Annals of Tropical Medicine & Parasitology

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Videomicroscopy of intralymphatic-dwelling Brugia malayi T. C. Case, M. H. Witte, D. L. Way, C. L. Witte, C. A. Crandall & R. B. Crandall To cite this article: T. C. Case, M. H. Witte, D. L. Way, C. L. Witte, C. A. Crandall & R. B. Crandall (1992) Videomicroscopy of intralymphatic-dwelling Brugia malayi, Annals of Tropical Medicine & Parasitology, 86:4, 435-438, DOI: 10.1080/00034983.1992.11812690 To link to this article: https://doi.org/10.1080/00034983.1992.11812690

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Annals of Tropical Medicine and Parasitology, Vol. 86, No.4, 435-438 (1992)

Videornicroscopy of intralyrnphatic-dwelling Brugia malayi In lymphatic filarial infections the adult nematode lives out its life span within the host lymphatic system hidden from view and seemingly impervious to immune surveillance. Even the microfilariae, although periodically disseminated in the bloodstream, are rarely seen alive in their intralymphatic birthplace. To elucidate

this aspect of the filarial life cycle we examined the intralymphatic phase in ferrets infected with Brugia malayi, using high-powered videomicroscopy (Ikegami video camera in series with a Mentor dissecting Olympus light or inverted Olympus light microscope at initial magnifications of x 25- x 66).

Fig. 1. Photographs with replica drawings (insets) of (A) highly motile adult Brugia malayi in a dilated ferret hindlimb lymphatic in vivo, and (B) adult worm, within an excised but intact lymphatic, traversing an intralymphatic valve (extreme right). 0003-4983/92/040435 + 04 $08.00/0

© 1992 Liverpool School of Tropical Medicine

Fig. 2.

Fig. 3.

VIDEOMICROSCOPY OF B. MALAY!

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Fig. 4. (A) Comparative size of lymphatic-dwelling microfilaria (arrow) ( ~ 250--300 11m x 7 11m) and adult male worm ( ~45 x 0·1 mm); (B) male with coiled tail and paired spicules; (C) and (D) adherence of rat polymorphonuclear leucocytes to worm head and mouth and sporadically to body cuticle (arrows) following incubation in media.

Several months after bilateral subcutaneous injection of B. malayi third-stage larvae into the groin, as described by Crandall eta!. (1982), an ectatic hindlimb lymphatic was surgically exposed between the femoral and popliteal lymph nodes after an intradermal hindpaw injection ofEvans blue dye. Nests of wriggling adult filarial worms were visible whipping

back and forth within the lymphatic lumen [Fig. l(A)]. After lymphatic isolation, double ligation and excision, the worms were filmed in situ, thrashing to and fro through intraluminal valves [Fig. l(B)]. After incision into the lymphatic wall, adult worms migrated out of their habitat into a Petri dish [Fig. 2(A)], where morphologic features were better delineated;

Fig. 2. (A) Live adult Brugia malayi (arrow) emerging from an excised lymphatic; (B) slightly bulbous male head and cross-striated cuticle; (C) entwined male and female worms; (D) and (E) female packed with embryonated ova. Fig. 3. Live Brugia malayi microfilariae within peripheral lymph showing (A) developing intraovarian form, (B) emergence from ova, (C) freely motile stage, and (D) adherence of rat polymorphonuclear leucocytes (arrows) following incubation in media (Rickenburg-like phenomenon).

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CASE ET AL.

these features included cuticle cross-striations and a slightly bulbous head [Fig. 2(B)], intertwined adult male and female worms [Fig. 2(C)], embryonated ova within the body segments of the female [Fig. 2(D) and (E)], microfilariae in various stages of development (Fig. 3), and a distinctly coiled tail with paired spicules in males [Fig. 4(B)]. Rat white blood cells added to the media adhered to microfilariae [Fig. 3(D)] and also to the cuticle, particularly at the head end, of adult filariae [Fig. 4(C) and (D)]. This so-called Rickenburg reaction (see Manson-Bahr, 1959) raises the speculation that the adult worm feeds on circulating lymphocytes, thereby averting host immunoresponsiveness and its own destruction. Videomicroscopy vividly captures on the screen the hidden intraluminal events in the life cycle of lymphatic-dwelling filarial worms, and allows direct visualization and documentation of interactions between host and parasite. This camera system may also permit direct observation (both in vivo and in vitro) of the capability of putative filaricidal drugs to kill the adult worm and its progeny. The authors express their appreciation to the photographer, Mr. Keven Siegert of Medical Television, Biomedical Communications, Arizona Health ACKNOWLEDGEMENTS.

Sciences Center, The University of Arizona, Tucson for his tireless assistance. The work was supported by Arizona Disease Control Commission Contract No. 8277000000-l-OAT6625 and ZB7492, and also in part by the UNDP/World Bank/WHO Special Programme for Research and Training in Tropical Diseases (ID No. 870051), and the National Institutes of Health No. Al19275 and NIAID Supply Contract No. Al-02642 US-Japan Cooperative Medical Science program.

T.

c. CASE

M.H. WITTE D.L. WAY C. L. WITTE Department of Surgery, University of Arizona College of Medicine, Tucson, Arizona, U.S.A. c. A. CRANDALL Department of Pathology, University of Florida, Gainesville, Florida, U.S.A. R. B. CRANDALL Department of Microbiology, University of Florida, Gainesville, Florida, U.S.A.

Received 12 August 1991, Accepted 14 May 1992

REFERENCES CRANDALL, R. B., McCREEVY, P. B., CoNNoR, D. H., CRANDALL, C. A., NELsoN, J. T. & McCALL, (1982). American Journal of Tropical Medicine and Hygiene, 31,752-759. MANsoN-BAHR, P. (1959). American Journal of Tropical Medicine and Hygiene, 62, 160-173.

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Videomicroscopy of intralymphatic-dwelling Brugia malayi.

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