BLOOD AND TISSUE NEMATODES IN MAN

• Filarial worms
• Arthropod transmitted/mosquito-borne parasites
• Infective stage to man: filariform/filiform/3rd stage microfilariae
• Infective stage to vector: microfilariae
• Diagnostic stages: Microfilariae: blood
Adult larvae: lymphatics
• Morphology:
1. Adult- threadlike, creamy, white, varies in length (2-50 cm)
2. larval stage- snake-like with a column of cells in the anterior to the posterior portion
3. Microfilaria-pre-larval stage, embryos produced by filarial worms usually found in the
blood or tissues of patients with filariasis, highly motile and threadlike
❖ Location of Microfilariae in man: peripheral blood and lymph spaces of the skin
• Sheath: a thin, translucent egg shell remnant covering the body of the microfilaria and
past the head and tail

SHEATHED Microfilariae
1. Wuchereria bancrofti
2. Brugia malayi
3. Loa loa

UNSHEATHED Microfilariae
1. Onchocerca volvulus
2. Dipetalonema perstans/Mansonella perstans
3. Dipetalonema streptocerca/Mansonella streptocerca
4. Mansonella ozzardi

PERIODICITY: rhythmical appearance of microfilariae in the peripheral blood

Types:
1. Periodic:
➢ Nocturnal – 10pm to 2am
➢ Diurnal – 10am to 2pm

2. Subperiodic:
➢ Nocturnally – peak count during night time
➢ Diurnally – peak count during day time

3. Non-periodic: during night time and day time

• Disease caused: Lymphatic Filariasis
• Clinical Manifestation:
a) HYDROCELE: swelling in the scrotum that occurs when fluid collects in the thin sheath
surrounding a testicle.
b) LYMPHEDEMA: swelling that generally occurs in one of the arms or legs
 c) ELEPHANTIASIS: condition characterized by gross enlargement of an area of the body,
especially the limbs
d) CHYLURIA: rare condition in which lymphatic fluid leaks into the kidneys and turns the
urine milky white

LABORATORY DIAGNOSIS FOR FILARIAE

1. MICROSCOPIC EXAMINATION
a) Wet smears & thick smears – microfilariae
➢ Stains: Wrights; Giemsa; Delafield Hematoxylin
➢Negative results: low intensity of infection; dead worms; obstructed
lymphatics
➢ DEC (Diethylcarbamazine) provocative tests (3mg/kg single dose)-
stimulates microfilariae to come out to the peripheral circulation

b) KNOTT’S METHOD
➢ 1 ml WB + 10 ml 2% formalin
➢ Centrifuge at 500 x for 10 minutes
➢ Sediment- thick and thin smears: Microfilariae & WBCs
➢ Centrifugation of the blood sample lyzed in 2% formalin (Knott’s
technique)

c) FILTRATION: Nucleopore or Millipore membrane (5 µm pore)

➢ 1 ml of fresh or anticoagulated blood is drawn up into a syring
➢ Lyze by adding 10 ml distilled water then pass through the Swinney filter
membrane
➢ Examine filter membrane

d) SKIN SNIPS: microfilariae of O.volvulus and M. streptocerca

➢ Use corneal-scleral punch, or a scalpel and needle
➢ Incubate sample for 30 minutes to 2 hours in saline or culture medium
then examine for microfilariae that would have migrated from the tissue to
the liquid phase of the specimen

2. CAPILLARY TUBE METHOD

➢ Use heparinized capillary tube; examine buffy coat layer

3. IMMUNOASSAY
➢ Antigen detection- circulating filarial antigens
➢ Molecular diagnosis using PCR- differentiation of filarial species and stage

4. IDENTIFICATION OF ADULT WORMS

➢ Ultrasonography- demonstrate live worms in the lymphatics
➢ Contrast lymphangiography & lymphscintigraphy using radiolabeled
albumin or dextran
➢ Tissue samples collected during nodulectomies (onchocerciasis)
➢ Subcutaneous biopsies or worm removal from the eye (loiasis)

TREATMENTt
1. DEC (Diethylcarbamazine)/Hatrazan
➢ A filaricidal piperazine derivative that kills both microfilariae and some adult
worms of Wuchereria bancrofti and Brugia malayi
➢ Periodic mass treatment: Single dose of DEC for 6 months
➢ 6 mg/kg body weight for 12 days (bancroftian filariasis)
➢ 3 – 6 mg/kg body weight up to a total dose of 36-72 mg/body weight
(brugian filariasis)
▪ Use of DEC fortified tablet salt ( 0.1% DEC or 0.3% DEC for 3-4
month

2. IVERMECTIN (single dose of 200-400 µg/kbw for 12 days)

▪ More effective if used in combination with DEC
▪ Relief of pain- cool the affected area
• Prevention
➢ Abaca workers-wear long sleeves shirts
➢ Use of mosquito repellants and/or mosquito nets; vector control (insecticides)

Wuchereria Brugia malayi Loa loa

bancrofti

Bancroft’s filarial Malayan filarial Eye worm, Loa

Common name worm worm worm

Bancroftian Malayan filariasis, Loasis, fugitive

Disease(s) filariasis, elephantiasis of swelling, calabar
caused elephantiasis of upper swelling(diameter:
lower extremeties 5-
extremeties, 10 cm); migration
lymphatics of rate under the
scrotum; skin:
wuchereriasis 1 inch/2 mins

Morphology MALE: 4 cm in MALE: 13-23 mm MALE: 30-34 mm

length in length by
by 0.1 mm FEMALE: 43-55 0.35 -0.43 mm
FEMALE: mm FEMALE: 40-70
viviparous, 8- mm
10 cm in length by by 0.5 mm
0.2-
0.3 mm

Development Development Maturation inside

 inside inside MOSQUITO:
MOSQUITO: 6-20 MOSQUITO: 2 10-12
days weeks days
VESSELS OF VESELS: 3-9 SUBCUTANEOU
MAN: 6 mons S
mons TISSUES: 1 year
LIFE SPAN: 5 LIFE SPAN: 1-15
years years
Habitat
(adult
worm)
Lymph vessels
and
lymph glands
(lower)
Lymph vessels
and lymph
glands (upper)
Subcut

Lymph vessels Lymph vessels Subcutaneous

Habitat and and lymph and
(adult lymph glands glands (upper) muscular tissues
worm) (lower)

blood blood blood

Specimen

Culex pipiens Mansonia spp Chrysops spp

Vector quinquefasciatus Anopheles spp (Deer fly /Mango
fly)

In the Philippines,
Endemicity the province
endemic for W.
bancrofti are:
Camarines Norte
& Sur, Bohol,
Samar, Albay,
Leyte, Sorsogon,
all provinces in
Mindanao,
Quezon, Mindoro,
Mt. Province,
Sulu,
Masbate,
Palawan,
Romblon
W. bancrofti
- Anopheles
 minismus var,
flavirostris – rural
type
- Aedes poecillus-
urban type of
bancroftian
filariasis (both
breed in the axils
of abaca &
banana)
B. malayi
- Mansonia
bonnae (in fresh
water)
-Mansonia
uniformis (rice
fields)

DIFFERENTIAL CHARACTERISTICS OF SHEATHED MICROFILARIAE

Wuchereria Brugia malayi Loa loa

bancrofti

200-300 µm 220-250 µm 250- 300 µm

Length

8 µm 6 µm 8 µm
Diameter

Stains slightly w/ Stains deeply w/ Almost colorless

Sheath Giemsa Giemsa w/Giemsa

Regular smoothly Irregular and Irregular and

Body curves curved twisted twisted same as
Graceful Stiff w/secondary B.
sweeping kinks malayi

Small/short (1:1) Large/longer (1:2) Larger/longer

Cephalic space (1:2)

Coarse/well Coarse, tend to Coarse tend to

Body nuclei separated overlap overlap

No nuclei pointed Nuclei present

Tail end tip 2 widely spaced rounded tip
nuclei blunt tip

nocturnal nocturnal diurnal

Periodicity
 UNSHEATHED FILARIAL WORMS

Onchocerca Mansonella Mansonella Mansonella

volvulus perstans streptocerca ozzardi

Common Blinding worm, Synonyms: Ozzard’s

name convoluted Dipetalonema,Acanthocheilonema
filaria

Disease Onchocerciasis dipetalonemiasis streptocerciasis Ozzard’s

cause , filariasis,
onchocercosis, mansonelliasis
river blindness ozzardi

Morphology MALE: 19-42 MALE: 45 mm in FEMALES: 65-81

cm length mm by 0.21-
by 130-210 µm FEMALES: 80 0.25 mm
FEMALE: 50 mm
cm in length
by 270-400 µm

Habitat Subcutaneous Inside body Inside body

tissues cavities cavities,
mesentery and
visceral fat

Specimen Skin snips blood blood blood

Vector Simulium Culicoides spp Culicoides spp Culicoides spp

damnosum (night biting)
(black flies;
buffalo fly)

Endemicity Found in Africa Found in Africa

DIFFERENTIAL CHARACTERISTICS OF UNSHEATHED MICROFILARIAE

Onchocerca Mansonella Mansonella Mansonella

volvulus perstans streptocerca ozzzardi

Length 250-300 µm 150-200 µm 180-240 µm 150-200 µm

Diameter 8 µm 4 µm 5 µm 4 µm

Sheath Absent Absent Absent Absent

Body curves Regular slightly Regular often Tail usually Regular slightly
twisted forms loops curved twisted
 Cephalic Large and Large Large Large
space bulbous

Body nuclei Coarse mostly Medium sized Fine mostly Fine mostly
separated tend to separated separated
overlap

Tail end No nuclei Nuclei present Nuclei present No nuclei

pointed tip rounded tip curved tip pointed tip

Periodicity Non periodic Non periodic Non periodic Non periodic