Sensitivity of Pollutant-In-Product Testing
Get some slides of Salmonellas and Shigellas and find some milk that tests positive to at least one. Make a dilution series of the milk up to bottle #14, being careful not to shake the bottles. Start with 2 drops of milk in bottle #1. Use an eye dropper to
deliver 2 drops to subsequent bottles. Begin testing at bottle #14, using the slide that tested positive. You will learn to search by frequency later. My sensitivity was routinely around bottle #12, for a variety of pathogens. It was the same for toxic elements starting with standard solutions, about 1000 mg/ml, showing this method is less sensitive than skin testing.
Purpose: To observe fluke stages in saliva and urine with a microscope.
a. A low power microscope. High power is not needed. A total of 100x magnification is satisfactory for the four common flukes, Fasciolopsis, sheep liver fluke, human liver fluke and pancreatic fluke.
b. Glass slides and coverslips.
c. A disposable eye dropper.
Fig. 84 Microscope, slides and coverslips.
d. For sanitation purposes (wiping table tops, slides, microscope and your hands) a 50% to 70% alcohol solution (not rubbing alcohol!) is best. Dilute 95% grain alcohol 7 parts alcohol plus 3 parts water. Vodka or 76% grain alcohol can be used undiluted.
e. Formaldehyde, 20%. Formaldehyde 37% is commonly available at pharmacies. Dilute this with equal parts of filtered water to get 18V%, which is close enough to 20%, for the purpose of “fixing” (killing) the specimens. store in a glass bottle in the garage, away from sunlight. Label. specimens that are fixed properly do not lose their life-like appearance.
f Iodine solution. This is only useful for the urine specimens. Lugol’s iodine and tincture of iodine are both useful. Ask a pharmacist to prepare Lugol’s Iodine Solution for you, as follows:
• 44 grams (P/^oz) iodine crystals
• 88 grams (3 oz) potassium iodide crystals Dissolve both in 1 liter (quart) filtered water. This may take a day of frequent shaking.
Method for saliva:
1. Pour the 20% formaldehyde into a small amber bottle or other receptacle to a depth of about 1/8 inch. Keep tightly closed.
2. The person to be tested is asked to salivate into the bottle so the organisms are immediately “fixed” without undergoing cooling first. The total volume should be about double the original amount of formaldehyde used. Make a mark on the container so the subject knows how much to produce. The resultant concentration of formaldehyde will be about 10%.
3. shake the bottle a few times. set it aside for 24 hours to settle (less if testing is urgent).
4. With a dropper, draw up some of the bottom settlings. Put one drop on a slide and apply a coverslip.
5. View under low power of microscope. Compare objects you observe with specimens obtained on slides from biological supply companies.
Note: Persons with HIV and moderate AIDS will show about one to ten parasite stages per slide. It requires several hours of searching. Persons with HIV and severe AIDS show 10 or more fluke stages per slide; this makes the task of finding them much easier. Persons with terminal untreated cancer have many more fluke stages than relatively well persons.
Method for urine:
1. Prepare bottles of formaldehyde fixative ahead of time. Put about Л to V2 inch of 20% formaldehyde in each. Keep tightly closed.
2. Add freshly voided urine from cancer or HIV sufferers to the formaldehyde in approximately equal amounts, resulting in a 10% formaldehyde solution. Shake immediately. Let settle several hours. The sediment has a higher number of fluke stages. Cancer victims with cervical or prostate cancer will show higher numbers of stages in urine than other cancer types.
3. Staining the slide is optional. It helps to outline fluke stages slightly. Prepare Lugol’s solution as described above.
Slides may be stained in either of these two ways:
• Put a drop of “fixed” urine on a slide. Add a drop of 50% Lugol’s (dilute 1:1 with filtered water). Apply coverslip.
• Put a drop of “fixed” urine on a slide. Apply cov — erslip. Add 1 to 3 drops of 50% Lugol’s to edge of coverslip and allow it to seep in.
Note: persons who have been treated for cancer or HIV using any of the known drugs may show only 1 to 2 fluke parasite stages per drop of saliva or urine. For this reason, you may need to search through 20 or more slides to find flukes. Very ill persons may show up to 10 parasites per drop (slide).
Taking Pictures Of What You See
You may be unsure of what you see even if you have the microscope slides of labeled flukes and their stages to study and compare. In real life, they vary so much in shape and size that absolute identification is difficult without experience. Unfortunately in a few hours, just as you are getting proficient, your magnificent specimens will be drying out and unfit for observation. To preserve them longer you can seal the edges by painting around the coverslip with fingernail clear enamel. Or dribble hot sealing wax along the edges and then place them in sealed plastic bags (one per bag). Melt sealing wax in a metal jar lid. Make an applicator from a piece of coat hanger wire bent in the shape of a small square to fit around the coverslip and a handle.
or take photographs. To take pictures of what you see under the microscope you will need a photomicrographic camera, which costs $200.00 and up (see Sources). It is easy to use. Remember to label your pictures so you know which slide they came from.
Even photographs do not scientifically prove identity of parasite stages, but it is very good evidence. Proof would require that the saliva or urine sample could be cultured and seen to produce the known parasite stages.