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सूक्ष्मजीवशास्त्र Microbiology

बी.एससी. सत्र ५ · ९ पाठ · ९ पाठांच्या नोट्स तयार

पाठ

  1. 1Food Microbiologyनोट्स
  2. 2Fermented Foods, Probiotics and Prebioticsनोट्स
  3. 3Dairy Microbiologyनोट्स
  4. 4Soil Microbiology and Biofertilizersनोट्स
  5. 5Plant Diseases Caused by Microorganismsनोट्स
  6. 6Water and Sewage Microbiologyनोट्स
  7. 7Industrial Fermentationनोट्स
  8. 8Production of Enzymes, Amino Acids and Vitaminsनोट्स
  9. 9Single Cell Protein and Mushroom Cultivationनोट्स

बी.एससी. सत्र ५ · सूक्ष्मजीवशास्त्र · पाठ 1

Food Microbiology

थोडक्यात सारांश

Food microbiology studies the microbes that spoil food, cause food-borne illness, and help make fermented foods. This topic covers factors affecting microbial growth in food, types of spoilage, food poisoning and food infection, methods of food preservation, and useful fermented foods including Indian examples.

महत्त्वाचे मुद्दे

  • Intrinsic factors (inside food): pH, moisture (water activity, aw), nutrients, oxidation–reduction potential, natural antimicrobials (e.g. lysozyme in egg white), biological structure (shells, skins).
  • Extrinsic factors (environment): storage temperature, relative humidity, gases (O₂, CO₂), presence of other microbes.
  • Most bacteria need aw above about 0.90; moulds can grow at lower aw (around 0.80 or less); dried foods resist spoilage.
  • Bread: moulds like Rhizopus stolonifer (black bread mould), Penicillium, Aspergillus; "rope" spoilage by Bacillus subtilis.
  • Fruits and vegetables: soft rot by Erwinia (now Pectobacterium); moulds.
  • Meat: putrefaction by Pseudomonas, Clostridium.
  • Canned food: swelling due to gas from Clostridium; flat sour spoilage (acid without gas) by Geobacillus stearothermophilus.
  • Spoilage signs: off-odour, sliminess, souring, gas, colour change, mould growth.
  • Food poisoning (intoxication): illness from a toxin already formed in food — Staphylococcus aureus (heat-stable enterotoxin; vomiting within a few hours), Clostridium botulinum (botulinum neurotoxin; paralysis; linked to poorly canned low-acid foods), Bacillus cereus (reheated rice).
  • Food infection: live pathogens eaten multiply in the gut — Salmonella (eggs, poultry), Shigella, E. coli O157:H7, Vibrio parahaemolyticus (seafood), Listeria monocytogenes (grows even at refrigerator temperature).
  • Preservation methods: high temperature (pasteurization, canning, boiling), low temperature (refrigeration about 4 °C, freezing −18 °C or lower), drying/dehydration, high salt or sugar (lowers aw: pickles, jams), chemical preservatives (sodium benzoate, potassium metabisulphite, sorbic acid), irradiation, modified-atmosphere packaging, fermentation, bacteriocins like nisin (from Lactococcus lactis).
  • Canning uses the 12-D concept: heat enough to reduce C. botulinum spores by 12 log cycles.
  • Fermented foods: curd/dahi, idli and dosa batter (lactic acid bacteria such as Leuconostoc mesenteroides and yeasts), bread (Saccharomyces cerevisiae), vinegar (Acetobacter), sauerkraut, soy sauce, cheese.
  • Probiotics: live beneficial microbes (e.g. Lactobacillus, Bifidobacterium) that improve health when eaten in enough amounts.

महत्त्वाच्या संज्ञा

  • Water activity (aw) – free water available for microbes (pure water = 1.0).
  • Intoxication – illness due to a pre-formed toxin.
  • Food infection – illness due to growth of live pathogens in the body.
  • Bacteriocin – antibacterial protein made by bacteria.
  • Hurdle technology – combining several mild preservation methods.

महत्त्वाची माहिती

DiseaseOrganismTypical food
Staphylococcal food poisoningStaphylococcus aureusMilk products, sweets handled by people
BotulismClostridium botulinumImproperly canned foods
SalmonellosisSalmonella spp.Eggs, poultry, meat
Emetic food poisoningBacillus cereusReheated fried rice
ListeriosisListeria monocytogenesSoft cheese, cold meats

५ महत्त्वाचे प्रश्न

Q1. What is water activity?

The ratio of the vapour pressure of water in a food to that of pure water at the same temperature. It shows how much free water is available for microbial growth; lowering it (drying, salting) preserves food.

Q2. Differentiate food intoxication and food infection.

Intoxication is caused by eating toxin already formed in food (e.g. S. aureus, C. botulinum); symptoms start quickly. Infection is caused by live microbes multiplying in the body (e.g. Salmonella); symptoms take longer.

Q3. Explain intrinsic and extrinsic factors affecting microbial growth in food.

  • Intrinsic: pH, aw, nutrients, Eh, antimicrobial constituents, biological structures.
  • Extrinsic: temperature, humidity, gas atmosphere, competing microbes.
  • Example: low pH of pickles and low aw of jams resist bacteria.
  • These factors decide shelf life.

Q4. Describe methods of food preservation.

  • Heat: pasteurization, sterilization, canning (12-D).
  • Cold: refrigeration, freezing.
  • Drying, smoking.
  • Salt/sugar (osmotic effect).
  • Chemicals: benzoates, sorbates, sulphites.
  • Irradiation, nisin.
  • Fermentation, packaging.

Q5. Write a note on botulism.

  • Caused by Clostridium botulinum, Gram-positive, anaerobic, spore-forming rod.
  • Toxin formed in low-acid, anaerobic foods (badly canned vegetables, meat).
  • Neurotoxin blocks acetylcholine release → flaccid paralysis, double vision, breathing failure.
  • Treatment: antitoxin, supportive care.
  • Prevention: proper canning (12-D), boiling suspected food (toxin is heat-labile), discard swollen cans.

परीक्षेसाठी टीप

Learn organism – disease – food as a table; questions on food poisoning almost always ask for examples.

बी.एससी. सत्र ५ · सूक्ष्मजीवशास्त्र · पाठ 2

Fermented Foods, Probiotics and Prebiotics

थोडक्यात सारांश

Fermented foods are foods changed by the controlled action of microorganisms, which improves taste, texture, shelf life and nutrition. India has a rich tradition of them — curd, idli, dosa, dhokla, jalebi batter, pickles. Probiotics are live beneficial microbes, prebiotics are the food that feeds them, and synbiotics combine both. This topic covers important fermented foods and the health role of probiotics.

महत्त्वाचे मुद्दे

  • Lactic acid bacteria (LAB) — Lactobacillus, Lactococcus, Leuconostoc, Pediococcus, Streptococcus thermophilus — are the main fermenting organisms. They are Gram-positive, non-spore-forming, catalase-negative and make lactic acid, which lowers pH and stops spoilage organisms.
  • Yeasts (Saccharomyces cerevisiae) make CO₂ and ethanol (bread, beverages); moulds (Aspergillus oryzae, Rhizopus) are used in Asian foods like soy sauce and tempeh.
  • Idli and dosa: soaked rice and black gram (urad dal) are ground and fermented overnight. Leuconostoc mesenteroides starts the fermentation and produces CO₂ that raises the batter; Enterococcus faecalis, Pediococcus and yeasts join later. The fermentation increases B vitamins and digestibility.
  • Dhokla (Bengal gram batter), jalebi batter, kanji (black carrot drink), bhatura dough and dosa are other Indian fermented foods.
  • Sauerkraut (fermented cabbage): heterofermentative Leuconostoc mesenteroides first, then homofermentative Lactobacillus plantarum; 2–3% salt draws out juice.
  • Pickles (cucumber, mango) and olives are also LAB fermentations under salt.
  • Bread: baker's yeast ferments sugars to CO₂ (raises dough) and ethanol (evaporates on baking).
  • Vinegar: two steps — yeast changes sugar to ethanol, then Acetobacter (Gluconobacter) oxidizes ethanol to acetic acid (about 4–8%).
  • Soy sauce (Aspergillus oryzae koji, then LAB and yeasts), tempeh (Rhizopus oligosporus), kimchi (Korean, LAB).
  • Probiotics: "live microorganisms which, when administered in adequate amounts, confer a health benefit on the host" (FAO/WHO, 2001). Examples: Lactobacillus acidophilus, L. rhamnosus GG, Bifidobacterium spp., Saccharomyces boulardii (a yeast).
  • Qualities of a good probiotic: survives stomach acid and bile, sticks to the gut lining, is safe (GRAS), has proven benefit, stays alive during storage.
  • Benefits: prevent and treat diarrhoea (including antibiotic-associated diarrhoea), help lactose digestion, compete with pathogens, strengthen gut barrier and immunity, may lower cholesterol.
  • Prebiotics: non-digestible food ingredients that feed good gut bacteria — inulin, fructo-oligosaccharides (FOS), galacto-oligosaccharides (GOS); found in onion, garlic, banana, chicory.
  • Synbiotics: probiotic + prebiotic together. Postbiotics: non-living microbial products or cell parts that give health benefits.

महत्त्वाच्या संज्ञा

  • Starter culture – known microorganisms added to start a controlled fermentation.
  • Homofermentative – LAB that make mainly lactic acid from glucose.
  • Heterofermentative – LAB that make lactic acid plus CO₂ and ethanol or acetic acid.
  • Probiotic – live microbe that gives health benefit in adequate amounts.
  • Prebiotic – non-digestible food that selectively feeds beneficial gut microbes.

महत्त्वाची माहिती

FoodRaw materialMain microorganisms
Idli / dosaRice + black gramLeuconostoc mesenteroides, Enterococcus faecalis, yeasts
Curd (dahi)MilkLactococcus lactis, Lactobacillus spp.
SauerkrautCabbageLeuconostoc, Lactobacillus plantarum
BreadWheat flourSaccharomyces cerevisiae
VinegarFruit juice / molassesYeast, then Acetobacter
Soy sauceSoybean + wheatAspergillus oryzae, LAB, yeasts

५ महत्त्वाचे प्रश्न

Q1. What are prebiotics? Give two examples.

Prebiotics are non-digestible food ingredients that pass to the colon and selectively feed beneficial bacteria like bifidobacteria. Examples: inulin and fructo-oligosaccharides.

Q2. Why are lactic acid bacteria important in food fermentation?

They produce lactic acid that lowers pH, preserves the food, gives sour flavour, and some make bacteriocins (e.g. nisin) and vitamins. They are safe (GRAS).

Q3. Describe the microbiology of idli fermentation.

  • Ingredients and soaking, grinding, overnight fermentation at room temperature.
  • Role of Leuconostoc mesenteroides (CO₂, acid), later Enterococcus, Pediococcus, yeasts.
  • Changes: leavening, pH fall, increased B vitamins and protein digestibility.
  • Flow chart.

Q4. Explain probiotics: definition, properties and health benefits.

  • FAO/WHO definition and examples.
  • Selection criteria.
  • Mechanisms: competitive exclusion, acid and bacteriocin production, immune modulation.
  • Health benefits and products in market (probiotic curd, capsules).

Q5. Write notes on vinegar and sauerkraut production.

  • Vinegar: alcoholic fermentation then acetic fermentation; Orleans (slow) and generator (quick) methods.
  • Sauerkraut: shredded cabbage, salt, anaerobic, sequence of LAB.

परीक्षेसाठी टीप

Learn the organism for each fermented food in table form; "idli fermentation" and "probiotics vs prebiotics" are frequent short notes in Maharashtra universities.

बी.एससी. सत्र ५ · सूक्ष्मजीवशास्त्र · पाठ 3

Dairy Microbiology

थोडक्यात सारांश

Milk is a highly nutritious food and so a good medium for microbes. This topic covers the composition of milk, sources of contamination, types of milk spoilage, pasteurization and its tests, milk quality tests like the MBRT, and fermented dairy products such as curd, yoghurt, butter and cheese.

महत्त्वाचे मुद्दे

  • Composition of cow milk (approximate): water about 87%, lactose about 4.5–5%, fat about 3.5–4%, proteins about 3.3% (mainly casein, plus whey proteins), minerals about 0.7%.
  • Milk inside a healthy udder is nearly sterile, but it gets contaminated from the teat canal, udder skin, milker's hands, utensils, air, water and fodder.
  • Normal milk flora: Lactococcus, Lactobacillus, Micrococcus, coliforms, yeasts and moulds.
  • Milk-borne diseases: tuberculosis (Mycobacterium bovis), brucellosis (Brucella), Q fever (Coxiella burnetii), typhoid, streptococcal infections.
  • Souring – lactic acid bacteria ferment lactose → lactic acid → curdling.
  • Gassy fermentation – coliforms, Clostridium produce CO₂ and H₂.
  • Proteolysis – protein breakdown, bitter taste (Bacillus, Pseudomonas).
  • Lipolysis – fat breakdown → rancid odour.
  • Ropy (slimy) milk – capsule-forming bacteria like Alcaligenes viscolactis.
  • Colour changes – blue (Pseudomonas syncyanea), red (Serratia marcescens), yellow (Pseudomonas synxantha).
  • Types of spoilage:
  • Pasteurization: LTLT/holding method — 63 °C for 30 min; HTST — 72 °C for 15 s; UHT — about 135–150 °C for a few seconds (long shelf life in sealed packs).
  • Coxiella burnetii is used as the index organism for pasteurization, as it is among the most heat-resistant non-spore-forming pathogens in milk.
  • Phosphatase test: alkaline phosphatase is destroyed by proper pasteurization; its presence shows under-heating or mixing with raw milk.
  • MBRT (methylene blue reduction test): methylene blue is decolourized by microbes as they use oxygen; faster decolourization = poorer quality.
  • Other tests: standard plate count, coliform count, resazurin test, clot-on-boiling test.
  • Fermented products: curd/dahi (Lactococcus lactis, Lactobacillus), yoghurt (Streptococcus thermophilus + Lactobacillus delbrueckii subsp. bulgaricus), cheese (rennet + starter culture; ripening with moulds like Penicillium roqueforti in blue cheese, P. camemberti in Camembert), butter and buttermilk, kefir.

महत्त्वाच्या संज्ञा

  • Starter culture – selected microbes added to start a fermentation.
  • Rennet – enzyme (chymosin) that coagulates milk protein in cheese making.
  • Pasteurization – heat treatment that kills pathogens but not all microbes.
  • Mastitis – infection of the udder (Staphylococcus aureus, Streptococcus agalactiae).
  • Index organism – organism used to set a process standard.

महत्त्वाची माहिती

MBRT grading (common textbook scale):

Decolourization timeMilk quality
5 hours or moreVery good
3–4 hoursGood
1–2 hoursFair
30 minutes or lessPoor

(Exact time bands vary slightly between textbooks and standards.)

५ महत्त्वाचे प्रश्न

Q1. What is the phosphatase test?

A test for proper pasteurization. Alkaline phosphatase, a natural milk enzyme, is destroyed at pasteurization temperatures; if it is still active, the milk was under-pasteurized or mixed with raw milk.

Q2. What is ropy milk?

Milk that becomes slimy and forms threads because of capsule-producing bacteria such as Alcaligenes viscolactis.

Q3. Describe the types of spoilage of milk.

  • Souring by lactic acid bacteria.
  • Gassy fermentation by coliforms, clostridia.
  • Proteolysis (bitter milk).
  • Lipolysis (rancid milk).
  • Ropiness.
  • Abnormal colours (blue, red, yellow) with organisms.

Q4. Explain the methods of pasteurization and their testing.

  • LTLT: 63 °C, 30 min.
  • HTST: 72 °C, 15 s, followed by rapid cooling.
  • UHT: very high temperature for seconds.
  • Purpose: kill pathogens (index Coxiella burnetii), increase shelf life.
  • Tests: phosphatase test, standard plate count, coliform test.

Q5. Describe the MBRT and its significance.

  • 10 mL milk + 1 mL methylene blue solution in a sterile tube.
  • Incubate at 37 °C in a water bath.
  • Note time taken for the blue colour to disappear.
  • Microbes use dissolved oxygen and reduce the dye.
  • Shorter time → more microbes → poor milk.
  • Quick, cheap test used at collection centres.

परीक्षेसाठी टीप

Remember the yoghurt pair (S. thermophilus + L. bulgaricus) and both pasteurization temperatures — they are asked often.

बी.एससी. सत्र ५ · सूक्ष्मजीवशास्त्र · पाठ 4

Soil Microbiology and Biofertilizers

थोडक्यात सारांश

Soil is full of microorganisms — bacteria, actinomycetes, fungi, algae and protozoa — that keep it fertile by breaking down organic matter and recycling nutrients. This topic covers soil microflora, the rhizosphere, the nitrogen cycle, nitrogen fixation, phosphate solubilization, and biofertilizers used in Indian agriculture.

महत्त्वाचे मुद्दे

  • Soil microbes: bacteria (most numerous), actinomycetes (e.g. Streptomyces, give the earthy smell due to geosmin), fungi, algae (including cyanobacteria) and protozoa.
  • Rhizosphere: narrow zone of soil around plant roots, rich in microbes because roots release sugars and amino acids (root exudates). R:S ratio compares microbial numbers in rhizosphere and non-rhizosphere soil (usually more than 1).
  • Nitrogen fixation: N₂ → NH₃.
  • Ammonification: organic nitrogen (proteins) → NH₃ (many decomposers).
  • Nitrification: NH₄⁺ → NO₂⁻ (Nitrosomonas) → NO₃⁻ (Nitrobacter).
  • Denitrification: NO₃⁻ → N₂ (Pseudomonas, Paracoccus) under anaerobic conditions.
  • Nitrogen cycle steps:
  • Symbiotic: Rhizobium in root nodules of legumes (gram, soybean, groundnut); Frankia in non-legumes like Casuarina; Anabaena azollae in the water fern Azolla.
  • Free-living (non-symbiotic): Azotobacter (aerobic), Clostridium (anaerobic), Beijerinckia, cyanobacteria like Nostoc and Anabaena.
  • Associative: Azospirillum with roots of cereals and grasses.
  • Biological nitrogen fixation uses the enzyme nitrogenase, which is damaged by oxygen and needs much ATP.
  • Leghaemoglobin in nodules (pink colour) binds oxygen and protects nitrogenase.
  • Phosphate-solubilizing microbes (PSM) like Bacillus megaterium, Pseudomonas, Aspergillus niger release organic acids that dissolve insoluble phosphates.
  • Mycorrhiza: fungus–root partnership; VAM/AM fungi (e.g. Glomus) increase phosphorus and water uptake.
  • Biofertilizers: preparations of living microbes that improve nutrient supply to plants. Types: nitrogen-fixing (Rhizobium, Azotobacter, Azospirillum, blue-green algae, Azolla), phosphate-solubilizing, potassium-mobilizing, mycorrhizal.
  • Carrier: lignite or peat powder is commonly used to carry the microbes.
  • Application: seed treatment (seed coated with slurry), seedling root dip (e.g. rice), soil application.
  • Advantages: cheap, renewable, eco-friendly, improve soil health; reduce chemical fertilizer use. Limitations: slower action, short shelf life, sensitive to heat and chemicals, need specific strains for specific crops.
  • Composting and vermicomposting use microbes (and earthworms) to turn organic waste into manure.

महत्त्वाच्या संज्ञा

  • Rhizosphere – soil zone influenced by roots.
  • Nitrogenase – enzyme complex that reduces N₂ to NH₃.
  • Root nodule – swelling on legume roots where Rhizobium fixes nitrogen.
  • Inoculant – biofertilizer culture applied to seed or soil.
  • Humus – dark, stable organic matter formed by decomposition.

सूत्रे

  • Nitrogen fixation (overall): N₂ + 8H⁺ + 8e⁻ + 16 ATP → 2NH₃ + H₂ + 16 ADP + 16 Pi
  • Nitritation: 2NH₄⁺ + 3O₂ → 2NO₂⁻ + 4H⁺ + 2H₂O (Nitrosomonas)
  • Nitratation: 2NO₂⁻ + O₂ → 2NO₃⁻ (Nitrobacter)
  • Denitrification: NO₃⁻ → NO₂⁻ → NO → N₂O → N₂

५ महत्त्वाचे प्रश्न

Q1. What is the rhizosphere effect?

The increase in the number and activity of microorganisms in the soil around roots, because roots release nutrients (exudates). It is expressed as the R:S ratio.

Q2. Name two symbiotic and two free-living nitrogen fixers.

Symbiotic: Rhizobium (legumes), Frankia (Casuarina). Free-living: Azotobacter, Clostridium (also cyanobacteria like Nostoc).

Q3. Describe the nitrogen cycle with the role of microbes.

  • Fixation: Rhizobium, Azotobacter, cyanobacteria.
  • Ammonification: decomposers.
  • Nitrification: Nitrosomonas, Nitrobacter.
  • Assimilation by plants.
  • Denitrification: Pseudomonas.
  • Draw cycle diagram with organisms on arrows.

Q4. What are biofertilizers? Explain their types, application and advantages.

  • Definition.
  • Types: N₂-fixing, P-solubilizing, mycorrhizal, K-mobilizing, BGA/Azolla for paddy.
  • Production: mass culture, mixing with carrier (lignite), packing.
  • Application: seed treatment, root dip, soil application.
  • Advantages and limitations.

Q5. Explain the formation of root nodules in legumes.

  • Root releases flavonoids; Rhizobium makes Nod factors.
  • Root hair curls around bacteria.
  • Infection thread forms and carries bacteria into the cortex.
  • Cortical cells divide → nodule.
  • Bacteria become bacteroids; leghaemoglobin protects nitrogenase.
  • NH₃ supplied to plant; plant gives carbohydrates.

परीक्षेसाठी टीप

Draw the nitrogen cycle with organism names written on each arrow; it fetches full marks in a 5-mark answer.

बी.एससी. सत्र ५ · सूक्ष्मजीवशास्त्र · पाठ 5

Plant Diseases Caused by Microorganisms

थोडक्यात सारांश

Plants suffer from diseases caused by fungi, bacteria, viruses, phytoplasmas and nematodes, and these cause huge crop losses in India every year. Plant pathology (phytopathology) studies the causes, symptoms, spread and control of these diseases. This topic covers how pathogens attack plants, the main symptoms, important bacterial, fungal and viral diseases of Maharashtra and Indian crops, and methods of control.

महत्त्वाचे मुद्दे

  • Disease triangle: a disease needs a susceptible host, a virulent pathogen and a favourable environment (humidity, temperature, rain).
  • Fungi cause most plant diseases (roughly 70–80%), then viruses, bacteria and others.
  • How pathogens enter: directly through the cuticle (fungi with appressoria), through natural openings (stomata, hydathodes, lenticels) and through wounds (most bacteria).
  • Weapons of pathogens: cell wall-degrading enzymes (pectinases, cellulases – cause soft rot), toxins (e.g. victorin, tabtoxin), growth regulators (auxin in crown gall), and blocking of xylem (wilts).
  • Common symptoms: leaf spots, blights, wilts, rots, cankers, galls, mosaic, yellowing (chlorosis), stunting, rusts, smuts, mildews.
  • Citrus canker – Xanthomonas citri; raised corky lesions on leaves and fruits (important for Nagpur oranges and Vidarbha citrus).
  • Bacterial blight of rice – Xanthomonas oryzae pv. oryzae; leaf tips dry and turn yellow-white.
  • Bacterial wilt of tomato, brinjal and potato – Ralstonia solanacearum; sudden wilting, bacterial ooze from cut stem.
  • Crown gall – Agrobacterium tumefaciens; tumours from transfer of T-DNA of the Ti plasmid (used in plant genetic engineering).
  • Soft rot of vegetables – Pectobacterium (formerly Erwinia) carotovorum.
  • Bacterial diseases:
  • Rust of wheat – Puccinia graminis tritici (black/stem rust); rust-coloured pustules.
  • Late blight of potato – Phytophthora infestans (an oomycete, a fungus-like organism); caused the Irish famine of 1845–47.
  • Blast of rice – Magnaporthe oryzae (Pyricularia oryzae).
  • Wilt of cotton and pigeon pea (tur) – Fusarium spp.
  • Powdery mildew (grapes, peas) and downy mildew (grapes, bajra).
  • Red rot of sugarcane – Colletotrichum falcatum.
  • Fungal diseases:
  • Viral diseases: tobacco mosaic (TMV, mechanical spread), leaf curl of tomato and papaya ring spot (whitefly or aphid vectors), banana bunchy top (aphid), yellow vein mosaic of okra (whitefly).
  • Phytoplasma diseases: grassy shoot of sugarcane, little leaf of brinjal, sandal spike (spread by leafhoppers).
  • Cultural: crop rotation, resistant varieties, removal of infected plants, clean seed.
  • Chemical: fungicides (Bordeaux mixture – copper sulphate + lime, discovered by Millardet in 1882; mancozeb, carbendazim), seed treatment, antibiotics like streptocycline for bacterial diseases.
  • Biological: Trichoderma viride, Pseudomonas fluorescens, Bacillus subtilis.
  • Legal: quarantine to stop entry of new pathogens.
  • Integrated disease management (IDM) combines all.
  • Control methods:

महत्त्वाच्या संज्ञा

  • Pathogen – organism that causes disease in a plant.
  • Symptom – visible change in a diseased plant (spot, wilt).
  • Appressorium – flattened tip of a fungal germ tube that presses into the host surface.
  • Vector – organism (insect) that carries a pathogen from plant to plant.
  • Bordeaux mixture – copper sulphate + lime fungicide.

महत्त्वाची माहिती

DiseaseCropPathogenType
Citrus cankerOrange, limeXanthomonas citriBacterium
Bacterial blightRiceXanthomonas oryzaeBacterium
Crown gallMany dicotsAgrobacterium tumefaciensBacterium
Late blightPotato, tomatoPhytophthora infestansOomycete
Stem rustWheatPuccinia graminisFungus
Red rotSugarcaneColletotrichum falcatumFungus
MosaicTobaccoTMVVirus
Grassy shootSugarcanePhytoplasmaWall-less bacterium

५ महत्त्वाचे प्रश्न

Q1. What is the disease triangle?

It shows that a plant disease develops only when three factors meet: a susceptible host, a virulent pathogen and a favourable environment. Removing any one prevents the disease.

Q2. Name the causal organism and one symptom of citrus canker.

Xanthomonas citri (Gram-negative rod). It forms raised, brown, corky lesions with a yellow halo on leaves, twigs and fruits.

Q3. Describe the mechanisms by which pathogens attack plants.

  • Entry: direct, natural openings, wounds.
  • Enzymes degrading cuticle and cell wall.
  • Toxins and growth regulators.
  • Vascular blocking in wilts.
  • Defence of plants (brief).

Q4. Describe two bacterial diseases of plants with symptoms and control.

  • Citrus canker and bacterial blight of rice (or crown gall).
  • Pathogen, symptoms, spread, control (copper sprays, streptocycline, resistant varieties, pruning).

Q5. Explain the methods of controlling plant diseases.

  • Cultural, chemical, biological, legal (quarantine), host resistance.
  • Integrated disease management.
  • Examples from Maharashtra crops.

परीक्षेसाठी टीप

Learn one bacterial, one fungal and one viral disease fully (pathogen, symptoms, control). Crown gall links to genetic engineering — mention the Ti plasmid.

बी.एससी. सत्र ५ · सूक्ष्मजीवशास्त्र · पाठ 6

Water and Sewage Microbiology

थोडक्यात सारांश

Water can carry disease-causing microbes, so testing and purifying drinking water is a key public-health job. This topic covers water-borne diseases, indicator organisms, bacteriological tests of water (MPN and membrane filter), water purification, and how sewage is treated by primary, secondary and tertiary methods.

महत्त्वाचे मुद्दे

  • Water-borne diseases: cholera (Vibrio cholerae), typhoid (Salmonella Typhi), bacillary dysentery (Shigella), hepatitis A and E (viruses), polio, amoebic dysentery (Entamoeba histolytica), giardiasis (Giardia).
  • Testing for every pathogen is not practical, so we test for indicator organisms that show faecal pollution.
  • Coliforms (Gram-negative, non-spore-forming rods that ferment lactose with acid and gas within 48 h at 35–37 °C) are the usual indicators. Faecal (thermotolerant) coliforms, especially E. coli, grow at 44.5 °C and show recent faecal contamination.
  • Other indicators: faecal streptococci (enterococci), Clostridium perfringens (shows older or intermittent pollution, as spores survive long).
  • An ideal indicator: present in faeces in large numbers, absent from clean water, survives slightly longer than pathogens, easy and quick to detect, does not multiply in water.
  • Presumptive test: water inoculated into lactose broth (or MacConkey broth) tubes with Durham's tubes; acid and gas = positive.
  • Confirmed test: positive tubes subcultured onto EMB or Endo agar (or BGLB broth); E. coli gives colonies with a green metallic sheen on EMB.
  • Completed test: colonies checked by Gram stain (Gram-negative rods) and gas in lactose broth.
  • MPN (multiple tube fermentation) test — three stages:
  • The MPN (most probable number) of coliforms per 100 mL is read from statistical MPN tables.
  • Membrane filter technique: known volume (e.g. 100 mL) filtered through a 0.45 µm membrane; membrane placed on selective medium (e.g. m-Endo); colonies counted directly. Fast and precise.
  • IMViC tests (Indole, Methyl red, Voges–Proskauer, Citrate) separate E. coli (+ + − −) from Enterobacter aerogenes (− − + +).
  • Water purification (municipal): sedimentation → coagulation/flocculation (alum) → filtration (slow or rapid sand filters) → disinfection (chlorination, ozone, UV).
  • Primary: screening, grit removal, sedimentation — removes solids.
  • Secondary (biological): aerobic microbes oxidize organic matter — activated sludge process, trickling filter, oxidation ponds; anaerobic digestion of sludge produces biogas (mainly methane and CO₂).
  • Tertiary: removes remaining nitrogen, phosphorus, and pathogens (chemical precipitation, filtration, chlorination).
  • Sewage treatment:
  • BOD (biochemical oxygen demand): oxygen used by microbes to oxidize organic matter in water in 5 days at 20 °C (BOD₅). High BOD = more pollution. COD uses a chemical oxidant (dichromate) and is usually higher than BOD.

महत्त्वाच्या संज्ञा

  • Indicator organism – organism whose presence shows faecal pollution.
  • Potable water – water safe to drink.
  • Activated sludge – flocs of microbes recycled in aeration tanks.
  • Residual chlorine – chlorine remaining after disinfection that keeps protecting water.
  • Eutrophication – excess nutrients causing algal blooms and oxygen loss.

सूत्रे

  • D₁ = dissolved oxygen (DO) of diluted sample at start (mg/L)
  • D₂ = DO after 5 days at 20 °C (mg/L)
  • P = decimal fraction of sample used in the dilution
  • BOD (mg/L) = (D₁ − D₂) ÷ P
  • Example: D₁ = 8.0 mg/L, D₂ = 3.5 mg/L, P = 0.02 (2% sample) → BOD = (8.0 − 3.5) ÷ 0.02 = 4.5 ÷ 0.02 = 225 mg/L.

५ महत्त्वाचे प्रश्न

Q1. Why is E. coli used as an indicator of faecal pollution?

It lives only in the intestine of humans and warm-blooded animals, is passed in large numbers in faeces, does not normally multiply in clean water and is easy to detect. So its presence shows recent faecal contamination.

Q2. Define BOD.

Biochemical oxygen demand — the amount of dissolved oxygen used by microorganisms to break down organic matter in a water sample during 5 days at 20 °C. It measures organic pollution.

Q3. Describe the MPN test for coliforms.

  • Presumptive: lactose broth tubes with Durham's tubes (e.g. 3 × 10 mL, 3 × 1 mL, 3 × 0.1 mL sample), 35–37 °C, 24–48 h; acid + gas = positive.
  • Confirmed: streak on EMB/Endo; typical colonies (metallic sheen).
  • Completed: Gram stain, lactose fermentation.
  • Read MPN/100 mL from tables.
  • IMViC to identify E. coli.

Q4. Explain secondary treatment of sewage.

  • Biological oxidation of organic matter by aerobic microbes.
  • Activated sludge: aeration tank, flocs of bacteria and protozoa, settling, sludge recycled.
  • Trickling filter: sewage sprinkled over stones covered with microbial film.
  • Oxidation ponds: algae–bacteria partnership.
  • Sludge digestion: anaerobic, produces methane-rich biogas.
  • Lowers BOD greatly.

Q5. Calculate BOD: D₁ = 7.8 mg/L, D₂ = 4.2 mg/L, 5 mL sample diluted to 300 mL.

  • P = 5/300 = 0.0167
  • D₁ − D₂ = 7.8 − 4.2 = 3.6 mg/L
  • BOD = 3.6 ÷ 0.0167 ≈ 216 mg/L (exactly 3.6 × 60 = 216 mg/L)
  • Indicates a heavily polluted sample.

परीक्षेसाठी टीप

Draw the three stages of the MPN test as a flowchart, and the sewage treatment plant as a block diagram (primary → secondary → tertiary).

बी.एससी. सत्र ५ · सूक्ष्मजीवशास्त्र · पाठ 7

Industrial Fermentation

थोडक्यात सारांश

Industrial microbiology uses microbes on a large scale to make useful products — alcohol, antibiotics, organic acids, enzymes, vitamins and vaccines. This topic covers the fermenter (bioreactor) and its parts, types of fermentation, strain improvement, media, downstream processing, and production of key products like penicillin, ethanol and citric acid.

महत्त्वाचे मुद्दे

  • In industry, "fermentation" means any large-scale culture of microbes to make a product, aerobic or anaerobic.
  • Primary metabolites are made during the growth (log) phase and are essential for growth (ethanol, amino acids, organic acids). Secondary metabolites are made mostly in the stationary phase (antibiotics, pigments).
  • Fermenter (bioreactor) — usually stainless steel. Parts: vessel, impeller/agitator (mixing), sparger (air bubbles), baffles (prevent vortex), cooling jacket or coils, probes for pH, temperature, dissolved oxygen and foam, inlet/outlet ports, sampling port.
  • Types of fermenters: stirred tank (most common), airlift, bubble column, packed bed.
  • Batch, fed-batch (nutrients added gradually, e.g. penicillin, baker's yeast) and continuous fermentation.
  • Submerged (liquid) and solid-state fermentation (on moist solids like wheat bran, e.g. some enzymes).
  • Upstream processing: strain selection and improvement (mutation, recombination, genetic engineering), media formulation, sterilization, inoculum development.
  • Industrial media use cheap raw materials: molasses (sugar), corn steep liquor (nitrogen, used in penicillin production), whey, starch.
  • Downstream processing: separation of cells (filtration, centrifugation), cell disruption (if product is inside), extraction, purification (precipitation, chromatography), concentration, drying, packaging.
  • Antifoam agents (silicone oils, vegetable oils) control foam.
  • Penicillin: Penicillium chrysogenum (high-yield strains); fed-batch, aerobic, 25–26 °C; precursor phenylacetic acid added to get penicillin G; corn steep liquor in medium.
  • Ethanol: Saccharomyces cerevisiae on molasses (sugarcane industry, important in Maharashtra); anaerobic; product distilled. Zymomonas mobilis is another producer.
  • Citric acid: Aspergillus niger, aerobic, on molasses or sucrose, with low iron/manganese and low pH.
  • Other products: vinegar/acetic acid (Acetobacter aceti), lactic acid (Lactobacillus delbrueckii), glutamic acid (Corynebacterium glutamicum), vitamin B₁₂ (Propionibacterium, Pseudomonas denitrificans), amylase and protease (Bacillus, Aspergillus), streptomycin (Streptomyces griseus).

महत्त्वाच्या संज्ञा

  • Bioreactor – vessel for growing microbes under controlled conditions.
  • Inoculum – starter culture added to the fermenter.
  • Precursor – compound added that becomes part of the product.
  • Downstream processing – recovery and purification of product.
  • Scale-up – moving a process from flask to pilot to industrial size.

महत्त्वाची माहिती

ProductOrganismRaw material / note
PenicillinPenicillium chrysogenumCorn steep liquor, lactose; phenylacetic acid precursor
EthanolSaccharomyces cerevisiaeMolasses; anaerobic
Citric acidAspergillus nigerMolasses/sucrose; low pH, low trace metals
Glutamic acidCorynebacterium glutamicumBiotin limitation
VinegarAcetobacter acetiEthanol → acetic acid
StreptomycinStreptomyces griseusSoybean meal medium

Equations:

  • Ethanol: C₆H₁₂O₆ → 2C₂H₅OH + 2CO₂
  • Vinegar: C₂H₅OH + O₂ → CH₃COOH + H₂O

५ महत्त्वाचे प्रश्न

Q1. Differentiate primary and secondary metabolites.

Primary metabolites are formed during active growth and are essential for the cell (ethanol, amino acids). Secondary metabolites are formed mainly in the stationary phase and are not essential for growth (antibiotics, toxins).

Q2. What is the role of the sparger and baffles in a fermenter?

The sparger releases sterile air as fine bubbles for aeration. Baffles are metal strips on the wall that break the vortex and improve mixing.

Q3. Describe the design of a typical stirred-tank fermenter.

  • Vessel of stainless steel with headspace.
  • Agitator and impellers; motor.
  • Sparger; air filter.
  • Baffles.
  • Cooling jacket/coils.
  • Probes: pH, temperature, DO, foam.
  • Ports: inoculation, sampling, harvest, acid/base and antifoam addition.
  • Draw and label.

Q4. Explain industrial production of penicillin.

  • Strain: high-yield P. chrysogenum.
  • Inoculum: spores → seed tanks.
  • Medium: corn steep liquor, lactose/glucose feed, phenylacetic acid.
  • Fed-batch, aerobic, pH about 6.5, 25–26 °C, several days.
  • Recovery: filter mycelium, solvent extraction at low pH, back extraction, crystallization as sodium/potassium salt.
  • Semi-synthetic penicillins from 6-APA.

Q5. Describe production of ethanol by fermentation.

  • Raw material: molasses diluted, pH adjusted (about 4–5), nutrients added.
  • Yeast inoculum (S. cerevisiae).
  • Anaerobic fermentation at about 30 °C for 2–3 days.
  • Distillation gives rectified spirit (about 95%); further dehydration for absolute alcohol/fuel ethanol.
  • By-products: CO₂, fusel oils.
  • Uses: fuel blending, solvents, beverages, pharmaceuticals.

परीक्षेसाठी टीप

Learn product – organism – key condition as a table, and practise drawing the fermenter with at least 10 labels.

बी.एससी. सत्र ५ · सूक्ष्मजीवशास्त्र · पाठ 8

Production of Enzymes, Amino Acids and Vitamins

थोडक्यात सारांश

Besides antibiotics and alcohol, microorganisms are used to manufacture enzymes, amino acids and vitamins on an industrial scale. Microbes are preferred because they grow fast on cheap media, can be improved genetically and give products in large amounts. This topic covers the production of amylase and protease, glutamic acid and lysine, and vitamin B₁₂ and riboflavin, along with strain improvement.

महत्त्वाचे मुद्दे

  • Why microbial enzymes? Fast growth, easy scale-up, no seasonal dependence, easy genetic improvement, and many are extracellular (secreted into the medium), making recovery simple.
  • α-Amylase – Bacillus amyloliquefaciens, B. licheniformis (heat-stable, used in starch liquefaction), Aspergillus oryzae.
  • Proteases – Bacillus licheniformis (subtilisin; alkaline protease used in detergents), Aspergillus.
  • Glucose isomerase – Streptomyces spp.; converts glucose to fructose for high-fructose syrup.
  • Pectinase – Aspergillus niger; clears fruit juices.
  • Lipase, cellulase (Trichoderma reesei), rennet substitute (microbial chymosin made in recombinant microbes).
  • Main industrial enzymes and producers:
  • Production steps for an enzyme: strain selection → medium (starch, corn steep liquor, soybean meal) → submerged or solid-state fermentation (wheat bran for fungal enzymes) → remove cells (filtration/centrifugation) → concentrate (ultrafiltration) → precipitate (ammonium sulphate or alcohol) → dry or formulate → immobilization for repeated use.
  • L-Glutamic acid (sold as monosodium glutamate, MSG) – Corynebacterium glutamicum (discovered by Kinoshita, Japan, 1957).
  • Key trick: biotin limitation (or adding penicillin or surfactants) makes the membrane leaky so glutamate is excreted.
  • Aerobic, pH about 7–8, 30–37 °C; ammonium supplied as nitrogen source.
  • L-Lysine – auxotrophic or regulatory mutants of C. glutamicum (Brevibacterium flavum) that escape feedback inhibition; used in animal feed.
  • Amino acid production:
  • Vitamin B₁₂ (cyanocobalamin) – made only by microbes; Pseudomonas denitrificans and Propionibacterium shermanii; needs cobalt and DMB (5,6-dimethylbenzimidazole); cyanide added during recovery converts it to cyanocobalamin.
  • Riboflavin (vitamin B₂) – fungi Ashbya gossypii and Eremothecium ashbyii, and Bacillus subtilis.
  • Vitamin C – partly microbial (Reichstein process step using Gluconobacter, or two-step fermentation).
  • Vitamin production:
  • Regulation tricks: use of auxotrophic mutants and feedback-resistant mutants so that cells overproduce the product.
  • Strain improvement methods: mutation (UV, chemicals) and screening, protoplast fusion, recombinant DNA, site-directed mutagenesis.
  • Uses: detergents (proteases, lipases), food (amylase in baking, pectinase in juice), textiles (amylase for desizing), leather (protease for dehairing), medicine (streptokinase, asparaginase), feed (lysine), flavour (MSG).

महत्त्वाच्या संज्ञा

  • Extracellular enzyme – enzyme secreted out of the cell.
  • Immobilized enzyme – enzyme fixed on a support so it can be reused.
  • Auxotroph – mutant that cannot make a nutrient and needs it in the medium.
  • Feedback inhibition – end product inhibits the first enzyme of its pathway.
  • Solid-state fermentation – growth of microbes on moist solid substrate with little free water.

महत्त्वाची माहिती

ProductOrganismMain use
α-AmylaseBacillus licheniformis, Aspergillus oryzaeStarch processing, baking, textiles
Alkaline proteaseBacillus licheniformisDetergents, leather
Glucose isomeraseStreptomyces spp.High-fructose syrup
Glutamic acidCorynebacterium glutamicumMSG flavour enhancer
LysineC. glutamicum mutantsAnimal feed
Vitamin B₁₂Pseudomonas denitrificans, PropionibacteriumMedicine, feed
RiboflavinAshbya gossypiiFood and feed supplement

५ महत्त्वाचे प्रश्न

Q1. Why is biotin kept limited in glutamic acid fermentation?

Biotin is needed for fatty acid synthesis. Under biotin limitation the cell membrane becomes weak and leaky, so glutamic acid leaks out of the cell into the medium instead of being used inside, giving a high yield.

Q2. Name two microorganisms used for vitamin B₁₂ production.

Pseudomonas denitrificans and Propionibacterium shermanii (P. freudenreichii). Cobalt must be added to the medium.

Q3. Describe the industrial production of amylase.

  • Organism and its features.
  • Medium, fermentation conditions (submerged or solid-state).
  • Downstream recovery and formulation.
  • Applications in starch, baking, textile and brewing industries.

Q4. Explain the production of L-glutamic acid by fermentation.

  • Corynebacterium glutamicum; medium (glucose/molasses, ammonia, minerals, limiting biotin).
  • Conditions: aerobic, pH, temperature, duration about 2–3 days.
  • Mechanism of excretion.
  • Recovery as MSG by crystallization.
  • Flow chart.

Q5. Write a note on microbial production of vitamins.

  • B₁₂: organisms, cobalt, DMB, recovery as cyanocobalamin.
  • Riboflavin: Ashbya gossypii, flavinogenic fungi.
  • Uses and importance.

परीक्षेसाठी टीप

Remember organism–product pairs from the table; examiners often ask "name the organism used for …". Draw one simple flow chart for any production process.

बी.एससी. सत्र ५ · सूक्ष्मजीवशास्त्र · पाठ 9

Single Cell Protein and Mushroom Cultivation

थोडक्यात सारांश

Single cell protein (SCP) is the dried cells (biomass) of microorganisms such as algae, yeasts, fungi and bacteria, used as a protein source for humans or animals. Mushrooms are edible fleshy fruiting bodies of certain fungi grown on agricultural waste. Both turn cheap or waste materials into protein-rich food, which matters for a country like India with protein malnutrition and plenty of crop residue.

महत्त्वाचे मुद्दे

  • The term single cell protein was coined by Carroll L. Wilson (MIT) in 1966.
  • Advantages of SCP organisms: very fast growth (bacteria double in 20–120 min, yeasts in about 1–3 h), high protein content, grow on cheap or waste substrates, need little land and water, not affected by season, and can be genetically improved.
  • Protein content (dry weight, approximate): bacteria 50–80%, yeasts 45–55%, algae 40–60%, moulds 30–50%.
  • Algae: Spirulina (cyanobacterium; about 60–70% protein, grown in alkaline ponds), Chlorella, Scenedesmus.
  • Yeasts: Saccharomyces cerevisiae, Candida utilis (grows on sulphite waste liquor, molasses), Kluyveromyces on whey.
  • Fungi: Fusarium venenatum (Quorn mycoprotein, UK), Trichoderma, Chaetomium on cellulose wastes.
  • Bacteria: Methylophilus methylotrophus on methanol (ICI's "Pruteen", animal feed); Pseudomonas on hydrocarbons.
  • Organisms and substrates:
  • Production steps: substrate preparation → sterilization → fermentation (continuous, aerated) → harvesting (filtration/centrifugation) → washing → reduction of nucleic acids → drying.
  • Problems / limitations: high nucleic acid content (bacteria up to about 15%) — excess raises uric acid and can cause gout and kidney stones (reduced by heat treatment that activates the cells' own RNases); possible toxins or contaminants; poor taste and digestibility of cell walls; high cost; public acceptance.
  • Mushroom biology: mushrooms are mostly Basidiomycetes. The fruiting body has a stalk (stipe), cap (pileus) and gills bearing basidiospores. The underground mycelium is the vegetative part.
  • Button mushroom – Agaricus bisporus; temperate (about 14–18 °C for fruiting); grown on composted straw with a casing layer. Main producer areas include Himachal Pradesh and Haryana; also grown in climate-controlled units.
  • Oyster mushroom (dhingri) – Pleurotus spp.; grows at about 20–30 °C; easiest for Maharashtra farmers; grown on chopped, soaked and pasteurized paddy or wheat straw in polythene bags.
  • Paddy straw mushroom – Volvariella volvacea; tropical, 30–35 °C.
  • Milky mushroom – Calocybe indica; suited to hot regions.
  • Common cultivated mushrooms in India:
  • Steps of oyster mushroom cultivation: spawn preparation (mycelium on boiled wheat/sorghum grains) → substrate (straw) chopped, soaked and pasteurized (hot water about 80 °C or chemical) → spawning (layering spawn in bags, about 2–5% of wet substrate weight) → spawn run in dark (about 2–3 weeks, white mycelium covers straw) → bags cut, high humidity (80–90%), light and ventilation → cropping of fruiting bodies in flushes.
  • Nutritional value: mushrooms are rich in protein (about 20–35% dry weight), B vitamins, minerals and fibre, and low in fat; they are good for diabetics and heart patients. Wild mushrooms can be poisonous (e.g. Amanita phalloides).
  • ICAR–Directorate of Mushroom Research, Solan (Himachal Pradesh), leads mushroom research in India.

महत्त्वाच्या संज्ञा

  • Single cell protein – dried microbial biomass used as protein food or feed.
  • Mycoprotein – protein-rich food made from fungal mycelium.
  • Spawn – pure mushroom mycelium grown on grain, used as "seed".
  • Spawn run – period when mycelium spreads through the substrate.
  • Casing – layer of soil-like material placed on compost to induce fruiting of button mushrooms.

महत्त्वाची माहिती

MushroomScientific nameFruiting temperatureSubstrate
ButtonAgaricus bisporusAbout 14–18 °CCompost + casing
OysterPleurotus spp.About 20–30 °CPaddy/wheat straw
Paddy strawVolvariella volvaceaAbout 30–35 °CPaddy straw
MilkyCalocybe indicaAbout 30–35 °CStraw + casing

५ महत्त्वाचे प्रश्न

Q1. Why is the high nucleic acid content of SCP a problem?

Humans break nucleic acids down to uric acid. Too much uric acid in blood can cause gout and kidney stones, so the RNA content of SCP for human food must be lowered.

Q2. What is spawn?

Spawn is the pure culture of mushroom mycelium grown on sterilized cereal grains (wheat, sorghum). It is used like seed to inoculate the substrate.

Q3. What is SCP? Give its advantages and limitations.

  • Definition and term origin.
  • Organisms with substrates (table).
  • Advantages: fast growth, high protein, waste use, no land.
  • Limitations: nucleic acids, toxins, digestibility, cost, acceptance.

Q4. Describe the production of SCP from algae or yeast.

  • Spirulina: open raceway ponds, alkaline medium (sodium bicarbonate), harvesting by filtration, drying.
  • Or yeast on molasses/whey: continuous aerated fermentation, centrifugation, RNA reduction, drying.
  • Uses as food supplement and feed.

Q5. Explain the cultivation of oyster mushroom.

  • Spawn preparation.
  • Substrate preparation and pasteurization.
  • Spawning and bagging.
  • Spawn run and cropping conditions.
  • Yield, harvesting and nutritional value.

परीक्षेसाठी टीप

Write the cultivation steps as a numbered flow chart. For SCP, always mention the nucleic acid problem — it is the most common short question.