Introduction and Background
In assessing the impact of microbes in the New South Wales (NSW) health system, it was discovered after laboratory testing evaluating Part A and B categories different types of colonies were discovered, for laboratory 1 evaluations. In laboratory 11specimens used in this experiment were blood, urine, cerebrospinal fluid; sputum, swabs from and
Mucous membranes.
Bacteria colony morphology:-. E Coli. This bacterium appeared to be creamy, shinny and rose to a level of 2-3 mm. M Luteus. This one was yellow shinny circular and rose to 1-2 mm. Fungal colony morphology; Saccharomytes cerevsaie ( yeast) and Aspergillus nidulans ( Mould ). Precautions must be taken while culturing moulds because highly resistant pores are produced and can be transferred by currents (Cohen-Gonsaud et al, 2005). Cellular Morphology- Bacteria Cell Characteristics M-luteus purple, E-coli pink. Gran Strain type Gram + M luteus, Gran –E- coli; Cell shape- M-Luteus- coccus, E-coli rod. Cell size discovered 1 um diameter for M- luteus, E-coli and 3-4 for E- coli. Cell patterns diplococcic for M- luteus and none for E-coli. Cell Characteristics for fungus are Saccharomytes cerevsaie ( yeast) , the shape was oval and size – 14un diameter.
Diagnosis of infectious diseases followed a fixed pattern. Clinical symptoms are the first step towards investigating an infectious disease. Then withdrawal of specimen from suspected areas. This is followed by microscopic examination of the specimen and a culture and sensitivity to determine what drugs the organism will respond to for treatment of the patient host. (Johnston, 2003).
As it pertains to laboratory 11 testing many genera were found to be opportunistic organisms in the hospital environment (Thomas et. al, 2007). In assessing the impact of microbes in the New South Wales (NSW) health system knowledge regarding antibiotic susceptibility, it was pertinent to understanding ways of improving aseptic techniques as well as nursing intervention regarding disease control and prevention.
Aims
Aims and purposes of conducting these experiments in Lab 1 are to gain first hand experience of how to articulate aseptic sampling techniques as well as culture of microorganisms in an agar plate. While gaining expertise of execution in the process the main objective is to examine both bacterial and fungal morphology.
For Lab 11 the aim was to offer students an understanding of how microbiological testing techniques are used to diagnose illnesses. A main objective is to expose students to the underlying principles pertaining to specimen collection. In lab 111 the aim was to examine several genera of bacteria with the objective of determining antibiotic susceptibility gaining a clear understanding between susceptibility and resistant strains.
Methods
Lab 1- Aseptic Technique- Sampling
The microbiological method encompassed working in groups of consisting of 3-4 students. Specifically, the task entailed using aseptic sampling as well as streak plating techniques to inoculate plates.
This process allowed students to loose the agar culture vessel and hold culture with left hand at the end of the vessel. Taking a mounted wire loop in the left hand sterile the equipment utilizing Bunsen flame until the wire gives a red glow. Allow to cool for 15-20 seconds.
Now pass the mouth of the vessel through the flame removing a small portion of the agar culture. As the loop is held quickly flame the neck of the vessel replacing the cap before inoculating culture.
Inoculation
When holding loop remove cap from glass bottle for inoculation and flaming. Touch the agar closest to the bottom of the bottle removing loop spirally. Flame bottle neck and sterilize loop once more.
Preparing a streak plate
After applying sampling techniques described above sample a small culture using the wire loop. Raising the lid of Petri dish with loop from left to right, make four parallel streaks. Lower lip and re-sterilize. After cooling raise and skim sterile over surface of four parallel streaks created previously. Repeat the procedure tow more time cross loop over the previous streaks.
Lab 11- Microbiological diagnosis
This method of specimen collection follows distinct steps which must be given adherence every time they are removed from the body of a client/ patient.
Specimens must be taken from sites pathogen seem to b most concentrated and normal flora contamination is minimal.
After withdrawal they must be transported to the medical laboratory immediately.
If specimen cannot be transported immediately they must be refrigerated to maintain temperature and obstruct rapid growth.
Also they must be collected before any antibiotic treatment is administered.
Precautionary measures must b taken to prevent cross infection while taking and transporting specimen.
Withdrawing blood specimen
Remove via sterile techniques and refrigerate immediately if the specimen cannot be transported.
Withdrawing urine
Clean area around urethra thoroughly
Allow patient/ client to void and collect the mid stream flow of urine
If a specimen is withdrawn via a Foleys catheter the same midstream principle applies Testing must be done half an hour after withdrawing or 5 hours if refrigerated
Cerebro- spinal fluid
Spinal taps are performed by the physician, but the nurse’s responsibility is to secure the specimen for transpiration to the laboratory
Specimen must be processed immediately due to the virulence of neisserria meningitis.
Withdrawing sputum
Collect in a sterile container
Instruct patient not to contaminate sputum with saliva
Bronchial aspiration is an alternative undertaken by the physican.
Withdrawing sputum
Collect in a sterile container
Instruct patient not to contaminate sputum with saliva
Bronchial aspiration is an alternative undertaken by the physician
Skin and mucus membrane swabs
Sterile polyester swabs are used to take specimen
Avoid touch uninfected areas
Except for a gonococci swab specimen they can be refrigerated
Lab 111—Antibiotic susceptibility testing
Bauer- Kirby method has been adopted for this project (University of Pennsylvania, 2012)
Equipment
2 Agar plate
Paper disks
Sterile swab sticks
Broth cultures
Antibiotic disks
5 sterile forceps
Procedure
Six different antibiotics will be used three for each lawn plate
Preparation of Lawn plate
Loose cap off from broth culture bottle and open swab stick
While holding bottle with left hand at the bottom take a swab stick with the right hand.
Remove cap of bottle with fingers and palm of right hand
Pass mouth of bottle through flame than dip swab stick into broth culture
Flame neck of bottle while holding swab stick.
Replace cap
Open agar plate lid
Beginning from the top touch swab stick to agar surface
Sweep over whole surface using smooth horizontal strokes.
Turn plate around 90 degrees using the same swab stick
Repeat process ensuring the entire surface is evenly covered with broth culture
With sterile forceps apply 3 antibiotic disks on each plate.
Combination
Plate 1 –gentamycin’ chloranohenical; carbenicillin
Plate 11- penicillin; vanomycin and celpalexcin
Summary of results
Lab 1
Experiment 1, 11, 111 & 1V
Table 4.1 – Microorganisms in the environment
Source
Numbers of different colonies
Appearance of tow most colony types
Air
3 type b s
Yellow five different color
Unwashed fingers
1 type
Washed fingers
none
Mouth Nose or ear ((which)
P type
Round white diploccocci1
Fomite
1 type
staphylococci
Lab 11
Epithelial cells seen per view- revealed casual none; clean catch- it prevents germs from penis or vagina from entering specimen Difference- casual cloudier due to suspended cells. Specimen noted from case study on dip slide 27 – 40. Colonies present on each side of dip slide- less than 300 yellow red and ph acid. Do colonies observed indicate infection? –No. Why – they are less than 300 Examination of unknown dip slide- Lactose fermentor turned to yellow.
CLED – Lactose fermentor shows yellow colonies while non lactose shows blue. MacConkey showed Lactose fermentor pink colonies; non lactose colorless. CLED supports growth of most UTI while MacConkey inhibits non enteric. With reference to table 4.2-Determination of properties causing UTI in specimen drawn indicated E coli organism.
There were highly resistant strains of Ps. Aeruginosa. Penicillin and vanomycin were detected as narrow spectrum being gram + only; while carbenicillin and gentamycin are broad spectrum displaying gram positive and negative properties respectively. Cell wall structure of gram negative differs from that of gram positive.
E- Coli showed resistance to penicillin and vanomycin. Pseudomonas Aeruginosa showed resistance to all antibiotics except gentamycin. Proteus mirabilis showed the same sensitivity with being susceptible only to the significance of these results indicates that with appropriate aseptic techniques such as hand washing; exposure to pure air and fomites, the growth of bacteria and fungus can be controlled (Thomas et .al, 2007) Importantly, the basic nutrient used was agar with no selectivity. In answering the questions posted for discussion the significant number of colonies appeared in open air and high use fomites. Bacteria thrive when conditions are favorable. These include adequate sunlight; presence of inorganic and organic properties. The accelerated and diminished growth patterns indicate either poor aseptic techniques or very good ones (Greenblatt, et al, 2004)
Subsequently, the effect of hand washing revealed that numbers varied and becoming less in concentration too. Evidently two factors which significantly increased airborne organisms were sneezing or coughing and shaking bed linen (Thomas et .al, 2007). To differentiate whether the bacteria cultured in the experiment was human E coli or another species, features displayed were observed to be gram stain; selective media through biochemical testing. Precisely, it is Gran negative and rod shaped as against others which could spiral or circular (Fotadar et.al, 2005).
Discoveries from exercise performed in lab 11 indicate that when withdrawing specimens all precautionary measures must be taken to avoid contamination as well as attempts at receiving the most accurate reading when from the laboratory. Specimen sampling and examination were the focus of this laboratory experience. Students were exposed to four types of sampling specimens frequently taken in routine interactions on clinical areas. Blood seems to be the most popular specimen then urine (Cohen-Gonsaud et al, 2005)
It must also be noted that when CLED or MacConkey are applied to urine cultures the reaction are not the same. Nurses ought to bear in mind that while both tests devices are very useful mechanism for transporting and culture they are not the same but there are distinct difference in the appearance of organisms in each instance.
Regarding lab 111 antibiotic susceptibility testing is significant to twenty first century North Southern Wales health care system because it alleviate the costs of using antibiotics which do not work on a particular infection (Mathew et.al, 2007).
It is a wonderful evidence based practice that identifies scientific nature of nursing and more importantly the medical profession. A study conducted by Kerr (2005) reiterated the importance of antibiotic susceptibility testing to highlight that it is a sure way of monitoring bacterial resistance to popular antibiotic therapies. He also pointed out that studies should be continuous since new strains of organism are being formed daily (Kerr, 2005).
The three foregoing laboratory tests were significant towards creating a better understanding for students’ implementation of aseptic techniques in their roles as nurses. Understanding how infection is transferred is vital in guarding against cross- infection not only on clinical areas, but generally as a world culture. Knowledge that all antibiotics do not cure diseases is information the public needs to have and educated regarding ways in which to avoid bacteria resistance when engaging in indiscriminate use of antibiotics.
The following experiments outlined the importance of aseptic techniques in nursing practice. This report offers significant evidence based information to suggest changes in the way it is practiced on clinical areas because cross infection as well as growth of harmful organisms could be detrimental to nurses and patients admitted for care (Saladin, 2007).
In relation to lab 11 some vital conclusions which can be made from the exercises are that there is evidence for application daily of safety measures in withdrawing specimens form patient transporting to the laboratory. Storage at the correct temperature to avoid misrepresentations of culture and sensitivity testing is vital to transparency in nursing intervention. Even though specimens such as cerebro spinal fluid or brachia aspiration may be conducted by the doctor, the nurse’s responsibility is to have it stored properly before and during transportation to the laboratory. Laboratory 111 testing report declares that both gram + and -bacteria put up resistance to antibiotics. In experiment, Proteus mirabilis and Pseudomonas Aeruginosa are the most resistant strain of bacteria presently (Michael et. al, 2009).
The experiment regarding practice of aseptic techniques in microbiological culture is indicative of advancing more measures for disease control within the NSW health care system. It means following the centre for disease control and protection guidelines more closely to avoid cross infection and accelerated growth of harmful microorganism ( Mukamolova et al., 2006).
Also, it is quite clear that the nursing implications pertaining to microbiological diagnosis lies in applying techniques that would ensure an uncontaminated specimen reaching the laboratory technician for testing so that the most accurate culture and sensitivity reading could be obtained.
Conclusively, nurses should have a sound knowledge of antibiotic susceptibility as well as resistance to be more articulate during nursing intervention as they participate in team management of clients on clinical areas
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