MALARIAL PARASITES

Transmitted by females anopheles mosquito

Falciparum and Vivax- responsible for 90 percent of cases of human malaria
o LIFE CYCLE
EXO ERYTHROCYTIC LIFE CYCLE
Sporozoites will be injected in human host and will infect the liver cells. They will
form shizonts. Schizonts will give birth to merozoites. Once the schizonts rupture,
merozoites enter the circulatory system
ERYTHROCYTIC CYCLE
Ring to troph to schizonts. Schizonts would rupture releasing the merozoites and
infecting adjacent erythrocytes.
Merozoites will develop to ring form. Ring form will develop to gametocytes.
(Macrogametocytes-female, microgametocytes-male)
SPOROGONIC CYCLE
Mosquito take blood meal taking gametocyte form. Gametocyte give birth to
ookinete. A group of ookinete will be called oocyst. Oocyst rupture and will release
sporozoites.
One antigenic feature of merozoites is the presence of PVM or parasitophorous vacuolar membrane
o PVM causes alteration in the morphology/antigenic properties of the malarial parasite that is why it
is hard to make a vaccine against malaria

PATHOGENESIS THE PATHOGENESIS OF MALARIA

Malaria will post no absolute diagnosis in clinical feature because signs and symptoms of malaria is similar to
other diseases (fever, nausea, epigastric pain)
PRE PATENT PERIOD injection of sporozoites until signs and symptoms are observed
o Plasmodium falciparum- 11 to 14 days
o Plasmodium vivax- 11 to 15 days
o Plasmodium ovale 14 to 26 days
o Plasmodium malariae 3 to 4 weeks
INCUBATION PERIOD injection of sporozoites until parasites can be demonstrated on body fluids (blood)
o Plasmodium falciparum- 8 to 15 days
o Plasmodium vivax- 11 to 20 days
o Plasmodium ovale 11 to 16 days
o Plasmodium malariae 18 to 40 days

Malarial infection causes regular paroxysm or episodes

CLASSICAL MALARIAL PAROXYSM
1. COLD STAGE
There is a strong or violent shaking
The patient will have core temperature that is high
There is intense peripheral vasoconstriction which causes the rise in temperature
Last up to 15 to 60 minutes
Start of having a fever
2. HOT STAGE
Also known as flash face
The patient have reddening of face and the temperature would reach as high as 40 to
41 degree celsius
Patient could experience headache, palpitations, tachypnea, epigastric discomfort,
thirst, nausea, and vomiting
Last up to 2 to 6 hours
Fever episode of malarial infection
 3. SWEATING STAGE
Include defervescence(lowering of temperature) and diaphoresis(profuse sweating)
Symptoms would diminish
Temperature lowers over 2 to 4 hours
CLASSIC PERIODICITY
Develop only if the patient is untreated and it corresponds to the length of erythrocytic cylcle
1. TERTIAN FEVER devided into two: benign and malignant
Benign tertian malaria
o Plasmodium ovale and Plamosdium Vivax
o It is benign because the fever would recur on the third day
Malignant tertian malaria
o Plasmodium falciparum
o The fever doesnt recur in the third day. Fever can sometimes recur in the
second day.
o More severe
2. QUARTAN FEVER
Plasmodium malariae
Fever on the fourth day

RECRUDESCENCE vs RELAPSE
1. RECRUDESCENCE
renewal of parasitemia or clinical features arising from persistent undetectable
asexual parasitemia in the absence of an exo erythrocytic cycle
blood has not been cleared of parasite
Plasmodium falciparum and Plasmodium malariae
2. REPLASPE
This is renewed asexual parasitemia following a period in which the blood contains
no detectable parasites
Plasmodium vivax and Plasmodium ovale

AGE OF ERYTHROCYTE INFECTED

Plasmodium falciparum- all ages of RBCs
Plasmodium vivax- young RBCs, may cause enlargement of RBCs
Plasmodium ovale- young RBCs, may cause enlargement of RBCs
Plasmodium malariae- old RBCs

PLASMODIUM PROTEINS contribute to the antigenic property of malarial parasites

1. PLASMODIUM FALCIPARUM ERYTHROCYTE PROTEIN MEMBRANE 1 (PfEMP1)

Most adhesive protein
Specific only for P. falciparum
Helps P. falciparum invade the immune response
2. ROSETTINS
Ligands for rosette formation
Most specifically, this would now help the schizont for its rosette formation
Helps the malarial parasite to adhere to RBCs and platelets
*Rosettins together with PfEMP1 Induce cytoadherence which results to fever
3. HISTIDINE-RICH PROTEINS (HRP)
Make adhesion more effective
4. RIFFINS
Digests hemoglobin forming hematin. Hematin is the pigment produced by malarial parasites
Mediate inflammation
Induce cytokines from monocytes and macrophages
 Most commonly released when schizonts rupture
5. GLYCOSYLPHOSPHATIDYLINOSITOL (GPI)
Surface antigens
Act like endotoxins of bacteria(acts the same as lipopolysaccharide of bacteria)
Stimulates monocytes to release TNF or cathepsins which results to fever

DEVELOPMENTAL STAGES

1. PLASMODIUM FALCIPARUM
May have marginal form. Marginal form has the capability to be present on the surface of RBCs.
Ring form very delicate ring and most commonly, 2 chromatin dots are seen
We can see 2 or more ring forms on one RBC
Trophozoite in the cytoplasm is a little thicker. There is present of basophilic stipplings (Maurer's dot)
Gametocyte- banana or crescent shape
2. PLASMODIUM VIVAX
Cause enlargement of RBCs
Ring- thicker than falciparum but is still delicate
Trophozoite- ameboid form
Gametocytes the same with plasmodium malariae and P. ovale
Basophilic stipplings - Schffner's dots
3. PLASMODIUM MALARIAE
Course ring
Trophozoite have the capability to have a band form trophozoite
Basophilic stippling - Ziemann dots
4. PLASMODIUM OVALE
Comet form trophozoites
Basophilic stipplings James dots
5. PLASMODIUM KNOWLESI
The same with the early ring forms of Plasmodium falciparum
Have two chromatin dots for its ring form
Has the capability to enlarge the RBC (2.75 increase in surface area)
Basophilic stipplings- Mulligan's dots

MALARIA PARASITES LABORATORY DIAGNOSIS

1. MICROSCOPIC EXAMINATION
A. Gold standard in identification of malarial parasites
B. Specimen can be taken anytime
C. Thick and thin blood smear
D. Usually done every 6 to 8 hours
E. Different stains that can be used: fields stain, wrights stain, giemsa stain
F. Romanowsky stain : use methanol as fixative
i. THICK FILM
A. 15 x 12 mm
B. Not fixed
C. For patients with low parasitemia
ii. THIN FILM
A. One cell thick
B. For specification of morphology and condition of RBCs
C. Fixed before staining
2. QUANTITATIVE BUFFY COAT METHOD
A. Use fluorescent microscopy
 B. Use to increase the yield of finding the parasite
C. Use the stain acridine orange
D. Observed under UV light source

E.

3. POLYMERASE CHAIN REACTION (PCR)

A. Use small amount of blood (10 microliters)
B. Enhances microscopic diagnosis
C. Can be used in cases of low parasitemia and case of mixed infection
D. Detect nuclei acid of malarial parasites
4. MALARIAL RAPID DIAGNOSTIC TEST
A. Principle would be immunochromatography in which we detect malarial antigens
B. It is very specific for P. falciparum
C. Uses whole blood
i. MALARIAL ANTIGENS
A. HRP II
HISTIDINE-RICH PROTEIN 2
Water soluble protein produced by trophozoites in young gametocytes of
Plasmodium falciparum
Sensitivity: 92.5%
Specificity: 98.3%
B. PLDH
PLASMODIUM LACTATE DEHYDROGENASE
Produced by sexual and asexual stages of Plasmodium falciparum only
Sensitivity: 88.5%
Specificity: 99. %
C. PLASMODIUM ALDOLASE
Can be produced by all the stages of plasmodium spp
Enzyme found in the parasite glycolytic pathway
5. SEROLOGIC TESTS
A. IHA- Indirect Hemagglutination Assay
B. IFAT - Indirect Fluorescent Antibody Test
C. ELISA Enzyme-Linked Immunosorbent Assay
Experiment 7a NEMATODA
NEMATODA
cylindrical, elongated, have tapery ends
tripoblastic: derived from 3 embyonic cell layers
ectoderm, mesoderm and endoderm
PARTS

1. BODY WALL
Outer covering
Outer cuticle
Sometimes being molt by the parasite (like a snake shedding skin)
Often bears cuticular setae. Setae are hair like structures surrounding the outer cuticle
Thin hypodermis and is muscular, andis composed of an alae and a bursa
1. Alae-longitudinal ridges derived from the cuticle
2. Bursa- located posterior of the parasite and is used for copulation. It is used by the
males to grasped the females
2. HYPODERMIS
Located beneath the cuticle
Cellular
Bulges known as Chords are seen in the body cavity
One dorsal, one ventral, and two lateral
Nuclei are seen inside the chords
3. MUSCULATURE (MUSCLE LAYER)
Beneath the hypodermis
Composed of longitudinal smooth muscles
There is no circular smooth muscles
Innervated by nerve trunks
4. PSEUDOCOELOM
Lacks epithelial lining
Space in which fluid may pass
Space between the muscle layer and the viscera organs
Serve as their circulatory system
Waste pass through diffusion and is carried by hemolymph direct to the anal region.
Hemolymph is the fluid like substance in the pseudocoelom. It is also responsible for the
distribution of nutrients to the body of the nematoda
BODY SYSTEMS

1. ALIMENTARY CANAL (DIGESTIVE SYSTEM)

Mouth surrounded by lips bearing sensory papillae. Sometimes the mouth of nematode would have
amphids. Amphids are chemosensory organs and lined by sensillae. Sensillae are hair like structures
surrounding amphids
*Ascaris lumbricoides have three lips- 1 dorsal and 2 ventral
Esophagus- musculature that pumps food into the intestine. Sometimes the structure of the
esophagus help in the nomenclature of the parasite
Intestines- tubular structures made up of single layer of columnar cells with prominent microvilli
2. EXCRETORY SYSTEM
Used for osmoregulation
Collects waste from the outer cuticle to the hypodermis until the muscular layer. And it will now
move the waste directly to anus
Sometimes the excretory tube will have an H-shaped.
There are two excretory tube anterior of the parasite and two excretory tubes posteriorly located
 3. REPRODUCTIVE SYSTEM
For the reproductive system, it is Gonochoristic or sometimes termed as bisexual meaning the male
and female reproductive system is separated.
Males
Relatively smaller than females
Have curved posterior end
Some is with copulatory spicules, and/or bursa (some may have copulatory spicules, some
may have bursa and some may have both)
1 to 2 testes into the seminal vesicle
For the gubernaculum, it aids in the descent of the gonads
Females
Start with the ovaries to the oviducts to the ootype then down to the uterus. The uteri will
now join to form the vagina and opens to exterior via the vulva
Mode of reproduction: copulation
Sperm is transferred to the vulva
Fertilization membrane is the covering of the young parasite that is being formed by
copulation. Fertilization membrane is called as the chitinous shell.
Beneath the chitinous shell is the second membrane impenetrable to all substances except
the CO2 and O2. CO2 and O2 are nutrients helpful for the growth of the parasite
Most of the products of copulation are eggs. These eggs are deposited in the feces of
humans. Those deposited eggs may contain uncleaved zygote, group of blastomeres(one of
the layers of the parasite) or complete larvae
Exception:
STRONGYLOIDES STERCORALIS
It may produces through parthenogenesis meaning it is capable of producing viable
eggs without fertilization, meaning it can produce its own parasite without male
parasite
Some Parasites would not lay their eggs onto the feces, rather the eggs are hatched in the
uterus and will release the larvae into the feces. This parasites are called VIVIPAROUS
Applicable in fillariae spp and trichinella spiralis

EXPERIMENT 7B
APHASMIDS
No lateral excretory canal
Esophagus is cylindrical in shape in which the esophageal glands forming stichosome
Eggs are discharged unsegmented with plug on either pole or hatched in uterus
Exception: Trichinella spiralis hatched their eggs inside the uterus yielding to larvae

Trichinella spiralis

Common name: biblical worm, trichina worm

Disease: trichinosis
Infective stage: encysted larvae
Habitat: inside skeletal muscle, as well as the small intestines
Has small, nonpapillated mouth with protrusible stylet
It is largely filled with stichosome
Intestine is a single tube extending to the cloaca or anus

ADULT MALES
Smaller than the female
 Have delicate anterior end with cephalic papillae
Posterior end is bluntly rounded with 2 lobular caudal appendages ( 2 lobular caudal
appendages help in copulation)
Reproductive system have a single coiled testes
ADULT FEMALE
It is viviparous meaning the eggs are hatched inside the uterus
Yield low larvae : 500 larva per month
3.5 x 6 mm
LARVA
Larva has spear like burrowing tip at anterior end and has a tapering anterior end (narrow
anterior end)
Encysted in the skeletal muscle and small intestine
Common muscles infected: Diapghragmatic, intercostal, lingual, mesenteric, lingual, deltoid,
biceps, laryngeal, extraoccular, gluteus, gastrocnemius, pectoral
ENCAPSULATION
Complete encapsulation: 21st day (formation of nurse cell)
Produces by the body in response to the presence of the larva
A capsule surrounding the larva would become a nurse cell giving nutrients to the larva
Nurse cell will undergo calcification after/in 30 years
Permanent capsule is produced in 3 months
There are 2 layers or coined as mantles. The inner mantle is composed of basophilic
degenerative muscle and fibroblasts and epitheloid cells. The outer hyaline layer is the
sarcolemma of the infected skeletal muscle fiber
TRICHINELLA SPIRALIS DIAGNOSIS

1. CPK (CREATINE PHOSPHOKINASE) and LACTATE DEHYDROGENASE would be muscle markers. Once
increased, they indicate destruction of muscle cells
2. EOSINOPHILIA increases as disease progresses
3. SEROLOGIC TESTING (BENTONITE FLOCCULATION AND LATEX AGGLUTINATION)
bentonite flocculation use for rheumatoid arthritis but since they have the same manifestation with
muscle destruction, it can be a supplement testing for diagnosis
latex agglutination
4. BACHMAN INTRADERMAL TEST
can be an indicator for recent infection
reagent: inactivated larvae antigen
Injected in the presumed infected host
Positive: white swelling surrounded by unraised irregular wheal up to 15 cm in diameter. The results
would show within 15 to 20 minutes
5. SKELETAL MUSCLE BIOPSY
Considered as the gold standard in the identification of trichinella spiralis
Identify encapsulated larvae

Trichuris trichura

Also known as the whip worm

Flesh colored
Male is smaller than the female
Posterior end resemble handle of the whip
Male have a curved posterior end with a conspicuous spicule

EGGS
Barrel-shaped or Japanese lantern shape with bipolar plugs
 Can be unembryonated or embryonated
A female adult would lay 1000 eggs per day
The eggs consider moisture as an ideal environment
TRICHURIS TRICHURA DIAGNOSIS

1. DIRECT FECAL SMEAR

2. KATO-THICK SMEAR
More reliable
Light infection average of 10 eggs per smear
Heavy infection average of 50 eggs per smear
Massive infection (TNTC) too numerous to count
3. PROCTOSCOPY
Done by physicians

Capillaria philippinensis

Also known as Pudoc worm

Originated from Ilocos Sur
Width is uniform from posterior to anterior
Body is divided into two equal parts. The first part is the digestive system and the second half is the
reproductive system.

MALE ADULT
Conspicuous spicule
Spicule sheath surrounds the spicule
FEMALE ADULT
Ovoviviparous can lay eggs if it is in its typical form
Typical form: fish eaten
Atypical female adult: viviparous
EGGS
Diagnostic stage
Peanut shape
Have bipolar plugs
Have striations
Develop inside the fishes
LARVA
Infective stage
CAPILLARIA PHILIPPINENSIS DIAGNOSIS

Stool exam
Demonstration of the eggs or the adult form in stool exams
The larva is not found in stool because they reinvade the small intestine