In vitro acaricidal activity of Bobgunnia madagascariensis Desv. against Amblyomma variegatum (Fabricius) (Acari: Ixodidae).

The objective of the study was to determine the acaricidal properties of Bobgunnia madagascariensis (Desv.) J.H. Kirkbr. and Wiersema (Leguminosae) against adult Amblyomma variegatum (Fabricius) ticks, using Tephrosia vogelii Hook.f. (Leguminosae) as a positive control. Plant extracts of both were prepared using methanol, acetone and chloroform as extraction solvents. Methanol leaf extracts of T. vogelii (0.014 g) and methanol fruit extracts of B. madagascariensis (0.0062 g) gave the highest mean extraction weights among the plant parts and solvents used. In free contact bioassays, only methanol extracts of the bark and leaf material of T. vogelii and methanol fruit extracts of B. madagascariensis produced 100 % mortality of A. variegatum ticks in 24 h. The acaricidal activity of methanol leaf extracts of T. vogelii persisted for up to 8 days while that of fruit extracts of B. madagascariensis persisted for only 6 days. In topical application bioassays, the toxicity of T. vogelii and B. madagascariensis extracts was found to be significantly different at 95 % confidence level, with B. madagascariensis extracts (LD50 0.030 w/v) being more toxic than T. vogelii extracts (LD50 0.555 w/v). This study has shown that plant extracts of B. madagascariensis and T. vogelii extracts have significant in vitro acaricidal activity against A. variegatum ticks and can thus be considered as alternatives for tick control. Further research is however required on persistence, safety and the required application rates.

Resistance status of ticks (Acari; Ixodidae) to amitraz and cypermethrin acaricides in Isoka District, Zambia.

This study was designed to obtain data on the farmer's approach to tick control and to determine whether Rhipicephalus appendiculatus Neuman, Amblyomma variegatum (Fabricius), and Rhipicephalus (Boophilus) microplus (Canestrini) were resistant to amitraz and cypermethrin acaricides, in Isoka District, Zambia. Prevailing tick control practices were documented by administering a semi-structured questionnaire to 80 randomly selected smallholder livestock farmers from four agricultural camps (Longwe, Kantenshya, Kapililonga, and Ndeke) in Isoka District. Modified larval packet test (LPT) bioassay experiments were used to determine the resistance status of the common tick species against amitraz and cypermethrin acaricides. Fifty percent of respondents practiced chemical tick control with amitraz (27 %) and cypermethrin (23 %) being the acaricides in use, and were applied with knapsack sprayers. Less than 3 l of spray wash per animal was used which was considerably lower than the recommended delivery rate of 10 l of spray wash per animal. No significant susceptibility change to amitraz at 95 % confidence level was observed in R. appendiculatus and A. variegatum against amitraz. However, a significant change in the susceptibility of R. (Bo.) microplus tested with amitraz was detected at 95 % confidence. The test population had a lower susceptibility (LD50 0.014 %; LD90 0.023 %) than the reference population (LD50 0.013 %; LD90 0.020 %). The results indicated that resistance to amitraz was developing in R. (Bo.) microplus. For cypermethrin, no significant susceptibility change at 95 % confidence was observed in any of the three species and thus resistance to this chemical was not observed.

Lymphatic Filariasis Elimination Status: Wuchereria bancrofti Infections in Human Populations after Five Effective Rounds of Mass Drug Administration in Zambia.

Lymphatic filariasis (LF), also commonly known as elephantiasis, is a neglected tropical disease (NTD) caused by filarial parasites. The disease is transmitted via a bite from infected mosquitoes. The bites of these infected mosquitoes deposit filarial parasites, Wuchereria or Brugia, whose predilection site is the lymphatic system. The damage to the lymph system causes swelling in the legs, arms, and genitalia. A mapping survey conducted between 2003 and 2011 determined LF as being endemic in Zambia in 96 out of 116 districts. Elimination of LF is known to be possible by stopping the spread of the infection through large-scale preventive chemotherapy. Therefore, mass drug administration (MDA) with diethylcarbamazine citrate (DEC) (6 mg/kg) and Albendazole (400 mg) for Zambia has been conducted and implemented in all endemic districts with five effective rounds. In order to determine whether LF prevalence has been sufficiently reduced to levels less than 2% antigenemia and less than 1% microfilaremia, a pre-transmission assessment survey (pre-TAS) was conducted. Therefore, post-MDA pre-TAS was conducted between 2021 and 2022 in 80 districts to determine the LF prevalence. We conducted a cross-sectional seroprevalence study involving 600 participants in each evaluation unit (EU) or each district. The study sites (sentinel and spot-check sites) were from districts that were the implementation units (IUs) of the LF MDA. These included 80 districts from the 9 provinces. A total of 47,235 people from sentinel and spot-check locations were tested. Of these, valid tests were 47,052, of which 27,762 (59%) were females and 19,290 (41%) were males. The survey revealed in the 79/80 endemic districts a prevalence of Wb antigens of 0.14% and 0.0% prevalence of microfilariae. All the surveyed districts had an optimum prevalence of less than 2% for antigenaemia, except for Chibombo district. The majority of participants that tested positive for Wuchereria bancrofti (Wb) Antigens (Ag) were those that had 2, 3, and 4 rounds of MDA. Surprisingly, individuals that had 1 round of MDA were not found to have circulating antigens of Wb. The study showed that all the surveyed districts, except for Chibombo, passed pre-TAS. This further implies that there is a need to conduct transmission assessment surveys (TASs) in these districts.

Sarcoptes mite epidemiology and treatment in African buffalo (Syncerus caffer) calves captured for translocation from the Kafue game management area to game ranches.

BACKGROUND: In Zambia, translocation of wildlife from National Parks to private owned game ranches demands that only animals free of infectious diseases that could adversely affect the expansion of the wildlife industry should be translocated to game ranches. Sarcoptes mange (Sarcoptes scarbiei) has been involved in the reduction of wildlife populations in some species. RESULTS: Sarcoptes mange (Sarcoptes scarbiei) was detected and eradicated from two herds of African buffalo (Syncerus caffer) calves captured in the Kafue GMA in July 2004 and August 2005. The overall prevalence was estimated at 89.5% (77/86). Sex had no influence on the occurrence and severity of the disease. Of the 86 calves used in the study, 72.1% had good body condition scores, 20.9% were fair and 7.0% were poor. Of the 77 infected calves, 53.2% were mildly infected, 28.6% were moderately and 18.2% were severely infected. Body condition score was correlated to the severity of the infection (r = 0.72, p < 0.000, n = 86) at capture. Eradication of Sarcoptes mites from the entire herd using ivermetcin was dependant on the severity of the infection. The overall ability of ivermectin to clear the infection after the first treatment was estimated at 81.8% (n = 77). It increased to 94.8% and 100% after the second and third treatments respectively. CONCLUSION: This is the first report on the epidemiology and treatment of Sarcoptes mange in African buffaloes in Zambia. This study improves our understanding about Sarcoptes scabiei epidemiology and treatment which will have further applications for the safe animal translocation.

Reprint of "Modelling the influence of temperature and rainfall on malaria incidence in four endemic provinces of Zambia using semiparametric Poisson regression".

Although malaria morbidity and mortality are greatly reduced globally owing to great control efforts, the disease remains the main contributor. In Zambia, all provinces are malaria endemic. However, the transmission intensities vary mainly depending on environmental factors as they interact with the vectors. Generally in Africa, possibly due to the varying perspectives and methods used, there is variation on the relative importance of malaria risk determinants. In Zambia, the role climatic factors play on malaria case rates has not been determined in combination of space and time using robust methods in modelling. This is critical considering the reversal in malaria reduction after the year 2010 and the variation by transmission zones. Using a geoadditive or structured additive semiparametric Poisson regression model, we determined the influence of climatic factors on malaria incidence in four endemic provinces of Zambia. We demonstrate a strong positive association between malaria incidence and precipitation as well as minimum temperature. The risk of malaria was 95% lower in Lusaka (ARR=0.05, 95% CI=0.04-0.06) and 68% lower in the Western Province (ARR=0.31, 95% CI=0.25-0.41) compared to Luapula Province. North-western Province did not vary from Luapula Province. The effects of geographical region are clearly demonstrated by the unique behaviour and effects of minimum and maximum temperatures in the four provinces. Environmental factors such as landscape in urbanised places may also be playing a role.

Modelling the influence of temperature and rainfall on malaria incidence in four endemic provinces of Zambia using semiparametric Poisson regression.

Although malaria morbidity and mortality are greatly reduced globally owing to great control efforts, the disease remains the main contributor. In Zambia, all provinces are malaria endemic. However, the transmission intensities vary mainly depending on environmental factors as they interact with the vectors. Generally in Africa, possibly due to the varying perspectives and methods used, there is variation on the relative importance of malaria risk determinants. In Zambia, the role climatic factors play on malaria case rates has not been determined in combination of space and time using robust methods in modelling. This is critical considering the reversal in malaria reduction after the year 2010 and the variation by transmission zones. Using a geoadditive or structured additive semiparametric Poisson regression model, we determined the influence of climatic factors on malaria incidence in four endemic provinces of Zambia. We demonstrate a strong positive association between malaria incidence and precipitation as well as minimum temperature. The risk of malaria was 95% lower in Lusaka (ARR=0.05, 95% CI=0.04-0.06) and 68% lower in the Western Province (ARR=0.31, 95% CI=0.25-0.41) compared to Luapula Province. North-western Province did not vary from Luapula Province. The effects of geographical region are clearly demonstrated by the unique behaviour and effects of minimum and maximum temperatures in the four provinces. Environmental factors such as landscape in urbanised places may also be playing a role.

Filarial infections in domestic dogs in Lusaka, Zambia.

Filariae are common parasites of dogs in many parts of the world, but little is known about the status of these infections in sub-Saharan Africa. A study was carried out to determine the occurrence and species of filariae among 272 dogs in Lusaka, Zambia. Giemsa stained blood smear and Knott's concentration methods revealed microfilariae in 16 (5.9%) of the dogs. PCR confirmed that most of these dogs had Acanthocheilonema reconditum infection. Ten (4.0%) of the examined dogs were positive for Dirofilaria immitis circulating antigen (by DiroCHEK(®) test), but D. immitis microfilariae were not identified in any of the dogs and the status of this infection remains unclear. Further studies are needed to explore the occurrence of filariae in Zambian dogs and the zoonotic potential for humans.

Mapping the geographical distribution of lymphatic filariasis in Zambia.

BACKGROUND: Past case reports have indicated that lymphatic filariasis (LF) occurs in Zambia, but knowledge about its geographical distribution and prevalence pattern, and the underlying potential environmental drivers, has been limited. As a background for planning and implementation of control, a country-wide mapping survey was undertaken between 2003 and 2011. Here the mapping activities are outlined, the findings across the numerous survey sites are presented, and the ecological requirements of the LF distribution are explored. METHODOLOGY/PRINCIPAL FINDINGS: Approximately 10,000 adult volunteers from 108 geo-referenced survey sites across Zambia were examined for circulating filarial antigens (CFA) with rapid format ICT cards, and a map indicating the distribution of CFA prevalences in Zambia was prepared. 78% of survey sites had CFA positive cases, with prevalences ranging between 1% and 54%. Most positive survey sites had low prevalence, but six foci with more than 15% prevalence were identified. The observed geographical variation in prevalence pattern was examined in more detail using a species distribution modeling approach to explore environmental requirements for parasite presence, and to predict potential suitable habitats over unsurveyed areas. Of note, areas associated with human modification of the landscape appeared to play an important role for the general presence of LF, whereas temperature (measured as averaged seasonal land surface temperature) seemed to be an important determinant of medium-high prevalence levels. CONCLUSIONS/SIGNIFICANCE: LF was found to be surprisingly widespread in Zambia, although in most places with low prevalence. The produced maps and the identified environmental correlates of LF infection will provide useful guidance for planning and start-up of geographically targeted and cost-effective LF control in Zambia.

Lymphatic filariasis in Luangwa District, South-East Zambia.

BACKGROUND: Past case reports and recent data from LF mapping surveys indicate that LF occurs in Zambia, but no studies have been carried out to document its epidemiology and health implications. The present study assessed infection, disease, transmission and human perception aspects of LF in an endemic area of Luangwa District, South-East Zambia, as a background for planning and implementation of control. METHODS: Two neighbouring rural communities were registered and a questionnaire survey undertaken. Clinical examination, and sampling of blood for circulating filarial antigens (CFA; marker of adult worm infection) and antibodies to Bm14 antigen (marker of exposure to transmission), were carried out during the daytime. Blood from CFA positive individuals was examined for microfilariae (mf) at night. Vector surveys were carried out in selected households, using light traps. RESULTS: 985 individuals aged ≥ 1 year were registered. The CFA prevalence increased with age from 1.2% in age group 1-14 years to 20.6% in age group 50+ years (overall 8.6%). Wuchereria bancrofti mf were identified in 10.9% of CFA positive individuals (corresponding to a community prevalence of 0.9%). Prevalence and intensity of Bm14 antibodies were much higher in individuals ≥ 30 years than in younger individuals (57.2 vs. 19.3%; 0.594 vs. 0.241 OD-values). Elephantiasis and hydrocele were well known clinical manifestations in the area, but only one case of hydrocele was detected in the study population. Identified potential vectors were Anopheles funestus and An. gambiae. CONCLUSION: The study confirmed that LF was endemic in the study communities, but infection and disease prevalence was low. Several indications, including a marked recent decline in CFA prevalence, suggest that transmission in the area is on the decrease, perhaps because of intensive application of malaria control measures targeting the Anopheles vectors. It is recommended that mass drug administration is initiated to accelerate this positive trend of decline in LF transmission in the area.

Phylogeography and demographic history of Amblyomma variegatum (Fabricius) (Acari: Ixodidae), the tropical bont tick.

The genetic diversity of Amblyomma variegatum (Fabricius) from four Caribbean islands and five African countries was compared by analyzing the sequences of three gene fragments, two mitochondrial (12SrDNA and D-Loop-DL), and one nuclear (intergenic transcribed spacer 2 [ITS2]). Genetic variability of the ITS2 DNA fragment consisted of only uninformative single nucleotide mutations, and therefore this gene was excluded from further analyses. Mitochondrial gene divergences among African populations and between Caribbean and African populations were very low. Nevertheless, the data suggest that A. variegatum is divided into distinct East and West African groups, the western group including all Caribbean samples. Phylogenetic analyses of the 12SrDNA and DL gene sequences showed that the West African A. variegatum clustered in a well-supported monophyletic clade, distinct from eastern paraphyletic lineages. Sequences of A. variegatum from the Caribbean were embedded in the West African clade, which supports the known West African historical origin for these ticks.

Detection of Trypanosoma brucei in field-captured tsetse flies and identification of host species fed on by the infected flies.

The prevalence of trypanosome infections in tsetse flies in the Chiawa area of Lower Zambezi in Zambia, with endemic trypanosomosis, was determined by a polymerase chain reaction (PCR) method that allowed the detection of trypanosome DNA and determination of the type of animal host fed on by the tsetse fly Glossina pallidipes, using tsetse-derived DNA extracts as templates. Ninety G. pallidipes (82 females and 8 males; 18.3%) of the 492 flies captured by baited biconical traps tested positive for the presence of Trypanosoma brucei species genomic DNA. Of the 90 T. brucei-positive flies, 47 (52.2%) also tested positive for vertebrate mitochondrial DNA. Sequence analysis of the vertebrate mitochondrial DNA amplicons established that they originated from 8 different vertebrate species, namely, human (Homo sapiens), African elephant (Loxodonta cyclotis), African buffalo (Syncerus caffer), waterbuck (Kobus ellipsiprymnus), roan antelope (Hippotragus equinus), greater kudu (Tragelaphus strepsiceros), warthog (Phacochoerus africanus), and goat (Capra hircus). Furthermore, to investigate the prevalence of trypanosome infections in domestic goats in the same area where trypanosomes had been detected in tsetse files, a total of 86 goats were randomly selected from 6 different herds. Among the selected goats, 36 (41.9%) were found to be positive for T. brucei species. This combined detection method would be an ideal approach not only for mass screening for infection prevalence in tsetse populations, but also for the prediction of natural reservoirs in areas endemic for trypanosomosis.

Two serine protease inhibitors (serpins) that induce a bovine protective immune response against Rhipicephalus appendiculatus ticks.

We have previously undertaken preliminary characterization of two Rhipicephalus appendiculutus serine protease inhibitors (RAS-1 and -2) as anti-tick vaccine candidates. In this study, to clarify this hypothesis, we generated and further characterized recombinant RAS-1 and -2 (rRAS-1 and -2) and tested their potency as a cocktail anti-tick vaccine in cattle. RT-PCR analysis showed that RAS-1 and -2 mRNA transcripts are expressed during all life cycle stages of ticks, independent of sex. As judged by SDS-PAGE rRAS-1 and -2 migrated as a molecular weight of around 64 and 60 kDa protein, respectively, considering that the expression vector produced a recombinant protein fused with 18-22 kDa TRX protein. RAS-1 and -2 were found not to be secreted into the bite site as determined by the reactivity of anti-tick saliva sera to rRAS-1 and -2, suggesting that both proteins are concealed antigens. Vaccination of cattle with a combination of rRAS-1 and -2 conferred significant protective immunity against ticks, resulting in 61.4% reduction in nymph engorgement rate, and in 28 and 43% increased mortality rate in adult female and male ticks, respectively. This is the first report on an anti-tick vaccine trial using a combination of two different serpins derived from R. appendiculatus, and using cattle as a natural host.