Collection and Identification of Plant Parts and Dental Caries Pathogens.
Plant Materials Collection
The small branches of locally available Z. zanthoxyloides and T. glaucescens were collected from local Ogbete main market Enugu and were authenticated by Prof. Okigbo R.N of the department of Botany of Nnamdi Azikiwe University, Awka. These plant materials were dried under the sun for two weeks and also cut into pieces of approximately 15cms and transferred to the oven set at 45°C for 20-30mins before it was reduced to fine powder with the aid of mechanical grinder. The powder plant materials were collected and stored in a tightly covered glass jar for further studies.
Collection and Recovery of Caries Sample
The dental extracts from 20 (twenty) patients with clinical features of dental caries were collected from School of dental technology and therapy clinic, Trans-ekulu, Enugu. The patients comprised of 8 males and 12 females with ages ranging from 16-40 years.
The samples were collected under strict aseptic conditions. Prior to the collection of dental caries sampling, patient was made to rinse the tooth with water. The tooth and the surrounding field were cleaned with 3% hydrogen peroxide and then decontaminated with a 2.5% sodium hypochlorite solution. The food debris on the chewing surface was removed using a dental excavating instrument .The tooth was then extracted by a clinician and then introduced into the 20ml broth of Brain Heart Infusion (BHI) in appropriate sterile screw cap bottles. The dental caries sample was collected from the extracted tooth using an excavator under aseptic conditions. The clinical samples were mixed well using a magnetic stirrer before incubation. The samples were then inoculated using the streak plate technique on to nutrient and sabouraud dextrose agar under various culture conditions- aerobic, microaerophilic, and anaerobic culture conditions for each patient sample (Holding and Colee, 1971).
The organism isolated was identified on the basis of morphological, cultural and biochemical characteristics according to standard procedures (Holding and Colee, 1971).
2.3 PREPARATION AND EXTRACTION OF PLANT MATERIALS.
Ethanol Extraction
20g of fine-powder stem of Z. zanthoxyloides and T. glaucescens was weighed and soaked in 200mls of ethanol in a conical flask and kept at room temperature (25°C) in a rotary shaker for 48 hours. After 48 hours, filtered through Whatman No1 filter paper; solvent was allowed to evaporate and stored at room temperature until when required for use.
2.5 IDENTIFICATION OF THE ISOLATES
The isolated organisms were identified using sub-culturing, gram staining technique and biochemical test like catalase test, indole test, coagualase test, methyl red test, oxalase test, sugar fermentation test.
Table 6 shows the results of the biochemical tests. The isolate can obtain its energy by means of aerobic respiration but not fermentation. In the Oxidation-Fermentation test, a yellow color change was produced only under both aerobic conditions, indicating that the EI can oxidize glucose to produce acidic products. In addition to glucose, the EI can also utilize lactose and sucrose, and this deduction is based on the fact that the color of the test medium broth changed to yellow in all three Phenol Red Broth tests. These results are further supported by the results of the Triple Sugar Iron Agar test. Although the EI does perform fermentation of these three carbohydrates, it appears that this bacterium cannot perform mixed acid fermentation nor 2,3-butanediol fermentation due to the lack of color change in Methyl Red and Vogues-Proskauer
After 5 days of growth each slant was tested using the gram staining technique to confirm the complete isolation of the bacteria. Both isolations were completely successful. Then each sample of bacteria was subjected to a series of tests for identification.
Streak plate technique was used to isolate pure culture for each bacteria (2). The Gram stain was used to determine Gram reaction and morphology of each bacteria (2) Selective and differential media such as, salt agar, MacConkey agar and blood agar were used for bacterial identification (2). Gelatin deeps were inoculated to detect production of gelatinase (2). Starch Agar plate were inoculated to detect amylase (2). Ocular reticle used to determine bacteria size (2). Motility deeps were inoculated to detect motility on bacteria (2). Thioglycollate broth used to determine oxygen requirements (2). Carbohydrate fermentation
The results of these tests prove that the unknown organism is Citrobacter freundii hereby referred to as C. freundii. C. freundii is a member of the Enterobacteriaceae family, like all the other unknowns given in this test. The species is a facultative anaerobic and is a gram-negative bacilli. C. freundii is a non-spore forming, motile bacteria that are long rod-shaped with a typical length of 1-5 μm.
The purpose of this laboratory is to learn about cultural, morphological, and biochemical characteristics that are used in identifying bacterial isolates. Besides identifying the unknown culture, students also gain an understanding of the process of identification and the techniques and theory behind the process. Experiments such as gram stain, negative stain, endospore and other important tests in identifying unknown bacteria are performed. Various chemical tests were done and the results were carefully determined to identify the unknown bacteria. First session of lab started of by the selection of an unknown bacterium then inoculations of 2 tryptic soy gar (TSA) slants, 1 nutrient broth (TSB), 1 nutrient gelatin deep, 1 motility
The bacterium’s colonies on trypticase soy agar had an irregular shape, yellow pigment, umbonate elevation, and entire margin. Through Gram staining, I microscopically examined that the bacterium was Gram positive, rod shaped, and arranged in chains. There was presence of bubbles after testing for catalase, therefore it was a catalase positive. The bacterium was an Endospore positive because after a close examination under a microscope, I observed that it had Subterminal endospore as the its position. The Acid from mannitol test resulted as positive because the phenol mannitol red broth became yellow orange. Finally, the V-P test was negative because the media turned yellow. Overall, these results means that the unknown bacteria were Bacillus
Firstly, samples were taken out carefully. The frozen tissues of sea cucumbers were thawed in the sink with running tap water followed by multiple washes using distilled water to remove the foreign particles. The surgical blades, surgical blade-holder, and labelled sample tubes were prepared. Then, the tissue samples were cut-sliced from each samples followed by storage in the properly labelled tubes. Each sample was stored in separate tube. Different blade was used for each sample. The obtained in-tube-samples were stored in -20 oC freezer for the next step of DNA extraction.
Experiment #3: The purpose of this experiment to test the chromatography of plant pigments the alcohol test strip test will be used.
Children are often sweet and adorable, but many times they may be eating too many sweets. According to the Channel Four News, it has been shown that the number one disease in children is the reoccurrence of caries, also known as cavities. Could it be too many sweets or could it be the lack of flossing and brushing teeth? For instance, many children go to bed or wake up without brushing their teeth and go throughout the day eating all sorts of food and candy. The result of eating and not brushing could lead to plaque build up and decaying of teeth. This leaves a child extremely unhappy and in excruciating pain. So who is going to fix this pain? Who is going to help prevent decay in millions of people’s mouths? Today, the world is lucky to have a well-studied field of dentistry.. Dentistry has much history, various specialties, advancements, and an irresistible salary.
I identified the genus and species of an unknown bacterial culture, #16, and I applied the following knowledge of morphologic, cultural and metabolic characteristics of the unknown microorganism according to the laboratory manual as well as my class notes and power point print outs. I was given an incubated agar slant labeled #16 and a rack of different tests to either examine or perform myself; the tests are as follows: Gram Stain; Nutrient Gelatin Test; Carbohydrate Fermentation; Dextrose, Lactose and Sucrose; IMVIC tests; Citrate, Indole, Mythel-Red and Vogues Proskauer test; as well as a Urease and TSI Test.
Janick. J. (2011). Center for New Crops & Plant Products - Department of Horticulture and
Infection control is a central concept to every practice of health care providers. Its main objective is to prevent the transmission of infectious diseases from both patients and health personnel (Martin et al., 2010). In dental clinic, infection control is a continuous concern for its professionals. They have to contact patients routinely and be exposed to their blood, saliva, dental plaque and pus that may contain infectious pathogens. It is important for the dental professionals to treat these fluids as if they are infectious and special precautions must be taken to handle them. In this essay, I will highlight the scope of infection control practices in dental clinics and the ways through which infectious microorganisms are transmitted in the dental clinic. Also, I will talk about some infection control guidelines implemented in dental clinics and how they meet the needs of the patients. Finally, from a personal perspective, I will mention some factors that affect the implantation of infection control guidelines and procedures.
However, on the a recent visit dated 23/12/13 the patient’s gingival condition had deteriorated, presenting with an increased plaque scores of 34% and bleeding scores to 63%. Intra oral examination also showed generalised oedema and erythema throughout in the mouth in response to this increase in plaque bacteria. The presence of supra-gingival calculus on lower anterior teeth and both sites of upper buccal molars and the patients BPE now reads 212 /121, putting the patients caries risk at a ‘High’ status.
All practicing dentists, dental associates, and laboratories follow standard precautions and recommendations specified by the Center for Disease Control (CDC). The standard precautions, previously known as the universal standard precautions, focus on the perception that all blood and bodily fluids, regardless if they contain blood, such as saliva, may be contaminated and should be considered infectious. (Bebermeyer). The infection control methods that are practiced in dental offices were established by the CDC in 2003 with Guidelines for Infection Control in Dental Healthcare Settings (Kohn). These guidelines include the use of protective barriers, personal protective w...
Culture plates of yeasts strains: S41, a pet 1 and M240, conical flasks containing Yeast Extract Potassium Acetate (YEPA), Yeast Extract Peptone Dextrose (YEPD) and Yeast Extract Palm Olein (YEPPO) media, pH indicator, inoculation loop, microscope, methylene blue, Bunsen burner and incubator.