Biology
Updated: 2008-07-02
Please send comments to: J.Elfick@uq.edu.au

9.196 Rhizopus nigricans, mouldy bread, Phylum Zygomycota
See diagram 9.196: Rhizopus, | Rhizopus nigricans, bread mould | See diagram 9.19.1: Rhizopus oryzae
Rhizopus genus have stolons, pigmented rhizoids, with sporangiophores from nodes above the rhizoids, dense white to brown cotton-like growth
Rhizopus nigricans, bread mould, stolonifer black bread mould
Rhizopus oryzae, R. arrhizuz) fermentation of foods, Japanese sake
1. Mount a small fragment of mould from old bread or decaying fruit and examine under the low power. Note the filaments that are the vegetative body of this fungus.
2. Invert a smaller flat transparent dish in the bottom of a larger flat transparent dish. Stand a piece of bread on the small dish. Add tap water to the large dish until it is half way up the side of the small dish. Cover the larger dish with its lid and leave the assembly to stand at room temperature. Under these moist chamber conditions, off-white, furry masses of mould (Mucoraceen and Aspergillacae) develop after a few days on the bread. Later these form grey-black or green sporophores.

9.197 Phytophthora infestans, Phylum Heterokontophyta
potato blight, die back, root rot, cocoa black pod
This fungus caused the potato famine in Ireland 1845. Examine the leaf of a potato plant infected with potato blight. Look for the lighter patches, or if the disease is advanced they will appear dark brown, where the leaf is attacked by the fungus. Obtain a prepared microscope slide of a section of an infected leaf and observe dark blotches on leaf tips and white mould under the leaves. Study prepared slides showing the method of reproduction.

9.198 Pythium, root rot, damping-off fungus, Phylum Oomycota
Pythium blight, cottony blight is a fungal disease of grass, especially turf grass and greenhouse seedlings. It causes small patches of blighted grass with shrivelled leaves during warm, wet periods. Observe Pythium under the low and high powers. Study prepared slides of the asexual and sexual methods of reproduction.

9.199 Mucor, Phylum Zygomycota, cheese making
See diagram 9.202
Place some damp bread under a large jar, and leave it for a few days until it becomes mouldy. Take a small portion of the mouldy bread and examine the fungus responsible with the high power. Look for spore-producing bodies of Mucor. Study prepared slides showing stages in sexual reproduction. Examine a culture of the two strains (+ and -).

9.200 Eurotium, Phylum Ascomycota
Examine a mycelium of Eurotium under high power. Note the branched, septate hyphae. Look for conidiophores producing chains of conidia. Observe perithecia attached to the mycelium by thicker hyphae of a darker colour. Gently press the coverslip to burst the wall of sterile hyphae so that the small asci will be extruded. Examine one ascus and look for the eight ascospores.

9.201 Agaricus, mushroom, Phylum Basidiomycota
See diagram 9.201: Agaricus
Agaricus bisporus, white mushrooms, Swiss brown mushroom, edible mushrooms
Examine the reproductive portion of a mushroom. Choose a ripe fructification and remove the stipe and place the umbrella-shaped part, the pileus, on a piece of paper with the gills downwards. Leave for a few days, then gently lift the pileus. See the arrangement of the gills traced on the paper by thousands of small, black, reproductive spores that have been shed. Study a prepared slide of the longitudinal section of a gill to observe its detailed structure and the means of basidiospore production.

9.202 Penicillium, Penicillin antibiotic, Phylum Ascomycota
See diagram 9.202: Penicillium | See diagram 9.202.1: Penicillium with branching conidiophores | Penicillium, penicillin antibiotic
Penicillium, soil mould
P. purpurogenum (formerly P. rubrum) plant pathogen
P. candida, P. camemberti, P. glaucum, P. roqueforti, cheese making, P. chysogenum (formerly called P. notatum), antibiotic penicillin
P. italicum, P. olivaceum, fruit parasites
Different species produce antibiotics, and are essential for production of certain cheeses, e.g. Roquefort and Camembert. Antibiotics are excreted by micro-organisms to interfere with the growth or metabolism of other micro-organisms. In the original penicillin, the R-group was a mixture and may be varied by adding molecules to the nutrient solution in which the mould is growing. Penicillin interferes with the building up of the cell wall of the cells of Gram positive bacteria.
Gram-positive bacteria appear blue or violet under a microscope from the crystal violet dye in Gram stain. Gram-negative bacteria appear pink-red. However, some bacteria have developed the enzyme penicillinase, that can destroy penicillin.

9.203 Bathroom and kitchen mould
Moulds may grow in the "grout" between bathroom and kitchen tiles. In Australia these moulds are usually Penicillium, Rhizopus, Phoma, Phialophora, and Fusarium. They are mostly harmless although Phoma may cause allergies. They can be cleaned off with bleach solutions.

9.204 Yeast population, bakers' yeast Saccharomyces cerevisiae, Phylum Ascomycota
See diagram 9.204: Yeast cell forming bud
The simplest form of asexual reproduction is budding, in which a protuberance grows out from the parent cell and finally cutting itself off to become an independent cell. Budding occurs in yeasts. Natural sources of yeast include the wax-like coatings on smooth skinned fruits, especially grapes. However, bakers' yeast is usually obtainable. It reproduces rapidly, making it a good subject for observing population changes under varying conditions.
1. Set up tubes of sugar, molasses or honey solutions and a water control. Put a quarter cake of crumbled yeast in each tube. Compare the results. Put a one-hole stopper with glass tubing in the sugar yeast solution and allow the gas produced to bubble through a tube of clear limewater to detect the presence of carbon dioxide. Test the gas produced by the sugar yeast solution.
2. Fill a test-tube half full of 10% sugar solution, drop in a piece of bakers' yeast, plug the test-tube with cotton wool and allow to stand at room temperature. After 12 to 24 hours, transfer a drop from this culture on to a microscope slide using a glass rod, put a coverslip over it and examine the preparation under a microscope. Look for cells with protrusions or asexual buds. If you can see nuclei in the cells, look for their presence in the buds.
9.205 Sampling yeast populations
1. Start a culture each day for 10 days using one grain of yeast for each culture. On the tenth day sample and count all cultures with a microscope. Use a special slide for counting blood cells, but it is not essential. If the population on a given day is too large to count, dilute a sample by adding 1 mL of sample and 9 mL of water. Multiply the count by 10 to get the actual sample size. If one dilution is not enough, further dilution may be done until it becomes easy to count the number of organisms. The multiplication factor for two dilutions is 10 by 10 or 100: for three dilutions it is 10 by 10 by 10 or 1, 000. Note that each successive dilution is started from part of the previous dilution, not from the original sample. Graph the data obtained from cultures for analysis. Time is the independent variable and population size is the dependent variable.
2. An effective method of studying population growth with micro-organisms is to start a culture each day and, on the final day, to sample and count all cultures. For example, a new yeast culture can be started each day for 10 days, using one grain of yeast for each culture. On the tenth day samples are taken from each culture and counted with a microscope. A special slide for counting blood cells is desirable, but not essential. If the population on a given day is too large to count, dilute a sample by adding 9 parts water to 1 part of sample. Use 1 ml of sample and 9 ml of water. Multiply by 10 to get the actual sample size. If one dilution is not enough, make further dilutions until it becomes easy to count the number of organisms. The multiplication factor for two dilutions is 10 by 10 or 100: for three dilutions it is 10 by 10 by 10 or 1, 000. Note that each successive dilution is started from part of the previous dilution, not from the original sample. Graph the data obtained from cultures for analysis with time as the independent variable and population size is the dependent variable.

9.206 Find wild yeasts in flowers
See also 4.1.2: Enrichment of wild yeast strains
Put a few freshly picked flowers of deadnettle, woundwort or nasturtium, in a beaker. Cover over with a suitably sized transparent plastic bag and leave the flowers to stand in this "humidity chamber" at room temperature. After two days remove some flowers that are already open. Pull off the petals and squeeze the nectar from the nectaries onto a microscope slide. Put a coverslip over the nectar and examine it under a microscope. Look for nectar yeast cells. Note the shape of the nectar yeast cells and the size of the buds. Compare the nectar yeast cells with the bakers' yeast cells. Observe the cross form or star form of the branching chains formed by the wild yeasts on budding. Use agar syrup to culture the yeasts and moulds.

9.207 Lichens
A lichen is the symbiosis of a fungi and algae. The photosynthetic partner in the lichen thallus is called a photobiont. Each type of lichen has a characteristic shape. Crustose lichens form a thin crust. Foliose lichens are like leaves. Fructicose lichens are stalked. Squamulose (loosely attached thallus) lichens have a primary and a secondary thallus. The fungi are mainly Ascomycetes. Lichens may be nitrogen-fixing. Lichens are found in most places in the world and are very susceptible to air pollution in the atmosphere. They may be damaged by sulfur dioxide or other harmful gases, and may die. So lichens are sensitive plant detectors of air pollution. The fungi are mainly Ascomycetes. Example of common lichens include the following: Acarospora, a crustose lichen that forms a thin crust, Cladonia, cup lichen, reindeer "moss", a squamulose, loosely attached thallus lichen with a primary and a secondary thallus, Evernia, oak moss lichen, Hypogymnia, a foliose lichens like a leaf, Lobaria, lungwort, a nitrogen-fixing lichen, Peltigera, dog lichen, a nitrogen-fixing lichen and herbal medicine, Stricta, a nitrogen-fixing lichen, Usnea, a fructicose lichen that has a stalk.
1. Examine a specimen of a common lichen. Study a prepared slide showing a transverse section of the thallus revealing the presence of alga and fungus. Look for the penetrating haustoria.
2. Attach 10 lichens on a board. Then put them in three places, e.g. near a factory, in the central city and far from the factory. Observe any change in the lichens after one mouth. Count the percentage of damaged lichens. Lichens indicators are used as indicators of air pollution and species diversity.

9.208 Slide culture preparation to identify fungi
See diagram 9.208: Simple agar block method
1. To identify many fungi you must observe the conidiophores, i.e. the specialized hyphae where asexual spores cut off at the ends, and the way in which spores are produced. This method allows you to study fungi with little disturbance. Use one agar plate of nutrient agar, e.g. potato dextrose agar.
2. Use a sterile blade to cut out an agar block, 7 x 7 mm, small enough to fit under a coverslip.
9. Flip the agar block up onto the surface of the agar plate. Inoculate the four sides of the agar block with spores or mycelia fragments of the fungus to be grown.
4. Place a flamed coverslip centrally upon the agar block. Incubate the plate at 26oC until growth and spore-forming have occurred. Remove the coverslip from the agar block.
5. Apply a drop of 95% alcohol as a wetting agent. Gently lower the coverslip onto a small drop of lactophenol cotton blue on a clean glass slide. Leave the slide overnight to dry and later seal the edges with a coat of clear fingernail polish followed by one coat of red coloured polish.

9.209 Rhizobium in legumes
See also 3.30: Nitrogen-fixing bacteria | See diagram 9.209: T.S. Root nodule | See diagram 9.72: Root nodules
Rhizobium bacteria can enter the roots of legumes and cause lumps, root nodules, where they live. These bacteria can use the nitrogen gas from the air. We say Rhizobium can "fix" nitrogen from the air. Very few other living things can fix nitrogen. Some nitrogen goes into the stems and leaves of the legume plant. When the leaves fall off, some nitrogen compounds are added to the soil that other plants can then use. When the legume plants die, the nitrogen fixed by the Rhizobium can still be available to other plants. Some legumes, e.g. cowpea and lablab bean, are grown until the flowering stage and then dug into the soil as "green manure". When these plants rot, they leave nitrogen compounds in the soil both from the leaves and root nodules. Different crop plants require different strains of Rhizobium. Farmers can purchase commercial innoculum containing the strain appropriate for nodulation of their particular crop. Dig up different legume plants and count the number of nodules and note the sizes of the nodules. Cut open some nodules. The inside of a nodule that is fixing nitrogen is a pink-red colour. Inactive nodules are brown or green inside.

9.210 Grass in water
Put dead grass or fresh hay in a jar, cover with water and leave it uncovered to stand at room temperature for some days. Do not use mouldy hay because it may contain pathogenic fungi. After a few days, a skin forms on the surface of the water, the mould pellicle. After eight days, use a glass rod to transfer a part of the pellicle and some fluid to a microscope slide. Spread out the pellicle evenly and apply a coverslip. Examine the pellicle under the microscope. Observe rod-shaped or spherical unicellular bacteria. Observe unicellular organisms moving very rapidly through the field of view, e.g. Paramecium.

9.211 Teeth scrapings
Put white scrapings from the teeth on a microscope slide. Examine the scraping under a microscope and look for any bacteria. You may have to seek approval to work with samples from the human mouth because diseases may spread. Wear safety glasses and disposable surgical gloves.

9.212 Ginger beer plant
The Ginger beer "plant" is the yeast Saccharomyces florentinus (S. pyriformis) and the bacterium Lactobacillus hilgardii (Brevibacterium vermiforme). Add 5g of bakers' yeast to 25 ml of water and 1/2 teaspoon of ground ginger and sugar. Put in a screw-top jar and add half a teaspoon of sugar and some ground ginger every day for a week to keep the fermentation going. Dissolve 50g of sugar in 50 ml of boiling water. Strain the mixture and add the sugar solution and lemon juice to the liquid. Ginger beer soft drink is made from ginger powder and does not contain alcohol.

9.213 Viruses
See also 10.9.0: Sexually transmitted infections, STIs, HIV and AIDS
A virus is a strand or strand of nucleic acid covered by protein and sometimes a membrane. Viruses cause infected cells to produce progeny viruses. Retroviruses use the enzyme reverse transcriptase to copy the viral RNA (ribonucleic acid) into DNA (deoxyribonucleic acid). Plant viruses can usually be recognized by marks on leaves, e.g. mosaics, leaf streaks, and ring spots. Viruses are not affected by antibiotics. A bacteriophage is similar to a very small virus. It infects bacteria.
The classification of viruses can be based on the type and arrangement of the genetic material.
Group 1. dsDNA Double-stranded DNA viruses include oral herpes, genital herpes, chickenpox viruses, cold sore, Herpes simplex virus, types 1 and 2, HSV-1 and HSV-2, Adenoviruses human adenoids, tonsils, Human Papilloma Virus, HPV causes genital warts Condylomata acuminata, Molluscum Contagiosum Virus
Group 2. ssDNA Single-stranded DNA viruses include some of the smallest viruses.
Group 9. dsRNA Double-stranded RNA include viruses responsible for diarrhoea in children.
Group 4. positive sense ssRNA Single-stranded RNA viruses include influenza, hepatitis A virus, hepatitis C virus, HCV
severe acute respiratory syndrome SARS, foot-and-mouth disease, yellow fever, rubella viruses and most plant viruses
Group 5. negative sense ssRNA Single-stranded RNA viruses include influenza, measles, mumps, rabies, Ebola virus, foot-and-mouth disease
Group 6. ssRNA Diploid single-stranded RNA viruses that use reverse transcriptase, retroviruses, include HIV virus.
Group 7. ds DNA-RT Circular double-stranded DNA viruses that use reverse transcriptase, include hepatitis B Virus, HBV
Human Immunodeficiency Virus, HIV, and AIDS, acquired immune deficiency syndrome

9.213.1 HSV-1 and HSV-2
HSV-1, herpes simplex virus causes cold sores, painful blemishes of the mouth. It can become dormant for years, when drugs cannot affect it, then, years later, be revived by excessive sunlight or fever to cause a cold sore in the same place as before.
HSV-2 causes painful genital sores and chicken pox virus that can return late in life as shingles.

9.214 Aspergillus
See diagram: 9.203: Conidial head
Aspergillus flavus produces an aflotoxin, common soil saprophyte, pathogen of humans and animals
Aspergillus niger, black mould on food, common infection outside ear drum, common laboratory contaminant
Aspergillus oryzae

9.215 Candida (formerly Torulopsis)
Candida genus has yeast-like cells, blastoconidia, that reproduce by budding. All members of the genus occur naturally on humans. Candida albicans, forms cream coloured, smooth surface waxy colonies on an agar plate. It is a commensal of the gastrointestinal tract. However it may cause candidiasis (candidosis), a common yeast infection, especially of the vagina, where it is called thrush. Candida tropicalis is part of the normal mucocutaneous flora but may cause causes septicaemia and candidiasis. It has been isolated from polluted water, soil, and air contaminated by human excreta.
Candida albicans commensal on mucous membranes and gastrointestinal tract, pathogenic, causes thrush (candidosis)
Candida glabrata common yeast species on human body surface, pathogenic

9.216 Cryptococcus
C. gattii causes mass lesions in the lung and brain. C. neoformans causes fungal meningitis.
9.217 Fusarium
Most Fusarium species are soil fungi. Some are plant pathogens causing stem rot, root root and fruit rot.
F. oxysporum causes Panama disease of bananas, fusarium wilt and dangerous infections in human burns victims.
F. graminearum infects barley

9.218 Microsporum
M. audouinii
and M. ferrugineum cause non-inflammatory infections of the scalp, tinea capitis, especially in children.
M. canis causes ringworm, especially in children. The infection comes from cats and dogs.
M. gallinae causes "white comb" lesions in chickens.
M. nanum causes chronic non-inflammatory lesions in pigs. Infected pigs may infect humans.

9.219 Trichophyton, ringworm disease
Many strains of Trichophyton as medical fungi exist but they are generally classified as T. rubrum, downy type, and T. rubrum granular type. The downy type have slender microconidia and no macroconidia. They cause chronic infection of the skin and nails. The granular type have microconidia and cigar-shaped macroconidia. They cause tinea corporis in South-East Asian and in aborigines in Northern Australia. Also, it causes tinea pedis, athlete's foot, in feet in Europe and North America, favus, a chronic scarring form of tinea capitus, a type of ringworm of the scalp, and dhobie itch, tinea cruris, that affects the groin and nearby regions. Some strains can invade human hairs and cause the "black dot" tinea capitis.

9.220 Puccinia
Wheat rust, white pine blister rust, apple cedar rust, hollyhock rust, asparagus rust