Significant decline in lymphatic filariasis associated with nationwide scale-up of insecticide-treated nets in Zambia.

Lymphatic filariasis (LF) is a mosquito-borne disease, broadly endemic in Zambia, and is targeted for elimination by mass drug administration (MDA) of albendazole and diethylcarbamazine citrate (DEC) to at-risk populations. Anopheline mosquitoes are primary vectors of LF in Africa, and it is possible that the significant scale-up of malaria vector control over the past decade may have also impacted LF transmission, and contributed to a decrease in prevalence in Zambia. We therefore aimed to examine the putative association between decreasing LF prevalence and increasing coverage of insecticide-treated mosquito nets (ITNs) for malaria vector control, by comparing LF mapping data collected between 2003-2005 and 2009-2011 to LF sentinel site prevalence data collected between 2012 and 2014, before any anti-LF MDA was started. The coverage of ITNs for malaria was quantified and compared for each site in relation to the dynamics of LF. We found a significant decrease in LF prevalence from the years 2003-2005 (11.5% CI95 6.6; 16.4) to 2012-2014 (0.6% CI95 0.03; 1.1); at the same time, there was a significant scale-up of ITNs across the country from 0.2% (CI95 0.0; 0.3) to 76.1% (CI95 71.4; 80.7) respectively. The creation and comparison of two linear models demonstrated that the geographical and temporal variation in ITN coverage was a better predictor of LF prevalence than year alone. Whilst a causal relationship between LF prevalence and ITN coverage cannot be proved, we propose that the scale-up of ITNs has helped to control Anopheles mosquito populations, which have in turn impacted on LF transmission significantly before the scale-up of MDA. This putative synergy with vector control has helped to put Zambia on track to meet national and global goals of LF elimination by 2020.

Prevalence and source of trypanosome infections in field-captured vector flies (Glossina pallidipes) in southeastern Zambia.

The prevalence of trypanosome infections in tsetse flies, Glossina pallidipes, collected from Chiawa and Chakwenga in Zambia with endemic trypanosomosis was assessed by polymerase chain reaction (PCR). Out of the 550 G. pallidipes, 58 (10.5%) flies were found to harbor trypanosome DNA. Infection rates of tsetse with Trypanosoma vivax universal, Trypanosoma congolense savannah, T. congolense forest and T. congolense kilifi were 4.2% (23/550), 4.7% (26/550), 1.1% (6/550) and 1.6% (9/550), respectively. To determine the mammalian hosts of T. congolense and T. vivax infections from the tsetse flies, mammalian mitochondrion DNA of blood meal in these flies were analyzed by PCR and subsequent gene sequence analysis of the amplicons. Sequence analysis showed the presence of cytochrome b gene (cyt b) of 7 different mammalian species such as human, elephant, buffalo, goat, warthog, greater kudu and cattle. Goats which were main livestock in these areas were further examined to know the extent of its contribution in spreading the infection. We examined the prevalence of trypanosome infections in the domestic goat population in 6 settlements in Chiawa alone. Of the 86 goats sampled, 4 (4.6%), 5 (5.8%), 4 (4.6%) and 4 (4.6%) were positive for T. vivax universal, T. congolense savannah, forest and kilifi, respectively. These findings showed that the host-source of trypanosome infections in vector fly give a vital information about spread of infection. The result of this study will certainly contribute in elucidating more the epidemiology of trypanosomosis.

Studies on the economics of ticks in Zambia.

Two herds of experimental Sanga cattle were maintained under traditional savanna grazing management for three years in the Central Province of Zambia. One herd was kept free of ticks by regular acaricide treatment, while the other was given no tick control. Milk production, growth rate, fertility and mortality were monitored throughout the trial. The tick-free herd performed significantly better than the tick-infested herd, but the value of the additional production was much less than the cost of the acaricide used. It is concluded that there is no economic justification for intensive tick control under these conditions. However, strategic tick control would be justified if the quantity of acaricide used could be reduced by 50% without any major reduction in benefits. The results indicate that treating calves below 45 days of age reduces their performance, as does treating cows during periods of very low tick challenge. Therefore, it seems likely that economically beneficial strategic tick control policies could be developed.

The role of Rhipicephalus appendiculatus and R. evertsi evertsi males in inducing resistance in laboratory animals: preliminary studies.

Guinea-pigs infested with male ticks of the species Rhipicephalus appendiculatus, and rabbits infested with R. evertsi evertsi, acquired immunity to conspecific female ticks. The hosts were first infested with male ticks and thereafter were challenged with males and females of the same species. The mean weight of the engorged females of R. appendiculatus fed on guinea pigs previously infested with male ticks was 509.0 (+/- 41.4) mg compared with that of females fed on control guinea pigs (651.2 +/- 31.8 mg). Similar weight differences were observed for R.e. evertsi females which fed on rabbits previously infested three times with male ticks. The mean weight of the female ticks which fed on these rabbits was 520.1 (+/- 29.8) mg compared with 640.7 (+/- 30.2) mg of R.e. evertsi females which fed on control hosts. The concentration of gammaglobulins in the sera of rabbits was monitored at various intervals after the first infestation. It was found, for the first time, that infestation of laboratory animals with male ticks conferred immunity, but to a lesser degree than infestation with both sexes. It was also shown that the level of gammaglobulins increased from 3.4 +/- 0.28 g l-1 to 7.3 +/- 0.24 g l-1 in sera of rabbits hosts as a result of the feeding activity of males, but to a lesser extent than in sera of rabbits on which both sexes had fed (10.8 +/- 2.4 g l-1).

Monitoring of naturally acquired and artificially induced immunity to Amblyomma variegatum and Rhipicephalus appendiculatus ticks under field and laboratory conditions.

The ability of rabbits, goats and cattle to acquire immunity to the ixodid ticks Amblyomma variegatum and Rhipicephalus appendiculatus was studied under laboratory and field conditions. Rabbits were successfully immunized with crude salivary gland extract (SGE) and midgut extract (ME) obtained from flat or partly fed female R. appendiculatus ticks. The lowest numbers of larvae were produced by females fed on rabbits immunized with unfed midgut extract. Similar reductions in larval production could be induced after three infestations of rabbits with adult R. appendiculatus. Also, successive feedings of nymphs of R. appendiculatus on rabbits resulted in significantly reduced engorgement weights. Skin testing with SGE induced delayed-type hypersensitivity reactions, which could be correlated with immunity to R. appendiculatus in rabbits. Moreover, circulating antibodies were detected in rabbits with an ELISA using SGE of R. appendiculatus. Immunity to A. variegatum nymphs could be induced in rabbits by repeated infestations, but this failed in goats. Immunization of goats with midgut extract from adult A. variegatum did not protect against subsequent nymphal challenge, but strong skin reactions were noticed when adults ticks fed on immunized goats. Sodium dodecyl sulfate polyacrylamide gel electrophoresis of SGE and ME from A. variegatum revealed the presence of 48 protein bands in SGE and 29 bands in midgut extract. Western blotting employing serum from a rabbit immune to R. appendiculatus recognized a number of bands in SGE from R. appendiculatus, but also in SGE of A. variegatum. Immunity acquired by cattle to ixodid tick infestations under field conditions was monitored by skin testing with SGE and western blot analysis. In general, cattle with the lowest tick numbers manifested the strongest delayed-type hypersensitivity responses. Finally, western blot analysis employing sera from tick-infested and tick-naive cattle could not be related to actual immune status.

Delayed-type hypersensitivity test for assessing tick-immune status of cattle in Zambia.

Delayed-type hypersensitivity skin reactions were used to assess the tick resistance status of Tonga calves in Zambia. The antigen used in the tests was a homogenate of unfed nymphal Rhipicephalus appendiculatus which had been shown to give protective immunity in guinea pigs to adult female R appendiculatus. There was a significant negative correlation between the intensity of the reactions and the total number of ticks (Amblyomma variegatum, R appendiculatus, Hyalomma truncatum, Boophilus decoloratus and Rhipicephalus species) on the animals.

Morphogenetic diapause in Amblyomma variegatum (Acari: Ixodidae).

In southern Africa, Amblyomma variegatum Fabricius is characterized by a strict seasonal activity. Experiments were carried out to determine whether a diapause mechanism regulates this seasonality. Engorged A.variegatum females were exposed to controlled laboratory conditions or natural field conditions at different times of the year. Females exposed in a natural environment in September-October (short day) had significantly longer pre-oviposition periods than females exposed from November to March. The season in which the previous instar fed had no apparent effect on the engorgement or pre-oviposition periods of the females. Furthermore, artificial changes in photoperiod during and after female engorgement had no significant effects on pre-oviposition periods. It is tentatively concluded that the unfed female is the responsive stage to photoperiodic changes which induce diapause. Diapause could be terminated and oviposition induced by exposing females to a short period of chilling (18 degrees C for 48 h). It is concluded that a morphogenetic diapause mechanism exists in A.variegatum, which is probably induced by short day responses and terminated following rainfall and a concomitant decrease in soil temperature. The diapause, which occurs in females which fed early in the season, causes a delay in oviposition and therefore effectively synchronizes the life-cycle to ensure that eggs and larvae occur at a climatically favourable period.

Biology of Rhipicephalus appendiculatus and R. zambeziensis and production of a fertile hybrid under laboratory conditions.

In some areas of Eastern and Southern provinces of Zambia Rhipicephalus appendiculatus and R. zambeziensis are sympatric, and numerous intermediate forms as well as typical R. appendiculatus and R. zambeziensis are found. Experiments were carried out in order to clarify the taxonomic status of the intermediate forms. Biology and hybridization capacities of two strains of R. appendiculatus (A and D) and one strain of R. zambeziensis (H) were studied. Infestations on rabbits and calves were performed at the beginning and at the end of the Zambian rainy season. The adults of both species showed preference for the bovine host and performed better at the end of the rainy season. A recently isolated strain of R. appendiculatus had notably higher larval production per unit engorgement weight than the strain that had been maintained for prolonged periods under laboratory conditions. R. zambeziensis produces fewer larvae and had longer development periods than R. appendiculatus. The two species cross-breed under laboratory conditions. A fertile hybrid is produced by R. zambeziensis females and R. appendiculatus males, whereas a sterile hybrid is a result of a reciprocal cross. Morphology of adults of the fertile hybrid (H/A) is indistinguishable from the maternal species, whereas adults of the sterile hybrid (A/H) are of maternal, paternal and intermediate forms. Larval production indices (LPI) vary from 0.4 to 3.7 for the H (females) X A (males) cross, approximately three-fold less than in the maternal species. For the reciprocal cross, LPI ranges from 0.1 to 2.7, five- to thirty-fold less than in the maternal species. The fertile hybrid has a higher LPI than both first crosses, and under the most favourable climatic conditions LPI is even higher than for the maternal species. Adults of the sterile hybrid rarely formed pairs. Only partially engorged female ticks detached, which produced no viable eggs. Both hybrids may occur in areas where the two species are sympatric. However, detailed field studies will have to be performed in order to gain knowledge on population dynamics, interbreeding and the vector role of the two species and their hybrids under field conditions.

Bovine babesiosis (Babesia bovis infection) in Zambia.

Two cases of Babesia bovis, a parasite associated with the tick Boophilus microplus, are reported for the first time from the central part of Zambia. It is concluded that infected B. microplus ticks are occasionally introduced into central Zambia by tick-infested cattle from the north-eastern part of the country where B. bovis is endemic. The spread of B. microplus in Southern Africa in a westward direction is discussed and related to the epidemiology of bovine babesiosis in Zambia.