Use of Polyclonal Antibody for the Diagnosis of Human African Trypanosomiasis

Human African trypanosomiasis (HAT), commonly known as sleeping sickness is one of the neglected tropical diseases (NTDs), which is fatal if left untreated. Its diagnosis is a challenge since the signs and symptoms of the primary phase are not specific, the existing diagnostic methods have low sensitivity and specificity, and the available drugs have some toxicity. New, robust, and cost-effective techniques are needed for the early identification of parasites. This study aimed to assess the sensitivity and specificity of two different types of polyclonal antibodies against T. b. gambiense using antigen detection ELISA. Polyclonal antibodies against the expressed proteins Tbg I2 and Tbg I17 were produced using New Zealand white rabbits. The antibody titer measured was greater than 32 g/L after the 3rd immunization for the expressed protein Tbg I2. For the expressed protein Tbg I17, the antibody titer measured was greater than 32 g/L after the 4th immunization. The sensitivity and specificity of the Tbg I2 polyclonal antibody confirmed with Polymerase Chain Reaction (PCR) as gold standard were respectively 89.5% and 80.6%, while for the Tbg I17 polyclonal antibody, the sensitivity and specificity were respectively 92.1% and 88.9%. The area under the curve for the Tbg I2 polyclonal antibody was 0.90 ± 0.032, while for the Tbg I17 polyclonal antibody, the area under the curve was 0.92 ± 0.0. The Tbg I17 polyclonal antibody produced in New Zealand white rabbits has good sensitivity and good specificity;it can be successfully used in the diagnosis of HAT.

Compounds from African Medicinal Plants with Activities Against Selected Parasitic Diseases:Schistosomiasis,Trypanosomiasis and Leishmaniasis

Parasitic diseases continue to represent a threat on a global scale,particularly among the poorest countries in the world.This is particularly because of the absence of vaccines,and in some cases,resistance against available drugs,currently being used for their treatment.In this review emphasis is laid on natural products and scaffolds from African medicinal plants(AMPs)for lead drug discovery and possible further development of drugs for the treatment of parasitic diseases.In the discussion,emphasis has been laid on alkaloids,terpenoids,quinones,flavonoids and narrower compound classes of compounds with micromolar range activities against Schistosoma,Trypanosoma and Leishmania species.In each subparagraph,emphasis is laid on the compound subclasses with most promising in vitro and/or in vivo activities of plant extracts and isolated compounds.Suggestions for future drug development from African medicinal plants have also been provided.This review covering 167 references,including 82 compounds,provides information published within two decades(1997-2017).

High Field MRI in Human African Trypanosomiasis (HAT)

Human African Trypanosomiasis (HAT) or sleeping thickness is a forest and rural disease;where agriculture is the main activity. It is a chronic and lethal disease without treatment. HAT is caused by two parasites;Trypanosoma Brucei Gambiense (gTB) and Trypanosoma Brucei Rhodesiense (rTB) transmitted to humans by the tsetse fly. It is endemic condition in Africa between the 15&deg north latitude and the 20° south latitude. It is reported outside this area in travelers who stayed in endemic zone. Infection by gTB is wider and more frequent (98%) than that by rTB (2%). The Democratic Republic of Congo is the most affected country with more than 75% of reported cases. The geographical distribution is not homogeneous. There are more affected regions in a zone called “foci” which represents areas favorable to the development of the vector. Its diagnosis and treatment are very important because of its social and economic impact at both the individual and community levels. Promising molecules including fexinidazole are currently undergoing testing. Nowadays populations move more and more easily but the discovery of this disease in daily neuroradiological practice is exceptional. We propose in this paper through two observations, reminders on epidemiological, clinical and MRI features of HAT. It typically performs the edematous, bilateral and diffuse encephalitis. It is important to distinguish these aspects from the arsenic-induced encephalitis that may occur during treatment. Only vector control allows eradicating this disease. WHO has set targets elimination of HAT as a public health problem for 2020 deadline.

Human African Trypanosomiasis Successfully Treated with Melarsoprol in Pregnancy in a Niger Delta Rural Hospital

Human African trypanosomiasis (HAT) commonly known as sleeping sickness occurs in about 36 countries in sub-Saharan Africa and results in a large number of deaths and considerable illness. The drugs used in the treatment of HAT are very toxic and therefore might not be safe in pregnancy. Few published data exist on the treatment of HAT in pregnancy. We describe a case of T. brucei gambiense infection occurring in a pregnant woman that was successfully treated with Melarsoprol with no toxic effect to mother and the baby after 2 years of follow-up.

Human African trypanosomiasis in endemic focus of Abraka,Nigeria

Objective:To investigated the prevalence of human African trypanosomiasis(HAT),a neglected tropical disease caused by Trypanosoma brucei gambiens in an endemic focus of Nigeria,as it relates to age,sex and occupational differences.Methods:A total of 474 human subjects were screened using card agglutination test for trypanosomiasis kit.Positive samples were further investigated for parasite positivity in blood/serum and cerebrospinal fluid(CSF).Results:Of the 474 screened,44(9.3%) were seropositive with seroprevalence of 22(9.6%) in Urhouka,14(9.5%) in Umeghe and 8(7.9%) for Ugonu.The number of seropositives,observed for weakly,moderately and strongly positives for the three communities were 4,7 and 11 in Urhouka,4,5 and 5 in Umeghe and 3,2 and 3 in Ugonu respectively.Among the 16 volunteers with detected parasite in their blood,4 of them were weakly positive,5 of them were moderately positive and 7 of them strongly positive.4 volunteers from Urhouka community were found parasites in their CSF and they were all strongly positive.The difference between the seroprevalence of males and females was not statistically significant(OR=1.14,95%CI=0.37-3.4,P】0.05).The prevalence difference between age group 21-30 years old and the youngest and oldest age groups was statistically significant(OR=3.5,95%CI= 1.08-12.57,P【0.05) but not significant for other age categories (P】0.05),It was observed that farmers had significantly higher prevalence of HAT infection as well as greater risk of Trypanosoma brucei gambiense infection than inhabitants with other occupations (OR=3.25,95%CI=0.99-11.79,P【0.05).Conclusions:Human activities such as farming and visits to the river have been identified as major risk factors to HAT.Also the breakdown of HAT control program has been advanced for the rise in HAT in Abraka,an endemic focus in Nigeria.

Role of chemokines and cytokines in the neuropathogenesis of African trypanosomiasis

Trypanosoma brucei spp. cause human African trypanosomiasis(HAT) or sleeping sickness in humans and nagana in animals. The early stages of the disease have no specific symptoms; however, the late stage of the disease involves neurological signs of the disease, including disturbance of sleep patterns from which the disease derives the name sleeping sickness. During the late stage of African trypanosomiasis parasites, increased numbers of white blood cells and levels of cytokines and/or chemokines are found in the brain parenchyma and/or cerebrospinal fluid of animal models and HAT patients. In this mini review, contemporary findings on how chemokines and cytokines are thought to play an important role in the central nervous system invasion by the parasites, inflammation and the neuropathology of the disease are discussed. The levels of various cytokines and chemokines, such as interferongamma(IFN-γ), interleukin-1 beta(IL-1β), IL-6, IL-10, tumor necrosis factor-alpha(TNF-α), C-C motif chemokine 2(CCL2), CCL3, C-X-C motif chemokine 8(CXCL8, IL-8) and CXCL10, in the cerebrospinal fluid(CSF) of HAT patients correlate with the severity or stage of the disease. Thus, these molecules are possible candidates for differentiating between early and late stage HAT. The role of cytokines and chemokines in parasite invasion of the central nervous system is also being eluci-dated. IFN-γ, TNF-α and CXCL-10 are some of the cytokines and chemokines now known to facilitate parasite penetration of the brain parenchyma. Interestingly, they also constitute some of the candidate molecules with potential to differentiate between stage 1 and 2 of HAT. The increased levels of cytokines, such as IL-1β, IL-6, IFN-γ and TNF-α, as well as prostaglandins, during African trypanosomiasis might contribute to the neurological dysfunctions that occur during HAT.

HLA-G 3’UTR 14 bp Insertion Is Associated with a Decreased Risk of Developing Human African Trypanosomiasis in the Côte d’Ivoire Population

Human African trypanosomiasis (HAT), or sleeping sickness, caused by Trypanosoma brucei gambiense, is associated with diverse clinical outcomes. Host’s genetic factors involved in immunity are potential factors that can regulate infection. Genetic polymorphisms within HLA-G could influence the level of HLA-G expression and therefore play a critical role in infection outcomes. The goal of our study was to investigate the association of 14 bp Indel HLA-G polymorphism with the susceptibility/resistance to HAT. DNA samples were collected from 119 cases, 221 controls and 43 seropositive individuals living in Ivorian HAT foci. The 14 bp Indel polymorphism was determined by PCR. Homozygous individuals for 14 bp insertion had a lower risk of progressing to active HAT (p = 0.012, OR = 0.27, 95% CI: 0.09 - 0.8). Moreover, the frequency of 14 bp insertion homozygous genotype was higher in the seropositive group (11%) than in the HAT cases group (3%) (p = 0.043, OR = 0.27, 95% CI: 0.07 - 0.99), which suggested a protective effect of 14 bp insertion homozygous genotype. Genetic polymorphisms in HLA-G may be associated with a variable risk to develop HAT. The 14 bp insertion appears to favour the occurrence of long-lasting T. b. gambiense latent infections.

Sero-Prevalence Study of Camel Trypanosomiasis in Selected Villages of Galkayo, Somalia

A cross-sectional study was carried out aimed to estimate sero-prevalence of camel trypanosomiasis and to investigate the related risk factors of the disease in Four Selected Villages of Galkayo, Mudug region from 14th March 2016 up to 20th April 2016. Blood samples were collected from 69 randomly selected camels of the four study villages, and samples were allowed to clot at room temper to detect the sero-prevalence of trypanosome using Card Agglutination Test for Trypanosomiasis (CATT). The results indicated that the overall sero-prevalence of trypanosomiasis in camels that 15.9% (11) samples were positive for trypanosome evansi (T. evansi) was recorded. Higher infection was found in female (19.4) as compared to male (11.1). However, there is no statistically significant difference in sero-prevalence between sex categories (P > 0.05). High Test infection of sero-prevalence was noted 20% in young age (10), and there was statistically significant difference (P < 0.05) in susceptibility among age groups. These results seem to indicate that T. evansi infection is high in the study area. There is need of further control of camel trypanosomiasis through the uses of curative and prophylactic drugs to avoid the various problems.

Environmental Mapping of Potential Habitats for Anthropod Vectors of Trypanosomiasis in Northern Nigeria

This paper presents analysis of a cost effective methodology using remotely sensed data analysed within a geographical systems (GIS) environment for mapping out potential habitats of anthropod vectors responsible for trypanosomiasis in northern Nigeria. These geographical locations are areas with high population of livestock in Nigeria. Animal Trypanosomiasis is considered as an arthropod-borne viral disease which is endemic in about 37 countries of the sub-Saharan Africa and in particular northern Nigeria. This anthropod-borne viral disease remains a threat to both humans and livestock in many communities, and the outbreak of such diseases is shown to relate to fluctuations in the changing climate mostly experienced whenever there are changes in global precipitation which also relates to changes in sea surface temperatures otherwise known as “El Niňo Southern Oscillations” (ENSO). Monthly Satellite imageries in the form of Normalised Difference Vegetation Index (NDVI) at 250 meters spatial resolution obtained from NASA-MODIS/CMD were subjected to principal component analysis utilizing the standardized principal components within a GIS environment and supplemented with Digital Elevation Model (DEM) data in the analysis. Results from the maps showed that pockets of probable habitats of these anthropod vectors responsible for trypanosomiasis mostly located around forest islands characterized by dry woodland and savanna, and in some cases around gallery forests and few lowland and riverine areas. This study provides baseline information for policy makers in Nigeria and other stakeholders as a cost effective measure for mapping potential habitats for anthropod vectors responsible for trypanosomiasis in Northern Nigeria. Further studies are encouraged so as to clearly understand the magnitude and actual locations of the habitat of this vector and find ways of targeting their locations for minimizing or even eradicating these vectors.

Performance of clinical signs and symptoms,rapid and reference laboratory diagnostic tests for diagnosis of human African trypanosomiasis by passive screening in Guinea:a prospective diagnostic accuracy study

Background Passive diagnosis of human African trypanosomiasis(HAT)at the health facility level is a major component of HAT control in Guinea.We examined which clinical signs and symptoms are associated with HAT,and assessed the performance of selected clinical presentations,of rapid diagnostic tests(RDT),and of reference laboratory tests on dried blood spots(DBS)for diagnosing HAT in Guinea.Method The study took place in 14 health facilities in Guinea,where 2345 clinical suspects were tested with RDTs(HAT Sero-K-Set,rHAT Sero-Strip,and SD Bioline HAT).Seropositives underwent parasitological examination(reference test)to confirm HAT and their DBS were tested in indirect enzyme-linked immunoassay(ELISA)/Trypanosoma brucei gambiense,trypanolysis,Loopamp Trypanosoma brucei Detection kit(LAMP)and m18S quantitative PCR(qPCR).Multivariable regression analysis assessed association of clinical presentation with HAT.Sensitivity,specificity,positive and negative predictive values of key clinical presentations,of the RDTs and of the DBS tests for HAT diagnosis were determined.Results The HAT prevalence,as confirmed parasitologically,was 2.0%(48/2345,95%CI:1.5–2.7%).Odds ratios(OR)for HAT were increased for participants with swollen lymph nodes(OR=96.7,95%CI:20.7–452.0),important weight loss(OR=20.4,95%CI:7.05–58.9),severe itching(OR=45.9,95%CI:7.3–288.7)or motor disorders(OR=4.5,95%CI:0.89–22.5).Presence of at least one of these clinical presentations was 75.6%(95%CI:73.8–77.4%)specific and 97.9%(95%CI:88.9–99.9%)sensitive for HAT.HAT Sero-K-Set,rHAT Sero-Strip,and SD Bioline HAT were respectively 97.5%(95%CI:96.8–98.1%),99.4%(95%CI:99.0–99.7%)and 97.9%(95%CI:97.2–98.4%)specific,and 100%(95%CI:92.5–100.0%),59.6%(95%CI:44.3–73.3%)and 93.8%(95%CI:82.8–98.7%)sensitive for HAT.The RDT’s positive and negative predictive values ranged from 45.2–66.7%and 99.2–100%respectively.All DBS tests had specificities≥92.9%.While LAMP and m18S qPCR sensitivities were below 50%,trypanolysis and ELISA/T.b.gambiense had sensitivities of 85.3%(95%CI:68.9–95.0%)and 67.6%(95%CI:49.5–82.6%).Conclusions Presence of swollen lymph nodes,important weight loss,severe itching or motor disorders are simple but accurate clinical criteria for HAT referral in HAT endemic areas in Guinea.Diagnostic performances of HAT Sero-K-Set and SD Bioline HAT are sufficient for referring positives to microscopy.Trypanolysis on DBS may discriminate HAT patients from false RDT positives.

Clinicopathological alterations in wild mammals from the reservoir system of Trypanosoma cruzi:a scoping review

Trypanosoma cruzi is the etiologic agent of Chagas disease.This flagellated protozoan is transmitted to humans as well as different species of domestic and wild animals via vectors from the Reduviidae family(known as"kissing bugs").Despite the fact that hundreds of species of wild mammals are part of the reservoir system,the morphologi-cal changes and clinical manifestations resulting from the pathogenesis of the infection have been largely neglected.The aim of this review is to systematically compile the available information regarding clinicopathological altera-tions in wild mammals due to natural infection by T.cruzi.Information was obtained from six online bibliographic data search platforms,resulting in the identification of 29 publications that met the inclusion criteria.Mortality was the most common clinical manifestation,cardiac damage was the main finding at necropsy,and lymphoplas-macytic inflammation was the most frequent microscopic injury.Thus,regardless of its role as a reservoir,T.cruzi has the potential to affect the health status of wild mammals,a situation that highlights the need for further research to analyze,measure,and compare its effects at both the individual and population levels.

Evaluating the impact of targeting livestock for the prevention of human and animal trypanosomiasis,at village level,in districts newly affected with T.b.rhodesiense in Uganda

Background:Uganda has suffered from a series of epidemics of Human African Trypanosomiasis(HAT),a tsetse transmitted disease,also known as sleeping sickness.The area affected by acute Trypanosoma brucei rhodesiense HAT(rHAT)has been expanding,driven by importation of infected cattle into regions previously free of the disease.These regions are also affected by African Animal Trypanosomiasis(AAT)demanding a strategy for integrated disease control.Methods:In 2008,the Public Private Partnership,Stamp Out Sleeping Sickness(SOS)administered a single dose of trypanocide to 31486 head of cattle in 29 parishes in Dokolo and Kaberamaido districts.This study examines the impact of this intervention on the prevalence of rHAT and AAT trypanosomes in cattle from villages that had(HAT^(+ve))or had not(HAT^(-ve))experienced a recent case of rHAT.Cattle herds from 20 villages were sampled and screened by PCR,pre-intervention and 6-months post-intervention,for the presence or absence of:Trypanosoma brucei s.l.;human infective T.b.rhodesiense;Trypanosoma vivax;and Trypanosoma congolense savannah.Results:Post-intervention,there was a significant decrease in the prevalence of T.brucei s.l.and the human infective sub-species T.b.rhodesiense in village cattle across all 20 villages.The prevalence of T.b.rhodesiense was reduced from 2.4%to 0.74%(P<0.0001),with the intervention showing greater impact in HAT^(-ve) villages.The number of villages containing cattle harbouring human infective parasites decreased from 15/20 to 8/20,with T.b.rhodesiense infection mainly persisting within cattle in HAT^(+ve) villages(six/eight).The proportion of T.brucei s.l.infections identified as human infective T.b.rhodesiense decreased after the intervention from 8.3%(95%CI=11.1-5.9%)to 4.1%(95%CI=6.8-2.3%).Villages that had experienced a recent human case(HAT^(+ve) villages)showed a significantly higher prevalence for AAT both pre-and post-intervention.For AAT the prevalence of T.vivax was significantly reduced from 5.9%to 0.05%post-intervention while the prevalence of T.congolense increased from 8.0%to 12.2%.Conclusions:The intervention resulted in a significant decrease in the prevalence of T.brucei s.l.,human infective T.b.rhodesiense and T.vivax infection in village cattle herds.The proportion of T.brucei s.l.that were human infective,decreased from 1:12 T.brucei s.l.infections before the intervention to 1:33 post-intervention.It is clearly more difficult to eliminate T.b.rhodesiense from cattle in villages that have experienced a human case.Evidence of elevated levels of AAT in livestock within village herds is a useful indicator of risk for rHAT in Uganda.Integrated veterinary and medical surveillance is key to successful control of zoonotic rHAT.

Preventing the transmission of American trypanosomiasis and its spread into non-endemic countries

American trypanosomiasis,commonly known as Chagas disease,is caused by the flagellate protozoan parasite Trypanosoma cruzi.An estimated eight million people infected with T.cruzi currently reside in the endemic regions of Latin America.However,as the disease has now been imported into many non-endemic countries outside of Latin America,it has become a global health issue.We reviewed the transmission patterns and current status of disease spread pertaining to American trypanosomiasis at the global level,as well as recent advances in research.Based on an analysis of the gaps in American trypanosomiasis control,we put forward future research priorities that must be implemented to stop the global spread of the disease.

Update of transmission modelling and projections of gambiense human African trypanosomiasis in the Mandoul focus, Chad

Background:In recent years,a programme of vector control,screening and treatment of gambiense human African trypanosomiasis(gHAT)infections led to a rapid decline in cases in the Mandoul focus of Chad.To represent the biology of transmission between humans and tsetse,we previously developed a mechanistic transmission model,fitted to data between 2000 and 2013 which suggested that transmission was interrupted by 2015.The present study outlines refinements to the model to:(1)Assess whether elimination of transmission has already been achieved despite low-level case reporting;(2)quantify the role of intensified interventions in transmission reduction;and(3)predict the trajectory of gHAT in Mandoul for the next decade under different strategies.Method:Our previous gHAT transmission model for Mandoul was updated using human case data(2000-2019)and a series of model refinements.These include how diagnostic specificity is incorporated into the model and improvements to the fitting method(increased variance in observed case reporting and how underreporting and improvements to passive screening are captured).A side-by-side comparison of fitting to case data was performed between the models.Results:We estimated that passive detection rates have increased due to improvements in diagnostic availability in fixed health facilities since 2015,by 2.1-fold for stage 1 detection,and 1.5-fold for stage 2.We find that whilst the diagnostic algorithm for active screening is estimated to be highly specific(95%credible interval(CI):99.9-100%,Specificity=99.9%),the high screening and low infection levels mean that some recently reported cases with no parasitological confirmation might be false positives.We also find that the focus-wide tsetse reduction estimated through model fitting(95%CI:96.1-99.6%,Reduction=99.1%)is comparable to the reduction previously measured by the decline in tsetse catches from monitoring traps.In line with previous results,the model suggests that transmission was interrupted in 2015 due to intensified interventions.Conclusions:We recommend that additional confirmatory testing is performed in Mandoul to ensure the endgame can be carefully monitored.More specific measurement of cases,would better inform when it is safe to stop active screening and vector control,provided there is a strong passive surveillance system in place.

Integrating innovations:a qualitative analysis of referral non-completion among rapid diagnostic test-positive patients in Uganda’s human African trypanosomiasis elimination programme

Background:The recent development of rapid diagnostic tests(RDTs)for human African trypanosomiasis(HAT)enables elimination programmes to decentralise serological screening services to frontline health facilities.However,patients must still undertake multiple onwards referral steps to either be confirmed or discounted as cases.Accurate surveillance thus relies not only on the performance of diagnostic technologies but also on referral support structures and patient decisions.This study explored why some RDT-positive suspects failed to complete the diagnostic referral process in West Nile,Uganda.Methods:Between August 2013 and June 2015,85%(295/346)people who screened RDT-positive were examined by microscopy at least once;10 cases were detected.We interviewed 20 RDT-positive suspects who had not completed referral(16 who had not presented for their first microscopy examination,and 4 who had not returned for a second to dismiss them as cases after receiving discordant[RDT-positive,but microscopy-negative results]).Interviews were analysed thematically to examine experiences of each step of the referral process.Results:Poor provider communication about HAT RDT results helped explain non-completion of referrals in our sample.Most patients were unaware they were tested for HAT until receiving results,and some did not know they had screened positive.While HAT testing and treatment is free,anticipated costs for transportation and ancillary health services fees deterred many.Most expected a positive RDT result would lead to HAT treatment.RDT results that failed to provide a definitive diagnosis without further testing led some to question the expertise of health workers.For the four individuals who missed their second examination,complying with repeat referral requests was less attractive when no alternative diagnostic advice or treatment was given.Conclusions:An RDT-based surveillance strategy that relies on referral through all levels of the health system is inevitably subject to its limitations.In Uganda,a key structural weakness was poor provider communication about the possibility of discordant HAT test results,which is the most common outcome for serological RDT suspects in a HAT elimination programme.Patient misunderstanding of referral rationale risks harming trust in the whole system and should be addressed in elimination programmes.

Estimating the economic and social consequences for patients diagnosed with human African trypanosomiasis in Muchinga,Lusaka and Eastern Provinces of Zambia(2004-2014)

Background:Acute human African trypanosomiasis(rHAT)caused by Trypanosoma brucei rhodesiense is associated with high mortality and is fatal if left untreated.Only a few studies have examined the psychological,social and economic impacts of rHAT.In this study,mixed qualitative and quantitative research methods were used to evaluate the socio-economic impacts of rHAT in Mambwe,Rufunsa,Mpika and Chama Districts of Zambia.Methods:Individuals diagnosed with rHAT from 2004 to 2014 were traced using hospital records and discussions with communities.Either they,or their families,were interviewed using a structured questionnaire and focus group discussions were conducted with affected communities.The burden of the disease was investigated using disability adjusted life years(DALYs),with and without discounting and age-weighting.The impact of long-term disabilities on the rHAT burden was also investigated.Results:Sixty four cases were identified in the study.The majority were identified in second stage,and the mortality rate was high(12.5%).The total number of DALYs was 285 without discounting or age-weighting.When long-term disabilities were included this estimate increased by 50%to 462.The proportion of years lived with disability(YLD)increased from 6.4%to 37%of the undiscounted and un-age-weighted DALY total.When a more active surveillance method was applied in 2013-2014 the cases identified increased dramatically,suggesting a high level of under-reporting.Similarly,the proportion of females increased substantially,indicating that passive surveillance may be especially failing this group.An average of 4.9 months of productive time was lost per patient as a consequence of infection.The health consequences included pain,amnesia and physical disability.The social consequences included stigma,dropping out of education,loss of friends and self-esteem.Results obtained from focus group discussions revealed misconceptions among community members which could be attributed to lack of knowledge about rHAT.Conclusions:The social and economic impact of rHAT on rural households and communities is substantial.Improved surveillance and strengthening of local medical services are needed for early and accurate diagnosis.Disease prevention should be prioritised in communities at risk of rHAT,and interventions put in place to prevent zoonotic disease spill over from domestic animals and wildlife.Supportive measures to mitigate the long-term effects of disability due to rHAT are needed.

Diminazene aceturate modified nanocomposite for improved efficacy in acute trypanosome infection

Objective: To investigate the improved antitrypanocidal activity and toxicity of diminazene aceturate modified Nano drug in experimental rats. Methods: Aqueous leaf extract of Hyptis suaveolens was used to reduce gold tetrachloride to its nanoparticle size and this was characterized and formulates with naturally synthesized polyhydroxybutyrateas a Nano carrier. A total of thirty (30) albino rats were group into 6 (A-F) of 5 rats each & infected intraperitoneally with 0.2 mL of the inoculum containing about 1×103 Trypanosoma brucei brucei parasites per 0.2 mL of blood. Groups A and B were treated with 3 and 6 minutes released orange PHB, Groups C and D were treated with 15 and 30 minutes released mango PHB formulated tablet while Groups E and F were negative (untreated) and standard drug (Dininazene aceturare) respectively. Results: The free drug and modified orange synthesized polyhydroxy butyrate shows antitrypanocidal activities by reducing the replicating rate of the parasite as compared to infect untreated. While the modified- mango synthesized shows increasing order of replication. There were significant increases in all the haematological parameter evaluated in the infected treated groups compared to infect untreated. But no significant difference (P<0.05) observed in the Catalase activity in the serum and liver of all the groups whereas, the modified orange synthesized shows significant decrease in other enzymes activities evaluated when compared with the free drug, mango synthesized and the infected untreated groups. Conclusion: Orange synthesized modified diminazene aceturate show efficacy as free drug with limited toxicity that can enhance the therapeutic.

Improved Detection of Sleeping Sickness Cases by LED Fluorescence Microscopy: Evidence from a Prospective Multi-Centric Study in the Democratic Republic of the Congo

Background: Confirmatory diagnosis of Trypanosoma brucei gambiense human African trypanosomiasis (HAT) is based on demonstration of parasites by microscopy. However, the sensitivity of routine microscopy methods is very low, and many cases are missed and left untreated. A clinical study was conducted in the Democratic Republic of the Congo to evaluate the accuracy of improved microscopy methods in diagnosis of HAT. These included examination by fluorescence microscopy (FM) of acridine orange (AO) stained smears of whole blood and smears made following a new procedure for concentrating trypanosomes by selective lysis of red blood cells (RBC). Methodology/Principal Findings: Venous blood was collected from 213 HAT cases, 101 HAT suspects and 95 controls and used to determine the accuracy of four microscopy methods: bright field microscopy of Giemsa-stained thick blood smears, FM of AO-stained thick blood smears, FM of AO-stained thick blood smears prepared after RBC lysis and concentration, and FM of AO-stained thin blood smears prepared after RBC lysis and concentration. The sensitivity of FM using thick blood smears stained with AO was 3 times higher than bright field microscopy using Giemsa-stained thick blood smears [19.7% (95% CI: 14.9% - 25.6%) versus 6.1% (95% CI: 3.6% - 10.2%)]. When the RBC lysis and concentration procedure was included, sensitivity of the test was further enhanced to 23.0% (95% CI: 17.9% - 29.1%) with thick blood smears and 34.3% (95% CI: 28.2% - 40.9%) with thin blood smears. Specificity of all four microscopy methods was 100% (95% CI: 96.1% - 100.0%). However, the miniature anion exchange chromatography technique (mAECT) and capillary tube centrifugation (CTC) method remained more sensitive. Conclusions: These new methods have practical advantages, including shorter staining time, ease of demonstration of parasites, and the possibility of archiving slides. They could, therefore, be alternative methods to improve case detection where concentration procedures such as mAECT or CTC are not performed.

Antiprotozoal assessment and phenolic acid profiling of five Fumaria(fumitory) species

Objective:To explore some Fumaria species which were recorded to be traditionally used against malaria and other protozoal diseases.Methods:Consequently,in the current study,antiprotozoal effect of the ethanol extracts obtained from five Fumaria species(Fumaria densiflora,Fumaria cilicica,Fumaria rostellata,Fumaria kralikii,and Fumaria parviflora)was investigated against the parasites;Plasmodium falciparum(malaria)and Trypanosoma bruceirhodesiense(human African trypanosomiasis)at 0.81 and 4.85μg/mL concentrations.Results:Among them,Fumaria densiflora extract exerted the highest antiplasmodial(93.80%)and antitrypanasomal effect(55.40%),while the ethanol extracts of Fumaria kralikii(43.45%)and Fumaria rostellata(41.65%)showed moderate activity against Plasmodium falciparum.Besides,phenolic acid contents of the extracts were analyzed using high performance liquid chromatography(HPLC)and trans-cinnamic(4.32 mg/g)and caffeic(3.71 mg/g)acids were found to be the dominant phenolic acids in Fumaria densiflora.Conclusions:According to our results,Fumaria densiflora deserve further study for its promising antiprotozoal activity.