Bacterial characterization

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Microbiology Assignment3
Page 1 of 5
Bacterial Characterization
Submitted to: Sir Sanabil Yaqoob
Submitted by: Zakia Akram
L1F19BSFT0115
UNIVERSITY OF CENTRAL PUNJAB
FACULTY OF LIFE SCIENCES
YEAR 2020

Microbiology Assignment3
Page 2 of 5
BACTERIAL CHARACTERIZATION
Bacteria Examples Staining
method
Mode of
Nutrition
Mode of
reproducti
on
Growth sequence Culture media
Cocci
 Streptococcus
 Staphylococcus
 Aerococcus
 Pediococcus
 Tetragenococcus
Gram Positive
+
Heterotrophic Binary
Fission
pH. Temp Time  Phenyl ethyl
alcohol agar
 Mannitol salt
agar
 Bile Esculin
Agar
Bacilli
 Bacillus thuringiensis
 Bacillus popilliae
 Bacillus subtilis
 Bacillus cereus
 Bacillus anthracis
Gram Positive
+
Heterotrophic Binary
Fission
 Tryptic soyagar
 Hektoen enteric
agar

Microbiology Assignment3
Page 3 of 5
Spiral
 Spirillum volutans
 Spirillum minus
 Treponema pallidum
 Helicobacter pylori
 Campylobacter jejuni
Gram
Negative
-
Heterotrophic
Autotrophic
Chemotrophic
Binary
Fission
 Nutrient agar
 Xylose lysine
deoxycholate
Vibrio  Vibrio cholerae
 Vibrio
parahaemolyticus
 Vibrio anquillarum
 Vibrio vulnificus
 Vibrio alginolyticus
Gram
Negative
-
Heterotrophic Binary
Fission
 Xylose lysine
deoxycholate
 Thiosulfate‐
citrate‐bile
salts‐
sucroseagar
Spirochete
 Spirochaeta
 Borrelia burgdorferi
 Brachyspira pilosicoli
 Borrelia garinii
 Borrelia afzelii
Gram
Negative
-
Heterotrophic Binary
Fission
 Xylose lysine
deoxycholate
 Borrelia-
specific
Barbour-
Stoenner-
Kelly media

Microbiology Assignment3
Page 4 of 5
Actinomycetes
 Actinomycetes
 Thermoleophilia
 Bifidobacteriatales
 Streptomyces
venezuelae
 Nocardia asteroides
Gram Positive
+
Heterotrophic Conidia
(spore
formation)
 Mannitol salt
agar
 Starch
caesin media
Mycoplasma
 Mycoplasma genitalium
 Mycoplasma hominis
 Mycoplasma pneumoni
a
 Ureaplasma parvum
 Ureaplasma urealyticum
Gram Positive
+
Heterotrophic Binary
Fission
Budding
 Mannitol salt
agar
 Oxoid Mycoplasm
a Agar Base

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The genus Veillonella was first isolated by Veillon and Zuber in 1898. Prevot further described these bacteria in 1933. Morphology: • Gram-negative anaerobic cocci , 0.3-0.5 um in diameter • Non motile • non spore forming Habitat: • mouth • upper respiratory tract • Intestine • vagina Characteristic: • pH Optimum pH, 6.5-8.0. • Temperature Optimum temperature, 30-37`C. • usually isolated mixed with other bacteria (These organisms are hypothesised to form a cooperative metabolic system in which a waste product, lactic acid, produced by S. mutans is utilised by V. dispar.) • Chemoautotrophs (Chemoautotrophs obtain their energy as well as their carbon from inorganic sources; they do this by the oxidation of inorganic molecules such as sulphur or nitrite.) • Non-fermentative. • produce a highly endotoxic lipopolysaccharide • are generally resistant to tetracycline Species: Veillonella atypica, V. dispar are found in humans V. parvula V. caviae V. criceti in rodents. V. ratti V. rodentium Diseases: Dental caries Abscesses of teeth and gums. sinusitis Osteomyelitis Endocarditis Laboratory diagnosis:  Are small, spherical, gram-negative cocci appearing as pairs, masses, and short chains.  Non hemolytic  Growth may be improved by adding 1% sodium pyruvate and 0.1% potassium nitrate to the basal medium.  Colonies grow slowly on normal laboratory media, their growth being encouraged by factors such as lactate, pyruvate and nitrate.  enriched thioglycollate broth  Some form catalase.  Small white colonies.  Colony produce of a red fluorescence at a wavelength of 365 nm. This fluorescence is better observed in a Veillonella selective medium (Rogosa medium) and disappears in contact with oxygen.  Growth is usually visible after 48 hours in appropriate anaerobic conditions.  glucose or any other carbohydrate is not fermented;  indole -ve  nitrate is reduced.  H2S is produced  propionic and acetic acids, C02, and H2 are produced from lactate during growth. Lactate 2 propionate + acetate + CO2  Oxidase-negative. Catalase-negative, but some species produce an atypical catalase lacking porphyrin.

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CULTURE METHODS Indications for culture - Isolate bacteria in pure cultures. Demonstrate their properties. Obtain sufficient growth for preparation of antigens & for other tests. Typing bacterial isolates. Antibiotic sensitivity. Estimate viable counts. Maintain stock cultures. Streak culture or surface plating Lawn or carpet culture Stroke culture Stab culture Pour plate method Anaerobic methods of culturing bacteria Streak Culture Routinely employed for isolation Platinum / Nichrome loops LAWN OR CARPET CULTURE Uniform surface growth Bacteriophage typing Antibiotic sensitivity testing Preparation of bacterial antigens & vaccines STOKE CULTURE Tubes containing agar slopes For slide agglutination & other diagnostic tests. STAB CULTURE By puncturing a suitable medium with a long, straight charged wire. For gelatin liquefaction, stock cultures & motility POUR PLATE METHOD 1 ml of appropriately diluted inoculum is added to 15 ml of molten agar and poured on petridish. Colonies appear through out the depth of medium. Used to estimate viable count, recommended method for quantitative urine cultures. BROTH CULTURE Inoculated by a charged loop, pipette or syringes. For blood cultures & sterility testing.

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Pitha‟ is one of the most popular traditional foods made up from various kinds of cereal grains like rice, wheat‟s, flours etc. The microbiological quality of pitha on retail sale in the street of Dhaka city was studied with thirty six different samples by detecting the range of total viable count (TVC) of bacteria, indicator microorganisms, such as Escherichia coli, Enterobacteriaceae and the common food pathogens Staphylococcus aureus, Bacillus cereus., Salmonella Spp. and Pseudomonas spp. In our study, the majority (67.5%) of the microbial contaminants were gram negative rods. The TVC of bacteria was ranged from (9.0x103) cfu/g to (1.2x106) cfu/g. Total coliform, fecal coliform and E. coli were detected from 30.56%, 22.2% and 13.8% of the samples respectively indicating unsatisfactory results. For Salmonella spp. 16.6% of the samples were unable to reach the acceptability levels. Besides, according to the ICMSF (International Commission on Microbiological Specifications for food), in terms of Pseudomonas spp. 33.3%, for Staphylococcus aureus 12.2% and for Bacillus cereus 11.1% of the samples were not suitable for the consumption. All these phenomena indicate poor hygiene practices. This study emphasizes the need to improve production hygiene.

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Common carp (Cyprinus carpio) is one of the commercially important and commonly cultured fish. In the hatchery intensive incubation leads to microbial overgrowth in C. carpio eggs that hamper egg development, hatchability and larval survivability. The aim of this study is to find out causes of mass mortality in C. carpio eggs during peak- breeding season between March to May 2015 at Mafatema fish hatchery, Chanchra, Jessore sadar upazilla. In the present study three disinfectants with three different concentrations in each such as methylene blue 1, 3 and 5mg/L., malachite green 1, 3 and 5mg/L., sodium chloride 1, 2 and 3g/L were used to observe the hatching rate of fertilized eggs and survival rate of larvae. Bacterial load of culture water was examined during the induced breeding of C. carpio with mycological examination of egg samples with different disinfectants. The total bacterial count fluctuated from 3.4 x 108 CFU/ml to 32.7 x 108 CFU/ml during the period of fertilization to 4days of hatching. The fertilized eggs infected by Saprolegnia spp. were appeared as tuft hairy like balls with a white cottony envelop. Among all the treatment 1mg/L methylene blue, 3mg/L malachite green and 1g/L sodium chloride showed significantly better (P<0.05) hatching rate 95·33±2·08, 88.00±2.64 and 92.33±4.04% respectively. The same concentration of methylene blue, malachite green and sodium chloride showed significantly better (P<0.05) better survival rate 95·00±4.35, 75.00±3.00 and 87.00±6.24% respectively. Finally among all the treatment 1mg/L of methylene blue showed significantly better (P<0.05) hatching and survival rate 95·33±2·08% and 95·00±4.35 % respectively. So 1mg/L of methylene blue is the best disinfectant for C. carpio fertilized egg treatment.

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