Material needed in phytobacteria studies

1. Equipment for A standard Phytobacteriology
Introduction
A wide range of laboratory instruments, glass ware and plastic goods is available from scientific equipment
supplies, but unfortunately not always suitable for microbiological work. The best is not always the most
expensive, but rarely the cheapest. Basic equipment that has been found practicable in Phytobacteriology
(i.e. for isolation, cultivation, storage and testing of Phytobacteriology bacteria).
I. Glassware, plastics goods etc
a. Petri Dishes: 10 cm plastic & glass petri dishes. Glass Petri dishes may be cheaper, because on the average one
dish can be used ca. 100 times.
b. Test tube and bottles: Rimless test tube of heavy quality are made for bacteriological work. For storage and
shipping screw caped tubes are often preferred.
c. Erlenmeyer flask : 50,100,300,500,100 ml volume

2. Equipment for A standard Phytobacteriology…continued
I. Glassware, plastics goods etc…continued
d. Beakers : 50, 100, 250, 500,1000 ml
e. Measuring cylider 100, 500, 100 ml
f. Graduate pipettes (0.1 to 25 ml)
g. Pastuer Pipettes

3. Equipment for A standard Phytobacteriology…continued
I. Glassware, plastics goods etc…continued
g. Durham’s tube (25-30 x 5-6 mm)
h. Pluggs: cotton wool, metal or plastic cap, soft synthetic sponge bungs
i. pipetting aide: rubber bulbs, syringe like machins that holds pipettes more rigidly.
j. Spreader from glass rod in L shape =Drigalski Spatula
k. Inoculation loops and wires: usually made of 25 SWG Nicro wire,

4. Equipment for A standard Phytobacteriology…continued
I. Glassware, plastics goods etc…continued
l. Forceps, Knives scalpels dissecting needle, syringes
with hypodermic needles,thermometer(0-100 oC
m. Mortar and pestle in different sizes
n. Racks and baskets: Test tube and culture bottle
racks should be made of polypropylene (as wire
baskets) of metal covered with polypropylene or
nylon so that they can be autoclaved.

5. Equipment for A standard Phytobacteriology…continued
II. Instrument and equipment
a. Bunsen burner
b. Whirl mixer for test tube ( e.g. Mixomat)
c. Magnetic stirrer, preferable with heating plate
d. Eppendorf pipette diveces variable volume
e. pH meter
f. Sterile filtiration
g. Colony counter
h. laboratory precision balances (operation at 1 mg-100g) e.g Sartorius type
i. Spectrophotometer ( eg. Spectronic 20, Bausch and Lomb)
j. Spray gun, special glass atomizer for plant inculation
k. Blender and grinders
l. Centrifuges: The ordinary laboratory centrifuge capable of exerting a force up to 3000g. To deposit bacteria
within a reasonable time. For general microbiology , a centrifuge capable of holding 15 and 50- ml buckets and
working at speed of 6000 rev/minute is adequate

6. Equipment for A standard Phytobacteriology
II. Instrument and equipment…continued
m. Incubator : usually operated at 15-50 0C
n. water bath: The content of a test tube placed in water bath are
raised to the required temperature much more rapidly than in an
incubator.
o. Gyratory shares: for liquid cultivation of bacteria
p. Refrigrators, deep freezers
q. Glassware washing machine
r. Distillation apparatus or ion-exchange rrsin
s. Dry heat oven
t. Autoclaves

7. Temperature of moist heat required to kill bacteria
Vegetative cells of microorganisms or spores Moisture temperature exposure for 15 minutes
Phytopathogenic bacteria >50 0C
Saprophytic bacteria >600C
Fungi >600C
Fungal spores >800C
Bacterial spores >1200C

8. Supplied and consumable Items
1. Ingredients for preparation of Media (Agar, peptone, antibiotics,
enzymes etc)
2. Disinfectant: ethanol, clorox
3. Plastic bags
4. Paper bags
5. Writing inks
6. Stickers (Labeles)
7. Filter paper
8. Tissue paper

9. Table. List of some laboratory Chemicals and Reagents for
Phytobacteriology Laboratory
No Chemical/Reagent Name Formula Remark
1 Agar, bacteriological
2 Agar, Oxoid
3 Ammonium chloride NH4Cl
4 Ammonium di-phosphate NH4H2PO4
5 Ammonium oxylate
6 Beef extract
7 Bromothymol blue
8 Calcium carbonate CaCO3
9 Calcium nitrate Ca (NO3)2)
10 Casein hydrolysate

10. Table. List of some laboratory Chemicals and Reagents for
Phytobacteriology Laboratory…continued
11 Cellobiose Carbon source
12 Crystal violet
13 Dextrose
14 D-Glucose
15 Dulcitol Alcohol
16 Ethanol 95% Alcohol
18 Glucose
19 Iodine
20 Iron sulphate (Ferrous
sulphate)
FeSO4.7H2O

11. Table. List of some laboratory Chemicals and Reagents for
Phytobacteriology Laboratory…continued
21 Lactose
22 Magnesium Sulphate MgSO4.7H2O
23 Malt extract
24 Maltose
25 Manganese sulphate MnSO4.7H2O
26 Manitol
27 Methyl green
28 Methyl violet B
29 Nutrient broth
30 Peptone, bacteriological (oxoid)

12. Table. List of some laboratory Chemicals and Reagents for
Phytobacteriology Laboratory…continued
No Chemical/Reagent Name Chemical formula Remark
31 Phenol Red
32 Potassium bromide KBr
33 Potassium chloride KCl
34 Potassium iodide KI
35 Potassium phosphate (Dibasic) K2HPO4
36 Potassium di phosphate KH2PO4
37 Potassium hydroxide KOH
38 Potassium Nitrate KNO3
39 Proteose peptone (Difico No.3)
40 Safranin

13. Table. List of some laboratory Chemicals and Reagents for
Phytobacteriology Laboratory…continued
41 Sodium chloride NaCl
42 Sodium di hydrogen
phosphate
NaH2PO4
43 Sodium hydrogen
phosphate
Na2HPO4
44 Sodium hydroxide NaOH
45 Sorbitol
46 Sucrose
47 Starch (hydrolyzed)
48 Yeast Extract
49 Antibiotics
50 Bacitracin

14. Table. List of some laboratory Chemicals and Reagents for
Phytobacteriology Laboratory…continued
51 Cycloheximide
52 Cephalexin
53 Chloramphenicol,
54 D-methionine
55 Gentamycin
56 Kasugamycin
57 Nitrofurantoin
58 Polymyxin B-sulphate
59 Trimethoprim
60 Vancomycin

15. Glasshouse or Climate chamber
 Modest glasshouse or growth chamber to keep
experimental pots and growing on tests.

16. Safety Precautions and Principles in Phytobacteriology
Laboratory
 Safety precautions have to followed during microbiological work, because toxic
substances( antibiotics, bactericides, enzyme inhibitors etc) are used and
becuasesome plant pathogenic bacteria are also asssociated with human disease
(eg. Pseudomonas aerugina, Pseudomonascepacia and Erwina herbicola).
Instruction in safety should be a part of the general training given to all laboratory
workers
Safety Practices
 Food, candy, gum and beverage for human consumption should not be stored or
consumed in the laboratory
 Smoking should not be permitted
 Proper overalls should be worn at all times and removed before leaving laboratory.
They should be kept apart from outdoor and other clothing.
 Keep hands away from the mouth, nose, eyes, face , and hair to prevent self
inoculation
 Keep personal items such as coats and shoes out of the labratory

17. Safety Precautions and Principles in Phytobacteriology
Laboratory…continued
Safety Practices…continued
 Wash hands frequently and always after removing soiled protective clothin, before
leaving the laboratory area, before eating.
 Keep laboratory and incubators doors closed
 Use mechnical pipetting device when you work with dangerous solutions and
suspension
 A supply of suitable disinfectant at used dilution should be available at every work
station
 Bench and work surfaces should be disinfecfed regularly and after spillages of
potentially toxic or infected materials
 All contaminated items should be autoclaved or otherwise disinfected as soon as
possible before use washing or re-using.
 Disinfect and autoclave all hypodermic syringes and needles before disposal.
 It is cardinal use rule that no infected material shall leave laboratory.

18. Limitations for proper supply and acquisitions of
laboratory chemicals/Reagents in Ethiopia
 Few or limited regular/specialized/authorized importer and supplier
 Expensive nature of the materials
 Existing supplier only import up on request, provided the required
items are in bulk
 Some reagents require special handling