Adult worms of the rural strain of Wuchereria bancrofti in Peninsular Malaysia obtained from a successful experimental transmission in an immunosuppressed Macaca fascicularis are described for the first time. Although the worms, especially females, were slightly smaller, they were similar in morphology to those of the periodic and non-periodic W. bancrofti previously described.
Although current programmes to eliminate lymphatic filariasis have made significant progress it may be necessary to use different approaches to achieve the global goal, especially where compliance has been poor and 'hot spots' of continued infection exist. In the absence of alternative drugs, the use of higher or more frequent dosing with the existing drugs needs to be explored. We examined the effect of higher and/or more frequent dosing with albendazole with a fixed 300 mg dose of diethylcarbamazine in a Wuchereria bancrofti endemic area in Odisha, India. Following screening, 104 consenting adults were randomly assigned to treatment with the standard regimen annually for 24 months (S1), or annually with increased dose (800 mg albendazole)(H1) or with increased frequency (6 monthly) with either standard (S2) or increased (H2) dose. Pre-treatment microfilaria counts (GM) ranged from 348 to 459 mf/ml. Subjects were followed using microfilaria counts, OG4C3 antigen levels and ultrasound scanning for adult worm nests. Microfilarial counts tended to decrease more rapidly with higher or more frequent dosing at all time points. At 12 months, Mf clearance was marginally greater with the high dose regimens, while by 24 months, there was a trend to higher Mf clearance in the arm with increased frequency and 800 mg of albendazole (76.9%) compared to other arms, (S1:64%, S2:69.2% & H1:73.1%). Although higher and/or more frequent dosing showed a trend towards a greater decline in antigenemia and clearance of "nests", all regimens demonstrated the potential macrofilaricidal effect of the combination. The higher doses of albendazole did not result in a greater number or more severe side effects. The alternative regimens could be useful in the later stages of existing elimination programmes or achieving elimination more rapidly in areas where programmes have yet to start.
The known filaricides, suramin and diethylcarbamazine citrate, were tested against subperiodic Brugia malayi infection in the leaf-monkey, Presbytis cristata. As expected, intravenous suramin at 10 mg/kg daily x 5 days or 17 mg/kg weekly x 5 weeks, did not show any microfilaricidal activity, but substantially reduced the recovery of live adult worms to 50.6% and 13.6% of controls respectively. Oral diethylcarbamazine citrate at 6 mg/kg daily x 6 or 10 days reduced final microfilarial counts to 30% of initial counts four weeks post-treatment and adult worm recovery was reduced to 4.5% and 0% of controls respectively. Although the antifilarial activity of these drugs against the infection in this non-human primate model appears to be similar to that seen in man, these results have to be confirmed using larger groups of animals.
Matched MeSH terms: Microfilaria/drug effects; Microfilaria/growth & development
The Presbytis cristata--Brugia malayi model, now established as a reliable non-human primate model for the experimental screening of potential filaricides, was monitored at monthly intervals for changes in the liver and renal function tests and also for alkaline phosphatase levels during infection. Animals infected with 200-400 infective larvae became patient at 50-90 days post-infection and geometric mean microfilarial counts were above 1000 per ml from the fourth month onwards. There were no significant changes in the biochemical parameters monitored throughout the period of observation. This is an important observation as any changes seen in these parameters during experimental drug studies can be attributed to drug reaction or toxicity and this will be invaluable in decision making as to drug safety.
Matched MeSH terms: Microfilaria/growth & development
BmR1 recombinant antigen has previously been shown to demonstrate high sensitivity and specificity in the serological diagnosis of brugian filariasis in humans. In this study, the pattern of recognition of antibody to BmR1 during Brugia malayi infection was investigated by employing Meriones unguiculatus as the experimental model. Thirty two gerbils were infected subcutaneously with 120 L(3); and two control groups each comprising 25 animals were employed. ELISA using BmR1 was used to detect filaria-specific IgG antibodies elicited by the gerbils; using sera collected from the day 1 until day 150 post-inoculation (p.i.). The results showed that BmR1 detected B. malayi infection in gerbils harboring adult worms irrespective of the presence of circulating microfilaria, and was exemplified by positive ELISA results in nine a microfilaraemic animals that harbored live adult worms. The initial time of the antibody recognition was at day 8 p.i. and the antibody titre showed some correlation with adult worm burden.
In Peninsular Malaysia, only Wuchereria bancrofti and Brugia malayi are reported to cause human filariasis. Brugia pahangi infects many of the same animal hosts as the zoonotically transmitted subperiodic B. malayi. There is a well-recognized need for improved diagnostic techniques for lymphatic filariasis. Parasite antigen detection is a promising new approach, and it will probably prove to be more sensitive and specific than clinical, microscopic and antibody-based serological methods. We recently generated monoclonal antibodies (MAb XC3) from in vitro culture products of adult B. pahangi (B.p. IVP). Filarial antigenemia was quantitated in various hosts including the sera from 6 Malaysian Aborigines with acute lymphatic filariasis. In hosts infected with brugian filariasis and dirofilariasis, antigenemia was scored ranging from 90 ng/ml to 960 ng/ml. None of the control animal and human sera had antigenemia above 90 ng/ml. In addition, MAb XC3 and B.p. IVP were applied in several seroepidemiological surveys among household cats in Kuala Selangor in order to correlate information gathered for future studies of possible cases of human infection. Out of the 81 cats surveyed, 10 (12.35%) and 5 (6.17%) were parasitologically positive for B. pahangi and B. malayi, respectively. However, 21 (25.92%) were antigenemia positive when serologically investigated with MAb XC3. Antifilarial antibodies to B.p. IVP by direct ELISA showed very high cross-reactivity with non-filarial gut worm infections. 16 (19.75%) cats had reciprocal titers ranging from 320 to 2,560. Only 1 (1.23%) cat from this group was antigenemic.
A control programme against subperiodic brugian filariasis was implemented in three villages, (Kg. Ampungan, Kg. Sebangkoi and Kg. Sebamban) in Sarawak, Malaysia. In Kampong Ampungan, the mass administration of diethylcarbamazine (DEC-citrate) combined with residual house spraying of pirimiphos-methyl reduced microfilarial rate to 8% of the pre-treatment level and microfilarial density (MfD50) to 44% of the pre-treatment level over a period of four years. In Kampong Sebangkoi and Kampong Sebamban, where only mass DEC therapy was applied, the microfilarial rate and MfD50 declined distinctly in the second blood survey but increased gradually in two subsequent follow-up blood surveys. In Kg, Ampungan, we observed a significant reduction of infective biting rate (88.3%), infection rate (62.5%) and transmission potential (88.1%) of Mansonia bonneae at the fourth spray round. The corresponding reduction rates in Kg. Sebangkoi and Kg. Sebamban were 35.3%, 26.7%, 42.2% and 24%, 30.8% and 15.4% respectively. The biting density of the vector was reduced by 79.8% indoors and 31.8% outdoors at the sprayed village, while only a slight decrease in densities (17.9% indoors and 12.4% outdoors) was observed at the unsprayed village. Bioassay tests revealed that pirimiphos-methyl had a substantial fumigant effect on the vector. The integrated control measure in controlling subperiodic brugian filariasis is discussed.
R. sabanus and R. muelleri are very common in the lowland forests of Malaysia. In nature they are infected with Breinlia sp. and D. ramachandrani. In an attempt to determine whether they are also susceptible to subperiodic B. malayi and thereby being potential reservoirs of infection of the disease, 24 R. muelleri and 17 R. sabanus were experimentally infected with the parasite. Results show that although they can support the full development of the parasite, they are poor hosts. This confirms the observation that in Malaysia natural infection of Rattus spp. with the parasite has not been seen. These rats therefore are probably not important in the zoonotic transmission of subperiodic B. malayi in Malaysia.
Presbytis cristata monkeys infected through the inoculation of between 200 and 400 subperiodic Brugia malayi infective larvae (L3) in the right thigh, in both thighs or in the dorsum of the right foot were followed up for varying periods of up to about 8 months after infection. All 148 inoculated animals became patent, with mean prepatent periods being between 66 and 76 days. In animals injected in the thigh, the patterns of microfilaraemia were similar, there being a rapid rise in the geometric mean counts (GMCs) of microfilariae during the first 10-12 weeks of patency, which then plateaued at levels of greater than 1000/ml. Adult worm recovery, expressed as the percentage of the infective dose, was significantly higher in animals injected with 100 L3 in each thigh, being 9.4% as compared with 2.8%-4.8% in other groups. It is therefore recommended that animals should be injected with 100 L3 in each thigh and that the testing of potential filaricides in this model be carried out during the phase of rapid increase in microfilaraemia to ensure that any microfilaricidal effect can easily be detected.
Quantitative understanding of the transmission dynamics of lymphatic filarial parasites is essential for the rational planning of control strategies. One of the most important determinants of transmission dynamics is the relationship between parasite yield, the success rate of ingested microfilariae (mf) becoming infective larvae in a mosquito vector, and mf density in the source of the human blood meal. Three types of relationship have been recognized in human filaria/mosquito couples--limitation, facilitation and proportionality; facilitation has hitherto been observed only in the couple Wuchereria bancrofti/Anopheles gambiae in Burkina Faso, in experimental studies on a high density mf carrier. The present paper demonstrates facilitation in W. bancrofti/An. gambiae and W. bancrofti/An. arabiensis in lower mf density carriers in The Gambia and Tanzania, and in W. bancrofti/An. funestus in Tanzania. Facilitation was not found in An. melas in The Gambia nor in An. merus in Tanzania. Analysis of published data shows limitation at low level mf densities in W. bancrofti/Culex quinquefasciatus in Sri Lanka, and in the same couple in India. Limitation also occurs in Brugia malayi/Aedes togoi in experimental cats; proportionality occurs in B. malayi/Mansonia bonneae in Malaysia. The epidemiological significance of these host/parasite relationships is discussed, and supporting evidence for its validity is presented from the published results of large-scale control programmes.
The filaria vector competence of Anopheles stephensi was compared with Brugia-susceptible Aedes aegypti Liverpool strain, An. gambiae Badagry Lagos strain and An. dirus Perlis Malaysia strain. An. stephensi ingested more Brugia pahangi microfilariae, had the highest infectivity rate and yielded more infective mosquitoes than the other two anopheline species. The overall vector competence of An. stephensi was 0.13 times that of Ae. aegypti, 0.62 times that of An. gambiae and 2.17 times that of An. dirus. However, heavy mortality among infected An. stephensi in the present investigation indicates that the filaria vectorial capacity of the mosquito might be limited epidemiologically. The relationship between filaria vector competence and mosquito foregut armature is discussed. It was observed that the relative vector competence of the three anopheline species tested was in the same order as their relative degrees of armature elaboration. The converse would be expected if foregut armatures really give partial protection to the mosquitoes against filarial infection. It is suggested that high host microfilariae density favours larval survival proportional to the degree of armature development in Anopheles (Cellia) species.
Hematological changes were monitored in the leaf-monkey, Presbytis cristata, infected experimentally with 200 subperiodic Brugia malayi infective larvae. Prepatent periods were 54-86 days and peak microfilarial geometric mean counts (GMCs) were 1324 per ml blood. Total leukocyte and differential counts were measured at pre-infection, and then at weakly intervals before and during patency. Blood eosinophil level increased to about thrice the initial level at 3 weeks post-infection and this was maintained for the next 13 weeks before it started to rise again, increasing to more than 5 times the initial level at 20 weeks post-infection. The observed pattern of eosinophilia is probably related to the level of microfilaremia and the destruction of microfilariae in the spleen. There was no significant change in the total leukocyte counts during the period of observation.
DEC or ivermectin (IVM) in combination with albendazole (ALB) has been the recommended strategy of the Global Programme to Eliminate Lymphatic Filariasis (GPELF) since 2000. Despite effective population coverage (> 65%) with several rounds of MDA with DEC or combination of DEC plus ALB, microfilariae persist in few individuals and they continue to be the source of infection for transmitting LF. We report an individual's variability in response to DEC by defining the response as complete absence of microfilaria (mf) (post-treatment mf count = 0) and non-response as presence of mf (post-treatment mf count ≥ 1). We analyzed follow-up data on individual's response to treatment from two randomized clinical trials in which 46 microfilaremic individuals were treated with single-dose DEC (6 mg/kg body weight). They were classified into low, medium, and high mf density categories based on their pre-treatment mf counts. Of the 46 individuals, 65.2% have not responded throughout the 12-month post-treatment period. Application of a logistic regression model with fixed (age, gender, mf density, post-treatment time, and their interactions) and random (individual's response over time) effects indicated that treatment response is independent of age, gender, and time. The overall treatment response increases in low and decreases in high mf density categories. Furthermore, the estimates for the random coefficients model showed that there is a greater variability in response between individuals over post-treatment time. The results substantiate that individual variation in response to DEC exists which indicate the importance of studying the parasite as well as host genetic factors associated with DEC action.
Histochemical demonstration of acid phosphatase activity in microfilariae gives sufficiently characteristic and consistent results for the differentiation of even closely related species. No difference could be detected among nocturnally periodic, nocturnally subperiodic and diurnally subperiodic Brugia malayi, but they could readily be distinguished from B. pahangi. Similarly, Dirofilaria repens could be readily distinguished from D. immitis and B. booliati from B. sergenti. The enzyme distribution pattern of a Malaysian rural strain of Wuchereria bancrofti was different from those of other regions.
Lymphatic filariasis is endemic in Asia. The infections persist as a major cause of clinical morbidity and a significant impediment to socioeconomic development. Its prevalence is increasing world wide, largely because of rapid unplanned urbanization in many endemic areas. It is estimated that at least 120 million people are infected. In our study on foreign workers, a total of 241 day time blood samples were collected. The countries represented were Bangladesh (134), Indonesia (103), Pakistan (3) and Myanmar(1). The tests conducted on blood samples were thick blood film for microfilaria and thin blood film for malaria and quantitation of eosinophiles using the Giemsa stain. Out of the 241 blood samples tested, one was positive for Wuchereria bancrofti and one other was positive for malaria (Plasmodium falciparum) each from Bangladesh and Indonesia respectively. As for the blood eosinophiles, 39 (16.18%) blood samples showed high eosinophilia. Fifteen (6.22%) were from Banglandesh and 24 (9.96%) were from Indonesia. The Bangladeshi male who was positive for Witcherrria bamuofti also showed eosinophilia of 22%. We believe that some of these cases with high eosinophilia, may be positive for microfilaria. We may have missed some cases because of the methodology we chose. Lymphatic filariasis is endemic in Bangladesh and Indonesia. In Malaysia W. brancrofti, especially in the cities have been eliminated. However their vectors for the transmission of W. bancrofti is rampant in the cities. With the influx of immigrants with W. bancrofti and in relation to their occupational nature, W. bancrofti may eventually be introduced into the community and change the whole facet of the disease in Malaysia.
Two out of six monoclonals (McAbs) produced against subperiodic Brugia malayi infective larva (L3) antigens impaired B. malayi L3 motility independently of human buffy coat cells. Scanning electron microscopy studies showed damage to L3 surface and loss of regular cuticular annulations. The two McAbs (BML 1a and BM1 8b) did not affect B. malayi microfilaria (mf). They were IFAT-positive with B. malayi adult and L3 antigens; other McAbs which did not affect mf or L3 motility were IFAT-negative. All six McAbs did not promote cellular adherence of normal human buffy coat cells to mf or L3.
Infective larvae of Brugia malayi subperiodic obtained by dissection of infected Aedes togoi were injected subcutaneously into the scrotal region of Mastomys natalensis. From altogether 58 infected male M. natalensis 81% showed consistently or intermittently detectable microfilaraemia, whereas in 19% of the animals no microfilaraemia could be detected at any stage. The mean prepatent period was 136 days; the microfilarial density varied from 1 to 535 per 20 c. mm blood. In those animlas with consistently detectable and in general higher microfilaraemia an average of 13.1 live adult worms were found, against an average of 6.4 adult worms in animals with intermittent detectable and in general lower microfilaraemia. An average of 1.5 worms was found in animals which at no stage showed detectable microfilaraemia. A correlation between worm burden and prepatent period could be observed in the individual groups. From the total of 520 live adult worms recovered at necropsy, 37% were found in the lungs, 29% in the parenchyma of the testes and 34% in the lymphatic system. 47% of live fertile female worms were found in the lymphatic system, whereas the majority, i.e; 52% of infertile female worms were detected in the lungs. In addition, 380 encapsulated dead worms were found, most of them (98%) in the lymphatic system. 61% of a total of 900 live and dead worms were found in the region of the lymphatic system.
Matched MeSH terms: Microfilaria/growth & development
Filariasis, a parasitic infection endemic in parts of India, Myanmar, islands of the South Pacific, West and East Africa and Saudi Arabia can be diagnosed from various types of cytopathological specimens. This case documents the detection of filarial infection from hydrocele fluid cytology in a 30-year-old Myanmar migrant worker in Malaysia.