Trends in Food Science & Technology 15 (2004) 318

Biotechnologies: past history, present state and future prospects

Joseph H. Hulse
Visiting Professor in Industrial Biotechnologies, UMIST, Manchester, UK and at CFTRI, Mysore, India and MS Swaminathon Research Foundation, India

The paper presents a chronological review of biotechnologies, ancient and modern. It outlines the discovery of naturally occurring drugs by Babylonians, Egyptians, Chinese, Greeks and Romans, and the evolution of extraction, preservation and transformation technologies. It describes how pharmaceuticals progressed from empiricism, through chemical identication and synthesis to modern advances in genomics, proteomics, bio-informatics and syntheses by cultured cells from various genetically modied organisms. While biotechnologies for drugs rst progressed through chemistry, until relatively recently food technologies evolved by mechanisation, the gradual replacement of human hands by machines. Present and predicted industrial demand for bioengineers exceeds supply. The cost and complexity of emerging biotechnologies call for signicant revision of curricula and reorganisation of acedemic departments related to life sciences and biotechnologies. Urgently needed is active interdisciplinary cooperation in research and development, both in universities and industries, cooperation involving biochemists, bioengineers, mathematicians, computational scientists, systems analysts and specialists in bioinformatics. Bioscientists and biotechnologists must acquire more sensitive awareness of civil societies concerns and the ability to communicate with private citizens, politicians and the media. Recognising the inexorably rising demand for reliable health services, for safe and adequate food supplies, present and future opportunities for employment in industries devoted to food and drug technologies have never been greater. # 2003 Elsevier Ltd. All rights reserved.
0924-2244/$ - see front matter # 2003 Elsevier Ltd. All rights reserved. doi:10.1016/S0924-2244(03)00157-2

The British physicist Lord Kelvin gave as his opinion that if you can dene precisely and measure exactly that of which you speak, your opinions can be counted as credible; if not, they must be deemed doubtful. Let me begin with a denition relevant to this discussion: Biotechnologies are processes that seek to preserve or transform biological materials of animal, vegetable, microbial or viral origin into products of commercial, economic, social and/or hygienic utility and value. Bioengineers are men and women qualied to design, develop, operate, maintain and control biotechnological processes. One could cite instances in which (i) Biotechnology is exclusively equated with genetic modications and transgenesis, (ii) Biotechnology denotes a bioscientic activity that has not progressed beyond the research laboratory. In one American dictionary biotechnology is dened as synonymous with ergonomics: the study of human work in relation to a prevailing environment. The name Biotechnology rst appeared in Yorkshire early in the 20th century. A Bureau of Biotechnology began as a consultant laboratory in Leeds which from 1899 provided advisory services in chemistry and microbiology to fermentation industries in the north of England. The two Manchester universities (soon to be fused into one) have long and distinguished records in fermentation biochemistry. In 1912, Dr Chaim Weizmann isolated a strain of Clostridium acetobulylicum which converted carbohydrate into butanol, acetone and ethanol, a discovery extensively used for industrial production of acetone and butanol. In 1923, Dr Thomas Kennedy Walker welcomed the rst students into his Department of Fermentation Industries, possibly the rst of its kind, in what is now the University of Manchester Institute of Science and Technology. Later the departmental name was changed to Industrial Biochemistry, semantically similar to biotechnology. The undergraduate course was an amalgam of bioscience and bioengineering. From 1923 until he retired 35 years later, Professor Walkers students advanced to senior positions in food, pharmaceutical and related bio-industries in very many countries.

The interrelation of food and drugs

This presentation assumes that most graduates in bioengineering will progress to senior positions in food,

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pharmaceutical and related biotechnological industries. Though their historical patterns of growth and development have diered, food and drugs and the industries that produce them have long been closely associated. Standards of quality and safety for foods and drugs are customarily administered by the same regulatory agency, the US Food and Drug Administration being typical. As is discussed later, modern food and pharmaceutical processors employ similar technologies and methods of product and process control. Among the earliest historical records (ca 2900 BCE), the Chinese proclaimed a close association of foods with medicines, both being essential to good health, both derived from plant and animal sources. The Chinese believe many ailments can be cured by diet. They were the rst to add burnt sponge, an aquatic source of iodine, for people suering from goitre. The Emperor Fu-Hsi and his successors advocate that health depends on two principles: Yin and Yang. Yin weakness comes from malfunctions among internal organs and is indicated by a hot condition, red tongue and weak pulse. Yang weakness results when internal organs fail to absorb essential nutrients, indicated by a cold condition, Chinese medicinal foods are classied as hot or cold, strong or weak. Among an impressive list of Chinese medicinal foods, some are no doubt eective; others of dubious credibility. Horn from deer antlers is claimed to relieve fatigue, impotence and skeletal deformities. Ginseng root (Panax schinseng) is claimed, with little reliable pharmacological evidence, to alleviate diabetes, cardiovascular, digestive, liver and other diseases. Analyses of dierent samples from similar ginseng remedies show signicant variance in composition. Oriental beliefs in therapeutic foods is attracting North Americans, one-third of whom are said to buy herbal remedies as alternatives to prescription drugs. Americans search for a nutritional Elixir vitae has been evident for over half a century. During the 1950s vitamin supplements were in fashion; during the 1960s proteins and amino acids were in favour; in the 1970s extensive publicity was given to essential fatty acids and cholesterol in relation to cardiovascular disfunctions; during the 1980s dietary bre was of paramount interest. At present the fashion is with functional foods (which beg the question: what are non-functional foods?) and nutraceuticals, foods believed to possess benecial pharmacological properties. It is not surprizing that the Chinese claim to be the originators of nutraceutical concepts.

the raw materials and processed products. Foods found to be acceptable and satisfying, and medicinals that cured or alleviated particular maladies were discovered by chance, trial and error, and painful experience. The history of food processing, in large part, is a history of bioengineering, the gradual replacement of human hands and energy rst by animals, later by machines. Industrial processes of fractionation and transformation used today were developed hundreds of years ago. What began as labour-intensive artisanal crafts were progressively mechanized. In addition to providing an immense diversity of food products, food industries have progressively reduced the human eort and energy needed in factories, restaurants and homes.

Food preservation
The basic principles of food preservation: control of (1) active water content, (2) ambient atmosphere, (3) temperature, (4) pH; and (5) thermal inactivation of microbial and biochemical sources of spoilage, were discovered empirically hundreds of years ago. Mediterranean, Asian and Amerindian people used sun drying to preserve milk, meat, sh, fruits and vegetables, into their sun-dried pemmican northern Amerindians added dried berries that provided ascorbic acid. Sliced potatoes were freeze-dried by early Amerindians, ice gradually subliming in the dry air and low atmospheric pressure of the high Andes. Stone age Britons dried grains over open res to prevent sprouting. Over 4000 years ago, the Chinese preserved sh by osmotic dehydration with salt. Republican Romans reduced water activity in meat and fruits by adding salt or honey. About 5000 years ago, Middle Eastern farmers stored grains in earthenware amphora each hermetically sealed by an impervious goat skin. All stages of insect metamorphosis were asphyxiated by respired CO2. Seneca described how Romans preserved prawns in snow from the Appenines. The frozen food industry developed after Clarence Birdseye, an American, observed how whale, seal and reindeer meat were naturally preserved during the cold Canadian winter. Modern canning, bottling and boil-in-the-bag were anticipated in Republican Rome where chopped spiced meats were sealed and boiled inside the cleaned womb of a sow or the body cavity of a squid. Fermentation and pickling of fruits and vegetables is an ancient practice among Asian and Mediterranean people. The Babylonians preserved their milk by lactic fermentation. Ethanol was distilled in China over 3000 years ago. Homer described wine as A gift from the gods.

Food and drugs: science and technology

A characteristic common to food, drugs and most other basic need industries is that technologies discovered by perceptive empiricism long preceded any scientic understanding of the biochemical properties of

Grain millingthe rst continuous process

Fractionation of cereal grains by pulverization, sieving and winnowing, and extraction of olive oil by

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pressing, began in Egypt and nearby Mediterranean countries 7000 years ago. Commercial bakeries and breweries existed in Babylon and Egypt 5000 years before Eduard Buchner and Emil Fischer discovered the enzymic conversion of carbohydrates. The history of grain milling illustrates how labourintensive artisanal processes were progressively mechanized. The primitive pestel and mortar gave way in Egypt to the saddle stone: where grain spread on a stationary smooth stone slab was pulverized by an upper stone pushed backwards and forwards by a kneeling slave. Later, the Greeks devised a shearing action by incising herring-bone grooves into the interfaces of the upper and lower stones. In the rotary quern, evident in several ancient Mediterranean countries, an upper stone was continuously rotated over a lower static stone. By cutting an eye-hole in the centre of the upper stone, grain was fed in a steady stream, the pulverized product being carried to the periphery by centrifugal force. Grain milling was the rst known continuous industrial process. Rotary querns were at rst driven by slaves walking on a treadmill, later by camels or donkeys. After the Romans invented the water wheel, throughout the Roman empire grain mills were built close by rivers or running streams. The Domesday Book, William Is inventory of the nations assets published in 1085, recorded over 5000 water mills in Britain. The rst windmills appeared in Persia (now Iran) in the 10th century CE. In 1784, an early version of James Watts steam engine was installed in a London our mill. Less than 100 years ago, in North America, more grain mills were powered by water wheels than by steam engines. The rst mill powered by electric motors came on stream in 1887 in Wyoming. Though more precisely engineered, modern roller mills with their incised steel break rolls operate on the same principles as the early saddle stone and rotary quern. Hand winnowing has its modern equivalent in the middlings purier, an enclosed vibrating gravity table with screens of diverse mesh size. Separated bran is removed by suction fans. A smart indigenous software programme enables modern wheat our mills in India to be operated from a desk top computer. At the same time, poor rural Indian women grind local grains by pestel and mortar or saddle stone, and extract oil from groundnuts by rotary querns.

the steam engine, was mechanized faster than food processes. In the British baking industry, mechanical dough mixers were not much in evidence until after 1920. Mechanization moved more rapidly after World War II. During the 1930s, in a typical Manchester bakery, six men working an 8 h shift produced about 2400 loaves. In 1990, three men working an 8 h shift could produce over 65,000 loaves: 400 versus 22,000 loaves per man. The rst signicant change came in the 1960s when British scientists replaced traditional long fermentation processes by high energy mixing of bread doughs containing ascorbic acid. Continuous malting in breweries began with the Wanderhaufen moving malt couch. Continuous fermentations, in which the substrate passes over an immobilized microorganism or biocatalyst, are now common among modern bioindustries. Humphrey Davys discovery of nely divided platinum as a catalyst led to the catalytic hydrogenation of vegetable oils to produce hard fats for shortening and margarine. About the same time, solvent extraction of vegetable oils competed with mechanical expression. In contrast to food processing, pharmaceutical industries advanced more from chemistry than engineering, starting in the 18th century in the German dyestus industries after von Hofmann was appointed Professor of Chemistry at the University of Berlin.

Pharmaceuticals in ancient times

Survival and health, the fate of the human soul and body after death, and the supernatural inuence of the sun, moon and stars intrigued many of our early antecedents. Primitive peoples searched for panaceas and palliatives to cure their diseases. Early Palestinians and Sumerians believed disease was a punishment for sin and could be mitigated by magical charms and drugs with supernatural powers. Shen-Nung (ca 2700 BCE) is acclaimed as the Chinese founder of acupuncture and drug therapy. He and his contemporaries described diabetes, smallpox, measles, cholera and various dysenteries. Their 1800 medical prescriptions included ephedrine, camphor, and cod liver oil. Arsenic and mercury compounds acted as bactericides. Respiratory diseases were treated by surrounding the patient in a pile of burning herbs. The Egyptian Ebers papyrus (ca 1550 BCE), discovered by 20th century archeologists, describes treatments for rheumatism, schistosomiasis, diabetes and intestinal parasites. It lists 875 drugs compounded from ca 500 substances: metallic salts, such vegetable extracts as gentian, senna, castor oil, vermiuge and henbane. Sumerian cuneiform tablets from Hammurabis reign describe hepatic diseases, fevers, gonorrhoea, various strokes and scabies. Drugs included hellebore (believed to cure madness), mandrake root and opium.

Mechanization of traditional biotechnologies

The patterns and pace of mechanization have progressed dierently among dierent industries. Though the transition from rural domestic spinning and weaving into large mechanized factories evolved over more than two centuries, in Britain the textile industry, stimulated by cheap coal carried on canal barges, and

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During the 4th and 5th centuries, BCE, the Greek school of Hippocrates, published over 70 treatises on medical theories and practices, and prescribed more than 300 remedies, most from plants, to be administered orally or via other orices. The Greeks were aware of potential dangers in drug therapy; the Greek word farmakon (Pharmakon) being translated as drug, medicine, poison or magic potion. The Greeks believed that health (eucrasia) resulted from a harmonic blend, disease (dyscrasia) from imbalance among four humours: black bile, yellow bile, phlegm and blood. A tri-humorous concept of air, bile and phlegm existed among Ayurvedic Indians. Some 300 years after Hippocrates, Dioscorides, a Greek physician, regarded as the father of Materia Medica, formulated over 600 remedies from plant and animal tissues. Dioscorides medicines were prescribed for more than 1500 years. Galen of Pergamon, a physician of the 2nd century CE, added more vegetable remedies, known as Galenicals, to Dioscorides collection. Until the middle of the 19th century, medicine and pharmacy were more magical and mystical than scientic. Plantagenet physicians treated fevers by burying victims up to the neck in a dunghill; gout was treated with asses hooves; wealthy patients aicted with ague, itch or erysipelas were dosed with nely ground amethysts, pearls and sapphires. It is dicult to discover what useful drugs the alchemists discovered in their pursuit of the Elixir vitae, the elusive substance that would ensure eternal life. Alchemists wrote their reports in cryptic codes and obscure symbols to confuse their competitors. What is comprehensible is more redolent of the kitchen than the laboratory. Alchemical substances included sugar of lead, butter of antimony, oil of vitriol, cream of tartar and milk of lime. Paracelsus, a Swiss alchemist of the 15th century, is sometimes regarded as the father of chemistry. He disputed Galens theories and developed the notion of iatrochemistry: examination of substances to detect possible medicinal potency. He proposed various medical prescriptions. The rst printed medical book: Laxierkalendera treatise on laxativescame from the Gutenberg presses in 1457. In 1564, the worlds rst Pharmacopoeia Augustina was published in Augsburg. In 1616, the Royal College of Physicians published the Pharmacopoeia Londonensis which listed drugs then permitted in Britain.

rational distinction. Though it needed Maxwells mathematical genius 40 years later to transform Faradays electromagnetic induction principles into electric motors and power generators, the 1850s mark the point from which new technologies based on scientic principles appeared alongside empirically discovered technologies used to process foods, textiles, drugs and ceramics. After his mentor Humphrey Davy discovered the anaesthetic nitrous oxide, in 1818 Michael Faraday demonstrated that ether was a more eective anaesthetic. But before von Liebig published his Organic Chemistry in its Application to Physiology and Pathology in the mid-19th century, studies on the eectiveness of drugs can best be described as blindly empirical. For many centuries in Europe, pharmacy was the business of apothecaries who extracted and compounded medicines from natural vegetable and mineral sources. In Ancient Greece, physicians and apothecaries were discrete professions (an apoyek was a shop that sold drugs). In 1617, King James I created the Society of Apothecaries, giving them responsibility for production and sale of drugs and some poisons. Benjamin Franklyn dened the respective roles of American physicians and apothecaries, with laws that licensed apothecaries to sell drugs, poisons and narcotics. The rst codied food and drug laws were enacted in 1860 in the United Kingdom. In the 19th century, British apothecaries worked with a Materia Medica cabinet containing 270 samples of roots, barks, leaves, seeds and chemicals. The British Pharmaceutical Society received a Royal Charter in 1843. A consolidated British Pharmacopoeia was published in 1864 and revised in 1898 and 1914. The 1864 edition described only four synthetic drugs; over 80 were listed in the 1914 edition, almost all imported from Germany. Before World War I, Britain had no synthetic pharmaceutical industry, only a few vaccines being processed.

From empiricism to science

From the mid-1800s analytical chemistry, microscopy and cytology made impressive progress. Chemotherapy was stimulated by identication of microbial pathogens and means by which they could be controlled. Wohlers conversion of ammonium isocyanate into urea showed that naturally occurring organic substances can be synthesized from non-biological chemicals. Pharmacology progressed through research begun in Strasbourg on specic actions of drugs on particular body tissues. The worlds rst Chair of Pharmacology was in Estonia. Discovery of hormones, extracted from endocrine and ductless glands and later synthesized, added an important dimension to therapeutic medicine and to development of pharmaceutical industries. Until the 20th century food processing progressed through engineering, pharmaceutical technologies

Pharmaceutical industries
In his book Brave New World, Aldous Huxley proposed that the history of economic and industrial development is of two periods: pre- and post-Henry Ford. I would argue pre- and post-Faraday as a more

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through chemistry. Ancient remedies and plant extracts were the rst raw materials of pharmaceutical industries. Several drugs in early pharmacopoeias were later declared ineective or dangerous. Active substances were dissolved in ethanol and/or water; compounded with diluents and pressed into pills coated with gelatin or sugar; or into tablets with polysaccharide gums as binders, and lubricants to permit release from tableting machines. For treatment of skin wounds and infections, antiseptic drugs were dispensed as ointments in lanolin or water-in-oil emulsions.

Drugs: natural and traditional

Though today roughly 20% of all commercial pharmaceuticals are derived from natural and genetically modied microorganisms, there is lively commercial interest in natural and traditional sources. The Pzer drug company was among the rst to collect and screen botanical specimens from tropical forests. Merck in cooperation with the national Institute of Biodiversity is screening plants, insects and microorganisms from Costa Rica. Ethnobotanical expeditions in the tropical forests of the Amazon have delivered more than 10,000 species for examination. From Colombia over 1500 species, reported by local people to be biologically useful, are being studied. The ethical issue of biopiracy is being raised where foreign companies and their agents, engaged in botanical collecting, are taking away biological materials and indigenous experience in traditional medicine without reimbursement to local people. As observed by an Asian scientist: We have the biodiversity, they [the auent nations] steal it to support their biotechnologies. In response to public interest in ancient and traditional medicines, in 1992 the United States National Institutes of Health established an Oce of Alternative Medicines. Data bases on natural medicinals exist at the Royal Danish School of Pharmacy in Copenhagen, and at the University of Illinois, Chicago School of Medicine. The latter, known as NARPALERT, is administered by Professor Norman Farnsworth. Across the planet, there exists a vast unexplored source of plants and microorganisms. Of over 100,000 identied species, fewer than 200 microorganisms produce substances used by food, pharmaceutical or other industries. The higher orders of terrestrial plants represent more than 65% of the worlds biomass but fewer than 6% of identied species are commercially cultivated. Of the 80,000 plants believed to be edible, fewer than 20 provide 90% of the worlds food calories.

substances found in medicinal plants. The rst proprietary drug Aspirinacetylsalicylic acidwas synthesized by reacting acetic anhydride with salicylic acid from willow bark (Salix spp). Later, codeine was produced by methylation of morphine. In the late 1900s, Paul Ehrlich observed how certain dyes injected into animals, stained specic tissues. Ehrlich explored whether similar dyes would stain and inactivate microorganisms. He unsuccessfully tested 500 dyes on 2000 mice inoculated with pathogenic trypanosomes. He then synthesized more than 600 arsenic compounds with chemical structures similar to diazo dyes. His 606th compound inactivated the trypanosomes without adverse eect on the mice. The eective compound, named salvarsan, contained an AsAs group analagous to the NN group in diazo dyes and showed anity with protein in the pathogen comparable to the anity of diazo compounds with protein bres in wool. Salvarsan and its successor neosalvarsan, eective against Spirochaeta pallida the pathogen that causes syphilis, laid the basis for chemotherapy. In 1919, Heidelberger and Jakobs in Germany discovered that some azo derivatives of sulphanilamide destroyed bacteria. In 1935, a scientist at the Bayer company found the red azo dye prontosil to be eective against Streptococci that caused puerperal and scarlet fevers. In the 1930s, scientists at May and Baker in Britain synthesized over 600 sulphanilamide derivatives. The 693rd, which eectively treated bacterial pneumonia, was named M&B693. May and Baker synthesized over 3000 related compounds, several being eective bactericides. In 1936, the British Medical Research Council dened Chemotherapy as medical treatment by synthetic chemical compounds that react specically with infective organisms. The process of synthesizing chemotherapeutic substances and determining potency in laboratory animals is expensive and time consuming. Between 1936 and 1960, one of Britains largest pharmaceutical companies tested over 45,000 synthetics out of which only 16 became marketable drugs. During World War II, Britain lost access to Peruvian bark, the natural source of the anti-malarial quinine. Antimalarials were urgently needed to protect armed forces men and women posted to humid tropical countries. The only two synthetics available caused undesirable side eects. Between 1942 and 1946, the ICI Pharmaceutical company tested ca 1700 synthetics before discovering proguanil hydrochloride, given the trade name Paludrine. Malaria (literally bad air), is also known as paludism or swamp fever (Latin palus=swamp).

Synthetic drugs and chemotherapy

During the late 19th century, encouraged by their success with synthetic dyes, the German companies Bayer, Hoechst and Merck began chemical synthesis of drugs, rst making analogues and derivatives of active

Antibiotics
While pursuing his microscopic studies, Pasteur suggested that microorganisms might be induced to attack one another. In 1928, Alexander Fleming, at London

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University, observed that a mould spore from Penicillium notatum inhibited growth in a bacterial culture which it infected accidentally. The therapeutic potential of this discovery was overlooked until re-examined in 1939 by Howard Florey and Ernst Chain at Oxford. From their results, penicillin was isolated and chemically characterized. Subsequent research in Britain and the USA identied other useful species and strains of Penicillium, synthesized penicillin derivatives, and developed systems of large scale culture, isolation and purication. Penicillin was but the rst of an impressive series of antibiotics extracted from various species of Actinomycetes and other microorganisms. Long before Flemings discovery, primitive Micronesian people were known to scrape moulds from trees which they rubbed into wounds to prevent festering.

and Germany. Under investigation is a synthetic hormone, gestogen, which restricts reproductive processes in male gonads.

Industrial biotechnologiespresent value

The earlier text outlined how food processing and pharmaceutical industries progressed over the past 6000 years. Food processing began with simple artisanal technologies, human hands being gradually replaced by machines. Not until the late 19th century did science become an inuential force in food and drug industries. The pharmaceutical industry evolved from medicines compounded by apothecaries, most from local plant extracts, into chemical isolation, identication and synthesis of pharmacologically active substances and their derivatives. Given their importance to humans, commercial livestock and domestic pets, it is not surprizing that food and drug industries constantly expand and diversify, to satisfy demands of expanding, auent and aging populations. The total world value of industrially processed foods is about $1750 billion USD. Sales value of commercial pharmaceuticals (not including veterinary medicines) is close to $450 billion USD, 49% being sold in the USA, 24% in the European Union, 16% in Japan, with barely 11% for the rest of the world. Food processing industries, with sales over $500B per annum, comprise the largest industrial sector in the USA. Food industries in the EU employ more than 2.5 million people, they process two-thirds of all farm produce with sales close to $400 billion. Indian food processors employ more than 2 million people; at least 200 million Indians frequently buy processed foods. In 2002 the value of Indian processed foods was over 1000 times the value in 1962. It is impossible to estimate the total value of foods sold direct from farmers to local markets, or the proportion of food produced that is spoiled or wasted.

Hormones
More than 100 years ago, Claude Bernard, a French physiologist, reported that certain critical bodily functions are regulated by centres of internal secretion. These were identied as endocrine and ductless glands that secrete hormones (Greek hormon=to urge on). Adrenaline, rst extracted from the suprarenal glands of animals, was chemically characterized in the 1920s and later industrially synthesized. In 1921, in Toronto, insulin was isolated from Langerhens Islets extracted from porcine pancreas. During the 1980s, Canadian scientists synthesized an insulin precursor by a genetically modied bacterium. More recently, pancreatic cells that synthesize insulin were cultured, isolated, microencapsulated and transplanted into the bodies of diabetic patients to produce insulin in vivo. Thyroxine, generated by the thyroid, was synthesized in 1926, cortisone was isolated from the cortex of suprarenal glands in 1935 and commercially synthesized in 1956. In the intervening years, other hormones have been synthesized by GM microorganisms including avian and bovine growth hormones which stimulate body weight gain in farm animals and cultured sh, and milk production in bovines. Gonadotropins synthesized by GM bacteria induce gravid female sh to deposit their eggs when held captive in aquaculture systems. The eggs are later fertilized by cryogenically preserved milt (male sh sperm). Synthetic oestrogen and progesteron steroids inhibit ovulation and/or fertilization in women. The 50 year history of oral contraceptives, and the related medical, social and religious issues are reviewed in two recent books: Sexual chemistry: a history of the contraceptive pill by Lara Marks (Yale Press), and This mans pill: reections on the 50th birthday of the pill by Carl Djerassi (Oxford University Press). Clinical trials on chemical contraceptives for males are in progress at the Human Reproductive Sciences Unit in Edinburgh, and by pharmaceutical companies in the Netherlands

Pharmaceutical industrieschanging patterns

Though several similarities between food and drug industries have been noted, there are divergent dierences. Pharmaceuticals are processed by relatively few large corporations, while food industries include such giants as Nestle and Unilever together with thousands of medium and small scale companies. Pharmaceutical companies invest between 9 and 18% of their revenues in research and development. The average R&D investment for some 3500 Canadian registered food processors is less than 0.15% of sales revenue. Most pharmaceutical companies began as divisions of, or spin-os from chemical industries and expanded through acquisitions and mergers. In 1953, Watson and Crick described the helical structure of DNA. In 1973, the rst gene was cloned, in 1974 cloned genes were expressed in a foreign bacterial species. In 1976, Genentech became the rst company in

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the USA created for research to explore and exploit DNA. Between 1981 and 1999, specialist bioscience companies in the USA grew from 80 to over 1270. Ernst and Young report 1180 such enterprises among EU member countries. Many evolved from university bioscience departments. Some were highly successful, others with insucient venture capital, and inexperienced management, did not survive. Academic scientists with ambition to own a specialist bioscience company should have access to deep cash pockets. Risks are high and protable innovations do not come quickly. The biomedical industry now consists of two interrelated entities: (1) large pharmaceutical corporations (2) specialist bioresearch enterprises, described as Second generation biotechnology companies. In 2001, total revenue of the six largest bioscience companies was ca $8 billion; research and development investment between 20 and 37% of revenue. They devise and develop new processes and products to pilot plant and preclinical stages. Pharmaceutical companies expand the processes and subject the products to in vitro and in vivo clinical trials to determine potency, reliability and safety. For a new drug to progress from the laboratory to nal approval may cost between $300 million and $800 million and take between 10 and 15 years.

improved by computer modelling. Diagnostic processes are enhanced and speeded up by molecular modelling, by DNA microchips, and by recent advances in genomics, a name coined in 1980. Drugs synthesized by GM organisms include vaccines, immune regulators, substances to control cardiovascular disorders and various hormones. Modern vaccines include (1) toxoids-inactivated toxins extracted from cultured pathogens (for tetanus and diphtheria); (2) attenuated pathogens (for pertussiswhooping cough); (3) isolated biochemically modied antigens of various novel applications. Vaccines from GM viruses include whole virions (poliomyelitis); split vaccines (inuenza); isolated antigens (hepatitis B). Recent additions to the biosciences lexicon include Genomicsstudy of genomes and DNA nucleotide sequences; Proteomicsrelated to specic proteins produced by genomes; Metabolomicsinuence of gene expression on metabolites; Transcriptomicsproling of gene expressions using DNA/RNA micro assays.

Bioengineering processes
The immense diversity of active products from biotechnologies includes whole viable or attenuated cells, metabolites within cells or diused into the culture medium. Typical industrial processes progress through several stages: i. Identication and isolation of cells to be cultured. ii. Determination of optimum culture and harvesting systems. iii. Scale-up to large batch or continuous bioreactors. iv. Down-stream processes for fractionation, extraction, purication and sterilization. v. Methods for process control and product quality. vi. Protocols to ensure safety and containment throughout development and production. The over-riding objective is to maximise economic yield of stable eective products. A bioengineer with many years of experience recently said: Even where genetic modications, laboratory and pilot plant trials are entirely successful, scale-up to an economically ecient industrial process is inevitably frustrating, more costly and time-consuming than was forecast. In addition to synthesis by microorganisms, developments are progressing with cells from higher plants, animals, insects and GM viruses. Bacteria and viruses are cultured for metabolite synthesis, and for use as vectors to transfer genes between organisms. Cells may be cultured in batch bioreactors or in continuous systems where the nutrient medium percolates through or over and is transformed by the immobilized cells. Simi-

Biotechnologies: future prospects

Over the past 20 years, biotechnologies have evolved from intellectually intriguing biosciences into diversifying industries that produce useful biologicals from biocatalytic reactions, genetically modied bacteria, funghi, viruses, plant, mammalian and insect cells. Some techniques modify genetic composition and expression; others accelerate and adjust metabolic processes. Of particular interest to bioengineers are reliable means to expand from laboratory to factory scale, and technologies for the isolation, purication and sterilization of end products. Equally critical are reliable systems of product quality and process control. Earlier processes of extracting, screening and chemically modifying natural biochemical substances are giving way to identication of how specic diseases are caused, how particular drugs act to prevent or cure them. More eective diagnostics, prophylactics and therapeutics are being designed and synthesized by molecular modelling and combinatorial biochemistry. In the past, an organic chemist might synthesize 50 new compounds in a year, computer-assisted modern biochemistry can generate several thousand. Computers devise molecules to be systematically compared with computer-stored molecular structures. One company screens a million compounds against a target protein every 6 months. Rapid biological screening makes use of membranes from human or animal organ cells grown in tissue culture. Immunogenicity of specic antibodies can be

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larly, metabolites may be synthesized by isolated, immobilized enzymes. Plant cell culture begins by propagation of a callus, a mass of undierentiated cells. To derive a new plant with shoot and root, cells from the callus must be cultured in dierent media. Desirable metabolites can be extracted from a callus without progression to a shoot and root. Plant cell culture seems better suited to synthesis of metabolites useful in foods, biopesticides and cosmetics than for biomedicals. Mammalian and insect cell cultures oer more interesting opportunities for biomedical applications. Sources of mammalian cells include kidneys from aborted embryos and ovarian cells from Chinese hamsters which replicate relatively rapidly. Insect cell cultures, in combination with GM virus vectors, produce recombinant proteins and viral insecticides. The baculo virus, which infects insect cells, genetically modied yields specic proteins in high-density insect cell culture. Mammalian cells generate metabolites of greater purity, potency and complexity than most microbial cultures but, being highly sensitive, require careful culture in relatively small bioreactors. Means to expand mammalian cell culture in batch or continuous systems presents an interesting challenge to bioengineers. Monoclonal antibodies, plasminogen activators, hormones to stimulate blood cell growth and Factor VIII to control blood clotting are among products from mammalian cell culture. In association with specic viruses mammalian cells will produce viral vaccines and recombinant proteins used in gene therapy. In 1997, Human embryonic stem cells (HESC) were isolated from discarded human embryos. It is postulated that pluripotent stem cells may be cultured into dierent cells with the capacity to replace or repair cellular tissues in various human organs. Whether embryo stem cells will realise their hypothetical potential seems to depend as much on legislation inuenced by religious belief as on bioscience.

fractionation. Supercritical gas/liquid extractions (SGE) are useful for substances sensitive to organic solvents or susceptible to oxidation. At pressures between 10,000 and 40,000 kPa, carbon dioxide is a benign solvent for essential oils, oleoresins, natural terpenoids, caeine, and other sensitive biochemicals. Unlike many organic solvents, SGE leaves no toxic residues. Membrane processing, reverse osmosis, ultraltration, microltration, nanoltration and electrodialysis are among other industrial fractionation technologies. Chromatographic systems include gel ltration, ionexchange and anity separations that use binding interactions between proteins and packing materials, with various ligands coupled into hydrophilic support matrices.

Preservation and sterilization

Foods are for healthy nourishment; drugs to diagnose, prevent or cure disease. It is critical that all foods and drugs be free from organisms that may cause insult or injury to those who consume and use them. Generally speaking, biological materials such as foods and pharmaceuticals can be preserved by any process that (1) inhibits, destroys or removes and prevents re-entry of pathogenic and microorganisms that cause spoilage; (2) restricts adverse biochemical and biophysical change. Degradation of food and other biological materials can be restricted by packaging under inert atmosphere, by reducing water activity and thermal sterilization. Freeze-drying eectively lowers water activity in sensitive biologicals. Rapid freezing in liquid nitrogen prior to freeze-drying (lyophilisation) restricts cell disruption by slow-growing ice crystals.

Thermal processes and alternatives

Thermal processing in hermetic containers (cans, bottles, laminated plastics) takes a long time for heat to be conducted throughout the material. Excessive heating of foods and other biologicals can cause adverse change in critical functional properties, nutritional quality, avour, physical structure and texture. The higher the temperature, the longer the time, the greater the degree of biochemical and biophysical change. Existing processes that reduce heat damage include spray-drying, tubular and scraped surface heat exchangers, and steam injection followed by aseptic packaging Several alternative means of preservation are in various stages of investigation and development.

Downstream processing
Downstream relates to all that follows bioreactor synthesis: the isolation, purication and sterilization of end products. Downstream processes are estimated to absorb ca 80% of production costs, indicating an urgent need for more economical downstream technologies and bioengineers competent to design and operate them.

Isolation
Synthesized substances are isolated from various bioreactor fractions: insulin from harvested cells, some vaccines from supernatant uids. intra-cellular metabolites are released by mechanical, chemical or enzymatic rupture of cell walls; antibiotics by liquid:liquid extraction; tolerant volatile substances by fractional distillation; heat-sensitive enzymes by aqueous phase liquid

Irradiation
Ionising radiations can inactivate microorganisms and kill insects. Radiation sources for food and pharmaceuticals include gamma rays from radioisotopes Co60 or Ces137, X-rays or electrons generated by machines. Absorbed radiation is measured in Grays or

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kiloGrays (kGy), 1 Gray being equivalent to 1 Joule per kg. In the USA, irradiation is permitted for microbial control in dehydrated enzymes (10 kGy), spices (30 kGyJ), poultry (3 kGy), various pharmaceuticals and other biological materials. Data in parentheses are maximum permitted doses. The higher the dose, the greater the inactivation of microorganisms. High doses can induce molecular disruption and generate highly reactive free radicals which in turn cause unpredictable biochemical modications. In general, higher doses are permitted in biologicals consumed in small quantities (e.g. spices) or in prescribed pharmaceuticals. A WHO 1997 report states that at legally permitted doses, irradiation of foods will not cause toxicological diculty or signicant nutrient loss. Apart from consumers suspicions, still evident, the main constraints to food irradiation are economic. Capital costs are high, emissions from radioactive isotopes cannot be switched o, so to derive maximum benet there must be a constant supply, 24 h every day, 365 days every year, of high value material to be processed. Irradiation processes call for skilled bioengineers and physicists to ensure safety of all workers who must come close to the equipment. In most cases, irradiation is uneconomic for grain disinfestation even at the relative low doses required.

tems. Quartz crystals facilitate controlled outputs ranging from 500 W to 50 kW with 8090% energy eciency. Computer modelling programmes determine optimum conditions for dierent purposes. Main constraints include relatively high capital costs and need for highly skilled engineers for operational control.

Ultra-high hydrostatic pressure (UHP)

Lethal eects on microorganisms of isostatic pressures between 500 and 10 k bar (50 kPa1 MPa) were discovered over a century ago. UHP food processing has been applied mainly to fruit juices and jams. Industrial equipment maintains pressures from 400 to 800 MPa. Biomaterials in exible or semi-rigid packages, evacuated before sealing, are immersed in a uid in a high pressure vessel. The UHP is transmitted through the uid to the biomaterial. In acidic products vegetative cells are inactivated at 400 MPa, bacterial spores after 30 min at 600 MPa. UHP minimizes loss of nutritional and functional properties. Constraints include high capital cost, precise engineering and skilled operational control.

Pulsed energy
Three forms of pulsed energy for microbial inactivation are under study: (1) Pulsed electric elds (PEF); (2) Pulsed light (PL); (3) Pulsed magnetic elds (PMF). With PEF, induced electric potential causes lethal irreversible polarization of cell membranes. Critical potential varies with species, cell morphology and ambient conditions. Vegetative cells are inactivated at eld strengths between 15 and 30 kV/cm, alternating polarity pulses being more eective than constant polarity. Pulsed energy is not yet eective against spores or degradative enzymes. Pulsed light activates an inert gas lamp to generate broad band light ashes, 20 000 times the intensity of sunlight at the earths surface. PL is eective against surface vegetative organisms. Pulsed energy systems bear high capital costs and need precise operational control.

Ohmic heating
When an electric current ows through a substance of suitable conductivity, heat is uniformly generated. Ohmic heating is eective for uids and particles suspended in uid media. The uid is pumped through a column between two electrodes between which the current passes. The sterilised product is rapidly cooled and passes aseptically into sterile containers. Heating is uniform and of short duration. Commercial models range from a 10 kW pilot scale that processes 100 kg/h, to 300 kW machines to process 3 t/h. Capital costs range between 375,000 and two million pounds sterling. Operating costs depend on the power consumed and properties of the products processed.

Ultrasonics (US) Microwave (MW) and radio frequency (RF) heating

MW and RF depend on electromagnetic energy generated from a magnetron to produce an electric eld that alternates at radio or microwave frequencies. Heat is generated in biological materials by rapid reversal of molecular polarization. MW and RF provide uniform, short-time heating with high internal temperatures. Most widely known are domestic MW ovens, industrially MW and RF processes are used in dehydration, microbial inactivation and cooking. International electromagnetic compatibility regulations limit industrial processes to specic frequency bands that do not interfere with communication sysUltrasonics use sound waves at frequencies higher than detected by human ears (20 kHz). Microbes in liquid suspension are inactivated by alternating pressures and cavitation. With mild heat, US inactivates vegetative cells and can remove dirt inaccessible to conventional cleaning. US is used industrially to accelerate or control crystallization, ltration, hydrogenation of lipids and aging of alcoholic beverages.

Process and product quality control (QC)

Simply dened, QC objectives are to ensure (1) the properties of raw materials and nal products comply with dened specications; (2) consistency of essential

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properties among all production runs. Specications are laid down by (1) international protocols, (2) government regulatory agencies, (3) customers, secondary processors and retailers; (4) processing company managers. Specications, analyses and assessments of foods and drugs are designed to ensure safety to consumers and eectiveness within the conditions prescribed for use. More than 8000 processed foods and 7000 approved drugs are commercially produced in North America and Europe, together with unknown quantities of traditional foods and drugs on other continents. A comprehensive account of all recommended and tentative QC procedures would ll many CD Roms. This paper oers only an overview of present trends and practices. Bioengineers must ensure that materials of construction are compatible with biological materials to be processed. Processing equipment must be easy to clean and, where necessary, to sterilize.

On-line systems
Analyses of random samples from nished products in a quality control laboratory is gradually being superseded by control systems that use on-line sensors, probes and monitors that continually assess critical properties. When a defective property is identied, a feed-back signal corrects the faulty processing parameter, all on-line determinations being recorded in a computer. More than 100 devices determine ow rates, apparent viscosities and various rheological properties. Others record temperature, pressure and RH gradients. Various critical properties are assessed by change in electrical conductivity or dielectric constant. Chemical sensors respond to changes in pH, specic ions, organic radicals and impurities. Biosensors employ immobilized bacteria, enzymes, antigen-antibody reactions and DNA probes. By multivariant analysis of responses to mixed aromatics, an electronic nose can detect desirable or obnoxious odours. Spectroscopic on-line methods are too many and diverse to be catalogued. Ultrasonics detect particle size distribution, emulsion breakdown and various adulterants. Near infra-red can be calibrated to determine moisture, protein, lipid and various other component concentrations. Magnetic resonance imaging is an advanced spectroscopic method based on dierent magnetic properties of atomic nuclei when placed in a magnetic eld. The eld induces dierent energy levels between protons aligned with and protons aligned against the eld. MRI, most widely used to diagnose defects in the human body, now is applied to detect infections in aseptically packaged foods and drugs. Being non-destructive, it oers 100% inspection of critical biological materials. Immunological methods attach enzyme labels to antibodies to react to specic pathogens, hence the

name Enzyme linked immunosorbent assays (ELISA). Automated ELISA systems are based on a dipstick technology originally developed for testing pregnancy in women. An ideal sensor must be accurate, reliably responsive, robust and tolerant to processing conditions, easy to install and maintain, inexpensive in relation to product market value. Magnetic Resonance Imaging (MRI) and other expensive systems are economic for pharmaceutical but generally too expensive for most industrial food processes. On-line control systems are as much the responsibility of production bioengineers as of quality control biochemists and microbiologists. Many on-line sensors and probes determine a reaction or response indirectly related to the property critical to product safety and eectiveness. Bioengineers must therefore comprehend the relation between the response recorded and the critical product property to be determined. Sensors and their responses should be systematically checked and correlated with direct methods of determination.

Quality control (QC) and genetically modied (GM) organisms

The use of living organisms to synthesize pharmaceuticals, the complexity of the process technologies, call for changing patterns of process and product control. In addition to ensuring product safety and eectiveness, control systems must characterize and monitor organisms used, cell culture conditions, reaction, recovery and purication processes. QC of biologicals produced by viruses, microbial, plant, insect and mammalian cells is more complicated than of pharmaceutical substances isolated from medicinal plants or synthesized by chemical reactions. Accurate analysis of novel proteins synthesized by rDNA in GM organisms is an important priority. Progress is evident in automation of electrophoresis, amino acid analysis and gene sequencing. HPLC coupled with mass spectrometry and immunochemistry is extending the frontiers of protein analysis. Robotics, though relatively slow, are useful for tedious activities such as isotope labelling. Of urgent need are reliable methods to determine picogram levels of possible oncogenic DNA in mammalian cell cultures.

The future of bioengineering

From the data provided by the Ernst and Young regional biotechnology studies, from other publications and discussions with biotech industry executives, it is clearly evident that the demand for bioengineers and biotechnologists exceeds present and predicted supply. One study forecasts that over the next decade industrial opportunities for bioengineers will rise by 80%, for research and development bioscientists by ca 60%. Since the mid-1970s, modern biotechnology industries have generated more than 100 new drugs and vaccines.

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In the year 2000, world-wide investment in biotechnologies amounted to $37 billion USD which, according to a recent Canadian study, is expected to increase by 30%/year over the foreseeable future. According to a McKinsey Company report, biotechnology research and industrial development appear attractive to venture capital of which, in Canada, 90% is invested in the health care sector. Whether or not these predictions of future growth will be precisely accurate, expansion and diversication of bioscience and bioindustries seems inevitable given that most governments among European Union and North American nations declare health care, food safety and environmental protection related to human and animal health as high priorities. At the time when pharmaceuticals were mainly derived from medicinal plants and chemical synthetics, factory engineers were mostly chemical engineers. It is now evident that classical chemical engineering is inadequate for modern and advancing techniques of biosynthesis, extraction, isolation and purication of biologicals produced by various cell cultures and genetically modied organisms. The academic qualications and experience needed in industrial bioengineers are rapidly changing. To provide the knowledge and skills industries need, universities must evolve from traditional unidisciplinary, narrowly specialised departments into integrated interdisciplinary units. The Manchester Interdisciplinary Biocentre [MIB] illustrates how academic bioscientic research and teaching must be organised in the future. In a newly designed facility, the MIB will be staed by biophysicists, biochemists, mathematicians, computation scientists bioengineers and systems analysts. Bioengineers, together with all other involved disciplines, must work cooperatively from the early synthesis through to factory-scale processes, the whole being integrated into an orderly organisation by mathematically-trained systems analysts. An academic evolution to active, organized interdisciplinary teaching, research and development is essential for all biosciences and biotechnologies: pharmaceuticals, food and all else on which healthy lives depend. Recognizing the rate at which bioscience is expanding and diversifying, university departments could protably emulate their schools of management colleagues by oering short, intensive courses to enable industrial biotechnologists and bloengineers continually to upgrade their knowledge and skills. To satisfy the evolving needs of modern bioindustries, universities must critically assess and, in some instances, restructure their departmental organizations, curricula and research programmes. An obiter dictum from Rene Descartes Discourse on Method seems apposite and appropriate. All that is at

present known is almost nothing in comparison with what remains to be discovered. . . . We could free ourselves from an innity of maladies of body and mind if we had knowledge of their causes and the remedies provided by nature.

Chronologies of biotechnologies for food and drugs

BCE (BC) 4th/3rd Millennium Egyptians developed grain milling, baking, brewing.

3rd Millennium Chinese Emperor Fu-Hsi proposed health food principles of Yin, Yang. Chinese Shen-Yung Originator of acupuncture and natural drug therapy. Prescribed 1800 biological and chemical remedies. Egyptians, Sumerians preservation of milk, vegetables by acid fermentations.

2nd Millennium Egyptians prescribed plant remedies for rheumatism, diabetes, schistosomiasis. Sumerians treated hepatic diseases, gonorrhoea, strokes and scabies. Chinese distilled ethanol. Burnt sponge (iodine) to alleviate goitre. 1st Millennium Natural, open-air freeze-drying of potatoes by Andean Amerindians.

5th/4th Century Hippocrates School of Medicine Hippocratic Oath; 70 medical treatises; 300 remedies.

4th Century Aristotle classied known plants and animals; Theophrastus wrote History of plants. Greek word Pharmacon means both medicine and poison.

2nd Century Dioscorides: Father of Materia medica; prescribed > 600 drugs and remedies. Romans invented the water wheel.

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CE (AD) 2nd Century Galen of Pergamon: Many drugs known as Galenicals. Alchemy began in Alexandria; continued until 16th Century. Alchemists inuenced by chromatic and morphological associations (e.g. red wine and red meat generated good blood; white wine and meat conducive to anaemia). Yellow saron prescribed for jaundice: lung-shaped leaves of lungwort for respiratory diseases.

10th/11th Century Persians invented the windmill. Avicenna translated Aristotle, Dioscorides, Galen other early Greeks into Arabic. Arabians Materia medica: medicinal plants, extracts from wood and tree bark.

Robert Hooke, (cf Hookes Law: relation between stress and strain in elastic materials) described cellular structure of cork and various plant tissues. Anton von Leeuwenhoek (Netherlands) invented a microscope (magnication 30) described blood corpuscles, cells in sh tissues and animalcules (protozoa) present in organic matter in ponds. 1664 Thomas Willis (Oxford) described human brain and cranial nerves. 1670 Kaspar Bauhin (Swiss) classied higher plants into genera and species. 1679 Hamm (Netherlands) discovered spermatozoa. 1690s First herbarium of North American plants by immigrant physician to Canada. First studies of medicinal plants used by native aboriginals.

15th/l6th Century First medical text book on Laxatives printed on the Gutenberg Press. Paracelsus practised alchemy and medicine, discovered laudanum and opium. Drugs classied by function: hypnotics (opium, poppy juice, atropine from nightshade); pain relievers, anti-pyretics (willow bark, laudanum, strychnine); laxatives (castor oil, senna); emetics (tartar emetic); diuretics (Digitalis later for cardiovascular disorders). Derivatives of copper, mercury, antimony and sulphurwonder drugs of the Renaissance. Mercury for syphilis later discovered to cause neurological toxicity. Worlds rst pharmacopoeia printed in Augsburg. Antimony (tartar emetic) a panacea; later shown to cause cardiovascular collapse.

17th Century King James I created British Society of Apothecaries. Digitalis discovered in UKfoxglove a rural folk remedy for dropsy. Thomas Sydenham, Birmingham physician, differentiated between scarlet fever and measles. William Harvey described heart as stimulus for blood circulation, measured blood ow. Peruvian bark, brought to Europe by Spanish Jesuits, therapeutic for malaria (Lit. bad air) later discovered to contain quinine.

18th Century Linnaeus (Swede) formulated taxonomic system of classication for plants and animals later rened by Antoine de Jussieu (for plants) and Georges Cuvier (for animals). Edward Jenner observed British milk maids immune to cowpox. Jenner developed rened method of vaccination with controlled doses of cowpox serum. Jenners process safer than older Asian processes [Vacca (Latin)=Cow]. 1763 Extract of English willow bark relieved arthritis and rheumatic pain (salicylic acid isolated a century later). French pharmacien Pelletier isolated an emetic from ipecacuanha, strychnine from an Indian tree (genus Strychnos), morphine from opium poppy seed, brucine from angostura, quinine from Peruvian bark, caeine from coee berries, veratrine (jervin) from hellebore. Spallanzani (Italian) sterilized foods and organic materials by heating in hermetic containers. Spallanzani demonstrated fertilization of eggs by spermatozoa. 1798 Thomas Malthus Essay on Population. Human population will exceed food supply. Pharmacy as discrete profession combining apothecaries arts, botany, chemistry. First chair of Pharmacology at Dorpat in Estonia to study eects of therapeutics, toxicology, physiology and botany.

19th Century Humphrey Davy discovered nitrous oxide as anaesthetic. Michael Faraday discovered ether as anaesthetic.

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Rudolph Virchow diagnosed and described leucaemia, thromboses, embolisms. Virchows hypothesis: cells ultimate units of all organic life and pathological disturbance. Virchows cellular theory of pathology coincident with atomic theory of physics. Virchows studies plus advances in analytical chemistry formed basis of pharmacology: chemical composition related to therapeutic function. Alkaloids rst therapeutics isolated: morphine (1806), strychnine (1818), quinine (1820) 1816 G. Cuvier (French) categorised animals in four classes: Vertebrata, Mollusca, Articulata, Radiata. 1832 J. von Liebig (German) demonstrated that chloral hydrate induces sleep. 1820 Bracconot (French) hydrolized gelatin to produce glycine, meat and woolleucine. 1838 Berzelius (Swedish) coined the name Protein (Greek roteosProteos: that which comes rst) for nitrogenous organic compounds. 1846 von Liebig isolated tyrosine. 18601900 Most other essential amino acids isolated. Last was threonine in 1930. 184050s J. von Liebig recognized proteins, lipids, carbohydrates and various minerals as essential to human and animal nutrition. 1854 Lawes and Gilbert (UK) demonstrated dierence in nutritive value among plant proteins fed to pigs. 1825 F. B. Raspall, using iodine as a dye, revealed the distribution of starch in plant cells. Regarded as the founding father of histochemistry. 1828 Friedrich Wohler (German) synthesized urea from inorganic ammonium isocyanate. 1827 K. E. von Baer (Estonian) described the mammalian egg. 1830 Robert Brown (Scotland) described nuclei in plant cells. 1838 M. J. Schleiden and Theodur Schwann (Germany) cooperatively recognised the similarity on nuclei in plant and animal cells. From early 1830s active interest in cytology. 184080 Progressive discovery of chromosomes naturally colourless, made visible by selective anity with chemical dyes. 1840s60s Several reports that organisms develop by repeated cell division organic cells consist of nuclei embedded in protoplasm, (Greek rst to be moulded). 1840s William Perkin (UK) trying to synthesise quinine from analine sulphate accidentally produced rst synthetic dye: mauveine, laid basis for the European dyestus industries.

1847 J. Y. Simpson (Edinburgh) nitrous oxide, ether and chlorophorm as anaesthetics. 1855 Nathaniel Pringsheim (German) reported fusion of male spermatozoa with female ovum and how resultant cell dierentiated into new orgnanisms. 1870s E. Strasburger and Oskar Herrwig (Germany) described (1) division of nuclei, (2) fusion of two nuclei one from each parent. 1858 A. R. Wallace (UK) Theory of evolution transmitted to Charles Darwin. 1859 Charles Darwin On the Origin of Species by means of Natural Selection. 1850s German chemical and pharmaceutical industries developed. Chemical synthesis of drugs. Among rst, amyl nitrate to treat angina pectoris. Alkaloids atropine, cocaine and nicotine synthesized. 1870 Veronal (phenobarbitol) synthesized by Emil Fischer. 1835 Agostino Bassi (Italian) reported disease in silk worms caused by microfungus. 1860s Casimir Davaine (French) identied bacteria in blood of animals infected with Anthrax bacteria. 1860s Louis Pasteur (French) proved microorganisms are the cause not the result of fermentation in decaying matter. 1860s80s Pasteur revealed the link between bacteria and disease by studying infected silkworms, other sources of pathogens. He showed bacteria as the cause of acidity in beer. Whitbreads UK breweryrst to use microscope for quality control. 1860s Robert Koch (German) devised selective staining to assist microscopic identication of microorganisms. 1882 Koch identied Mycobacierium tuberculosum as the infective cause of pulmonary tuberculosis. Kochs diagnostic procedure: (1) isolate and culture the organism; (2) infect laboratory mice with the cultured organism; (3) recover the organism from mouse tissue. 1867 Joseph Lister (UK surgeon) used phenol to disinfect wounds. 1870 C. J. Eberth (German) laid the basis for virology by ltering out Anthrax bacteria from blood from which Pasteur diagnosed diseases caused by microscopically invisible organisms that passed through microbial lters. Pasteur began diagnosis of viral diseases. 1866 Gregor Mendel identied inheritable characters in noticeably dierent pea varieties. Mendels results ignored until rediscovered by American scientist in 1900.

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1880 First pharmaceutical industries in UK. Most R and D in government laboratories cf Germany: most by private industry. Burroughs Welcome most research active in UK. Pharmaceutical companies treated with suspicion in USA, (American Society for Experimental Pharmacology excluded industrial pharmacologists until 1941). 1883 Johann Kjeldahl (Netherlands) Analytical method to determine nitrogen in proteins. 1890s Isolation of substances active in endocrines: suprarenin and thyroidin. 1890s Paul Ehrlich demonstrated anity of various cells for particular dyes permitting microscopic identication by staining of microorganisms and cells from various organs. Ehrlich demonstrated anities between particular cells and certain drugs. He synthesized salvarsan (arsphenamine) and neosalvarsan (neoarsphenamine) eective against Spirochaeta pallida the infective pathogen for syphilis. Little further advance in chemotherapy until sulphur drugs (1930s) antibiotics (1940s) Scientists at Edinburgh University determined composition of various alkaloids, and pharmacology of strychnine, codeine, morphine, atropine and derivatives. Chemotherapy began with discovery and therapeutic control of pathogenic organisms. Vaccines for cowpox, cholera, typhus and typhoid developed. In vivo synthesis and extraction of hormones from animal tissues. Extracts of thyroid gland relieved myxodemia; adrenal extracts raised blood pressure. Living tissue used to monitor therapeutic activity, and for quality control. 1895 X-rays discovered by Roentgen applied in diagnostics.

20th Century Recognition that Mendels theory of inheritance applies to all plants and animals. Chromosomes composed of genes that control sex and biological characters of organisms T. H. Morgan (USA) elucidated nature and function of genes by work on Drosophila spp. From 1900 More scientic progress in pharmacology, biosciences than in past 5000 years. Puried isolates, derivatives and synthetics replaced simple remedies. Animal experiments complemented in vitro and empirical studies from human patients. Early 1900s Emil Fischer (German) synthesised long-chain peptides form 18 amino acids.

1914 Thyroxine isolated. 1921 In Toronto: Insulin isolated from Langherens islets in pig pancreas. UK Large scale vaccination against diphtheria, tetanus, yellow fever, small pox. 19001930s Adaptation by mutation to ecological conditions by various organisms. 1927 H. J. Mueller accelerated mutations by X-rays; UV, radioctive emanations. Chemical mutations by ethyl methane sulphonate. Plant chromosomes doubled by colchicine, an alkaloid extracted from seeds and corms of the Meadow Saron. 1929 Alexander Fleming accidentally discovered penicillin (anthropologists had much earlier described how primitive Micronesians rubbed moulds scraped from trees into wounds to prevent festering). 1920s Dyestus as antimicrobials. 1935 Sulphur drugs rst developed by Bayer company in Germany. Diversied by May and Baker in UK. 1939 Cortisone isolated. 1939 Florey and Chain cultured P. notatum and isolated penicillin. Penicillin, subsequent antibiotics eective against pathogenic Streptococci, Staphyloccoci, Meningococci, Gonococci and Pneumonococci. 1930s/40s Chemical pesticides to attack malaria and typhus vectors. 1930s Frederick Hopkins reported rst vitamins. 1955 Salk Polio vaccine developed. 195070s Crossing of sexually incompatible species by somatic hybridization (fusion of totipotent cells), embryo rescue, cell and tissue culture. 1953 Watson and Crick discovered double helix of DNA. 1973 First gene cloned. 1974 Cloned gene rst expressed in a foreign bacterium. First hybridoma created. 1974 US Asilomar Conference proposed safety guidelines for rDNA research. 1976 Genentechrst specialist bioscience company to exploit rDNA research. 1978 Genetic Manipulation Advisory Group (GMAG) created by UK government. 1978 US Presidents Commission for Studies of Ethics in Medicine & Biomedical Research. 1980 US Supreme Court ruled microorganisms may be patented under US law 1980 Genentechs public stock oering recorded highest ever price per share increase on New York stock exchange. Price/share rose from $35 to $89 in 20 min. 1980 Name Genomics rst used. 1981 Eighty new specialist bioscilbiotech companies formed in USA.

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1980s/90s Rockafella Foundation and International Rice Research Institute devised transgenic means to transfer pest resistances between wild and cultivated Oryza spp. Other transgenic food crops created. 1982 First rDNA vaccine for colibacillosis approved in Europe. 1982 Human insulin by rDNA approved in US and UK. 1992 US NIH Oce of Alternative Medicine. 1994 US Dietary Supplement Education Act legally dened food/drug supplements: vitamins, minerals, herbs, amino acids, other plant metabolites and extracts. Late 1900s Increased understanding of (i) genomicsmolecular characterization of species, (ii) bioinformaticsdata banks and data processing for genomic analysis, (iii) genetic transformationsvarious transgenic techniques to transfer genes between unrelated species, (4) molecular breedingidentication and translocation of useful biological properties by marker assisted selection, (5) diagnosticsidentication of pathogens by molecular characterization, (6) improved immunology in humans, animals, plants and sh by recombinant DNA vaccines. 1998 Human embryonic stem cells rst isolate from aborted fetus.

21st Century 2000 Human genome sequenced. 2001 US Presidents Stem Cell Research Council. Estimated 1/3 of all Americans buy herbal remedies in place of prescription drugs. Herbal remedies oered for colds, inuenza, gastrointestinal disturbances, rheumatoid and osteoarthritis, diabetes, hypertension, atherosclerosis, asthma, depression, cancer and HIV/ AIDS. Diculties of herbal medicines: inadequate standardisation. Nutraceuticals: foods purported to contain substances pharmacologically benecial. No food and drug laws in USA specically relate to herbal remedies.

technology & Biological Sciences Research Council. Building long-term capability. (1996). Ottawa: Canadian Human Resources Study in Biotechnology Human Resources Development Canada. Leading in the next millennium. (1998). Ottawa: Rept of the National Biotechnology Advisory Committee Industry Canada. Annual reports of the NRC Biotech Research Inst. Ottawa: National Research Council of Canada. Biotechnology: opportunities & constraints. (1985). Ottawa: Intl Devt Res Centre (IDRCMR 110e). European Union Council Directives on biotechnology. Rue de la Loi 200, B-1049, Brussels: EU. Good Manufacturing Practice, Position Statement on Genetic Modications. British Inst of Food Sci and Tech, London: EU. Ismael Seraglio & Persley G J. Promethean Science: Agriculture, Biotechnology, Environment. CGIAR, World Bank, Washington DC. Ernst & Young. Focus on fundamentals: the biotechnology report. Ernst & Young L L P, NY. European Commission. Life sciences and biotechnology: A strategy for Europe. E C Publications, Luxembourg. Doyle, John J & Persley G J. Enabling safe use of biotechnology: EDS Series No 10. (1996). World Bank, Washington DC. FAO/WHO. Strategies for assessing the safety of foods produced by biotechnology. (1991). FAO, Rome. IFST(UK). Guide to food biotechnology. (1996). Inst Food Sci and Tech, UK. J of Pharmagenomics. Advanster Communicatiosn. Edison, NJ.

References to related technology

Biotechnology Ernst & Young. Annual regional biotechnology reviews. Oxford, UK: Oxford Business Publications. Bioscience Engineering: BBSRC review of biochem eng. (1999) Swindon, UK: (British) Bio-

Biotech journals Biotechnology & Bioengineering. NJ: John Wiley. Enzyme & Microbiology Technology. Elsevier Press. Trends in Biotechnology. Elsevier. Biochemical Engineering Journal. John Wiley. Journal of Chemical Technology & Biotechnology. Elsevier Science Press. Food & Bioproducts Processing. Transactions of the Institute of Chemical Engineers UK. J Biotechnology. Elsevier. Bioprocess & Biosystems Engineering. Springer. Biotechnology Progress. American Chemists Society. BioCanada 2000. Montreal: Montreal Business Magazines.

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Historical Gribbin, J. (2002). Science: a history. Penguin Books, UK. Mees, C.E.K. (1947). The path of science. New York: John Wiley. Boorstin, D.J. (1998). The seekers. New York: Random House. Boorstin, D.J. (1993). The creators. New York: Vintage Books, Random House. Boorstin, D.J. (1985). The discoverers. New York: Vintage Books, Random House. Rose, S. (1976). The chemistry of life. UK: Penguin Books. Dixon, B. (1976). What is science for? UK: Penguin Books. Jacob, H.E. (1954). Sechstausend Jahre Brot. Hamburg: Rowohit Verlag GMBH. Corran, H.S. (1975). A history of brewing. London: David and Charles. Walton, J., Barondess, J.A., & Lock, S. (Eds.). (1994). The Oxford medical companion. Oxford University Press. Dun, J. (1999). History of medicine. University of Toronto Press. Porter, R.W. (1997). The greatest benet to mankind: a medical history of humanity. New York: W.W. Norton & Co. Bynum, W.F., & Porter, R. (Eds.). (1993). Companion encyclopaedia of the history of medicine. London: Routledge. Sonnedekker, G. (Ed.). (1976). Kremers and Urdangs history of pharmacy Philadelphia: Lippincott.

De Burgh, W.G. (1947). The legacy of the ancient world. London: McDonalds & Evans. Tacitus, Cornelius. (1956). The annals of Imperial Rome. Penguin Classics. Livius, Titus. (1960). The early history of Rome. Penguin Classics. Tannahill, Reay. (1973). Food in history. New York: Stein & Day. Root, Waverly. (1980). Food. New York: Simon & Shuster. Porter, R., & Teich, M. (Eds.). Drugs and narcotics in history. Cambridge University Press. Magner, L.N. (1994). A history of the life sciences. New York: M. Dekker. Porkert, M., & Ullmann, C. Chinese medicine: its history, philosophy and practice. New York: W. Morrow.

By this author: Joseph H Hulse Science, Agriculture and Food Security. (1995). National Research Council of Canada. Ethical issues in biotechnologies and international trade. (2002). Journal of Chemistry, Technology and Biotechnology, 77, 607. Opportunities for industrial bioengineers. (2001). Food Science and Technology, 15, 34; Opportunities for industrial bioengineers. (2002). Food Science and Technology, 16, 34. Biotechnologies: new homes for an old dilemma. (1984). Journal of the Canadian Institute of Food Science and Technology, 17(3), iii.

Any Suggestions?
Articles published in TIFS are usually specially invited by the Editors, with assistance from our International Advisory Editorial Board. However, we welcome ideas from readers for articles on exciting new and developing areas of food research. A brief synopsis of the proposal should rst be sent to the Editors, who can provide detailed guidelines on manuscript preparation. Mini-reviews focus on promising areas of food research that are advancing rapidly, or are in need of re-review in the light of recent advances or changing priorities within the food industry. Thus they are shorter than conventional reviews, focusing on the latest developments and discussing likely future applications and research needs. Features are similar in style to mini-reviews, highlighting specic topics of broad appeal to the food science community. The Viewpoint section provides a forum to express personal options, observations or hypotheses, to present new perspectives, and to help advance understanding of controversial issues by provoking debate and comment. Conference Reports highlight and assess important developments presented at relevent conferences worldwide. TIFS also welcomes Letters to the Editor concerned with issues raised by published articles or by recent developments in the food sciences. All Review-style articles are subject to editorial and independent peer review by international experts in the appropriate eld.