Anti-onchocerca Metabolites from Cyperus articulatus:Isolation,In Vitro Activity and In Silico‘Drug-Likeness’

The aims of this investigation were to isolate active ingredients from the roots/rhizomes of Cyperus articulatus used as herbal medicine in Cameroon for the treatment of human onchocerciasis and to assess the efficacy of the metabolites on the Onchocerca worm.The antifilarial activity was evaluated in vitro on microfilariae(Mfs)and adult worms of the bovine derived Onchocerca ochengi,a close relative of Onchocerca volvulus.Cytotoxicity was assessed in vitro on monkey kidney epithelial cells.The structures of the active compounds were determined using spectroscopic methods and their drug-likeness evaluated using Lipinski parameters.Two secondary metabolites,AMJ1[containing mustakone(1)as the major component]and linoleic acid or(9Z,12Z)-octadeca-9,12-dienoic acid(2)were isolated.Both compounds were found to kill both the microfilariae and adult worms of O.ochengi in a dose dependent manner.The IC50s for AMJ1 were 15.7 lg/mL for Mfs,17.4 lg/mL for adult males and 21.9 lg/mL for adult female worms while for linoleic acid the values were,15.7 lg/mL for Mfs,31.0 lg/mL for adult males and 44.2 lg/mL for adult females.The present report provides the first ever evidence of the anti-Onchocerca efficacy of AMJ1 and linoleic acid.Thus,these secondary metabolites may provide a lead for design and development of new antifilarial agents.

Relationship between pro-and anti-inflammatory cytokines profiles and some haematological parameters in some Cameroonians infected with Onchocerca volvulus

Objective:To investigate the relationship between white blood cells,lymphocytes,monocytes, and Interleukin(IL)-1α,IL-6.IL-10 and IL-13 production in Cameroonians with Onchocerca volvulus(O.volvulus) infection.Methods:A lolal of 357 individuals from five sites at Upper Sanga.Lekkie.Nyoug.Kelle and Sanaga Maritime divisions and located along Sanaga valley of Sanaga River in South Cameroon were screened for the presence of O.volvulus using the skin snip.The levels of the interleukins(IL-) namely IL-1α,IL-6,IL-10 and IL-13 were evaluated using enzyme linked immunoabsorbent assay techniques.Haematological parameters were evaluated using standard laboratory automated analyser.Results:O.volvulus microfilariae were found in skin tissues of 85(23.81%) volunteers.The mean interleukin(IL-) levels in the 0,volvulus control and infected individuals were IL-1αin(1.65±0.79 and 2.31±0.5) pg/ml.:IL-6 in(278.36±55.34 and 20l.74±34.56) pg/ml.:IL-10 in(436.03±208.64 and 418.49±I47.88) pg/ml.and IL-13 in(8.98±7.28 and 3S.06±11.92) pg/mL.There was a negative correlation between monocyte counts and IL-10 concentration in positive individuals.A negative correlation of IL-6 with while blood cell and lymphocyte counts was observed(P【0.05).The level of IL-13 was positively associated with microfilaria]load(P【0.05).Conclusions:We observed depressed IL-6 and raised IL-13 concentrations in the sera of individuals with onchocerciasis which implicate these interleukins in the immunological responses of the disease.Therefore,these IL-6 and IL-13 are associated with O.volvulus infection among Cameroonians.

DPPH antiradical scavenging, anthelmintic and phytochemical studies of Cissus poulnea rhizomes

Objective: To investigate the phytochemical constituents, antioxidant and anthelmintic activities of the crude methanol extract of Cissus populnea(C. populnea) rhizomes. Methods: Phytochemical screening was performed using standard protocols, and column chromatography of silica gel was used for the compounds isolation. DPPH antiradical scavenging assay was performed in order to evaluate the antioxidant activity. Total phenolic content was evaluated using the Folin–Ciocalteu assay. The anthelmintic activity was screened on the bovine adult male forms of parasitic nematode Onchocerca ochengi, by the in vitro evaluation of the inhibition of adult worm motility and mortality. Worms were incubated in the presence of different concentrations of the plant extract and effects on survival were monitored after 24 and 48 h. Results: The preliminary phytochemical screening revealed the presence of phenolic compounds, saponins, steroids, tannins, and terpenoids. Bergenin and a mixture of phytosterol, β-sitosterol and stigmasterol were isolated from this extract and were identified by nuclear magnetic resonance, mass spectrometry and by comparison with published data. The crude methanol extract of C. populnea rhizomes showed a strong DPPH antiradical activity with a good amount of total phenolic content((20.69±2.13) g gallic acid equivalent/100 g of extract) and significant anthelmintic activity comparable to the standard drug ivermectin. Bergenin was found to be inactive even after 72 h of incubation. Conclusions: This study constitutes the first report on the anthelmintic activity of this plant and supports the traditional use of C. populnea as a natural antioxidant and anthelmintic.

Evaluation of Onchocerciasis: A Decade of Post Treatment with Ivermectin in Zainabi and Ririwai Doguwa Local Government Area of Kano State

Rapid Assessment Method (RAM) were carried out to assess the current situation of Onchocerciasis after repetition of annual community directed distribution of Ivermectin in Zainabi and Ririwai of Doguwa Local Government area of Kano State. Certain manifestations, like nodules, leopard skin and blindness, were used to measure the endemicity level in the community. The subjects of 30 - 50 years who are engaged in rural occupation, resident in that community, were examined for the presence of nodules, skin lesion and blindness. The common manifestation in both communities is nodules with 3 (3.40%) and 2 (3.44%). Leopard skin and blindness were found in Zainabi with 2 (2.27%) and 2 (2.27%). The manifestation of Onchocerciasis was found in older age groups of 49 - 70 and 50 - 69 respectively, which give an indication that the disease was eliminated in the community due to mass distribution of Ivermectin in the previously known endemic community. We recommend mass distribution of Mectizan in other identified endemic foci.

盘尾丝虫ASP1蛋白佐剂活性区不同标签融合表达与佐剂活性比较

为了探明纯化标签对盘尾丝虫活化相关分泌蛋白1(activation-associated secreted protein 1,ASP1)佐剂活性的影响,将其佐剂活性区PR-1编码序列分别与类弹性蛋白多肽(elastin-like polypeptide,ELP)、ELK16自聚肽或His标签进行融合表达,用相变循环、离心洗涤和镍亲和层析进行融合蛋白纯化;将ELP与传染性法氏囊病病毒(Infectious bursal disease virus,IBDV)VP2基因片段进行融合表达,用蛋白酶切除ELP标签后与不同标签融合PR-1免疫小鼠,两次免疫后不同时间采血分离血清,采用ELISA检测VP2特异IgG、IgG1和IgG2c滴度,用试剂盒检测免疫小鼠血清的IFN-γ、TNF-α、IL-6、IL-10浓度。结果显示,ELP-PR1、ELK-PR1、His-PR1和ELP-VP2融合蛋白在重组大肠杆菌中均获得正确表达,纯化蛋白纯度大于90%;3种标签融合PR-1均能增强免疫小鼠的抗原特异IgG、IgG1和IgG2应答,并能刺激小鼠产生IFN-γ、TNF-α、IL-6或IL-10细胞因子。在3种PR-1融合蛋白中,ELP-PR1的佐剂活性最强,ELK-PR1与不完全弗氏佐剂相当。本研究探索了不同纯化标签对PR-1免疫佐剂活性的影响,为传染性法氏囊病新型亚单位疫苗佐剂研制提供思路。

Impact of five annual rounds of mass drug administration with ivermectin on onchocerciasis in Sierra Leone

Background:Onchocerciasis is endemic in 12 of the 14 health districts of Sierra Leone.Good treatment coverage of community-directed treatment with ivermectin was achieved between 2005 and 2009 after the 11-year civil conflict.Sentinel site surveys were conducted in 2010 to evaluate the impact of five annual rounds of ivermectin distribution.Methods:In total,39 sentinel villages from hyper-and meso-endemic areas across the 12 endemic districts were surveyed using skin snips in 2010.Results were analyzed and compared with the baseline data from the same 39 villages.Results:The average microfilaridermia(MF)prevalence across 39 sentinel villages was 53.10%at baseline.The MF prevalence was higher in older age groups,with the lowest in the age group of 1-9 years(11.00%)and the highest in the age group of 40-49 years(82.31%).Overall mean MF density among the positives was 28.87 microfilariae(mf)/snip,increasing with age with the lowest in the age group of 1-9 years and the highest in the age group of 40-49 years.Males had higher MF prevalence and density than females.In 2010 after five rounds of mass drug administration,the overall MF prevalence decreased by 60.26%from 53.10%to 21.10%;the overall mean MF density among the positives decreased by 71.29%from 28.87 mf/snip to 8.29 mf/snip;and the overall mean MF density among all persons examined decreased by 88.58%from 15.33 mf/snip to 1.75 mf/snip.Ten of 12 endemic districts had>50%reduction in MF prevalence.Eleven of 12 districts had≥50%reduction in mean MF density among the positives.Conclusions:A significant reduction of onchocerciasis MF prevalence and mean density was recorded in all 12 districts of Sierra Leone after five annual MDAs with effective treatment coverage.The results suggested that the onchocerciasis elimination programme in Sierra Leone was on course to reach the objective of eliminating onchocerciasis in the country by the year 2025.Annual MDA with ivermectin should continue in all 12 districts and further evaluations are needed across the country to assist the NTDP with programme decision making.

Prevalence of epilepsy in the onchocerciasis endemic middle belt of Ghana after 27 years of mass drug administration with ivermectin

BackgroundIn onchocerciasis-endemic areas with high ongoing Onchocerca volvulus transmission,a high prevalence of epilepsy has been reported.This study aimed to determine the prevalence and clinical characteristics of epilepsy in the Bono Region of Ghana following 27 years of implementation of ivermectin mass drug administration(MDA).MethodsBetween October 2020 and August 2021,cross-sectional surveys were conducted in nine communities in the Tain District and Wenchi Municipality of the Bono Region of Ghana.In the first stage,a random door-to-door approach was used to screen the population for epilepsy using a pre-tested questionnaire.Persons suspected of having epilepsy were invited for a second-stage neurological examination for case verification.Community O.volvulus microfilarial infection status and Ov16 seropositivity were also determined.Ninety-five confidence intervals(95%CI)for prevalence values were calculated using the Wilson Score Interval.ResultsOf the 971 participants,500(51.5%)were females,and the median age(interquartile range)was 26(15‒43)years.Fourteen participants(1.4%,95%CI:1.0‒2.0)were diagnosed as having epilepsy with generalized seizures being the most frequent seizure type(85.7%,12/14).The overall microfilarial prevalence of O.volvulus was 10.3%(November 2020)and 9.9%(August 2021);the Ov16 seroprevalence was 22.2%(June 2021).Only 63.2%took ivermectin in the last round of MDA distribution in March 2021.ConclusionsThe 1.4%prevalence of epilepsy in the Bono region is similar to the median epilepsy prevalence in sub-Saharan Africa.However,the persistent microfilarial prevalence and low ivermectin study coverage call for the Ghana Onchocerciasis Elimination Programme to step up its efforts to ensure that the gains achieved are consolidated and improved to achieve the elimination of onchocerciasis by 2030.

The positive influence the Onchocerciasis Elimination Program for the Americas has had on Africa programs

A recent article Is onchocerciasis elimination in Africa feasible by 2025:a perspective based on lessons learnt from the African control programmesH in Infectious Diseases of Poverty claimed that undue influence on African programs by concepts developed by the Onchocerciasis Elimination Program of the Americas(OEPA)is detrimental to stopping mass drug administration(MDA)in Africa.This claim is made despite a record year for MDA stoppage in four African countries of>3.5 million treatments in 2018,far exceeding any past OEPA or African Program for Onchocerciasis Control(APOC)stop MDA success.

福斯盘尾丝虫病的流行病学调查及福斯盘尾丝虫线粒体COⅠ基因的序列分析

以内蒙古地区骆驼盘尾丝虫病流行病学调查为基础，基于细胞色素氧化酶工（COⅠ）基因的分子标记特点，进而为骆驼福斯盘尾丝虫传播媒介的研究奠定基础。在骆驼屠宰季节，进行骆驼福斯盘尾丝虫感染情况的检查，并分离虫体，提取DNA，用PCR方法扩增CO工基因，然后克隆和测序，并运用MEGA5．05和DNAStar7．1软件进行序列分析。结果显示，骆驼福斯盘尾丝虫主要寄生于骆驼的项韧带间隙，骆驼的平均感染率为92．2％，其中阿拉善左旗骆驼感染率高达100％，最大感染强度为每峰骆驼23个结节。对福斯盘尾丝虫COⅠ基因的序列分析结果表明，该基因长689bp，与GenBank中公布的同属线虫其他种的COⅠ基因的同源性介于89．8％～93．8％之间。对内蒙古地区骆驼感染福斯盘尾丝虫的状况进行了调查，并在国内外首次对福斯盘尾丝虫COⅠ基因序列进行了测序分析，为该虫种的分子标记及传播媒介的进一步研究奠定了重要基础。

骆驼福斯盘尾丝虫病组织病理学观察

为更有效地对骆驼盘尾丝虫病进行防治,利用流行病学调查和常规病理学观察的方法,对骆驼盘尾丝虫病进行研究。结果表明:内蒙古阿拉善左旗骆驼盘尾丝虫病平均感染率为85.4%,感染强度为1~15峰-1,虫体形成的结节分布在骆驼的颈项韧带处;共收集到209个结节,它们的形态和质地各不相同,软的结节内形成纤维组织囊腔,硬的结节内形成钙化灶;骆驼年龄段不同,结节的分布呈不同变化;病理组织学观察显示,囊腔周围有各种炎性细胞侵润,囊腔内有不同状态的雌虫虫体,骆驼盘尾丝虫病表现为骆驼的寄生虫性肉芽肿。