Efficacy of oral and transepidermal application of diethylcarbamazine citrate in Brugia malayi infected cats

This experimental study showed that daily oral administration of DEC at levels of 5 to 15 mg/kg/day with food for 3 weeks decreased the level of both developing and adult B. malayi in infected cats. There were no adverse reactions due to the medication. Topical application of 5% DEC in skin cream or in mineral oil appears to be effective in killing developing B. malayi in cats.     

Effect of diethylcarbamazine citrate on Brugia malayi infections in cats following daily, weekly, or monthly administration

Cats with patent infections of Brugia malayi were treated by intraperitoneal injection of diethylcarbamazine citrate (DEC) for 6 consecutive days, weekly for 6 consecutive weeks or monthly for 3 months. Each cat received a total of 100 mg DEC per kg. At necropsy 7 months after infection, no living worms were recovered from any of eight cats treated weekly and only one of nine cats treated daily had a single living Brugia. Five of nine cats treated monthly and six of eight untreated controls had one or more living worms. Cats treated weekly showed a larger decline in microfilariae than those of the other treated groups. The mean microfilariae level of untreated controls increased 2-fold. At necropsy, gross appearance of regional lymphatics in daily and weekly treated cats resembled those of uninfected controls more closely than those in cats treated monthly or untreated. Differences in degree of histological changes between groups of infected cats were not apparent. Weekly administration of DEC appeared to be the most effective regimen; monthly treatment was less effective.     

The effect of diethylcarbamazine citrate on incidence and recovery rates of Brugia malayi microfilaremia in Sabah, Malaysia

Mass drug administration via 3 modes of delivery reduced the incidence and prevalence rates and intensity of Brugia malayi infection in 3 rural villages in the Bengkoka Peninsula, Sabah, in 1982-1983. A dosage of 6 mg diethylcarbamazine citrate (DEC-C)/kg body weight was administered either daily or weekly (total of 6 doses, 36 mg/kg body weight), and impact on B. malayi cases were comparable in the 3 villages. A total of 384 people participated in the DEC-C regimens, and all pregnant women and children under 2 years were excluded from the study. Bekessy's method of estimation of incidence and recovery rates was applied to data on B. malayi microfilaremia before drug administration. Treatment with DEC-C by any of the 3 modes of delivery drastically reduced the number of episodes of patent microfilaremia, incidence and prevalence, and median microfilarial density. Reduction was sustained for at least 18 to 24 months after treatment.     

Effect of treatment with diethylcarbamazine on immune responses to filarial antigens in patients infected with Brugia malayi

We studied the effect of treatment with diethylcarbamazine (DEC) on immune responses to parasite antigens in humans infected with Brugia malayi. In vitro lymphocyte proliferative responses to microfilarial antigens increased in patients who became amicrofilaremic after treatment with DEC. No changes in reactivity were observed in amicrofilaremic individuals who were given DEC or in a small number of patients who remained microfilaremic after treatment. Reactions to other antigens (PPD and SKSD) were not affected by drug treatment. Serum titers of antibodies to the sheath of B. malayi microfilariae did not significantly change during the period of observation. These findings indicate that DEC partially reverses the state of cellular unresponsiveness to parasite antigens associated with patient filarial infections.     

Experimental attempts to induce ocular infection in cats with subperiodic Brugia malayi.

In an attempt to determine whether the finding of human ocular lesions due to B. malayi was due to the site of entry of the infective larvae, cats were infected via ocular instillation, subconjunctival inoculation, subcutaneous inoculation around and infective mosquito bites over the eyes. Although no conjunctival lesion was seen patent infections were produced via ocular instillation, subconjunctival and subcutaneous inoculation of infective larvae. Adult worms were also recovered from periorbital tissues. Adult worms were localized mainly in the lymphatic system of the head and neck regions of the cat. The results show that the conjunctival lesions seen in humans could be due to the site of bite of the mosquito and thus the entry of the infective larvae.     

The efficacy of a single-oral-dose administration of ivermectin and diethylcarbamazine on the treatment of feline Brugia malayi

The combination of ivermectin and diethylcarbamazine (DEC) have been shown to be superior to either drug alone for the suppression of Brugia malayi in humans, but their efficacy against infection with B. malayi in cats has never been investigated. Fourteen asymptomatic microfilaremic (1-200 microfilariae/20 microl blood) cats received oral doses of ivermectin (400 microg/kg body weight) and DEC (6 mg/kg body weight) as a single treatment. A two-month post-treatment examination revealed that 87-100% of the microfilariae in each subject had been cleared, with two of the subjects being amicrofilaremic. A further reduction in microfilarial levels was observed until the final follow-up, at 8 months post-treatment, when the mean clearance rate was 99% and 12 out of the 14 subjects (86%) were amicrofilaremic. The combination of ivermectin and DEC demonstrated a microfilaricidal effect superior to that of either drug used alone, both in the initial rapid clearance of microfilariae, and in sustaining the effect for 8 months. This finding has important implications for the control of brugian lymphatic filariasis in the cat reservoir.     

Efficacy of diethylcarbamazine medicated salt in interrupting Brugia malayi transmission in hill tribe settlements in Kerala State.

A filariasis survey carried out about eight years after achieving zero microfilaria (mf) rates following administration of diethylcarbamazine (DEC) medicated salt in the Kani hill tribe settlements in Quilon and Thiruvananthapuram districts of Kerala State revealed that there was no reappearance of Brugia malayi infection in the experimental areas. Mf rates were maintained at zero level in the experimental villages, while in the control villages, 2.9 per cent mf positives were observed. Mansonia (Mansonioides) uniformis dissected did not reveal filarial infection. It is concluded that DEC medicated salt regime in the experimental areas of Kani hill tribe settlements has been successful in effectively interrupting B. malayi transmission. Pilot studies in other B. malayi endemic areas of India using DEC medicated salt regime with the objective of eliminating B. malayi transmission are advocated, since the parasite has a restricted distribution in India and is already showing a declining trend.     

Apparent failure of ultrasonography to detect adult worms of Brugia malayi

Adult worms of Wuchereria bancrofti, or rather their characteristic movements (the 'filarial dance'), can now be detected in the scrotal lymphatics of microfilaraemic males, using ultrasonography. This ability has been used to delineate the lymphatic pathology of bancroftian filariasis, guide the surgical removal of the adult worms and, most importantly, assess the macrofilaricidal effects of antifilarial drugs. In the present study, the first report of the use of ultrasonography in brugian filariasis, 22 men (aged 18-62 years) with 60-2972 (median = 370) Brugia malayi microfilariae/ml blood were subjected to ultrasonography using a linear, 7.5-MHz probe. In addition, four other men (aged 19-35 years), with W. bancrofti microfilaraemia [28-524 (median = 234) microfilariae/ml], were similarly examined. Adult worms were not detectable in any of the patients with B. malayi parasitaemia but were detected in the scrotal lymphatics of two of the four individuals with W. bancrofti infection. The reasons for the failure to detect adult B. malayi and the limitations of ultrasound as a screening tool are examined. The results highlight the differences between the two species that cause most lymphatic filariasis and the need for rapid development of tools that can be used for the control of brugian lymphatic filariasis.     

In vitro development of Brugia pahangi and Brugia malayi in cultured mosquito thoraces.

In vitro cultivation of Brugia pahangi and subperiodic Brugia malayi one-day old larvae to infective stage larvae (L3) within thoraces excised from Aedes aegypti (Black Eye, Liverpool) and Anopheles quadrimaculatus was attempted. The mosquito thoraces were excised under aseptic conditions, 24 h after a blood meal on either B. pahangi- or B. malayi-infected jirds. The excised thoraces were washed aseptically and inoculated into a diphasic media. A nutrient agar base was overlaid with either Grace's insect cell culture medium or Schneider's Drosophila medium or a mixture (1:1) of these two media. Each overlay medium contained a 1 x concentration of antibiotic/antimycotic mixture plus 20% fetal bovine serum. The excised thoraces provided the intracellular milieu for development of Brugia larvae. In Grace's or Schneider's insect tissue culture medium alone, the filaria larvae of both species developed only to the second larval stage after 12 days; whereas, in a mixture (1:1) of Grace's and Schneider's media, some one-day old larvae of both Brugia species developed to the infective larval (L3) stage after 12 days. However, large numbers of both species of larvae developed to the infective larval stage when, prior to providing an infective blood meal, the mosquitoes of both species were fed 1 x concentration of antibiotic/antimycotic mixture in a 10% sucrose solution containing 0.1% p-aminobenzoic acid for 6 days. These results showed for the first time that if one-day old Brugia larvae are confined intracellularly in excised thoraces, they can then develop in insect tissue culture media without adding a feeder layer of mosquito cells or conditioning the media with mosquito cell lines.     

Detection of adult Brugia malayi filariae by ultrasonography in humans in India and Indonesia

In bancroftian filariasis, ultrasonography (USG) is a suitable tool to monitor infection by the detection of adult filariae in addition to antigen detection tests (ICT, Og4C3). However, in brugian filariasis, ultrasound examinations in humans have so far failed to detect adult worms and no antigen test is available to verify infections in patients who are carriers of adult worms but amicrofilaraemic. In this study, we describe the feasibility of detection of adult Brugia malayi filariae by USG. Worm nests were detected in 4 of 32 patients in India and Indonesia, located in the breast, the thigh, the calf and an inguinal lymph node. The study shows that adult filariae of B. malayi in humans can be detected by USG, but the technique is limited by the fact that worm nests seem not to be stable over time in humans, as is the case in bancroftian filariasis.     

The vectors of Brugia malayi in Southern Thailand.

Mansonia uniformis, with an infective rate of 0.02, was incriminated as the vector of periodic Brugia malayi in Pattani province. Mansonia bonneae and Ma. dives, with infective rates of 0.18 and 0.20 respectively, were the vectors of B. malayi in Narathiwat, where the microfilarial periodicity was the subperiodic form.     

Ultrasonography in filaria-infected rodents: detection of adult Litomosoides sigmodontis and Brugia malayi filariae

OBJECTIVE: To evaluate the usefulness of ultrasonography (USG) in the detection of adult filariae in rodents. Wuchereria bancrofti are frequently detected using USG in humans, whereas adult Brugia malayi have not been so far. METHODS: A Meriones unguiculatus with Litomosoides sigmodontis infection was examined to visualize adult filariae of a similar length as W. bancrofti. Similarly, three Mastomys coucha, infected with B. malayi, were examined using USG to verify whether the adult worms, which are far smaller than W. bancrofti and L. sigmodontis, can be located using USG in the animals. RESULTS: Adult L. sigmodontis were detected using USG in the pleural cavity of M. unguiculatus, and in M. coucha adult B. malayi were visualized in the hearts, lungs, axillary lymph nodes and scrotum. Ultrasound findings were verified by dissection of the rodents. CONCLUSIONS: Although adult B. malayi are far smaller than L. sigmodontis and W. bancrofti, they can be detected using USG in rodents. USG may serve as an adjunctive tool to support parasitological examinations and can add information on filarial infections at any time point of an observation period, particularly in cryptic infections and without the need for invasive measures or killing of the rodent. Thus, USG can support the early detection of macrofilaricidal activities of new compounds and can be used to determine the location of adult worms in the animals. It is possible to give a rough estimate of the number of adult worms, but determination of the exact numbers of adult filariae in various locations is impossible with USG.     

Experimental Brugia malayi infections in the rhesus monkey.

Twenty-eight rhesus monkeys in 3 groups were exposed to single (Group I), double (Group II), and multiple (Group III) inoculations with B. malayi infective larvae. Infections were monitored by microfilarial and blood counts, selected biochemical tests, IFA responses, and records of body temperature and lymphadenopathy before and/or after treatment with DEC. As a whole, the highest microfilaraemia levels were observed in Group II and lowest in Group III monkeys. Eosinophilia was a common occurrence but reached the highest mean levels in Group III. Intermittent fevers and lymph node enlargements were observed in all groups of monkeys and the occurrence of these appeared to be correlated. No definite pattern of antibody production was discernable among groups, but an inverse relationship existed between microfilaraemia and detectable microfilarial antibodies. Treatment with DEC produced a microfilaraemia-taxic effect within the initial half hour and responses to treatment varied according to individuals. Although post-treatment reinfection appeared to cause lymphoid responses and tissue eosinophilia, no substantial resistance to reinfection was observed.