Sunday, 13 December 2015

For Me, The Fun is Over....

I have retired.  I expect this to be my final post to ‘Fun With Microbiology’ (What’s Buggin’ You?).  For those interested, the preceding post explains my circumstances.  Some of you who may have stumbled upon my blog post entitled ‘Toys’ , about the photographic equipment used, may already know about how this blog came to be.  For those who don’t, perhaps I can offer a short summation here.

In 2006 I had a personal encounter with a microorganism – a non-work related infection which left me a paraplegic.  During my convalescence, my wife suggested I start a blog to pass my time.  I had no idea what a blog was but I soon learned by posting about my other interests; my canoeing adventures became ‘Tales from the Paddle’ while my attempts at building stringed musical instrument were documented in ‘Thunderhouse Instruments’. Stories and descriptions, each supported with photographs.

My employer held my position until I could return some two plus years later.  On my return I discovered that our lab had acquired digital photographic equipment to document interesting cases.  The Nikon camera came first, followed some time later by the DMD-108 digital micro-imaging device.  Surprisingly few cared to make use of this equipment.  With my love of photography, I was quite happy to take command and photographically document interesting specimens that passed through our lab.

With my disability came a change in transportation.  I exchanged my van for a power wheelchair.  Now held captive by the schedule of our regional disability transport service, the bus would frequently deliver me to my workplace over an hour before and pick me up an hour after working hours.  I began to occupy these extra hours before and after work by photographing interesting organisms and specimens that I or my colleagues had come across.  What to do with all these photomicrographs?  Why, start another blog!  Well, continue one actually.  While convalescing, I had toyed with the idea and uploaded a few ‘film’ photographs then both hastily and jokingly entitled the blog ‘Fun with Microbiology (what’s buggin’ you) –a title I have ever since regretted.
And so it started.  Slowly at first but soon found myself expanding the content.  I now look back at the earliest posts and wish I could go back and rewrite and re-photograph all those wonderful isolates.  I have always found frustration with textbooks that offer one small black and white photo or worse yet, only a line drawing tucked into a corner of the page.  I thought, if one photo is worth a thousand words, twenty photos must speak volumes.  These are fascinating organisms which develop over time and one small photo does not do them justice.  While the blog was to be about all microorganisms, I found that fungi were the most photogenic and so they came to occupy a disproportionate volume of posts within this blog.  I also discovered that pursuing microorganisms and their features with the camera was one of the best learning experiences one could have.  It beats the #@&% out of any three day, thirty pupil course.

There are so many more organisms I had hoped to cover…

 Rhodotorula rubra on Sabouraud Dextrose Agar

I have also been frustrated by finding many scientific papers are not freely available and must be purchased from the author or authorized distributor.  It is sad that scientific information cannot be freely shared by all interested, yet I do realize incurred costs must be covered.  All the photographs but one on my blog site were taken by myself and are my property.  I freely share all photographs within this blog with those who may find them of use.  I only ask that you not claim them as your own work and give this site or myself credit.

I cannot see any circumstances which would allow me to continue meaningful posts.  So the fun for me has ended.  I hope yours continues….

My grateful thanks to my colleague D.D. whose guidance, support and infectious enthusiasm made the contents of this blog possible.  Further thanks to all my colleagues for their friendship and support. You know who you all are. 
For my immediate colleagues, I am updating a DVD containing power point presentations of all the Fungal isolates presented on my blog.  I hope to have copies for you soon…

 Coming soon for my immediate colleagues

Lights out...
(I hope your fun with microbiology continues!)

*   *   *

The "Fun" Ends...

Well folks, it is over.  Everything must end, and so must this blog.  The fun has ended for ‘Fun With Microbiology (what’s buggin’ you?)’.  I was offered a package by my employer and I’ve decided to take early retirement.

The writing had been on the wall for some time.   Hospital operating costs outpace government funding, leaving most health care institutions financially stressed.

Hospital mergers were the first line of defense –circle the wagons…there is strength in numbers!  Ours did just that –merging with two other hospitals and pooling their resources.

The second line of defense was to divest ourselves of disciplines and tests in order to save even more money.  We watched our parasitology lab dismantled with specimens now sent to the regional Public Health Laboratory.  Tuberculosis testing also went to the PHL with an inevitable increase in turn-around time.  Hepatitis testing was sent to the Chemistry Department – how that saves money escapes me.  The ‘best’ tests were exchanged for ‘good’ tests – Enzyme Immunoassay tests (EIA) substituted for the more costly Polymerase Chain Reaction (PCR) molecular testing.  Still mycology remains…but for how long?

The third line of defense was to be attrition of staff.  A moratorium on hiring all but the most essential of employees came into effect.  Many positions now offered would be part-time or contract in order to retain flexibility with the additional advantage of not having to pay for benefits.

Yet I wonder how much money is saved when inadequately trained part-timers work in several institutions to make ends meet and confuse one for the other.  Tests often arrive improperly collected.  Incessant phone calls interrupt begging information as to how to do their job.  Work load increased while staff decreased.   Perhaps quality may suffer while overworked staff find vacation requests frequently denied due to workload.  Moral plummets!

The fourth line of financial defense?  Well, we all knew it the day “merger” was announced that staff were caught in the cross-hairs.  Employees are the most expensive asset of any hospital.  MRIs are bought and paid for once.  Laboratory instrumentation is often provided free of charge as long as the consumables (test kits) for the instrument continue to be purchased.  But staff, they keep earning their salary year after year, after year.

…and so for staff such as myself, longevity at the institution became a disadvantage.  My earnings were at the top of the scale and I was entitled to the most vacation time.   I could be replaced by an entry level technologist.  Offered a ‘package’ as an incentive, I knew that I could do no better.  I was burnt out anyways. 

After 37 years in the healthcare profession, I’m done.  I will miss the pure science tremendously, but I have come to hate the business!

I think I have also earned the right to sound off just a little; (nobody reads this anyways)

The Regulatory Collage:
When I first started, we had no government regulation, no collage oversight.  We had pride in our knowledge and ability and I have no doubt we did excellent work.  The regulatory Collage now looms overhead and carries a big stick to whack any member that fails to comply with its rules and regulations.  The dreaded ‘Audit’ to see if you have read enough papers,  attended enough seminars, taken enough courses to satisfy their authority.  Yet I've seen no evidence that those who know how to ‘jump through the hoops’ always become better and more knowledgeable technologists.  It has become a game.

The ‘Collage’ was first sold to us back in 1993 as an organization that would promote our interests and standing as healthcare professionals.  In reality, it is nothing more than a regulatory and disciplinary body hovering over our profession where my colleagues and I pay to protect the public from ourselves, the medical technologists.  Membership fees increase almost yearly.

Continuing Education:
The cost of many courses offered by various education institutes exceed what I’m willing to pay on my income.  Crying poverty, the days have long passed when my employer paid for or contributed to my education which ultimately benefits them.  These days the medical technologist must absorb the cost of further education with little or no possibility that the effort would be reflected in their income or advance their career.   While we remain eager to increase our knowledge, these days we pay…our employer benefits. 

The Workplace:
How the workplace has changed!  The days are gone when microbiology labs were well stocked with a variety of media and individual chemicals.  When an organism would not identify by standard methods, text books were consulted, dry media was reconstituted and QC’d, individual tests were added as necessary.  We had the luxury of a fully stocked media room complete with an autoclave. Every day, every organism was an ongoing learning experience –and we were given the freedom to apply our knowledge; to learn and explore.

These days, media is purchased from an outside supplier.  Only the most basic supplies and test kits are stocked.  If an organism is not identified immediately by laboratory instrumentation, an alternative test may be available, or just as likely, the organism is packaged to be sent to the regional lab for further testing.  The media room went long ago as did the autoclave.  Even sterile distilled water is now purchased.

I often felt like ‘Lucy on the Chocolate conveyor belt line’, where specimens pass by at a furious rate as you desperately do you best to keep up without making errors.

Standard Operating Procedures (SOPs) and Algorithms rule the lab.  Individual thinking has been replaced by uniform conformity.

Reflex Orders:
In the past, technologists at our institution could initiate ‘reflex orders’.  If one test did not directly give a useful result but strongly provided evidence for additional or alternative tests, we could personally order that test and fulfill it.  The answer might be on the doctor’s desk before he could ask the question.
‘Ghost cells’ in a sputum or pleural fluid may suggest acid fast bacilli.  Previously I could order tests for appropriate staining and culture.  No longer.

 A throat culture specifically intended for the detection of group ‘A’ Streptococci revealed that a child had a pure growth of Streptococcus pneumoniae, however my instruction was not to report it as “the test is only for Group ‘A’ Streptococci.  No other respiratory cultures were ordered for the child!  SOPs rule the day, and I had no SOP for that situation.  Where once I could report this directly or order the appropriate test, now a game of phone tag ensues, trying to get a verbal message to someone who would be willing and able to act on my word.  Where once we were professional enough to act on our results and place ‘reflex orders’ as necessary, these days only the physician can initiate the order.  

Quality Control:
If you were to find yourself in a hospital in need of serious medical attention, you would wish the institution adhered to well established rules and regulations to ensure utmost quality.  Hospitals are assessed periodically to ensure they meet established standards.  Hospital accreditation requires inspection by assessors who follow their own SOPs and algorithms.  No one denies that Quality Control (QC) is important.

In the laboratory, everything must be “controlled”.   At times it seems that the more graphs, charts, check-marks and signatures you can produce…the more professional you believe you are!  Of course quality control is important; however make it relevant, concise and applicable.  At times one loses sight of reality in the attempt to jump through the hoops and satisfy the inspectors and nonsense is the result.  Examples follow;

·         To QC LactoPhenol Cotton Blue Stain our SOP directs the technologist to stain a Penicillium specimen with the LPCB for the positive control and with saline for the negative control.  Never in my career has putting this blue dye on a fungus not turned it blue.  As for the negative control, what am I to observe to determine if this LPCB passes QC?  That saline doesn’t spontaneously turn the fungus blue?  I have never obtained an answer, yet we are directed to do the test.

·         To QC Germ Tube Media our SOP instructs us to place Candida albicans in an aliquot of horse serum as the positive control and Candida krusei in another aliquot as the negative control.  Again, I ask “what am I to observe in the QC aliquot containing Candida krusei to determine if the horse serum passes QC?”  This species does not produce germ tubes so it shouldn’t!  If it does, does the QC fail?  Or does it just mean the organism we thought was C.krusei, isn’t.  C.albicans is both positive and negative control!  In horse serum it passes if it produces germ tubes and fails if it doesn’t.  Done!  No, follow those SOPs and Algorithms – don’t deviate, don’t think!  What a waste of effort, time and supplies.

Another irritant is the use of commercial (read expensive) ‘single use’ vial or dispenser of reagents.  Identical lot numbers stored in an identical manner are QC’d each morning for use that day.  That single vial or dispenser of reagent is for use, not per person, but for the entire lab.  Shouts of “who’s got the catalase” or “anybody have the spot indole” as we leave our station to retrieve the reagent.  I try to imagine a construction site where one hammer or one drill is QC’d each work day followed thereafter by construction workers stumbling over each other as they retrieve the single tool they find themselves in need of.  Early in my career we took a trip to the hospital pharmacy to obtain a bottle of ‘off the shelf’ (read inexpensive) hydrogen peroxide and made up spot indole from ‘scratch’ in the media room.  We QC’d each reagent and once satisfied of their activity they were stored appropriately.  Technologists would obtain an individual aliquot for daily use and were observant to any deterioration of activity.
Price has increased while technologist’s involvement has decreased.

Technologist’s Changing Role:
Where once my role as a technologist was primarily to work on clinical specimens, diagnose infectious illnesses and liaise with other medical personnel, over the years the scope of required duties has changes significantly.   In addition to the duties listed above, we have now stock supplies; act as file clerks, answer continuously interrupting telephones –duties more akin to secretarial work.  While I do not feel above this sort of work and am happy to help where and when needed, I feel that this work, if part of my job description, is a waste of my education and the taxpayer’s dollar.  Changing roles and QC madness even has technologists checking the available paper in copy machines on a daily basis, and then signing off as having done so.  I would be surprised if the lowliest clerk in some law firm would have such a responsibility enforced.  When the paper runs out, you’ll know – and you’ll add paper…

Loss of Skills:
As a cost saving measure, our hospital as many others, are divesting themselves of particular microbiological laboratory disciplines such as Parasitology, Mycology and Tuberculosis testing.  These are particularly challenging skills to master and achieve competence.  I was particularly lucky to work in hospitals offering all these disciplines throughout my career.  Increasingly these tests are now being sent to a central laboratory for testing.  Most new graduates entering clinical microbiology will no longer have exposure to these disciplines after their studies.  The disciplines will be lost to all but those who work in the central reference laboratory or possibly huge ‘downtown’ hospitals.  The ‘well rounded’ technologist will be a thing of the past.  In the end, the work must be done, whether the hospital lab or the central reference lab and here in Canada, the taxpayer pays for both.  How that saves money is beyond my understanding.

Going Green:
I suppose that getting rid of the hazardous Bunsen burner which used that nasty fossil fuel, natural gas, in exchange for single use, disposable plastic loops that now fill the hazardous waste bin,  could be argued as having ‘gone green’.

Cultivation of a Poisonous Work Environment:
My employer adopted an new attitude towards its employees whereas longevity no longer has any bearing on advancement or remuneration (pay increases).  There is a point to be made that those who work harder and contribute more should receive the greatest rewards.  The downfall is that the evaluation can be quite subjective with those "sucking up" to the boss may get preferential treatment.

One can volunteer for every project and take on every task yet not produce quality work while others in the background may do more than their share without any fanfare.  Every workplace has one of those loud, boisterous, "look at me" go-getters that never misses an opportunity to impress the boss.

Computers now log every key-stroke and every error.  We now live in fear of how our human errors will be tabulated and now come back to haunt us.  My concern is that human nature may now try to hide errors rather than correct them which may be detrimental to the patient.  I can still hear my first manager's words concerning errors;  "bring any errors to our attention as our only concern is to correct, not punish".  My how the attitude has changed!

To further poison the atmosphere -in unmistakable terms we were instructed that we must keep an eye on each other and turn in anyone who arrives late, or leaves early.  Those that take a longer lunch or surf the internet for something other than work - we must run to the boss and squeal!  While life never runs on such a strict time clock, we are now obligated to turn in our colleagues and co-workers.  Working through lunch on a busy day goes unnoticed while those taking five minutes longer when the workload allows should be turned in.

It is a very clever concept.  It is a shift from 'the employees against the employer' to 'the employees against each other'.  We can no longer be upset at our employer if we don't get a raise - we should blame our colleagues!  What a poisonous atmosphere!

Those that turn in fellow co-workers and colleagues should realize that they will be shunned and ostracized should they be recognized as the source of the betrayal.

What does it do for the health and well being of the medical technologist?  Already in a stressful profession where the workload is ever increasing, we must now "walk on egg-shells" in fear of making any errors and look over our shoulder to see if a colleague is monitoring our work ethics.
The hospital has hired, and proudly promotes their Employee Assistance Program.  However this treats stress and burnout at the back end while I feel they should prevent it from every happening in the first place.

For me personally, I realized that Medical Laboratory Technology had become a Job and was no longer a Profession.  As such, I knew it was time for me to go...

Final Thoughts:
So, as you can see, many changes have taken place since I started as a medical technologist some 37 years ago.  Some changes for the good, some questionable, some of it just plain puzzling.

For the most part, I’ve enjoyed the profession though I find it would be hard to recommend it to anyone these days.  The workload, responsibility and accompanying stress is not for all.  It can be argued that the medical laboratory technologist is probably at the bottom of the health care professional hierarchy.  Since we work primarily within the laboratory and have no direct patient contact, the medical technologist usually finds themselves at both the bottom of the pay scale and in professional respect.  There is very little opportunity or positions available for advancement within the field.  Most move on to administrative duties, if that suits their life’s career path.

So that is it.  I’ve said my peace.  And once again I’ll say, I will miss the pure science tremendously, but not the healthcare business.

…after one final post,  off to enjoy my retirement!!

August 15th, 2016

As I had predicted, a former colleague informed me that the hospital laboratory had decided to divest itself of the Mycology Laboratory.  All specimens requiring fungal culture will be sent directly to the Provincial Laboratory.  The hospital laboratory I worked for physically merged with another right after my retirement.  While our laboratory had staff trained in mycology, most of the staff in the merging lab had no experience in culturing fungi.  By divesting itself of the mycology laboratory, the hospital lab saved money by having the Provincial Laboratory perform the culture and identification as well as not having to train incoming staff in the discipline.

Had I stayed on, I probably would have been unable to continue this blog.  This blog was about photography and while microbiology in itself is fascinating, fungi were the most photogenic organisms.  Mycology made my days enjoyable and rewarding.  Knowing this, I have no regrets in having taken my retirement.

My former hospital laboratory now strictly performs bacterial microbiology with a select number of rapid serological screens.  The day of the well rounded, all-inclusive microbiological hospital lab are probably gone unless, perhaps, in some very large city laboratory.  Sad...

I had considered purchasing my own personal microscope with colleagues at other facilities providing fixed slides for study, but it is a considerable expense for uncertain use and without photographs of the culture plates, I decided to let that idea die.

*   *   *

Unidentified Mould No. 2 -Challenge

I've had a tremendous amount of fun working with, and learning about the moulds posted here.  However, all things do come to an end as does this blog, and so I will leave this last one with you as a challenge.  Have some of your own "fun with microbiology" and let me know what you think the name of this mould should be.

It appeared as a plate contaminant on an un-inoculated plate of laboratory media.  While I took numerous photos, I did not follow through sufficiently to determining an identification.

Macroscopic Morphology:
-exhibited rapid growth on Sabouraud Dextrose Agar at 30oC
-the isolate developed a short, 'fuzzy' surface
-colour was a grey-brown to a dark brown with a lighter coloured outer fringe
-the reverse was dark brown 
-I did not test the growth at 37oC or above so this remains unknown.

 Unidentified mould - after 7 days on Sabouraud Dextrose Agar (SAB or SDA) at 30oC (Nikon)

Microscopic Morphology:
-isolate developed septate, branched hyphae
-conidia developed directly from the hyphae (sessile) or from short stalks that appeared somewhat 'inflated' where the attached to/supported the conidium
-these conidia (?) were rather large, up to 10 µm in diameter.  Some appeared intercalary and perhaps were chlamydospores (?)
-spiral hyphae were occasionally observed

 Unidentified mould - a first look at the edge of a slide culture does not reveal much detail.
(100X, LPCB, DMD-108)

 Unidentified mould - at a slightly higher magnification conidia closely attached to the parent hyphae are evident.  (250X, LPCB, DMD-108)

Unidentified mould - younger, immature conidia retain the lactophenol cotton blue (LPCB) stain while the maturing conidia begin to develop a dark pigment.
(400X, LPCB, DMD-108)

Unidentified mould - immature conidia (blue) & maturing (dark) conidia occur in about equal numbers in this photo.
(400X, LPCB, DMD-108)

Unidentified mould - darkly pigmented sessile conidia seen along the hyphae.  Some younger (blue staining) conidia still seen closer to the ends of the hyphae.
(400X, LPCB, DMD-108)

Unidentified mould - as above
(400X, LPCB, DMD-108)

Unidentified mould - large numbers of conidia produced.
(400X, LPCB, DMD-108)

Unidentified mould -as above
(400X, LPCB, DMD-108)

Unidentified mould -sympodial growth pattern (arrow) ?  Further explanation follows further below.
(400+10X, LPCB, DMD-108)

Unidentified mould -a round conidium attached to the hypha by a short stalk, wider at the conidium than at the hypha.  (1000X, LPCB, DMD-108)

Unidentified mould -large, immature conidia attached to hyphae
(1000X, LPCB, DMD-108)

Unidentified mould - rather large conidia measuring up to 10 µm in diameter.  Note the base still attached to the one free conidium.  (1000X, LPCB, DMD-108)

Unidentified mould -spiral hyphae were regularly seen.
(1000X, LPCB, DMD-108)

Unidentified mould -conidia at different stages of maturity.  Note the attachment of the dark conidium  near center left of the photo.  It appears to still be attached by its inflated base or stalk.
(1000X, LPCB, DMD-108)

Unidentified mould - not a good photo -Why did I include this?
(1000X, LPCB, DMD-108)

Unidentified mould - two conidia attatched to or growing within? the hypha
(1000X, LPCB, DMD-108)
Unidentified mould -septate, branching hyphae with terminal and sessile conidia.
(1000X, LPCB, DMD-108)

Unidentified mould -large terminal oval conidium (chlamydospore?) seen in center of photo.  Immediately behind it there is a branch with a basal septum supporting a slightly out of focus conidium.  (1000X, LPCB, DMD-108)

Unidentified mould - just left of center, is what appears to be an intercalary conidium (chlamydospore).  That is, it appears to be in the middle of, and continuous with the hypha and not attached to it.  (1000X. LPCB, DMD-108)

Unidentified mould -conidia attached directly to the hyphae (sessile), on short, somewhat inflated stalks, and one near center-right on a longer simple stalk (or branch?)  Septations in the hyphae clearly visible.  (1000X, LPCB, DMD-108)

Unidentified mould -another photo as above.  A couple of the conidia appear to have a concave dimple.  Curious.  (1000X, LPCB, DMD-108)

Unidentified mould -and because 'more is better', here is another photo showing different stages of maturity and attachment.  (1000X, LPCB, DMD-108)

Unidentified mould -septate hyphae with sessile conidia.
(1000X, LPCB, DMD-108)

Unidentified mould - growth pattern of hyphae suggests sympodial growth (see next slide).  Pigment or stain may be exuded by the hyphae.  (1000X, LPCB, DMD-108)

Growth pattern of Sympodial vs Monopodial

Unidentified mould -Sessile conidium - resting right on the hypha.
(1000+10X, LPCB, DMD-108)

Unidentified mould -you may remember this photo from earlier in this post, however, here is a closer look at the central dark conidium and the short inflated base attaching it to the broken hypha.  A younger, ellipsoidal conidium appears in the lower left.  Note too the rather thick-walled appearance of the round conidium in the upper left.
(1000+10X, LPCB, DMD-108)

Unidentified mould
(1000+10X, LPCB, DMD-108)

 Unidentified mould -a photo showing many of the features mentioned previously.  Conidia (chlamydospores) appear to be intercalary, withing the hypha.  Suggestions of sympodial growth pattern.  Large ellipsoidal terminal conidium appears to have a septum (arrow), dividing it into two compartments.  (1000X, LPCB, DMD-108)

*   *   *

Friday, 11 December 2015

Sarocladium kiliense

Sarocladium kiliense (Formerly Acremonium kiliense)

Note: Recent revision of the genus Acremonium has placed the mould formerly known as Acremonium kiliense into the genus of Sarocladium and therefore is now known as Sarocladium kiliense[i].  Sarocladium currently contains 16 species with Sarocladium oryzae as the ‘type’ species in the genus.

Sarocladium kiliense is a ubiquitous soil saprophyte commonly found in many environmental locations.

Immunocompetent hosts are rarely infected by Sarocladium kiliense.  Those with underlying immunological disorders may be predisposed to infection, whether local or progressively systematic.  Infection is usually through traumatic implantation of the mould leading to the development of a granuloma.  Endocarditis, CAPD-associated peritonitis and keratitis have been described in the literature.  Although rare, post-operative endopthalmitis has also been reported following cataract surgery[ii].   This emergent, opportunistic fungus may be resistant to common antifungal therapies[iii].

Sarocladium is known to contain a number of species pathogenic to plants.

Macroscopic Morphology:
The isolate presented here exhibited moderate growth, maturing in about 7 days on Sabouraud Dextrose agar at 30ᵒC.  Colony expansion was somewhat restricted.  The colony developed an off-white, greyish-white to light tan or pinkish colouration with fine, dry felt-like surface texture.  Delicate radial surface striations were evident.  Reverse appeared yellowish to light brown.

 Sarocladium kiliense -  10 days growth on Sabouraud-Dextrose Agar (SAB or SDA) at 30ᵒC (Nikon)

Microscopic Morphology:
Hyphae were hyaline, septate and narrow, often grouping together in cords (fascicled hyphae).  The delicate phialides appeared long (25 -45 µm), thin-walled and tapering.  The long phialides extending from the hyphae may have a basal septum, or if intercalary, they may produce a lateral phialidic (adelophialide) outgrowth of variable length, which lacks the basal septum.  Adelophialides may be found submerged in the medium.  Single-celled conidia are described as ellipsoidal to short cylindrical, (3 – 6 X 1.5 µm) in size.  Conidia accumulate in slimy masses around the tip of the conidiophore unless physically dispersed.  They, at times, may appear to “fall back” and accumulate along the sides of the conidiophore.  Cell arrangements are easily disrupted even when gentle care is taken with a slide culture.
Note:  The micron bar which appears in most photos may change scale between 100 µm to 50 µm without notice. When photographing at a magnification of 1000+10X, the bar extends past the borders of the screen at 100 µm.  It may or may not not be reset for subsequent photos at 1000X.

Sarocladium kiliense - long, thin and tapering phialides are appearant extending from the bundle of hyphae running diagonally through the photo.  (400X, LPCB, DMD-108)

 Sarocladium kiliense - again, the thin, delicate phialides extend from the parent hyphae with a bundle of conidia collected at the apex of each.  Single, free conidia also seen dispersed throughout the photo.  (400X, LPCB, DMD-108)

Sarocladium kiliense - and another photo.  Some phialides disappear out of the focal plane of the camera and only their bundle of conidia show where they re-emerge.
(400X, LPCB, DMD-108)

Sarocladium kiliense - perhaps a better shot of a row of phialides extending from the 'parent' hyphae with the conidia they have produced still clinging together at the apex. Many sources describe the accumulated conidia at the tip as being held together in "slimy" masses.  (400+10X, LPCB, DMD-108)

Sarocladium kiliense - a curious photo where the conidia seem to have 'fallen' down along side of the phailides which bore them.  Of course there is no gravity involved here, yet the tip of the phialide has no conidia which are found lined up along either side.
(400+10X, LPCB, DMD-108)

Sarocladium kiliense - often, these hyphae tend to form bundles or cords as they cling together.  Again, long thin, delicate phialides bear conidia at their tips.
(400+10X, LPCB, DMD-108)

Sarocladium kiliense - another view of the same.
(400+10X, LPCB, DMD-108)

Sarocladium kiliense - taken from the edge of a slide culture, this one phialide has a huge number of conidia accumulated around its tip.  Hard to say if this phialide is a 'hard working model employee', or just a 'hoarder', accumulating free conida drifting about that others had produced.  I quickly found out that the structures are very delicate and easily disrupted.
(400+10X, LPCB, DMD-108)

Sarocladium kiliense - at a higher magnification.
(1000X, LPCB, DMD-108)

Sarocladium kiliense - as above.  I'll add some more until you 'get the picture' or 'get bored'!
(1000X, LPCB, DMD-108)

Sarocladium kiliense - a bundle of hyphae running diagonally through the photo.  The delicate phialides are about 25 -45 µm in length, thin-walled and tapering.
(1000X, LPCB, DMD-108)

Sarocladium kiliense -   Single-celled conidia are described as ellipsoidal to short cylindrical, (3 – 6 X 1.5 µm) in size.  I'm not sure as to what structure my arrow points to at the base of the phialide as it doesn't really look like a conidium nor a chlamydospore.
(1000+10X, LPCB, DMD-108)

Sarocladium kiliense - nicely stacked conidia and hyphae arranged in 'cords' (fascicled hyphae).
(1000X, LPCB, DMD-108) 

 Sarocladium kiliense - phialides extending from the hyphae may have a basal septum (arrow).
(1000+10X, LPCB, DMD-108)

Sarocladium kiliense - as seen above, the long phialides extending from the hyphae may have a basal septum, or if intercalary, they may produce a lateral phialidic outgrowth
(adelophialide) of variable length which lacks a basal septum.
(1000+10X, LPCB, DMD-108)

Sarocladium kiliense - the shape of the conidia differs from the pointy-ended, curved shape of Fusarium species which also may be produced at the ends of phialides, similar to that shown here.
(1000+10X, LPCB, DMD-108)

Sarocladium kiliense - rather than a short phialide, this one (center-left) can probably be traced back under the bundle of hyphae.  (1000+10X, LPCB, DMD-108)

Sarocladium kiliense - stepping back a bit in magnification, the massive numbers of conidia produced is well demonstrated in this photograph.  (400X, LPCB, DMD-108)

Sarocladium kiliense - a first look at the fungus when it arrived in the lab in transport, it appeared as above.  (400X, LPCB, DMD-108)

Sarocladium kiliense - round structures were repeatably viewed amongst the mass of hyphae.
(1000X, LPCB, DMD-108)

Sarocladium kiliense - these ones measured at 2.8 µm  and  3.2 µm in diameter however the sources I consulted stated the mature chlamydospores of Sarocladium kiliense are 4 - 8 µm in diameter .
(1000X, LPCB, DMD-108)

Sarocladium kiliense - I cannot find any reference to the dimensions of the chlaymydospores that Sarocladium kiliense produces.  If not an artifact, I can only suspect that these structures are the chlamydospores sources mention in relation to this organism.
(1000+10X, LPCB, DMD-108)

Sarocladium kiliense - another photo as above.
(1000+10X, LPCB, DMD-108)

Sarocladium kiliense -intercalary chlamydospore (insert)
(1000X, LPCB, DMD-108)

Most Acremonium species are phenotypically similar and therefore other means, such as molecular techniques, are required to accurately speciate isolates.  Often it is sufficient to report the isolate simply as Acremonium species.  It was through molecular means that it was determined that the fungus previously referred to as Acremonium kiliense differed sufficiently to be reclassified as a Sarocladium.

One feature which is unique to this former Acremonium is that it frequently produces terminal and/or intercalary chlamydospores.  Therefore, if you have isolated a mould which looks in every other way like an Acremonium species, yet chlamydospores are present – you probably have a Sarocladium kiliense.

The isolate presented here grew well at ambient room temperature and at 30ᵒC.  It showed reduced growth at 35ᵒC and failed to grow at 37ᵒC, therefore would probably not be pathogenic for humans.
This isolate was urea positive.

[i] Acremonium phylogenetic overview and revision of Gliomastix, Sarocladium, and Trichothecium
Summerbell RC, Gueidan C, Schroers HJ, de Hoog GS, Starink M, Rosete YA, Guarro J, Scott JA. 2011.
Stud Mycol 68:139-162, 2011

[ii] Acremonium kiliense Endopthalmitis That Occurred after Cataract Extraction in an Ambulatory Surgical Center and was traced to an Environmental Reservoir.
Scott K. Fridkin, Frederic B. Kremer, Lee A. Bland, Arvind Padhye, Michael M. McNeil and William R Jarvis.
Clin Infect Dis: 22 (February), pg. 222 – 227, 1996

[iii] In Vitro Evaluation of Antifungal Drug Combinations against Sarocladium (Acremonium) kiliense, an
Opportunistic Emergent Fungus Resistant to Antifungal Therapies.
Fabiola Fernández-Silva, Javier Capilla, Emilio Mayayo, Deanna Sutton, Josep Guarro
Antimicrob. Agents Chemother. February 2014 vol. 58 no. 2 1259-1260

Return to Top (Most Recent Posts) 

*   *   *