Sociodemographic and Economic Factors Associated with Urinary Bilharzia in the Municipalities of Sô-Ava, Aguegues and N’Dali in Benin

Introduction: This research, conducted in the municipalities of Aguégués, Sô-Ava, and N’dali in Benin, focused on identifying socio-demographic and economic factors associated with urinary bilharzia in 415 school-aged children from 8 to 14 years. Using urine test strips and filtration techniques, the study found prevalences ranging from 11.88% to 34.53% across the municipalities. Method: Epidemiological data (urine) were collected and examined with test strips to assess the presence of terminal haematuria: the prevalence of infestation and parasite density was quantified using the filtration technique. Socio-demographic and economic factors were recorded using a questionnaire to assess the correlation with disease. Results: Prevalences of 34.53% (N = 48 out of 139), 13.53% (N = 18 out of 133) and 11.88% (N = 17 out of 143) respectively for the municipalities of Aguégués, N’dali and Sô Ava were calculated. The study showed that the variables “age”, “sex”, “religion” and “socio-professional” activity were not significantly correlated with bilharziasis (p > 0.05) and it appears that these factors are not related to bilharziasis in the surveyed households. While age, sex, religion, and socio-professional activity showed no significant correlation with bilharziasis, factors like agriculture, fishing, and place of residence were statistically significant in relation to the disease. Conclusion: The findings suggest that these socio-demographic and economic factors impede the elimination of schistosomiasis in the examined areas.

青海省海晏县犬粪样中贾第鞭毛虫的检测

从青海省海晏县采集10只牧羊犬粪样用于贾第鞭毛虫检测。所有样品经饱和盐水浓缩法和蔗糖密度梯度离心等方法处理后采用光学显微镜、免疫荧光试验(IF)和套式PCR方法进行检测。结果显示,其中1个样品为阳性(10%),PCR扩增片段长度为274bp,命名为QHGC201101株。测序结果与肠贾第鞭毛虫3VR株(HQ616609)核糖体小亚基RNA的同源性为100%,表明分离的虫种为肠贾第鞭毛虫,属于人兽共患型病原。

青海省隐孢子虫的分子特征

本研究中分别对来自西宁动物园的21种观赏动物,乌兰县的16头牦牛和42只山羊,用免疫荧光法(IFT)进行了隐孢子虫卵囊的调查。用套式PCR进行种和基因型的检测,并且用18SrRNA进行序列分析。用IFT分别在西宁动物园的17个观赏动物,乌兰县的2头牦牛和15只山羊中发现了隐孢子虫的卵囊。对IFT阳性样品进行PCR扩增,其中10份为PCR阳性。并且用黑豹(Panthera pardus)、黑颈鹤(Grus nigricollis),蛮羊(Ammotragus lervia),羚牛(Budorcastaxicolor),小熊猫(Ailurus fulgens)和白马鸡(Crossoptilon crossoptilon)等动物隐孢子虫卵囊的PCR序列分析结果跟与微小隐孢子虫鼠基因型进行了比较。本研究首次报道了青海省野生动物中隐孢子虫的感染情况,并且首次在北山羊上分离得到的隐孢子虫中发现了鹿源基因型,类似C.bovis基因型,以及在山羊和野牦牛上发现了新的Cryptosporidium spp。

Isolation and identification of antibacterial components in seed extracts of Argemone mexicana L.(Papaveraceae)

Objective:The column chromatographic fractions of chloroform(CH1,CH2,CH3) seed extracts of Argemone mexicana(A.mexicana) were screened for antibacterial activity and phytochemical analysis.Methods:CH3 fraction was isolated and identified by column chromatography,thin layer chromatography,spectral data analysis and phytochemical screening were used for analysis. Results:CH3 fraction was significantly active at 4 to 64 mg/mL against Staphylococcus aureus, Escherichia coli,Klebsiella pneumoniae and Pseudomonas aeruginosa with minimum inhibitory concentration of 1.5625 to 3.125 mg/mL.The active fraction(CH3) revealed presence of alkaloid with retention factor value of 0.44.The active antibacterial agent in the most potent fraction (CH3) was isolated and identified as N-demethyloxysanguinarine by thin layer chromatography (TLC) and phytochemical screening.The CH1 and CH2 fractions did not show inhibitory activity. Conclusions:The results support the ethnomedicinal use of seeds of A.mexicana for treatment of bacterial diseases.

Isolation and identification of bioactive antibacterial components in leaf extracts of Vangueria spinosa(Rubiaceae)

Objective:The column chromatographic fraction of ethyl acetate(EA1,EA2,EA3,EA4 and EA5) leaf extracts of Vangueria spinosa(V.spinosa) were screened for antibacterial activity and phytochemical analysis.Methods:EA3 fraction was isolated and identified by column chromatography,thin layer chromatography,spectral data analysis and phytochemical screening were used for analysis.Results:EA3 fraction was significantly active at 4 to 64 mg/L against Staphylococcus aureus,Escherichia coli,Klebsiella pneumoniae and Pseudomonas aeruginosa with minimum inhibitory concentration of 1.5625 to 3.1250 mg/mL.The active fraction(EA3) revealed the presence of flavonoid with retention factor value(R_f) of 0.39.The active antibacterial agent in the most potent fraction(EA3) was isolated and identified as flavonoid(-)-epicatechin-3-O-β-glucopyranoside by thin layer chromatography(TLC) and phytochemical screening.EA1 and EA2 show inhibitory activity at 4 to 64 mg/L against Staphylococcus aureus only where as fraction EA4 and EA5 do not shows any inhibitory activity within that range of concentration against any bacteria.Conclusions:The results support the ethnomedicinal use of leaf of V.spinosa for the treatment of bacterial diseases.

Epidemiology of cercarial stage of trematodes in freshwater snails from Chiang Mai province,Thailand

Objective:To investigate the epidemiological situation of cercarial trematodes infection in freshwater snails from different water resources in Chiang Mai province,Thailand.Methods:The snail specimens were collected from 13 districts of Chiang Mai province during April 2008 to February 2012.The prevalence of cercarial infection in snails was investigated using the crushing method.The drawing was done with the help of a camera lucida for the morphological study.Results:A total of 2479 snail individuals were collected and classified into 7 families,11 genera,and 14 species,Among them,8 snails species were found to be infected with an overall prevalence of 17.27%(428/2479),which infected with nine groups of cercariae;gymnocephalous cercaria,strigea cercaria,megalurous cercaria,monostome cercaria,parapleurolophocercous cercaria(Haplorchis cercaria),pleurolophocercous cercaria,furcocercous cercaria(Transversotrema cercaria),xiphidiocercaria,and virgulate cercaria.The parapleurolophocercous cercaria was found to be the dominant type among the cercarial infection in the snails(64.25%).Conclusions:The various species of snails found in the research location act as the intermediate hosts for the high prevalence of parasitic infection of many species of mammals.This work will provide new information on both the distribution and first intermediate host of trematodes.

Larvicidal activity of Dregea volubilis and Bombax malabaricum leaf extracts against the filarial vector Culex quinquefasciatus

Objective:To analyze the larvicidal activity of two plant leaf powder and leaf-extracts, Dregea volubilis and Bombax malabaricum against Culex quinquefasciatus.Methods:The larvicidal bioassay was done for powdered leaves of Dregea volubilis and Bombax malabaricum individually and their methanol extracts against first-,second-,third- and fourth-instar larval forms of Culex quinquefasciatus.Mortality rate was recorded after 24,48 and 72 h of postexposure. LC50 and LG90 values were calculated at different time intervals for third instar larvae. Results:All the graded concentration（0.1%,0.2%,0.3%,0.4%,0.5%） of powdered leaves showed significant（P 【0.05） larval mortality.The mortality rate was higher in 50 ppm doses of methanolic extracts of both the plants against Culex quinquefaciatus.The corresponding 【C50 values were 56.97 ppm and 48.85 ppm,respectively after 24 h of exposure.There is no mortality of non-target organism such as Chironomous circumdatus,Oreochromis niloticus niloticus and Diplonichus annulatum within 72 h of post exposure to 0.1%,0.2%,0.3%crude powdered leaves and methanolic extracts of both the two individual plants under the laboratory condition.The results of preliminary qualitative phytochemical analysis of both the plants revealed the presence of many bioactive principles such as steroids,tannins,flavonoids,triterpenoids,saponins,etc. that may be responsible for their bio-control potentiality.Conclusions:The results have shown potential and eco-friendly use of both plant extracts against larva of Culex quinquefasciatus for the first time.

Control of human filarial vector,Culex quinquefasciatus Say 1823(Diptera:Culicidae)through bioactive fraction of Cayratia trifolia leaf

Objective:To investigate the mosquito larvicidal activity of Cayratia trifolia(L.)Domin(Vitaceae:Vitales)(C.trifolia)which is distributed in many parts of India with medicinal properties as vector control is facing threat due to the emergence of resistance to synthetic insecticides.Methods:Young and mature leaves of C.trifolia were investigated for larvicidal activity against3rd instars larvae of Culex quinquefasciatus in different seasons throughout the year.The active fractions were extracted using six different solvents in a non-polar to polar fashion viz petroleumether,benzene,chloroform:methanol(1:1 v/v),acetone,absolute alcohol and distilled water.Dose dependent mortality was recorded against each solvent extract.Determination of LD_(50)and LD_(90)were executed through log-probit analysis using the most bioactive fraction.The fluctuations in mortality were statistically co-related through ANOVA analyses concerning different seasons and types of leaves as random variables.Justification of larvicidal activity was established through student's t-test.Costing effects were evaluated on the non-target water fauna under laboratory conditions.Thin layer chromatographic techniques were performed for phytochemical analysis and categorization of chemical personality of the active fractions using the most effective solvent extract following standard methods.Results:Significant variations in mortality rate were noted with respect to the type of leaves(mature and senescence),concentration of leaf extract and between seasons.The water extract among all the solvent extracts was found to induce cent percent mortality at 50 mg/L in test mosquito species within 24 h with a LD_(50)and LD_(90)value of 10.70 mg/L and 27.64 mg/L respectively.No significant mortality was recorded in non-target water population.Chromatographic analyses of the water extract revealed the presence of steroids,triterpene glycosides,essential oil,phenolics and diterpenes as secondary phytochemicals.Conclusions:Water extract of C.trifolia leaf promised as a cost effective and potent larvicidal agent against Culex quinquefasciatus.

Survey on gastrointestinal parasites and detection of Cryptosporidium spp. on cattle in West Java,Indonesia

Objective:To evaluate the presence of gastrointestinal parasites on cattle in Indonesia because the prevalence of parasites varies between counlries depending on the terrain surrounding livestock farms and investigations in Indonesia have never been performed.Methods:Fecal samples from cattle at 35 farms in 7 districts in West Java,Indonesia,has been examined using the floatation or sedimentation methods,and a immunofluorescence assay and experimentally inoculation to mice for Cryptosporidium or Giardia spp.Results:153 of 394 examined cattle(38.8%)were infected with gastrointestinal parasites.The prevalence of Eimeria spp.,Nematoda spp.(including Oesophagustomum and Bunostomum-like),Fasciola gigantica and Paramphistomum spp.was 22.4%,11.2%,12.5%and 3.8%,respectively.Cryptosporidium andersoni(C.andersoni)was also found in two samples.One isolate of this parasite was confirmed to be transmitted to mice,in contrast to the isolates from other countries.Conclusions:although this survey is preliminary,the results shows that the infection of gastrointestinal parasites in Indonesia was not high,but these infected cattle could be as a potential source leading to economic losses in livestock production.

Mosquito larvicidal activity of some common spices and vegetable waste on Culex quinquefasciatus and Anopheles stephensi

Objective:To investigate the larvicidal activities of crude and chlor of orm:methanol(1:1 v/ v) extracts of some common spices(Cuminum cyminum,Allium sativum,Zingiber offinale, Curcuma longa) and vegetable waste(Solanum tuberosum germinated tuber) against Anopheles stephensi and Culex quinquefasciatus mosquito larvae.Methods:Larval mortality of above mosquito species were observed after 24,48 and 72 h of exposure to five concentrations of aqueous extract(0.1%,0.2%,0.3%,0.4%and 0.5%) and four concentrations(25,50,75 ppm) of chlor of orm:methanol(1:1 v/v) extract.The lethal concentration of individual spices or vegetable waste was determined by log-probit analysis(at 95%confidence level) and effect of crude and chlor of orm:methanol(1:1 v/v) extracts were recorded on non target organisms.Results: Relative mortality rate of both larval mosquito species were recorded in the following sequences: Cuminum cyminum】 Allium sativum】Zingiber offinale,Curcuma longa 】 Solanum tuberosum germinated tuber for crude extract,and efficacy of chlor of orm:methanol(1:1 v/v) extract were as follows:Curcuma longa 】 Zingiber offinale 】 Solanum tuberosum germinated tuber 】 Cuminum cyminum】 Allium sativum.Conclusions:Crude and chlor of orm:methanol(1:1 v/v) extract of Cuminum cyminum,Allium sativum,Zingiber offinale,Curcuma longa and Solanum tuberosum germinated tuber can be recommended effectively in mosquito control programmes at very low concentrations.No mortality and other abnormalities were noticed on non target organisms and further studies are needed to investigate the chemical structure of active principal which are responsible for larvicidal activity.

Mosquito larvicidal effect of orthophosporic acid and lactic acid individually or their combined form on Aedes aegypti

Objective:To observe the effect of two common organic acids on the larvae of Aedes aegypti (Ae.aegypti)(L),the natural vector of dengue fever/dengue hemorrhage fever,chikugunya and allergic skin reaction especially in children.Methods:Two common organic acids(lactic acid and orthophosporic acid of gradually increasing concentration) were used against laboratory reared third instars larvae of Ae.aegypti in order to observe the rate of mortality after 8.16 and 24 h of post exposure respectively in laboratory.Results:Larval mortality rates recorded were in the following sequences:orthophosphoric acid and lactic acid at 1:1 combination 】orthophosphoric acid】lactic acid.Conclusions:These two organic acids may be used perfectly in combination (1:1) along with other conventional vector control methods to reduce the Ae.aegypti population, especially in those areas where surveillance and supervisory mechanism are poor or insufficient.

Mosquito larvicidal activity of Rauvolfia serpentina L.seeds against Culex quinquefasciatus Say

Objective:To establish the larvicidal activities,if any of solvent extracts of Rauvolfia serpentina (R.serpentina) L.seeds against Culex quinquefasciatus(Cx.quinquefasciatus) Say.1823 as target species.Methods:Seeds of R.serpentina were extracted with five solvents graded according to the polarity[viz.petroleum ether.benzene,ethyl acetate,acetone and absolute alcohol] continuing one after another with the same seeds.Results:Mortality rate with petroleum ether extract was significantly higher than other extracts.The mortality rates of late 3rd instar larvae wore 50.33±5.51.10.00±1.00.0.00±0.00.21.33±1.53 and 0.00±0.00 in 100 ppm concentration of petroleum ether,benzene,ethyl acetate,acetone and absolute alcohol respectively,after 24 h of exposure period.Conclusions:Results of this study show that petroleum ether extract of R. serpentina seed may be considered as a potent source of mosquito larvicidal agent.

Studies on seasonal fluctuation of different indices related to filarial vector,Culex quinquefasciatus around foothills of Susunia of West Bengal,India

Objective:To collect information on the seasonal activity of filarial vector around foothills of Susunia of West Bengal,India.Methods:Indoor-resting mosquitoes were collected from the human habitations of 16 villages.Collected mosquitoes were identified and examined for different parameters following standard methods recommended by World Health Organization and pioneer workers.Results:Overall man hour density,infection rate,infectivity rate,human blood index and daily mortality rate of the vector were assessed as 10.54%,6.31%,1.38%,77.33%and 13.00%, respectively,which were 12.17%,8.63%,2.26%,100.00%and 17.00%,respectively in the summer season.Average load of microfilaria,1st stage,2nd stage and 3rd stage larvae of Wuchereria bancrofti in infected vectors were 10.02.7.50,5.56 and 4.68,respectively,which were 11.58,9.90. 6.27 and 5.44,respectively in summer.Among the searched shelters 63.37%,8.16%and 2.08% were found to be invaded by any vector,infected vector and infective vector,respectively,which were 68.75%,12.76%and 3.91%,respectively in summer.Conclusions:Different indices related to the vector were much higher in summer,which indicates that,summer is the most favorable season for transmission of lymphatic filariasis in the study area.Available data will help to formulate an effective vector control measure.

Lymphatic filariasis in the foothill areas around Susunia of West Bengal in India

Objective:To investigate the filarial epidemiology in 16 foothill villages around Susunia hill,Bankura district, West Bengal,India.Indices studied were microfilaria rate,mean microfilarial density,filarial disease rate and endemicity rate.Other indices related to transmission were incrimination of vector species,man-hour density of the vector,vector infection and infectivity rates,human blood index of the vector etc.Methods:Examination of 20μL night blood samples by finger prick and clinical examination for filarial diseases of 3 737 people(2 241 male and 1 496 female) was done randomly covering nearly 22%population of the study area.Aspects related to vectors were dealt by regular collection and dissection of mosquitoes.Results:Overall microfilaria rate,mean microfilarial density,disease rate and endemicity rate were 6.10%,10.86%,20.20%and 25.58%,respectively. Causative parasite was identified as Wuchereria bancrofii and Culex quinquefasciatus was incriminated as the vector therein.Vector infection rate,infectivity rate and human blood index were assessed to be 6. 31%,1.38%and 77.33%,respectively.Conclusion:Present study is highly endemic for bancroftian filariasis. More than one fourth of the population under study were filarial victims indicating an overall alarming situation and immediate measure should be taken to rectify the situation.

Epidemiological situation and molecular identification of cercarial stage in freshwater snails in Chao-Phraya Basin, Central Thailand

Objective: To investigate the prevalence of cercarial trematode infection in snails and to examine the reconstruction of the phylogenetic relationship to explain the molecular system of cercarial stage trematodes to estimate the infection rate of in the definite host from the Chao-Phraya Basin.Methods: The snails were collected from 10 provinces of the Chao-Phraya Basin,Thailand by stratified sampling method. The snails were examined for cercarial infection by the crushing method. All DNA specimens were amplified with internal transcribed spacer 3(ITS3) and ITS4 primer based on PCR technique. The sequence data were aligned and used to reconstruct the phylogenetic tree by unweighted pair-group method with arithmetic means with 10 000 bootstraps.Results: The overall rate of cercarial infection was found to be 5.90%(122/2 067). Snails in the family Thiaridae were found to be in the highest prevalence followed by Lymnaeidae, Bithyniidae, Planorbidae, Viviparidae, and Ampullariidae, respectively, while the Buccinidae family(Clea helena) did not reveal any infections. The frequently found species of cercariae were parapleurolophocercous cercariae, cercariae and megarulous cercariae. The monophyletic tree separated the snails into five groups comprised of Heterophyidae, Strigeidae, Lecithodendriidae, Philophthalmidae and Echinostomatidae using the sequence of Angiostrongylus cantonensis as an out-group.Conclusions: This study was the first to report on cercarial infection in the Chao-Phraya Basin, Thailand. This revealed that a high variety of freshwater snails were infected by cercariae stage trematodes with a high prevalence. The sequence data of ITS2 can be used to investigate the phylogenetic relationships of trematodes at the family level and in each clade of different families separated by the definitive hosts.

Malaria vaccines:looking back and lessons learnt

The current status of malaria vaccine approaches has the background of a long and arduous path of malaria disease control and vaccine development.Here,we critically review with regard to unilateral interventional approaches and highlight the impact of socioeconomic elements of malaria endemicity. The necessity of re-energizing basic research of malaria life-cycle and Plasmodium developmental biology to provide the basis for promising and cost-effective vaccine approaches and to reach eradication goals is more urgent than previously believed.We closely analyse the flaws of various vaccine approaches,outline future directions and challenges that still face us and conclude that the focus of the field must be shifted to the basic research efforts including findings on the skin stage of infection.We also reflect on economic factors of vaccine development and the impact of public perception when it comes to vaccine uptake.

Cytokine profile in murine toxoplasmosis

Objective:To investigate which cytokines are produced after acute infection of mice with Toxoplasma gondii(T.Gondii) RH strain.Methods:Mus domesticus domesticus mice in infected group were inoculated with with highly virulent T.Gondii RH strain by intraperitoneally.Serum samples were obtained from infected and non-infected mice for cytokine levels for ELISA assay.Results:The concentrations of tumor necrosis factor-α,interferon-γ,interleukin(IL)- 10 and IL-12 in the cardiac blood sample of the infected mice were significantly higher than those in uninfected controls(P【0.05).The levels of transforming growth factor-1βdecreased in mice infected with T.gondii compared to those of the controls,the decrease was statistically significant(P【0.05).No significant difference was observed in levels of IL-4 between infected and healty control groups(P】0.05).Conclusions:According to our findings,immune response into T helper type 1 was predominant during acute T.gondii infection.Further characterization and purification of Toxoplasma molecule(s) implicated in the regulation of cytokines could lead to the development of new drug prospects to control Toxoplasma infection.

Multiplex PCR assay for discrimination of Centrocestus caninus and Stellantchasmus falcatus

Objective: To develop the multiplex PCR method based on the internal transcribed spacer 2 to discriminate the intestinal trematodes, Centrocestus caninus(C. caninus), and Stellantchasmus falcatus(S. falcatus).Methods: Four species of heterophyid trematodes including C. caninus, S. falcatus,Haplorchis taichui and Haplorchoides sp. were amplified and the specific primer was designed based on the internal transcribed spacer 2 region. Two specific primers were used to validate the optimized PCR conditions: the specificity test and the sensitivity test.Results: Both of these specific primers confirmed the specificity through multiplex PCR reaction which generated both PCR products(231 and 137 bp) in the mixed DNA template of C. caninus and S. falcatus with no cross-reaction with other heterophyid trematodes. The optimum annealing temperature of both primers was 54–59℃. The sensitivity test used the two-fold serial dilution DNA template, which was concentrated between 10 and 0.312 5 ng/mL. The lowest concentration of the DNA template of this multiplex PCR was 2.5 ng/mL.Conclusions: The technique described here proved to be a species-specific technique and was found to be a rapid method for the diagnosis of C. caninus and S. falcatus in terms of the larval and adult stages in intermediate and/or definitive hosts in the endemic area.

Epidemiology and molecular genotyping of echinostome metacercariae in Filopaludina snails in Lamphun Province,Thailand

Objective:To analyze the prevalence of echinostome metacercariae in Filopaludina dorliaris(F.dorliaris)and Filopaludina martensi martensi(F.martensi martensi)and genotype variation of echinostome metacercariae by using random amplified polymorphic DNA(RAPD)analysis.Methods:Filopaludina sp.snails were collected from eight localities of Lamphun Province,Northern Thailand and examined for echinostome metacercariae.RAPD-PCR was used lo analyze genotype variation of echinostome metacercariae.Results:A total of 3 226 F.dorliaris and F.martensi martensi snails were collected from eight localities.The overall prevalences of echinostome metacercariae in F.dorliaris and F.martensi martensi were 40.89%and 36.27%,while the intensity of infection was 20.37 and 12.04,respectively.The dendrogram constructed base on RAPD profiles,4 well supported domains were generated;(i)group of metacercariae from Ban Hong,Mae Ta,Meaung,Pa Sang,Toong Hua Chang,and Weang Nong that were clustered in the group of E.revolutum,(ii)Ban Thi,(iii)Lee,and(iv)3 adults of an out group.Condusioiis:This research demonstrated RAPD profiling has been a useful tool to detect DNA polymorphisms to determine genetic relationship between echinostome metacercariae in Lamphun Province,Northern Thailand.