volatility, thermal lability or high polarity. Phenylurea, carbamate, benzimididazole some pyrethroid,, organophosphorus, and quaternary ammonium derivatives are examples of such pesticides[6] . Chemiluminescence is defined as the emission of light as the result of a chemical reaction[7]. Chemiluminescence (CL) is one of the highest sensitive analytical techniques for the detection of pesticides, due to the high dilution of these substances in environmental samples and compounds in a great variety of matrices depending on their participation in a CL reaction as catalysts, precursors, oxidation, inhibitors and so on. Three Sensitive And Selective Methods Detecting Pesticides in Foods Using Chemiluminescence Determination of pesticides in water and vegetables by HPLC with post-column chemiluminescence using the luminol reaction .[8]. Chemiluminescence (CL) has been revealed as an excellent tool for the detection of pesticides.[9] .The best known example is the luminol (5-aminophthalylhydrazide), The Proposed mechanism of the luminol CL reaction shows in Fig 2, when (5-aminophthalylhydrazide) is oxidated in alkaline medium , it produces the excited 3-aminophthalate anion, which emits light when it is relaxed to the ground state using several oxidants such hexacyanoferrate (III), permanganat and hydrogen peroxide and so on [10]. The reaction presents a powerful application as detection system in flow injection analysis (FIA) or in liquid chromatography (HPLC), where luminol-type compounds can be used as a derivatization reagents allowing the analytes to be detected at very low levels. Fig. 2 Proposed mechanism for the luminol CL reaction. A scheme of the HPLC–UV–CL system is shown in Fig. 3. The reversed phase separation was carried out in a C18 column. A peristaltic pump was used to deliver luminol and potassium permanganate . After separation and UV/vis detection at wavelength of 220 nm, with the stream of potassium permanganate and luminol solutions. A mixing reactor (d1) linked both â€Å"T† connectors. Another mixing reactor (d2 ), was used to link the outlet of the inlet of the detection cell and the second connector. The resulting stream was passed to the detection cell and the CL emission from oxidation of luminol was recorded as the background blank signal (baseline). The quantification of carbamates was based on the increase in the CL intensity , calculated as A=[I/IIS], where A is the analytical signal, I is the net CL signal due to the presence of the NMCs in the luminol–KMnO4 system, IIS is the CL signal. due to the internal standard. Ethiofencarb was choosed to be the IS ,due to it is well differentiated from the rest of analytes in the selected chromatographic conditions and also it is a NMC which produced an enhancement of the CL emission from the luminol oxidation by potassium permanganate in alkaline medium. Fig, 3 Scheme of the HPLC-UV-CL manifold, d1, d2: mixing reactors Recovery experiments were carried out in vegetables to check the applicability of the proposed methodology ,to make sure if it can use in the detection of NMCs in the real sample. No NMCs were detected when a sample blank was analyzed using the proposed method. Three replicates were prepared at each concentration level, and each one was injected by duplicate. obtaining recoveries in the range of 107.7–88.6%. Chromatograms of cucumber sample are presented in Fig. 4. Fig.4 Chromatogram of a cucumber sample applying the proposed HPLC–CL method: (A) Blank sample; (B) sample spiked with 50ug kg−1carbofuran and methiocarb and 5ug kg−1 carbaryl. Peaks: 1, carbofuran; 2, carbaryl; 3, methiocarb; IS, internal standard. A sensitive and selective HPLC–CL method was used for the determination of NMCs in vegetables. Compared with other sensitive methodologies which use CL or fluorescence detection, this method is very simple because no derivatization process is needed. the great enhancement produced by these compounds on the CL emission of luminol using permanganate as oxidant makes the resolution of three carbamates (carbofuran, carbaryl, and methiocarb) in less than 14 min possible. Chemiluminescence Switching on Peroxidase Like Fe3O4 Nanoparticles for Selective Determination of Various Pesticides.[11] Due to the classical CL systems have very low efficiency which can not transform the chemical energy into light efficiently, so the enhancement of their CL efficiency to give intense emission intensity is one of the most important factors in quantitative analysis. From now on , a number of approaches have been explored for the enhancement of the efficiency. Such as developing intrinsically selective CL switching at the surface of Fe3O4 nanoparticles ,which make a great contribution to the sensitive detection and simultaneous determination of various pesticides. Fe3O4 nanoparticles with high peroxidase-like catalytic activity can catalyze the decomposition of dissolved oxygen to generate superoxide anions at their surface. The surface superoxide anions have high chances to be scavenged in the presence of ethanol. However, the scavenging effect can be effectively inhibited through the specific binding of target molecules on Fe3O4 nanoparticles. As shown in figure 5, water has no effect on the CL change but ethanol can lead to a significant CL quenching of the luminol−Fe3O4 system. EP is one of the widely used organophosphorus pesticides . there is no CL response of the EP aqueous solution in the luminol−Fe3O4 system by using pesticide ethoprophos (EP) as the target analyte. If we add EP ethanol solution ,which resulting a significant CL enhancement . we can abserved clearly. Fig 5 Pesticide Ethoprophos (EP) and the Mediums Produce the CL Response on Luminol (LUM)-Based CL Systems As shown in Figure 6A, With increasing the concentration of EP solution to the luminol−Fe3O4−ethanol system, the CL will significantly and continuously enhanced. Such high enhancing response has proved that EP molecules can inhibit the scavenging of superoxide anions efficiently. But not all organophosphorus have this function. As shown in Figure 6B, only EP and PF organophosphorus esters (containing P=O bond) with a phosphorus−sulfur bond (P−S bond) are able to light up the CL systerm in luminol−Fe3O4−ethanol solution. Fig 6 CL switching-on chemosensor for sensitively and selectivel (A) CL enhancement of the luminol−Fe3O4 ethanol system with increasing the concentration of EP . (B) The CL response selectivity to arious pesticides (0.1 mM): ethoprophos (EP), profenofos (PF), dylox (DL), and 2,4-dichlorophenoxyacetic acid (2,4-D). In order to make sense of the mechanism of CL switching, we choose the FeCl2 as source materials to study the interaction between EP molecules and Fe2+ ions. For the purpose of having better understanding of the coordinative groups, we do the theoretical energy calculations, as shown in Fig 7. Due to the important principle of energy minimum, the conformation was optimized as the coordination of sulfur atom instead of the oxygen atom. This result is reasonable because sulfur is more polarizable than oxygen and naturaly has a stronger coordinative bond formation. These calculations and discussions imply that EP molecules will have the strongest coordinative with Fe2+ ions, among the four different pesticides. Therefore, it is reasonable to suppose that the CL enhancement is attributed to the coordination of EP molecules on surface Fe2+ ions that inhibits the scavenging of superoxide anions. Fig 7 Optimized Conformations of the Complexes of (EP)2 -FeCl2 Based on the Theoretical Calculation of the Binding Energies (ΔE) for Different Coordination Formats The mechanism of CL switching is illustrated in Figure 8. Firstly decomposition of dissolved oxygen by the intrinsic peroxidase-like catalytic activity of Fe3O4 nanoparticles, we can get the superoxide anions, and due to the high surface-to-volume ratio of the nanoparticles, the resultant superoxide anions will absorb at the surface of the Fe3O4 nanoparticles (1).Then, ethanol reacts with superoxide anions through the radical scavenging process (2) there is no light emission after the addition of luminol (3) EP molecules are first added into the Fe3O4 colloidal solution before adding ethanol, (4). This molecular structure can effectively inhibit the scavenging of superoxide anions from ethanol (5) even if in the presence of ethanol, the addition of luminol still produces a strong light emission (6). The two CL response formats are combined to form a CL â€Å"switching-on† chemosensor (7). Fig 8 Mechanism of CL switching at the surface of Fe3O4 nanoparticles: The CL â€Å"turn-on† chemosensor was used to sensitively detect nonredox molecules specificitly. and has a a lot of application in lab experiment, because of it has the ability to detect molecules of interest in complexed and real samples after a simple magnetic separation.. The very simple and versatile strategy should open a new window of the development of CL chemosensors. And also make a great development in the application of magnetic nanoparticles (à ¤Ã‚ ¸Ã¢â‚¬ °) Analytical performances of validated chemiluminescent enzyme immunoassays to detect N-methylcarbamate pesticidesà £Ã¢â€š ¬Ã‚ 12à £Ã¢â€š ¬Ã¢â‚¬Ëœ Enzyme-linked immunosorbent assays (ELISAs) with chemiluminescent detection for the determination of carbaryl carbofuran and methiocarb were developed recently and the analytical parameters of these assays were compared with those of ELISAs with colorimetric detection. By using the chemiluminescent reagents for the detection of lower concentrations of horseradish peroxidase which can decrease the optimal antibody and conjugate concentrations and to reach better analytical parameters. Both in terms of accuracy and precision, results obtained by ELISAs correlated well. with those obtained by a liquid chromatography–electrospray mass spectrometry (LC/ESI/MS/MS) analysis, and also used as reference method to validate the immunoassays results. In order to fit into the linear part of the calibration curve and subsequently analysed by ELISA, each primary test sample was subjected to three independent dilutions .The dilution factors applied for the Nmethylcarbamate analysis are shown in Fig 9. Fig 9. Concentrations of immunoreagents and baby food sample dilutions applied in the ELISAs The reported results show that the chemiluminescent detection make a great improvement for the analytical performances of carbamates ELISAs. What’s more, the chemiluminescent ones with lower detection limits, can analyse various kinds of non-fatty samples simply and easily, just diluting them. This fact could avoid the extraction procedures and time consuming pre-treatment . Since the problems related with possible matrix effects can be easily overcome by using highly diluted samples. The time required to prepare the samples can be greatly reduced and their number increased significantly in each analytical session, as well as the reagents cost per assay is reduced. Conclusion : This review mainly introduce three Sensitive and Selective Methods Detecting Pesticides in Foods Using Chemiluminescence, each has advantages and disadvantages.(as shown in the following table). We can choose the appropriate method according to different situations. References: [1] Varsamis, D. C; Touloupakis, E.; Morlacchi, P.; Ghanotakis, D. C.; Giardi, M. T.;  Cullen, D. C. Talanta 2008, 77, 42–47. [2] Martà ­nez, E. J.; Barrales, P.; Cà ³rdova, M. L.; Medina, A. Anal Chimica Acta 2011, 684, 30–39. [3] Hua, L.; Mei, G.; Ping, H.; Yong, Z.; Long, L.; Hua, J.; Yong , M. Anal. Chem. 2012, 84, 255−261. [4] Alinorm 01/24A Appendix IX. Priority list of compounds scheduled for evaluation or reevaluation by JMPR. 2001 Joint FAO/WHO Meeting on Pesticide Residues (JMPR). [5] Xian , G.; Tao, C.; Guang, Y. Chromatography A. 2005, 1100, 160–167. [6] Lesueur, C.; Knittl, P.; Gartner, M.; Mentler, A.; Fuerhacker, M.; Food Control. 2008, 19 906–914. [7] Gracia, A. M.; CampaËÅ" na, J. J.; Chinchilla, J. F.; Pà ©rez, A.; Casado. Trends. Anal. Chem. 2005, 24, 927. [8] Fernando, J.; Pà ©rez, H.; Marà ­a, A.; Campa, N. Anal Chimica Acta.2008, 630,194–200 [9] Lopez, J. L.; Catala-Icardo, M. Anal. lett .2011,44,146-175. [10]Laura, G..G.; Ana, M.; GarcÄ ±Ã‚ ´a, C.; Jorge, J. ;Soto, C.;, Jose, F.; Huertas, F.; Gonza, L. C. Anal Chem,2005 ,24, 11, [11] Guan, G. J.; Yang, L.; Mei, Q. S.; Zhang, K.; Zhang, Z. P.; Han, M. Y. Anal. Chem. 2012, 84, 9492−9497. [12] Barbora, M.; Tomas, K.; Pavel, R.; Mar ´Ãƒâ€žÃ‚ ±a, J. M.; Antonio, A.; Angel, M.; Elida, F.; Fabiana, F.; Stefano, G.. Anal .Chimica. Acta .2005,528,243–248.

Ecology Lab Report: Creating an Ecosystem in a Bottle Essay -- essays

Objective: To create an ecosystem contained in a bottle with both terrestrial and aquatic environments sealed to the outside world. Hypothesis/Predictions: I predict the fish will last for 3 days. I predict the crickets will last for a week. Materials: 1. fish 2. Water 3. 2 plants 4. Soil 5. Rag 6. Graphite 7. 2 Liter Bottle (2) 8. Tape 9. 2 Snails 10. 3 crickets Procedure: Cut top off 2 liter bottle and bottom off another. Fill the one without a top with water and put a plant, 1 fish, and 2 snails in water. Put a rag through the top of other bottle and pile with graphite (rocks). Put dirt, plants, and anything else that is required or put in optionally in container. Data: Water Level 1- 10 Â ½ cm Plant height 1- 9 Â ½ cm 2- 10 Â ½ cm 2- 9 Â ½ cm 3- 10 Â ¼ cm 3- 9 Â ¾ cm 4- 10 cm 4- 10 cm 5- 10 cm 5- 10 Â ¼ cm 6- 10 cm 6- 10 cm 7- 10 cm 7- 10 cm 8- 10 cm 8- 10 cm 9- 98 mm 9- 10 cm Logs Day 1 5/22/07 The water has turned into a very light and transparent yellow. Everything in tank seems in order. Crickets and fish seem lively. Not much change. Day 2 5/25/07 The water is a medium shade of yellow. The fish and crickets s... ... yes the ecology project of course! Through the exciting endeavors of my experimentation I have discovered over time that my environment has changed dramatically! The mighty creatures of before have fallen to their graves. Only 50% of all life from the beginning of my project still lives. During the experiments I have discovered that both my early predictions came true. The crickets did live for a week and my fish lived for more then 3 days. My predictions came as true as Edward Cayce?s. During this experiment I learned that a tiny environment within a 2 liter bottle was possible and it could very easy be capable of life. It seems in the end this experiment has made me wiser and stronger in a sense and gave me strength to keep on marching toward a bright future in which the old days shall return and everyone shall be just as it once was many years ago?..

Fluke, or, I Know Why the Winged Whale Sings Chapter 35~36

CHAPTER THIRTY-FIVE Yeah, but You Can't Dance to It The Colonel was standing in the middle of the mother-of-pearl amphitheater when the whaley boys led Nate in. â€Å"You two go on now,† the Colonel said to the whaley boys. â€Å"Nate can find his way back.† â€Å"You came out of your lair,† Nate said. The Colonel looked older, more drawn than when Nate had seen him before. â€Å"I don't want to be in contact with the Goo for what I'm going to tell you.† â€Å"I thought it didn't get information that way,† Nate said. The Colonel ignored him. â€Å"I was hoping you would have had a brainstorm to solve my problem, Nate, but you haven't, have you?† â€Å"I'm working on it. It's more complex –  » â€Å"You've been distracted. I'm disappointed, but I understand. She's a piece of work, isn't she? And I mean that in the best sense of the word. Never forget that I chose to send her to you.† Nate wondered how much the Colonel knew about them and how he knew it. Reports from the whaley boys? From the Goo itself, through osmosis or some extended nervous system? â€Å"Distraction has nothing to do with it. I've thought a lot about your problem, and I'm not sure I agree with you. What makes you think the Goo is going to destroy humanity?† â€Å"It's a matter of time. That's all. I need you to carry a message for me, Nate. You'll be responsible for saving the human race. That should go some measure toward consoling you.† â€Å"Colonel, is there any chance you can be more direct, less cryptic, and tell me for once what the hell you're talking about?† â€Å"I want you to go to the U.S. Navy. They need to know about the threat of the Goo. One well-placed nuclear torpedo should do it. It's deep enough that they shouldn't have any problem justifying it to other countries. There won't be any fallout. They're just going to need someone credible to convince them of the threat. You.† â€Å"What about the people down here? I thought you wanted to save them.† â€Å"I'm afraid they're going to be a necessary sacrifice, Nate. What are five thousand or so people, most of whom have lived longer than they would have on the surface, compared with the whole human race, six billion?† â€Å"You crazy bastard! I'm not going to try to convince the navy to nuke five thousand people and all the whaley boys as well. And you're more deluded than I thought if you think they'd do it on my word.† â€Å"Oh, I don't expect that. I expect they'll send down their own research team to confirm what you tell them, but when they get here, I'll see to it that they get the message that the Goo is a threat. In any case you'll survive.† â€Å"I think you're wrong about the Goo finding us dangerous. And even if you were right, what if it just decides to wait us out? On the Goo's time scale, it can just take a nap until we're extinct. I'm not doing it.† â€Å"I'm sorry you feel that way, Nate. I guess I'll have to find another way.† Nate suddenly realized that he'd blown it – his chance to escape. Once he was outside Gooville, there would have been nothing to force him to do what the Colonel wanted. Or maybe there would be. Right then he wanted very badly to see Amy. â€Å"Look, Colonel, maybe I can do something. Couldn't you just evacuate Gooville? Drop all the people on an island. Let the whaley boys find somewhere else to live. I mean, if I reveal the Goo to the world, it's all sort of going to be out of the bag anyway. I mean –  » â€Å"I'm sorry, Nate, I don't believe you. I'll take care of it. Evacuation wouldn't make any difference to the people here anyway. And the whaley boys shouldn't exist in the first place. They're an abomination.† â€Å"An abomination? That's not the scientist I knew talking.† â€Å"Oh, I admit that they are fabulous creatures, but they would have never evolved naturally. They are a product of this war, and their purpose has been served. As has mine, as has yours. I'm sorry we didn't see eye to eye on this. Go now.† Just like that, this crazy bastard was going to plan B, and Nate had no idea how to stop him. Maybe that was what he was really brought here for. Maybe the Colonel was like someone who makes a suicide attempt as a cry for help, rather than an earnest attempt to end his life. And Nate had missed it. He started to back away from the Colonel, desperately trying to think of something he could say to change the situation, but nothing was coming to him. When he reached the passageway, the Colonel called out to him from the steps by the giant iris. â€Å"Nate. I promised you, and you deserve to know.† Nate turned and came a few steps back into the room. The Colonel smiled, a sad smile, resolved. â€Å"It's a prayer, Nate. The humpback song is a prayer to the source, to their god. The song is in praise of and in thanks to the Goo.† Nate considered it. A life's work contemplating a question, and this was the answer? No way. â€Å"Why only male singers, then?† â€Å"Well, they're males. They're praying for sex, too, aren't they? The females choose the mates – they don't need to ask.† â€Å"There's no way to prove that,† Nate said. â€Å"And no one to prove it to, Nate, not down here, but it's the truth. Whale song was the first culture, the first art on this planet, and, like most of human art, it celebrates that which is greater than the artist. And the Goo likes it, Nate, it likes it.† â€Å"I don't believe it. There's no evolutionary pressure for it to be prayer.† â€Å"It's a meme, Nate, not a gene. The song is learned behavior, not passed by birth. It has its own agenda: to be replicated, imitated. And it was reinforced. Have you ever seen a starved humpback, Nate?† Nate thought about it. He'd seen sick animals, and injured animals, but he'd never seen a starved humpback. Nor had he ever heard of one. The Colonel must have seen something in Nate's reaction. â€Å"There's your reinforcement. The Goo looks after them, Nate. It likes the song. I wouldn't be surprised if all of whale evolution – size, for instance – was accelerated by the Goo. We should have never started killing them. We wouldn't be at this juncture if we hadn't killed them.† â€Å"But we've stopped,† was all that Nate could think to say. â€Å"Too late,† the Colonel said with a sigh. â€Å"Our mistake was getting the Goo's attention. Now it has to end. The gene has had its three and a half billion years as the driving force of life. I suppose now the meme will have its turn. You and I will never know. Good-bye, Nate.† The iris opened, and the Colonel walked into the Goo. Nate ran all the way home, not sure how he had navigated through the labyrinth of tunnels, but found his way without having to backtrack. Amy wasn't at his apartment. His pulse was throbbing in his temples as he approached the buzzy, bug-winged speaky thing to try to call her, but he decided instead to go directly to her on foot. He checked at her place, and then at her mother's, then at every place they'd ever been together. Not only was Amy gone, but no one had seen her mother either. Nate slept fitfully, tortured by the notion of what the Colonel might have done to Amy because of his own stubbornness. In the morning he went searching for her again, asking everyone he encountered, including the whaley boys by the bakery, but no one had seen her. On the second day he went back through the corridors to the Colonel's mother-of-pearl amphitheater and pounded on the giant black iris until his fists were bruised. There was no response but a dull thud that echoed in the huge empty chamber. â€Å"I'll do what you want, Ryder!† Nate screamed. â€Å"Don't hurt her, you crazy fuck! I'll do what you want. I'll bring the navy down on this place and sterilize it, if that's what you want – just give her back.† When at last he gave up, he turned and slid down the iris facing the amphitheater. There were six killer-whale-colored whaley boys standing in the passageway opposite him, watching. They weren't grinning or snickering for once – just watching him. The largest of them, a female, let loose a quick whistle, and they crossed the amphitheater, walking in a crescent-shaped hunting formation toward him. Short of being a professional surfer or a bong test pilot for the Rastafarian air force, Kona thought he had found the perfect job. He sat in a comfortable chair watching sound spectrograms scroll across one computer monitor, while on another a program picked out the digital sequence in the subsonic signal and broke it into text. All Kona had to do was watch for something meaningful to come across the screen. Strange thing was, he really had started to learn about spectrographs and waveforms and all manner of whale behavior, and he was meeting the day feeling as if he was really doing something. He ran his hand over his scalp and shuddered as he read the nonsense text that was scrolling across the window. Auntie Clair had bought him four forties of Old English 800 malt liquor, then waited until he'd drunk them, before persuading him to let her cut his dreads down so they matched on both sides (because his true natural state should be one of balance, she said. She was tricky, Auntie Clair). The problem was, in jail his dreads had been almost completely torn off on one side, so by the time she finished evening things out, he was pretty much bald. Out of deference to his religious beliefs (to allow him a reservoir for his abundant strength in Jah, mon), Clair had left him a single dread anchored low on the back of his head, which made it look as if a fat worm was exiting his skull after a hearty meal of brain cells in ganja sauce. And speaking of the sacred herb, Kona was just on the verge of sparking up a bubbling smoky scuba snack of the dankest and skunkingish nugs when the text scrolling across the screen ceased being nonsense and started being important. He took a quick sip of bong water to steady his nerves, placed the sacred vessel on the floor at his feet, then hit the key that sent the streaming text to the printer. He stood and waited, bouncing on the balls of his feet for the printer to expectorate three sheets of text, then snatched the pages and dashed out the door to Clay's cabin. â€Å"I must be out of my mind,† Clay said. His suitcase was on the bed, and he was taking clothes out of the drawers and putting them into the case, while Clair was taking clothes out of the case, grouping them by a precise system he would never understand, and replacing them in the suitcase so that he would never find anything until he returned home and she helped him unpack. They had done this a lot. â€Å"I must be nuts,† Clay said. â€Å"I can't just go wandering around the oceans randomly looking for a lost friend. I'll look like that little bird in the book, the one that walks around asking everyone, ‘Are you my mother? ; â€Å"Sartre's Being and Nothingness?† Clair offered. â€Å"Right. That's the one. It's ridiculous to even leave port until we have something to go on – steaming around, burning up fifty gallons of fuel an hour. The Old Broad may have money stashed, but she doesn't have that kind of money.† â€Å"Well, maybe something will turn up in the whale calls.† â€Å"I hope. Libby and Margaret have a lot of sonic data streaming in from Newport, but it's still like looking for a needle in a haystack. Clair, she saw guys climbing into a whale –  » â€Å"So, baby, what's the worst that happens? You go to sea and do your best to find Nate and you fail? How many people ever did their best at anything? You can always sell the ship later. Where is it now anyway?† Just then the screen door fired back on its hinges and smacked against the outside wall with the report of a rifle shot. Kona came tumbling through the door waving pages of copy paper as if they were white flags and he was surrendering to everyone in the general Maui area. â€Å"Bwana Clay!† Kona threw the pages down on Clay's suitcase. â€Å"It's the Snowy Biscuit!† Clay picked up the pages, looked at them quickly, and handed one to Clair. Over and over the message was repeated: 41.93625S__76.17328W__-623__CLAY U R NOT NUTS__AMY Clay looked at Kona. â€Å"This was imbedded in the whale song.† â€Å"Yah, mon. Blue whale, I think. Just came in.† â€Å"Go back and see if there's more. And find the big world map. It's in the storeroom somewhere.† â€Å"Aye, aye,† said Kona, who had begun to speak much more nautically since Clay had purchased the ship, making his bid to go along on the voyage to search for Nate. He ran back to the office. â€Å"You think it's from Amy?† Clair said. â€Å"I think it's either from Amy or from someone who knows everything about what we're doing, which means it would have to be someone Amy talked to.† â€Å"What are the numbers?† â€Å"A longitude and a latitude. I'll have to look at the map, but it's somewhere in the South Pacific.† â€Å"I know it's a longitude and a latitude, Clay, but what's the minus six hundred and some?† â€Å"It's where pilots usually express altitude.† â€Å"But it's a minus.† â€Å"Yep.† Clay snatched the phone off of his night table and dialed the Old Broad as Clair looked quizzically at him. â€Å"Equipment change,† he whispered to Clair, covering the receiver with his hand. â€Å"Hello, Elizabeth, yes, things are going really well. Yes, they've picked up considerably. Yes. Look, I hate to ask this – I know you've done so much – but I may need one other little thing before we go to look for Nate and your James.† Clair shook her head at Clay's blatant playing of the missing-husband-shoved-up-a-whale's-bum card. â€Å"Yes, well, it may be a little expensive,† Clay continued. â€Å"But I'm going to need a submarine. No, a small submarine will be fine. If you want it to be yellow, Elizabeth, we'll paint it yellow.† After fifteen minutes of cajoling and consoling the Old Broad, making calls to Libby Quinn and the ship broker in Singapore (who offered him a quantity discount if he bought more than three ships in one month), Clay stood over a world map that was roughly the size of a Ping-Pong table, which Kona had spread out over the office floor, pinning the corners down with coffee cups. â€Å"It's right there, off the coast of Chile,† Clair said. She taught fourth-graders, and therefore basic world geography, so she could read a map like nobody's business. Kona placed a bottle cap on the spot where Clair was pointing. â€Å"We'll need nautical charts and the ship's GPS to be exact, but, basically, yep, that's where it is.† He looked at Kona. â€Å"Nothing else since that message?† â€Å"Same thing for five minutes, then just normal whale gibberish. You think the Snowy Biscuit is with Nate?† â€Å"I think she knew me well enough to know that I'd be thinking I was crazy to be looking. I also think that even if I believe the Old Broad's story about her husband, that doesn't explain how Amy was able to stay down for an hour on fifteen minutes' worth of air, so there was something going on with her that could be connected to this weirdness. She obviously knows more than we know, but – most important – we have nowhere else to look.† Kona looked at Clair, as if maybe she would answer his question. She nodded, and he resumed drinking his beer. Clay got down on his hands and knees on the map. â€Å"The ship broker says there's a deepwater three-man sub here, in Chuuk, Micronesia, that's about to finish up with some filming they're doing of deep shipwrecks.† Kona put a bottle cap on the atoll of Chuuk, Micronesia. â€Å"The owners will let me lease it for up to two months, but then a research team has it reserved for a deepwater survey in the Indian Ocean. The Clair is here, just north of Samoa.† Clay pointed. Kona put a third bottle cap just north of Samoa and did his best to drink off that beer while balancing the other two that he'd opened to get the caps. â€Å"So the Clair can probably be in Chuuk in three days. I'll fly in and meet them, pick up the sub, and then we can probably steam to these co-ordinates in four or five days if we cruise at top speed,† Clay said. â€Å"Now we're here –  » â€Å"We can't be, we can't be there,† said Kona. â€Å"Why not?† â€Å"Out of beers.† â€Å"So you get to that spot. Then what?† Clair asked. â€Å"Then I get in a submarine and see what there is to see six hundred and twenty-three feet down.† â€Å"So we're sure it's feet, not meters?† â€Å"No. I'm not sure.† â€Å"Well, I just want you to know that I am not comfortable with you doing this sort of thing, Clay.† â€Å"But I've always done this sort of thing. I sort of do this sort of thing for a living.† â€Å"So what's your point?† Clair asked. CHAPTER THIRTY-SIX Black and White and Red All Over Once, off the coast of California, Nate had followed a pod of killer whales as they attacked a mother gray whale and her calf. They first approached in formation to separate the calf from the mother, and then, as one group broke from the pod to keep the mother busy, the others took turns leaping upon the calf's back to drown it – even as the mother thrashed her great tail and circled back, trying to protect her calf. The whole hunt had taken more than six hours, and when it ended, finally, the killer whales took turns hitting the exhausted calf, keeping in a perfect formation even as they ripped great chunks of flesh from its still-living body. Now, in the amphitheater, as the killer whaley boys approached – their teeth flashing, the breath from their blowholes puffing like steam engines – the biologist thought that he was probably experiencing exactly what that gray-whale calf had during that gruesome hunt. Except, of course, that Nate was wearing sneakers , and gray whales almost never did. It was a big room. He had space to move. He just had to get around them. His sneakers squeaked on the floor as he came down the steps, faked right, then went left at a full sprint. The whaley boys, while amazingly agile in the water, were somewhat clumsy on land. Half of them fell for the fake so badly that they'd need a postcard to tell them how it all came out. They stooged into a whaley pile near the steps. The remaining three pursuers tried to fan out into a new formation, the alpha female coming the closest to getting between Nate and the exit. Nate was running in a wide arc around the amphitheater now, and by virtue of sheer speed he could tell he'd beat at least two of the remaining killers, but the alpha female was going to intersect with him before he got clear. She probably weighed three times what he did, so there was no going though her with a vicious body check. Maybe if he'd been on skates, he'd have tried it: pit his pure, innate Canadian skating force against her paltry cetacean hunting instinct and drive that bitch to the mother of pearl. But there were no skates, no ice, so at the very last second, as the female was about to slam him in a bone-breaking crunch against one of the benches that lined the walls, Nate pulled a spin fake, a move that was much more Boitano than Gretzky but nevertheless sent the big female tumbling over a bench in a tangle of black-and-white and i vory – like a flaccid piano botching the vaulting horse. Nate high-stepped the last twenty yards to the door, thinking, Yeah, three million years of walking upright not for nothing. Rookie. Meat. About the third step into his jubilation, Nate heard the sound of a great expulsion of air from his right, then a wet splat. Suddenly he saw his sneakers waving before his face. He felt the freedom of weightlessness, the exhilaration of flight, and then it was all gone as he slammed to the floor, knocking the wind out of himself. He slid to a stop in the huge loogie of whale spit that one of the trailing males had expectorated at his feet. Had he been able to breathe, he might have called a foul, but instead he struggled to get to his feet as the two males closed on him, showing dagger-toothed grins as they approached. Oh, my God, they're going to eat me! he thought, but then he saw that they both had unsheathed their long pink penises and were leading with a sort of a pelvic thrust. Oh, my God, they're going to fuck me! he thought. But when they got to him, one picked him up by the arms and bent him over forward, and he felt the great teeth scraping his scalp as his head slipped int o the whaley boy's mouth. No, they're definitely going to eat me, Nate thought. And in that suspension of time, right before the final crunch, amid the slow motion of an infinite last moment, clarity came to him, even as he screamed, and he thought, This is probably not going to go as well as the last time I was eaten. There's probably not going to be a girl at the end of this one. And then the female whistled shrilly, and the male stopped biting down just as his teeth were starting to cut into Nate's cheeks. The biting male pulled back and apologetically wiped saliva and blood from Nate's face, then propped him up and fluffed him a little, as if to show that he was good as new. Nate was still being held fast by the other male, but the biter was grinning sheepishly at the alpha female and making a squeaking noise that Nate, even with his limited understanding of whaleyspeak, understood as meaning â€Å"oops.† A half hour later they threw him into his apartment, and the alpha female grinned at him as she tore the stainless-steel doorknob out of the wall. The wall bled for a while after she left, then clotted over and rapidly began to heal. Nate stumbled into his bathroom and looked at himself in the mirror. There were bloody gashes down his forehead and cheeks. In another place and time, he realized, he would have gone to the emergency room to get stitched up. His hair was matted with blood, and he could feel at least four deep dents in his scalp where the whaley boy's teeth had broken the skin. There was a large knot at the back of his head where he'd hit the floor when he fell, and evidently he'd hit an elbow, too, because every time he bent his right arm, a sharp, biting pain shot all the way down to his fingertips. He pulled off his bloodstained clothes and climbed into the shower. Then, ignoring the strange fixtures that usually gave him pause, he leaned against the shower walls and let the water run over him until the bloody crust was gone from his hair and his fingers had shriveled with the moisture. He dried himself, then collapsed into his bed, wishing for a last time before he fell asleep that Amy was there, safe, next to him. He slept deeply and dreamed of a time when all the oceans were filled with a single living organism, wrapped like a cocoon around a single huge land mass. And in his dream he could feel the texture of every shore as if it were pressed against his skin. Nate awoke in the early hours before light came up in the grotto. He went into his living room and sat in the dark by the big oval picture window that looked out over the street and, ultimately, the Gooville harbor. There were shapes out there moving in the dark. Every now and then he'd catch the reflection of some dim light on a whaley boy's skin, but mostly he could tell they were out there by the sonar clicks that echoed around the grotto and by the low, trilling whistles of whaley-boy conversation. After an hour sitting there in the dark, he padded to the door and tried to open it. There was nothing but a smooth scar where the doorknob had been. The seal around the door was so tight it might have been part of the walls that framed it. In trying to work his fingers into the doorjamb, he realized that his elbow wasn't grating as it had been when he went to bed. He reached up to touch the gashes across his forehead and felt the scab flake away as easily and painlessly as dry skin. He immediately went to the bathroom and looked at himself in the mirror under the bright yellow bioluminescence. The gashes were healed. Completely healed. He brushed away the dried blood that had seeped after his shower to find new, healthy skin. It was the same with the dents in his scalp and the great goose egg at the base of his skull. He didn't even have a sore spot. He returned to the living room, fell into the chair by the window, and watched the light come up in the grotto. Outside, there was a lot of movement in the street and the harbor, and, watching it, Nate started to feel sick to his stomach, despite his miraculous healing. All the movement outside was that of whaley boys. There wasn't a single human out there anywhere. For two days he didn't see any other humans in Gooville, and even when he had screwed up his courage to use the buzzy, bug-winged speaky thing on the wall, he realized that he had no idea how to make it connect. By noon on the third day, he decided that he had to get out of the apartment. Not only couldn't he find Amy or do anything else while in here, but he was rapidly running out of food. He reasoned that the best time to make a break for it was in the middle of the day, when it seemed that the number of whaley boys out on the street was sparsest, because so many of them went down to the water at that time to swim. He dressed in long pants and sleeves for protection, then made the first attempt at the window. He tore one of the bone chairs from the floor in the kitchen, wiggling it first, as if loosening a baby tooth. He cast the chair at the center of the window with all his strength, preparing as he did to make the ten-foot leap to the street when it went though. But it didn't. It bounced back into the room. Next he looked for something sharp to try to puncture the window, but the only thing he could come up with were shards of the mirror in the bathroom, and although the mirror spider-webbed when he struck it, his fist wrapped in a towel, the shards stayed adhered to the bathroom wall, so all he'd really done was create a shiny mosaic. Finally, frustrated after three hours of ineffective attacks on the big window, he decided to hit it with the heaviest thing in the apartment: his body. He backed into the bedroom, sped through the living room, leaped into the air about halfway across, curled into a ball, and braced for impact. The window bulged out about three feet, until it appeared to the whaley boys outside that someone inside was trying to blow a giant bubble, and then it sprang back, trampolining Nate across the room into the far wall. At the bottom of the wall someone had installed a couch for just such an emergency, and Nate slid neatly into it with his newly flattened side down. â€Å"Well, that was just stupid,† he said aloud. â€Å"Boy, that was stupid,† Cielle Nuà ±ez said. She came into the living room and sat in a chair across from where Nate was piled onto the couch. â€Å"You want to tell me what in the hell you started?† â€Å"How did you get in? The knob is gone.† â€Å"Not on the outside. Come on, Nate, what did you do? Every human in Gooville has been locked down for the last three days. If I weren't the captain of a whale ship, I wouldn't have been able to come here either.† â€Å"I didn't do anything, Cielle, honestly. Where's Amy?† â€Å"No one knows. Believe me, that was the first place they went.† â€Å"Who?† â€Å"Who do you think? The whaley boys. They've taken over everything. Humans aren't even allowed near the ships. Ever since some of them heard you yelling about bringing the navy down here.† â€Å"I was. He has Amy, Cielle. I was just trying to get her back.† â€Å"Him? The Colonel? Why would he take Amy? She's one of the few who've ever even seen him. She's a favorite.† â€Å"Yeah, well no one is his favorite now.† Right then Nate made a decision. He wasn't going to get out of this place on his own, and the only person he could even consider an ally was sitting right there in front of him. â€Å"Cielle, the reason the Colonel called your ships back, the reason no one is allowed to leave the harbor, is that he wants you all here when the place comes down. He's got some plan to get the U.S. Navy, or somebody's navy, to attack Gooville with a nuclear torpedo. He thinks that the Goo is going to destroy the human race if he doesn't destroy it first. He wanted me to go to the navy. He thought I could convince them of the threat because of my scientific credibility, but I said no. That's when he took Amy.† â€Å"So all that yelling I heard you doing in the amphitheater – that wasn't you talking about bringing the navy here, that was just you trying to get Amy back?† â€Å"Yes. He's a loon, Cielle. I don't have any interest in bringing this place down. He thinks that there's some grand war going on between memes and genes, and that humans and the Goo are on opposite sides of it.† The whale-ship captain stood and nodded as if confirming something to herself. â€Å"Okay, then. That's what I needed to know. That's why he sent me here. I'll try to get them to send you some food.† â€Å"What? Help me get out of here.† Nate suddenly had a very bad feeling about this whole exchange. â€Å"I'm sorry, Nate. They have Cal. The whaley boys have him. You know how that feels. They told me I had to find out if you were plotting against the Colonel. Thank you for telling me. I think they'll let him go now.† She walked to the door, and Nate followed her. â€Å"Get me out of here, Cielle, at least –  » â€Å"Nate, there's nowhere to go. The only way out of here is a whale ship, and whaley-boy pilots are the only ones who can run them. They've been on notice not to let you on since we got here. Right now I couldn't leave if I wanted to.† She pounded on the door. â€Å"Open!† The door clicked open, and two all-black whaley boys stood outside waiting. They caught Nate by the shoulders and threw him back into the apartment as he tried to rush by them. â€Å"My own crew, Nate,† Cielle said. â€Å"See what you've done.† â€Å"He's going to kill you all, Cielle. Don't you see that? He's crazy.† â€Å"I don't believe you, Nate. I think you're the crazy one.† The door slammed shut. Back at Papa Lani, Clay was doing a final check on the equipment he was taking with him to meet his new ship. Diving and camera equipment lay spread out across the office floor. Kona was going through the checklist on the clipboard with a felt-tip pen. â€Å"So you tink the Snowy Biscuit going to be there?† â€Å"I'm going. I just wish that we could answer her. Tell her I'm on my way.† â€Å"You mean, like, put the digital in the whale sound and send it?† â€Å"Yeah, I know, we can't do it. Did you find a canister of soda lime for the rebreather's CO2 scrubbers?† â€Å"I can do that.† Kona held up the canister Clay was looking for and checked it off the list. â€Å"You can?† â€Å"I been looking at it long time. She not that hard to put that message back in the call. But how you going to send it? You need some gi-grandious big speakers under the water, mon. We don't have nothing like dat.† Clay stopped his inventory and pulled Kona's clipboard down so he could see his eyes. â€Å"You can put a message into the waveform so it would come out the same way we've been taking it out?† Kona nodded. â€Å"Show me,† Clay said. He went to the computer. Kona took the chair and pulled up a low-frequency waveform that looked like a jagged comb, and then he hit a button that took a small section and expanded it, which smoothed out the jags. â€Å"See, this part here. We know this a letter B, right? We just cut it and paste with other letters, make a goofy whale call. I got the all the letters but a Q and a Z figured.† â€Å"Don't explain, just do it. Here.† Clay scribbled a short message in the margin of Kona's checklist. â€Å"Then play it for me.† â€Å"I can play, but you won't hear it. It's subsonic, brah. Like I say, you going need some thumpin' speakers to send it. You know where we can steal some?† â€Å"We might not have to steal them.† While Kona pieced together the message, Clay grabbed the phone off his desk and dialed Cliff Hyland. The biologist answered on the second ring. â€Å"Cliff, Clay Demodocus. I need a favor from you. That big sonar rig of yours, will it broadcast subsonic frequencies?†¦ Good, I need you to take us out on your boat tonight, with your rig.† Kona looked at Clay. Clay grinned and raised his eyebrows. â€Å"No, it has to be tonight. I'm flying out for Chuuk in the morning. If I need to send out a signal, what can I plug in to it? Tape, disk recorder, what? Anything with a pre-amp?† Clay covered the receiver with his hand. â€Å"Can you put it on an audio disk?† â€Å"No problems,† Kona said. â€Å"No problem,† Clay said into the phone. â€Å"We'll meet you at the harbor at ten, okay?† Clay waited. He was listening, pacing in a little circle behind the surfer. â€Å"Yeah, well, we were just talking about that, Cliff, and we figured that if you said no, we'd just have to steal your boat and your rig. I could probably figure out how the rig works, right?† There was another pause and Clay held the phone away from his ear. Kona could hear an irritated voice coming out of the earpiece. â€Å"Because we're friends, Cliff, that's why I'd tell you in advance that I was going to steal your boat. Jeez, you think I'd just steal it like some stranger? All right, then, we'll see you at ten o'clock.† He hung up the phone. â€Å"Okay, kid, get this right. We have to have it ready and to the harbor by ten.† â€Å"But what you gonna do the bad guys get it?† â€Å"Even if they do, only Amy will know what it means,† Clay said. â€Å"Cool runnings, brah.† Kona was concentrating on putting the message together, his tongue curled out the corner of his mouth as an antenna for focus. Clay leaned over his shoulder and watched the waveform come together on the screen. â€Å"How did you figure this out, kid? I mean, it doesn't seem like you.† â€Å"How's a man supposed to work his science dub wid you yammerin' like a rummed-up monkey?† â€Å"Sorry,† Clay said, making a mental note to give the kid a raise if any of this actually worked.

What Neuroscience Tells Us About Morality - 1925 Words

Patricia S. Churchland s book entitled Braintrust: What Neuroscience tells Us About Morality, takes us on a walk through the scientific advances in evolutionary biology, genetics and neuroscience that have lead us to the question of wether morality can be explained or justified by science. The goal being an attempt at creating a framework for which to understand human morality. Churchland, a professor at the University of California San Diego in her book asks what is morality? Is it divinely inspired, or instinct, or an abstract set of rules? She argues, that a real understanding of morality begins with an understanding of the brain. Churchland begins by agreeing with the observations of Aristotle, Hume and Darwin that humans are social animal but extends them by asking what they mean in terms of brain physiology. However, David Hume s argument still remains, one cannot deduce what ought to be from something that is. 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