Toys

While I've always had an interest in photography, both macro and micro, inside and outside the laboratory setting, my talent has never measured up to my enthusiasm. Regardless I continue to try my best to document interesting specimens that I, or my colleagues encounter in the microbiology laboratory.

The photographs previous to this post were taken in entirety with a Leitz film camera head (right) mounted on the dual head Leitz microscope pictured below - all prior to the advent of digital imaging. Only the Tinea infected hair photo was taken using the same film camera mounted on a fluorescent microscope. Both Kodak Kodachrome & Ektachrome film of various speeds were previously employed.


In 2006 I suffered an injury which required prolonged rehabilitation and equally long absence from the microbiology laboratory. I started this blog out of boredom and posted older photos which I had on hand. On my return I found the lab had retired the old film camera and replaced it with a Nikon Coolpix 8400. The versatility of digital imaging was immediately realized particularly in allowing numerous shots with no waste of costly film & processing and instant feedback as to whether a particular feature of interest was adequately captured.


Above is the dual head Leitz microscope with Nikon Coolpix 8400 thatI've used for the majority of photos that follow in this blog. Prior to the lab's aquisition of the Nikon, this same microscope was outfitted with a Leitz film camera pictured top above right.

Microphotographs were fairly straight forward. The auto setting proved to be quite adequate for most stains and specimens. Flash was turned off. Photographs were captured in 300 dpi resolution and could be preserved or reduced in digital size or resolution as necessary. White balance could be corrected with photo editing software afterwords and cropped to focus attention on desired elements. Enhancements were kept to a minimum.

I quickly encountered several challenges when attempting macro photographs using the same Nikon 8400 in the laboratory setting, particularly in close-up photos of culture plates and media.

• Photos taken at close range could not be taken with a hand-held camera as it could not be held sufficiently steady so as not to blur the photograph. A tripod was found to be too cumbersome as it was always a challenge to set it to the appropriate height on a bench top without having the legs splay out too far. Tripod legs set closer together often became included in the critical area of the photograph and were difficult to crop out. A tripod became all but impossible to set up within a biological safety cabinet when photographing fungal cultures.
• Harsh overhead fluorescent lighting often produced both shadows and reflections from bench surfaces and clear plastic plate lids, frequently washing out the intended target.

As photography in our microbiology lab is almost exclusively of my own personal interest and the departmental budget does not currently permit purchase of a professional photo set-up, I improvised my own.
Modifying the spring-loaded articulated arm of an unneeded desk lamp, I constructed a mechanism that joined the lamp arm to the screw mount base of a spare tripod. The arm can be mounted using a clamp base attached to a workbench or when used in a biological cabinet, it is mounted in an alternate heavy movable metal lamp base. The spring-loaded arm can be swung into place without the base showing in the photo. The springs allow easy height adjustment, bearing the weight of the camera as it is raised or lowered in conjunction with the camera's zoom in order to obtain the ultimate combination. While the springs are too sensitive to motion to allow one to depress the trigger by hand, the Nikon 8400 comes with an infra-red remote controller. Once the camera is positioned, the spring-loaded suspending arm is released and vibration allowed to stabilize (just a couple of seconds) before snapping the photo. The movable base allows the arm to be moved a location in the lab with the best available lighting. The background can be selected to best contrast the item being photographed. All fungal plate photography is carried out in the biological cabinet so as not to contaminate the lab with fungal spores.

Above is my improvised spring-loaded lamp base with attached tripod screw mount allowing the camera position to be altered without getting the base into the photo. The heavy movable base allow the camera to be set up in a laminar flow hood for the safe photography of fungal cultures. A matt-black folder cover provides a disposable, non-reflective contrasting background for clear plates such as SAB. I rarely use the camera's flash as it tends to wash out images at the distances I require and reflections are hard to control.

New Toys (2010);
In the fall of 2010, our lab purchased a Leitz DMD-108 digital microscope. This headless (no occular) microscope utilizes LED illumination with the digital image being transmitted to a high-definition monitor. This unit allows for;

• immediate monitoring of images by multiple observers displayed real time on the high-definition screen.
• photography of any interesting field captured to a jump drive or other mountable storage medium.
• record of camera/exposure settings for each image captured.
• dictation of description of image features captured with built in microphone.
• real time transmission of image over internet for consultation.
• X & Y co-ordinates of image location for future orientation.
• Digital zoom in addition to magnification provided by standard objectives.
• White balance adjustment & colour correction for various common stains or custom settings.

While it has been exciting to obtain the Leitz DMD-108, I feel I have already encountered some limitations of this microscope. Immediately I noticed that it was extremely difficult to hold a field in focus when using the high oil immersion (1000X) objective. Perhaps the microscope still needs final set-up, however the scope was so sensitive that the image would drift out of focus immediately on release of the focus knob. This microscope was clearly better suited for lower magnifications where immersion oil was not employed. Oil immersion is most critical for the study of bacterial cells, limiting the usefulness of this instrument. Parasite species vary greatly in size and depending on which might require oil immersion or high-dry study. While permanent stained slides were easily viewed under high-dry power, wet preps of concentrates seemed to be difficult to follow as the suspension drifted across the field. Perhaps this requires some getting used to but suspended organisms seemed easier to follow on conventional microscopes. The same applied to fungal LPCB slide mounts however the larger size of conidiophorse and hyphae suspended in the more viscous LPCB reduced any focus and tracking challenges.
While an interesting instrument, it's usefulness in a microbiological setting is yet to be determined. The instrument seems better suited for histological/pathological specimens.

Above is a LPCB fungal specimen as seen on the high-definition monitor projected from the Leitz DMD-108 microscope seen on the right of the photo. Note the lack of occulars on this microscope.
In the future I will note any photos taken on this microscope. All others will continue to be taken using the Nikon 8400.

For anyone wishing to know more about the Leica DMD-108 Digital Imaging Microscope you can visit their site for specific information.

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Tinea capitis:

Fungus

Tinea capitis refers to dermatophytosis, a fungal infection of the scalp. Three types of in vivo hair invasion are recognized:

Ectothrix invasion is characterised by the development of arthroconidia on the outside of the hair shaft. The cuticle of the hair is destroyed and infected hairs usually fluoresce a bright greenish yellow colour under Wood's ultraviolet light. Common agents include M. canis, M. gypseum, T. equinum and T. verrucosum.

Single hair invaded by Tinea capitis (Calcofluor White Stain under Ultra Violet Microscope)

Magnification was not noted
(click on photo to enlarge for better viewing)

Endothrix hair invasion is characterized by the development of arthroconidia within the hair shaft only. The cuticle of the hair remains intact and infected hairs do not fluoresce under Wood's ultraviolet light which the physician might use in a presumptive diagnosis.. All endothrix producing agents are anthropophilic eg T. tonsurans and T. violaceum.

Favus usually caused by T. schoenleinii, produces favus-like crusts or scutula and corresponding hair loss.

Ditto

Tinea capitis infection of a single hair (Calcofluor White Stain under U.V. Scope)

The fungi that instigates tinea capitis thrives in moist and wet environments, and your risk increases if you have poor hygiene, prolonged skin wetness (such as that caused by sweating), and minor scalp injuries. Oral antifungal medication and medicated shampoos, such as those that contain selenium sulfide may be necessary to eliminate the infection.

The microphotographs above are of a single human hair with the arthroconidia visible within the hair shaft. They fluoresce because they were stained using Calcofluor White prior to being placed under a microscope cover slip and slide. Viewed under a fluorescent microscope at a magnification of x800.

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Neisseria menningitidis:

Bacterium


Neisseria meningitides, also known as meningococcus is classified as a gram negative diplococcus, meaning it stains red using the gram stain and appears as two round cells in pairs. This organism is best known for its ability to cause meningitis. Symptoms may include fatigue, fever, headache, neck stiffness to coma and death can occur in about 10% of the cases. There are several serotypes (can produce distinct antibodies in response to their invasion) and these my correlate with the geographical location one aquires the infection. Antibiotic treatment might include Cephalosporins, Erythromycin, Tetracycline or Ciprofloxacin.

The photo above is of a specimen obtained from a Lumbar Puncture (LP) where a fine needle is introduced into the spinal column in order to obtain some Cerebral Spinal Fluid (CSF). The fluid was processed in a cytospin centrifuge which concentrates the specimen onto a small spot on a microscope slide, flattening out any white cells present in the process while drawing off excess fluid. White cells may be polymorph nucleocytes or monocytes depending on the infecting agent. The white blood cells are the body’s immune response attempting to defend it from the invaders. The slide is then “fixed” to adhere the material to the slide and then stained by the “Gram” method. Neisseria meningitides will appear as single or “diplo” (two) red coloured cells, often within the white blood cells. x1000 Magnification under a light microscope

Helicobacter pylori:

Bacterium

Helicobacter pylori is a helical or spiral shaped bacterium which can infect the various areas of the stomach and duodenum. Infection can cause peptic ulcers, gastritis, duodenitis and perhaps even lead to cancer. Helicobacter pylori likes to live in the mucous layer of the stomach aiding it in surviving the stomach‘s acid environment. Helicobacter has enzymes which splits the urea molecule resulting in carbon dioxide and ammonia which further helps to neutralize and protect it from stomach acid.

There are several tests developed to detect the presence of Helicobacter pylori. The laboratory can look for antibodies against the organism in the blood, Helicobacter antigens in the stool itself, detecting ammonia generated by the bacteria in a “breath test”, but the definitive test is by gastric biopsy via an endoscope down the throat.

Today therapy usually is by an antibiotic such as amoxicillin, clarithromycin or metronidazole and the inclusion of a “proton pump” inhibitor such as omeprzole.

The above photomicrograph is of a gastric biopsy. A very small (<1mm dia) piece of gastric tissue is mashed onto a glass slide, fixed with heat, then stained with Basic Fuschin. Viewed under a light microscope x1000.

Histoplasma capsulatum:

Dimorphic Fungus


Histoplasma is a thermally dimorphic fungus found in nature. Soil contaminated with bird droppings or excrements of bats is the common natural habitat for Histoplasma. Although it is claimed to exist worldwide, tropical areas are where this fungus is more frequently encountered. Histoplasma capsulatum is the causative agent of a systemic mycosis called histoplasmosis. The spectrum of the disease varies from an acute benign pulmonary infection to a chronic pulmonary or fatal disseminated disease. Inoculation is primarily through inhalation.

For definitive identification of the fungus, (>35C) yeast-to-mould (<35c). style="font-weight: bold; color: rgb(51, 204, 0);" size="4">

Diphyllobothrium latum:

Parasite

June 2012:  Updated - Please visit my updated posting of Diphyllobothrium latum.

Diphyllobothrium latum is also known as the broad fish tapeworm. There are many different host species, mainly those carnivores which eat fish, and this includes man. These worms can reach to a length of 10 meters and can shed in the region of one million eggs per day. The eggs are ovoid with an operculum (cap) at one end and a knob at the other and measure 60 x 40 microns in size. When they are released into the intestine they are only partially embryonated and require from 8 days to several weeks for the infective coracidium to develop. When the eaten by the definitive host it passes through the stomach and the scolex becomes embedded in the mucosa of the small intestine and develops rapidly producing eggs within 10-14 days. People generally become infected when uncooked fish are eaten and it is particularly prevalent in those cultures which eat a lot of freshwater fish and prepare it by methods other than cooking.

In many cases human infections go largely unnoticed, because of the nonspecific symptoms such as intestinal discomfort, nausea, and diarrhea. However, in some cases pernicious anaemia develops which is related to the malabsorbtion of vitamin B12.

There are a large number of possible drugs available to treat this disease, the two main ones used are Niclosamide and praziquantel, both of which are highly effective.

Effective control measures include cooking fish properly or freezing the fish down below -12 C for a minimum of 24 hrs.

The photomicrograph above shows the operculated eggs of Diphylobothrium latum with the pointer pointing at the cap. A wet preparation of a concentrated faecal sample was made. x800. Fecal material and bacteria can be seen in the background.

Anaerobiospirillum succiniproducens

Bacterium


Anaerobiospirillum succiniproducens rarely isolated in the clinical laboratory. Septicemia appears to be the most frequently reported infection caused by Anaerobiospirillum followed by gastroenteritis. This strictly anaerobic, motile, gram-negative spiral bacterium is often mistaken for Campylobacter species when first encountered. Rapid identification is imperative as Erythromycin, effective against Campylobacter and antibiotics such as Penicillin or Metronidazole, frequently prescribed against anaerobic infections are ineffective against Anaerobiospirillum.

Entry into the body seems to be through the gastro-intestinal tract and may result in bacteraemia (bacteria in the blood). Gastrointestinal symptoms may include nausea, vomiting, abdominal pain, diarrhoea and rectal bleeding. Infection usually occurs when the patient already has some chronic underlying disorder.

The photomicrograph above is from a laboratory culture of the organism after it was initially isolated from the blood of a patient returning from overseas. A modified Leifson flagella stain was employed which stained the organisms red. The stain condensed around the very fine flagella making them visible under the light microscope. The flagella are whip like projections or tails which the organism uses to propel itself. Here you can see that there is a tuft of flagella on both ends of the bacterium. x1000 magnification.

Bipolaris specefara:

Fungus

Bipolaris is a dematiaceous filamentous fungus. It is cosmopolitan in nature and is isolated from plant debris and soil. Bipolaris can infect both immunocompetent and immunocompromised host. Bipolaris is one of the causative agents of phaeohyphomycosis. The clinical spectrum is diverse, including allergic and chronic invasive sinusitis, keratitis, endophthalmitis, endocarditis, endarteritis, osteomyelitis, meningoencephalitis, peritonitis, otitis media (in agricultural field workers) and fungemia as well as cutaneous and pulmonary infections and allergic bronchopulmonary disease. Bipolaris can infect both immunocompetent and immunocompromised host.

The hyphae are septate and brown. Conidiophores (4.5-6 µm wide) are brown, simple or branched, bending at the points where each conidium arises from. The conidia, which are also called poroconidia, are 3- to 6-celled, fusoid to cylindrical in shape and are light to dark brown in color.

Amphotericin B and ketoconazole are used in treatment of Bipolaris infections. Surgical debridement may be indicated in some cases, such as sinusitis

The photomicrograph above has the indicator pointed at one of the typical poroconidia which is attached to it‘s conidophore from where it has branched off the hyphae. It is of a wet preparation stained with Lactophenol Cotton Blue and viewed under x800.

Entamoeba coli:

Parasite

Enatamoeba coli is an amoeba which inhabits the human gut. It is considered to be an non-pathogenic commensal organism which along with thousands of other types of organisms makes up the normal gut flora. In the laboratory care must be taken in distinguishing the non-pathogenic Entamoeba coli from the pathogenic Entamoeba histolytica. Entamoeba can exhibit two forms, the trophozoite (troph) shown here and a cyst form which can exhibit up to 8 nuclei.

Treatment is not necessary but presence of this organism in a stool sample may indicate that a person has been exposed to a contaminated source bearing faecal matter and further investigation may be warranted.

The photomicrograph above is of an Entamoeba coli trophozoite in a concentrated faecal specimen stained by the Iron Haematoxylin method. A coarse cytoplasm is seen and the cell has a nucleus with dense unevenly distributed nuclear chromatin at it’s periphery and a central nucleolus. x1000.

Sporothrix schenckii:

Note:  I first posted this photo in 2008 while bedbound and recovering from a serious injury.  I had just heard about "bloging" and was just having fun playing, never thinking at that timeabout continuing this project as 'Fun with Microbiology'.  Sadly, this celluloid film print doesn't show much detail.  To remedy this, I have uploaded a new and improved Sporothrix schenckii complex blog post which can be found by clicking HERE.  I'm sure you will find the new and improved site much more informative than this lonely photo.  Y. (May 10th, 2015)
 
Dimorphic Fungus


Sporothrix schenckii is one of five species of thermal dimorphic fungi of medical interest. Dimorphic basically means “two forms” as the organism can express itself as a round cell yeast (>37C) or as a filamentous fungus (<37c>
In nature Sporothrix lives as a saprophyte on wood, decaying vegetation, Sphagnum moss, animal excreta and soil. It can grow on the thorns of roses and can be inoculated into the body by a prick of the thorn. The fungus can then morph to a yeast and grow in the lymph system, manifesting itself as lesions in the lymph nodes. It is often referred to as “Rose Handler’s disease”. Sporothrix can also cause lung infections.

Antifungals such as Ketoconazole may have some effect. Itraconazole shows some promise at low doses for several months. Intravenous Amphotericin B, usually the drug of last resort for fungal infections, works poorly and slowly.

The photomicrograph above shows the fungal mycelia with typical hyphae bearing conidia. The “wet preparation” is dyed with Lactophenol Cotton Blue stain for contrast. x800.

Cryptosporidium parvum:

Protozoan Parasite


Cryptosporidium is a coccidian protozoan parasite who’s oocysts can infect humans usually by the oral-fecal route. Contaminated water is often the source of infection. In recent reports improperly washed produce was implicated as the source of infection. In immunocompetent patients, cryptosporidiosis is an acute, self-limiting diarrhoeal illness lasting about 1 to 2 weeks. Symptoms may include frequent watery diarrhoea, nausea, vomiting, abdominal cramps with low grade fever. No safe and effective therapy for cryptosporidial enteritis has been successfully developed. Since cryptosporidiosis is a self-limiting illness in immunocompetent individuals, general, supportive care is the only treatment for the illness. In immunocompromised patients (AIDS, transplants, cancer patients, the illness may be much more serious. No effective treatment is available other than to prevent dehydration and let the illness run it’s course.

The above micro photograph is of a Cryptosporidium oocyst from a concentrated stool sample stained by Carbol Fuschin, then decolourized by acid-alcohol. The oocysts are “acid fast” and resist decolorization whereas other material loses the resulting red colour. The remaining material in the slide is fecal debris and normal intestinal bacterial. x1000.

Strongyloides stercoralis:

Parasite: Nematode


Strongyloides stercoralis is a human parasitic roundworm (Nematode) about 2.5 mm in length. The adult parasitic stage lives in tunnels in the mucosa of the small intestine. The genus Strongyloides contains 53 species and S. stercoralis is the type species. The infectious larvae penetrate the skin when there is contact with the soil. Some of them enter the superficial veins and ride the blood vessels to the lungs, where they enter the alveoli. They are then coughed up and swallowed into the gut, where they parasitize the intestinal mucosa. S. stercoralis has a very low prevalence in societies where faecal contamination of soil is rare.

Ivermectin is the drug of first choice for treatment. Thiabendazole was used previously, but owing to its high prevalence of side effects and lower efficay, it has been superceeded by ivermectin and as second line, albendazole.

The above photomicrograph is of the Strongyloides larvae in a wet preparation of concentrated fecal material. x800.

Scedosporium inflatum:

Fungus

Note: When I took this photograph, the only camera in my arsenal was a film camera.  Since then, changes have occured not only in photographic technology but in the nomencature of this genus.  This fungus previously known as Scedosporium inflatum was renamed as Scedosporium prolificans.  Please visit the new post within this blog by clicking on the link within this paragraph.  To learn more about the cameras used to take the photographs, please visit my blog post entitled 'Toys'.


Scedosporium inflatum is a filamentous fungus which is found in the severely immunodepressed including those with hematologic malignancy and recipients of allogeneic hematopoietic stem cell, heart-lung, lung, liver and renal transplants and occasionally in normal hosts as well. It can cause lesions in the sinuses, lungs, bones and central nervous system. The formation by S. inflatum of annelloconidia in wet clumps at the apices of annellides with swollen bases was found to be the most useful characteristic in differentiating this potential pathogen. Dissemination throughout the body might be prevented with combined itraconazole and fluconazole treatment. Variconazole is considered first-line treatment by some.

The above photo is of the hyphae and conidia stained with Lactophenol Cotton Blue and viewed x800.

E-coli & Ascorbic Acid Electron Micrographs

Back in my university days I found employment at University Hospital in London at which I studied the effects of the antibiotic Metronidazole (Flagyl) against various bacteria (1). Metronidazole has long been established as one of the premier antibiotics with activity directed against most gram negative as well as many gram positive anaerobic bacteria. More interestingly, our research unexepectantly found that Metronidazole was able to exhibit remarkable activity against the facultative organism Gardnerella vaginalis (2). This was found to be a result of the metronidazole molecule being metabolized in the liver to both an ‘acid’ and a ‘hydroxy’ metabolite. Further experimentation showed that the hydroxy metabolite exhibited significant activity against this organism.

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In the experiments using various anaerobic (oxygen hating) organisms, liquid tubed media was used to grow the bacteria and in which to dilute the antibiotic. In order to “reduce” the media, so that it was oxygen free, a variety of chemical ingredients could be added. We chose ascorbic acid and used the facultative anaerobic organism E-coli as a control. The test anaerobic organisms reacted as expected however significant changes were visible on the control E-coli bacteria as well, against which metronidazole should have no appreciable effect. The E-coli cells were visibly stressed and damaged, often showing uncharacteristic morphology such as incomplete cellular division, elongated cell forms and bifurcated ends. In a rather unexciting explanation, it was determined that this was due to the ascorbic acid content itself stressing the E-coli cells.

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Regardless of this outcome, the entire process served as a learning experience and afforded me a chance to work with the university’s electron microscope. I’ve included some of those electron micrographs here. In particular I never tire of marveling at the physical and molecular structure of these fascinating bacteria. Included here is an electron micrograph of an E-coli bacterium demonstrating the cell wall structure which is responsible for it’s gram staining properties. Other smaller photos show the stressed nature of the bacteria which should have appeared as rather uniform bacilli. Various magnifications were taken and if I recall, the electrondense negative stain was phosphotunstic acid (PTA).

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*all electron micrographs were taken by Yuri.

So, perhaps this is evidence that you should get your daily dose of ascorbic acid (vitamin C) as it may itself have a direct effect on bacteria in your system. You know what they say, “an apple a day keeps the doctor away” - perhaps with some truth to it.

(1) Edward D. Ralph & Yuri E. Amatnieks
Antimicrob Agents Chemother. 1980 March; 17(3): 379-382

(2) E.D. Ralph & Y.E. Amatnieks
Antimicrob Agents Chemother. 1980 July; 18(1): 101-104

Human Papilloma Virus (HPV) & Gardasil Vaccine

I rarely agree with anything the monkeys in suits that constitute our government do, but this rare case is the exception. The Government of Ontario with the Ontario Ministry of Health have put together a program to educate and immunize appropriate citizens against infection with the Human Papilloma Virus or HPV.

Some Science First;

Papillomaviruses were first identified in the early 20th century, when it was shown that skin warts or papillomas, could be transmitted between individuals by a filterable infectious agent. In 1935 Francis Peyton Rous, who had previously demonstrated the existence of a cancer-causing Sarcoma virus in chickens, went on to show that a papillomavirus could cause skin cancer in infected rabbits. This was the first demonstration that a virus could cause cancer in mammals. Since then over a 100 different human papillomaviruses have been identified. Some cause benign wart or papillomas, after which the virus is named, but some 30-40 of these strains can be transmitted through sexual contact. Many of these infecting virus strains don’t cause any noticeable symptoms until greater problems arise, most notably, cervical cancer.

The papillomaviruses are non-enveloped, 60 nanometer (nm) diameter capsids composed of 72 capsomer units, geometrically arranged in icosahedral symmetry which then covers a double-stranded circular DNA molecule of about 8000 base pairs in length. The L1 protein of the Capsomers is what the HPV vaccine utilizes in eliciting antibody response against the invading virus, thus destroying it.

The Immunization Program & Stats;

The Government of Ontario has just introduced a $39M voluntary program in which all 84,000 grade 8 girls are eligible to receive the anti-HPV vaccine (Gardasil) free of charge in three doses over a 6 month period. The vaccine has been approved by Health Canada for use in girls and women between the ages of 9 to 26. The vaccine protects against infection from four separate strains of HPV, which combined, cause 70 per cent of all cases of cervical cancer.
Those not eligible for this program can still receive the vaccine at their doctor’s office at a cost of around $400.
Cervical cancer is the second most common cancer for women aged 20 to 44, after breast cancer.
In Ontario, some 550 women are diagnosed with cervical cancer and 150 die every year.

Side Effects;

As with any vaccination, the expected side effects, if any, include redness, tenderness and swelling at the injection site and less commonly, fever, nausea, dizziness and headache. No foreign substance introduced into the body can be considered 100% safe however serious side effects are extremely rare and outweigh the benefits.

The Controversy;

In this day and age it is still mind-boggling that vaccinations or anything related to sexual transmitted diseases can stir up such public controversy. You would think that any loving parent would do whatever they could to best ensure and protect their child’s health. However, prudish attitudes, outdated values and misinformed beliefs still supersede logic in many households.

Those opposed to their child receiving the protective vaccine usually present an argument such as:
Since this virus is only transmitted by sexual contact, my pure little grade 8 darling couldn’t possibly get this disease as she is not sexually active (and will never be until the night of her wedding). The twisted logic that also follows is that because their child would be protected from sexual HPV infection, she is more likely to engage in sexual activity now that the fear of disease (and God) are removed from the equation. These arguments are more likely to be raised by parents whose child attends a Catholic School or practice other more conservative religions.

It may be news to many parents of girls attending Catholic schools but the rate of promiscuity is about the same in their institutions as for the general public of the same age group. Peer pressure, the desire to rebel and “spread your own wings”, simple curiosity, society’s ever more liberal morals, and pure raging hormones, all may supersede any cautionary words spoken by a parent. Sadly, as remote as it seems, rape still remains a reality in this world. The result is that teens are engaging in sexual activity and ever increasingly younger ages. It is beyond me how any intelligent, loving parent in today’s society would deny their child this protection on misguided ethical or moral grounds. Education, instruction and proper parenting will not be diminished by your child having this vaccine. In the end, which would really be worse, a pregnant child or one that grows to develop a possible fatal cancer?

A final thought. Will your "pure as the driven snow" daughter who has remained celibate remember to get, and be willing to pay the $400 (current cost) for the vaccine prior to her wedding day? Just remember that your future son in law "Johnny Angel" may not have led such an chaste life prior to this blissful day and may be harbouring the virus. One final wedding day present for "your little girl". Think ahead, then act responsibly NOW!

No vaccine is 100% protective and being vaccinated does not eliminate the need to see your doctor for regular checkups including checking for cervical cancer.

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