Trichinella spiralis

(Trichinellosis or trichinosis)
Unlike many parasites that demonstrate a high degree of host specificity, Trichinella spiralis,
the trichina worm, can be found in many species of carnivores and omnivores. Animals are
infected with T. spiralis when they ingest infective larvae (juveniles) in raw or undercooked
meat. The larvae mature into adults in the host's small intestine in a few weeks, and the
female worms give birth to larvae. (The males die after fertilizing the females, and the
females die after producing larvae.) The larvae enter the blood stream of the host and,
eventually, end up in the host's muscles. Here the larvae mature into infective larvae, and the
next host is infected when it eats these larvae. In the muscles the larvae cause a severe host
reaction that results in soreness and tenderness of the muscles (view diagram of the life
cycle). Although this parasite probably only rarely causes fatalities in humans, it can cause
extreme discomfort. Trichinosis is probably best known as a parasite that humans contract
from eating raw or undercooked pork. Through an aggressive program of meat inspection,
the incidence of trichinosis in pigs in the United States has been lowered to less than 1%, so it
is unlikely (but not impossible) that pork products purchased in your local supermarket will
contain Trichinella larvae. Most recent outbreaks of trichinosis in the United States have been
traced to pork products from pigs that have not been inspected and that have been slaughtered
privately. Because of its low host-specificity, almost any "wild" meat should be considered
suspect, and hunters should be careful when preparing meat from their kills. In particular, a
number of infections have been traced to contaminated bear meat.

A higher power magnification of the above image.

A larva in a "teased muscle" preparation.

Life Cycle of Trichinella spiralis

Adult Female

Adult Male

Infective Muscle Larva

Intravital microscopy has revealed the relationship of the Nurse cell to the surrounding tissues. The following
video (under construction) is an in vivo, living Nurse cell-parasite complex (we hope to make this video
available for download soon). The Nurse cell begins to form the moment the newborn larva penetrates the
striated skeletal muscle cell.

Schematic of a Larva Invading a Muscle Cell

BEGIN SPECULATION
The mechanism(s) by which the newborn larva enters the host cell is not known, but morphological evidence
gives hints as to the overall process. In muscle tissue, the larva penetrates out of the capillary and then braces
itself against the adjacent muscle cell. In doing so, it gains leverage enabling it with its anterior end to depress
the sarcolemmal membrane to the point of breaking. Then, it most likely brings its stylet into play,
poking a hole in the membrane, causing the host cell to "explode", witness the "ragged" edges of the
sarcolemmal membrane observed just moments after the larva entered the host cell. We envision the
overall process as similar to one of us poking our index finger straight into a fully inflated balloon, then
somehow managing to unsheathe a sharp finger nail, bursting the frail object. Therefore, we do not think
secreted enzymes are necessary to aid its passage from the blood stream to the intracellular environment of
the muscle cell. However, this last idea needs to be tested.

There are at least two distinct phases to the overall process:

1. De-differentiation of the muscle cell
2. Re-differentiation of the muscle cell into the Nurse cell
The morphological events resulting in the formation of the Nurse cell-parasite complex occur over a 14-16 day
period of time after the worm has entered the muscle cell. The diagram below summarizes the overall change.

The accompanying toluidine and methylene blue stained light microscope sections and electron micrographs are
from synchronously infected mouse muscle tissue embedded in plastic, and correspond to the time points in the
diagram. Click here to see these sections.

1. De-differentiation of the Muscle

[Back To Nurse Cell Formation]
[Back To Outline of Sections]
Gallery:

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De-differentiation EM Gallery

EM of the Stichocyte of Mature Muscle Larva

Mature Nurse Cell Cytoplasm

Granule Containing Cells of the Newborn Larva

The rate of increase in the worms volume is a crude measure of worm growth (see Growth Curve below), and
differentiation of its tissues, for example stichosome, is correlated with the growth and differentiation of the
Nurse cell. Only the rate of growth of the worm has so far been documented. The larva grows during the first
day after entering the muscle cell, doubling its volume. The parasite remains the same size over the next two
days, resuming growth on Day 4. During this pause, the host cell becomes disorganized with respect to its
contractile elements.

The first five days of the process of Nurse cell formation involves a loss of normal structures and contractile
functions. Actin and myosin filaments begin to disappear and the sarcolemmal membrane becomes separated
from the contractile elements. Click here for a series of EMs that document these observations. Muscle
mitochondria become vacuolated and ATP synthesis is uncoupled from aerobic metabolic pathways beginning on
Day 3, and remains so throughout the infection.

BEGIN SPECULATION:
We believe that mitochondrial dysfunction is essential to the life of the developing parasite, since the larva is an
anaerobe.
END SPECULATION
Thereafter, up to Day 19, the parasite increases its volume by 39% each day, and while growth continues, the
parasite achieves infectivity on Day 14-16.
BEGIN SPECULATION:
We hypothesize that the newborn larva induces at least some of the early events in Nurse cell formation, and
may do so by secreting substances into the milieu of the muscle fiber. The anterior half of the newborn larva
contains a row of cells inside of which are found numerous granules reminiscent of those seen in the fully
developed stichocytes of the infective larva. If these granules contain secretory proteins, then newborn
larvae most likely releases them into the newly penetrated muscle cell.
END SPECULATION

2. Re-differentiation of the Muscle

[Back To Nurse Cell Formation]
[Back To Outline of Sections]
Gallery:

Molecular Structure of Tyvelose

Epicuticle of T. spiralis contains tyvelosylated proteins

Most of the important changes related to down regulation of the fully differentiated host cell occur by Day 8.
Enlargement of host cell nuclei are at their maximum on that day (below), and it is at this point in the infection
that the stichocytes begin to produce and secrete tyvelose-decorated proteins into the milieu of the infected
host cell.

Hypothesis: Secretions of the larva induce host nuclei to enlarge:

Tyvelosylated parasite proteins (at least 4) are found on the epicuticular surface (Despommier and Kajima,
1969; Almond, et al), within the cytoplasm of all stichocytes, and within the cytoplasm and, most importantly,
the nucleoplasm of all nuclei of the developing Nurse cell. Nurse cell nuclei decrease in volume on Day 10 and
remain at that volume thereafter .
Begin Speculation:
The larva must influence the growth of the host cell. Are secreted proteins involved? If so, are any of them
related in function and/or structure to known mammalian cell growth factors? The enlargement of host cell

nuclei is correlated with the presence of secreted parasite proteins within the nucleoplasm of each enlarged
nucleus. The enlargement process results in very large nuclear pores. Does this then obviate the need for
nuclear localization signals from parasite secreted proteins so that they may enter and exit the nucleus at will?
End Speculation

2.a Collagen Capsule Formation

[Back To Re-differentiation]
[Back To Outline of Sections]
Gallery:

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Histochemistry and ImmunoHistochemistry of the Capsule

Invading cells breaking down a small portion of the collagen capsule

In Situ Hybridization using cRNAs for Collagen Type IV

In Situ Hybridization using cRNAs for Collagen Type VI
Northern Analysis

An acellular capsule begins to form outside the Nurse cell beginning on Day 10. It continues to thicken until Day
26, and is largely composed of collagen type IV and VI (Polvere, et al, 1998). Collagen synthesis occurs within
the cytoplasm of the developing Nurse cell and is directly correlated temporally with the thickening of collagen
along the outer surface of the Nurse cell.
BEGIN SPECUALTION:
The induction of the collagen outer capsule is a direct result of exposure to secreted proteins of the parasite.
END SPECULATION
Through the use of anti-collagen antibodies and specific RNA probes, the types of collagen present in the
capsule were determined. Two dominant collagen types were detected, type IV and type VI. Northern
analysis and in situ hybridization studies showed that type IV collagen synthesis begins on Day 10 and
ceases on Day 26, while collagen type VI begins on Day 10 and continues throughout the infection period.
BEGIN SPECULATION:
If collagen synthesis is up regulated by secreted proteins from the larva, then more than one parasite protein
must be involved, since each collagen gene family is regulated separately.
END SPECUALTION
Type IV collagen is a large molecule (Show structure and give MWT) and is found throughout the vertebrate
body, and most likely functions as a scaffolding element, holding cells in various tissues together. In contrast,
collagen type VI is a relatively short molecule (give structure and MWT), functioning to bind larger collagen
molecules together by cross-linking them, like the individual rungs of a ladder that together support the two
legs.
The Nurse cell cytoplasm is infiltrated with numerous, as yet unidentified, mononuclear cells (ems - 6 to 8 -,
and light microscopic photos). The number of different types of host cells that gain entrance to the Nurse cell
has yet to be determined. Their function(s) in Nurse cell formation and/or maintenance remains enigmatic.
Initial observations indicate that the process of cell recruitment into the Nurse cell is on-going.
BEGIN SPECULATION:
Careful visual inspection of the capsule by either light or electron microscopy does not suggest that there are

any obvious weak spots or holes in its outer surface, although we are not aware of any a systematic search for
such breaks. This aspect of capsule formation and maintenance needs to be thoroughly investigated. Since
trafficking of host cells in and out of the cytoplasm of the Nurse cell appears to occur continuously after Day 10,
the process most likely requires that invading cells break down at least a small portion of the collagen capsule
in order to facilitate their entrance. If this is a valid way of envisioning the process, then repair of the
capsule would, by necessity, also be an ongoing process. Our model would thus explain the need for continuous
synthesis of collagen type VI. In contrast, type IV is a more loosely packed molecule in the capsule, at least it
appears that way on electron micrographs. Thus, cells might be able to crawl in and out of the cytoplasm
without disturbing its lacy network of fibrils once they have disconnected type IV molecules from their crossconnecting type VI helpers.
END SPECULATION
Suggestions for experiments that explore some of the possibilities raised above:
a.

If various sub-sets of mononuclear cells from the host were first isolated from the blood of an infected
animal by cell sorting, then permanently labeled, they could then be injected iv, one sub-set at a time.
Labeled sub-sets of mononuclear cells arriving at the surface of the Nurse cell could then be identified
and tracked throughout the Nurse cell cytoplasm, based on their marker. The marker has to be
internal, since surface markers are likely to be shed during the act of diapedesis.

b.

Exposing an infected host simultaneously to labeled amino acid precursors of collagen type VI could
facilitate research into whether or not collagen type VI was involved in any repair mechanism using
autoradiography. Labeling of type VI collagen would be achieved by giving label after Day 30 of Nurse
cell formation, thus eliminating the chance of accidentally labeling type IV collagen. Patchy distribution
of labeled collagen type VI on the capsule surface of a mature Nurse cell would constitute positive
evidence for repair events, and may even allow for the identification of the cell that caused an event to
occur if marked cells were used simultaneously

c.

Nurse cells can be isolated and maintained for long periods of time in vitro (photos). The potential thus
exists for placing various mononuclear cell types in the presence of mature Nurse cells to explore the
possibility of observing the process of cells going in and out. Since cells loose their surface markers
while inside the cell that they are travelling through, by collecting mononuclear cells after they exit
from the Nurse cells in culture, one could possibly identify their surface markers after they are
regenerated using specific monoclonal antibodies.

Data Summary Table For Capsule Formation

Click here for the

Click here for the

histochemical stains

immunohistochemistry

Click here for the in situs

Click here for a

hybridizations

Northern Analysis
[Back To Re-differentiation]

2.b Circulatory Rete Formation

[Back To Re-differentiation]
[Back To Outline of Sections]

A network of circulation develops immediately adjacent to the collagen capsule (PHOTO of vascular casts and
diagram), beginning on Day 7, as evidenced by the induction of vascular endothelial growth factor (VEGF)
within the confines of the developing Nurse cell (in situ.). The rete consists of flattened, tortuous vessels, some
of which end blindly. They are generally wider than capillaries, and resemble vessels constituting sinusoids. The
presumed function of the circulatory rete is to enable the parasite to obtain nutrients and facilitate the transport
of small molecular wastes. This supposition needs to be documented by experimentation.

Suggestions for experiments to elucidate the function of the circulatory rete.

(see summary of inhibitors)
Interfering with the formation of the circulatory rete seems like a reasonable first order strategy to approach
the problem of the function of the rete for the parasite. Anti-angiogenic drugs and antibodies against essential
growth factors such as vascular endothelial growth factor (VEGF) and alpha V beta 3 integrin can be employed.
Antibodies directed against VEGF are effective in preventing the growth of capillaries towards the source of
VEGF. Examples of anti-angiogenic drugs include fumagillin, that inhibits division of vascular endothelial cells,
lavendustin-A, that inhibits tyrosine kinases flk, flt-1and flt-4, tricyclodecan-9-yl xanthanante that inhibits
collagen type IV synthesis (an essential scaffolding molecule for angiogenesis), and rIL-12. Interleukin 12
induces interferon gamma, which, in turn, induces protein IP-10, a known inhibitor of angiogenesis. The
mechanism of action of IP-10 is not known.
These approaches have all been clinically tested in humans and have shown promise as anti-tumor agents. Any
of these agents should show some activity against rete formation. Even partial inhibition should show some
effects on the growth curve of the larva, if the rete is essential for its nutrient acquisition. As new classes of
inhibitors are discovered and become available to the research community, a more comprehensive approach to
studying the circulatory rete will no doubt evolve.

Development of Trichinella spiralis in skeletal

muscles
Picture 1

Picture 2

Picture 3

TSLarvwandSK-ic
Migrated larvae

TSLarvcapsSK-ic
Encapsulated
larvae

TSLarvcaps1SK-ic
Calcified larvae

spir7.ic
Trichinella spiralis: Electron

Atlas de Parasitologa On Line

Trichinella spiralis

VOLVER AL INDICE

Diseo: Dr. Eduardo Luis Freggiaro

FORO BIOQUIMICO

Trichinella spiralis in muscle of dog

(Armed Forces Insitute of Pathology)

Slide 15 of 41

Slide 19 of 41

Trichinella spiralis is a tissue nematode

humans acquire by ingesting improperly
cooked meat, particularly pork. After
ingestion, the larvae enter the intestinal
mucosa and mature into adults. The adults
mate and larvae are produced. The larvae
make their way into body tissues,
particularly active muscles. The larvae then
encyst reaching lengths of 0.8 -1 mm. This
image is from a muscle tease-prep.

Home > > > (Trichinella spiralis)

. spiralis
engl.: pork worm

Trichinella spiralis (Histologie) [83]

(OWEN 1835, RAILLIET 1895) die kosmopolitische, relativ kleine Trichine (weibl. < 4,
mnnl. < 1,5 mm), deren geschlechtsreife Formen Dnndarmparasiten (Mukosa, Submukosa,
Lymphbahnen, s.a. Abb.) zahlreicher Suger sind (s.a. Darm-T.) u. deren Larven sich im
Muskelgewebe einkapseln (Muskel-T.). Verbreitung unter Tieren durch Fressen infizierter Tiere bzw.
Fleischabflle; Infestation des Menschen (Trichinose) durch rohes oder schlecht gekochtes
infiziertes (Schweine-)Fleisch: Im Magen frei gewordene lebensfhige Larven werden in der
Dnndarmwand in wenigen Tagen geschlechtsreif; die von befruchteten weibl. zahlreich geborenen
Larven gelangen ber Lymph- u. Blutgefe (Bluttrichine) in die Muskulatur (v.a. Zwerchfell) u.
kapseln sich dort ein (Trichinellenzyste = eingerollte T., umgeben von einer vom Perimysium
gebildeten Kapsel, an den Polen durch Granulationsgewebe verschlossen).

** Image Library! Center for Disease Control! Gov. ~ Robbins, S. Textbook

of Pathology, W.B. Saunders Co. Phila. 1957, 373.

Trichinella spiralis
Diseases | Sites and Sources | Diagnostic Factors | Virulence Factors | Treatment and
Prevention | Commentary
Category
pork worm
Classification

helminth, nematode

Diseases
Trichinosis (Trichinellosis)
fever

gastroenteritis

abdominal pain

nausea

vomiting

constipation

diarrhea

myalgia

eosinophilia

dyspnea

cardiovascular changes

petechial hemorrhage

pericardial effusion

periorbital edema

neurologic changes

CNS damage

Sites and Sources

animals, source

livestock, source

wild animals, source

wild hogs, source

hogs, source

pork, source

undercooked meat, source

intestine, pathogen

GI tract, pathogen

heart, pathogen

CNS, pathogen

muscle, pathogen

multiorgan, pathogen

Diagnostic Factors
muscle biopsy

eosinophilia

serology

Virulence Factors
life cycle

Treatment and Prevention

proper animal husbandry

meat inspection

proper food preparation

mebendazole

thiobendazole

steroids

Commentary

Trichinella can infect virtually all mammals. The adult worm lives in the
gastrointestinal tract, while the infectious larval form is present in cysts in
the striated muscles of these animals. Trichiosis is a zoonotic infection
transmitted by eating undercooked meat which contains the Trichinella
cysts. It is most often associated with pork although other animals are
potential sources. This disease was likely one cause of the religious
injunctions against eating pork. Infection is widely distributed with an
estimated 10-15 million human infections in North America. In this country
insufficiently cooked pork, or raw hamburger contaminated with pork, and
occasionally wild game are sources of human infection. Polar bears and
walruses have been implicated in outbreaks in the Arctic. After ingestion of
cysts in the meat, the organism develops to an adult in the intestine. The
female produces larvae which migrate throughout the body and encyst in
striated muscle. This causes inflammation and muscle pain. The larvae can
also migrate to the brain and cause a variety of symptoms. In fact, the
larvae can invade almost any part of the body producing inflammation and
disease which can mimic many different clinical syndromes. The disease
varies considerably in severity, depending on the number of organisms
present and the location of the migrating larvae. Patients with 10 or less
larvae are likely to be asymptomatic. 100 will cause significant disease
while 1000 to 5000 can cause death. Mild infections are characterized by
diarrhea and flu-like symptoms. With more severe disease there is
persistent fever, gastrointestinal distress, muscle pain, periorbital edema
and eosinophilia, amoung other things. Neurological symptoms appear in
the most severe cases. Laws against feeding uncooked garbage to hogs
resulted from an effort to control trichinosis, as well as some viral
infections in hogs.