High Parasitemia Research Articles

Assessment of Babesia ovis pathogenicity in goats: implications for transmission dynamics and host resistant

Babesia ovis, commonly associated with ovine babesiosis, poses a significant threat to sheep health, often resulting in severe clinical manifestations and high mortality rates. However, the impact of B. ovis on goats has remained uncertain, prompting us to investigate its pathogenicity in caprine hosts. Experimental infections using B. ovis-infected blood inoculation and infected tick infestation, were conducted on spleen-intact (n=5) and splenectomized (n=5) goats. The experimental infection was performed using fresh blood obtained from a B. ovis-infected splenectomized sheep. One spleen-intact sheep served as a control for the experimental infection with B. ovis-infected Rhipicephalus bursa ticks. While all experimentally infected sheep (#501, #575) displayed severe clinical symptoms and high parasitemia, goats exhibited resistance, showing no significant clinical manifestations or sustained parasitemia. Notably, B. ovis was detected in two spleen-intact goats via nested PCR, prompting further investigation into their role as reservoirs for tick-borne transmission. These goats were then infested with Babesia spp.-free R. bursa larvae (0.1 gr) and adults (50 females and 50 males) for transstadial and transovarial transmission experiments respectively. Results indicated that chronically B. ovis-infected spleen-intact goats are not significant sources for maintaining the tick-borne transmission cycle of the parasite. These findings highlight the differential susceptibility of goats to B. ovis infection compared to sheep and their limited role as reservoirs for parasite transmission. Understanding the role of goats in B. ovis transmission and their resistance mechanisms can inform effective control measures and reduce economic losses in affected regions. Further research into caprine babesiosis and host immunological responses is essential to fully elucidate their possible role as reservoirs of the parasite, and underlying mechanisms of host susceptibility and parasite pathogenesis.

High malaria parasitemia in an Italian man and severe delayed hemolytic anemia.

Parenteral artesunate has become the choice treatment of cases of high parasitemia malaria. We report the case of an Italian man with a high malaria parasitemia who promptly improved with intravenous artenusate, but who developed a delayed severe hemolytic anemia associated with intravenous artesunate treatment, requiring blood trasfusions.

Avian malaria in a feral-pet pigeon: a case report

BackgroundAvian malaria is caused by diverse parasite species of the genus Plasmodium, and it affects various bird species. The occurrence of this disease in some wild bird species is sparsely documented due to the scarce availability of samples. Hence the pathogenicity in some hosts is not completely known. In addition, feral birds may act as reservoirs bridging the transmission cycle from wild migratory birds to domestic and zoo-kept bird species.Case presentationAn owner of pigeons adopted a feral pigeon (Columba livia forma domestica) and housed it together with his other pet-pigeons. The bird died unexpectedly a few weeks after a surgical procedure and necropsy revealed a severely anaemic carcass, with pale organs and hydropericardium. Histopathologic analysis revealed inflammatory infiltrates in the lung and liver, and monocytes and Kupffer cells contained haemozoin pigment indicative of phagocytosis of Plasmodium-infected erythrocytes. A high erythrocytic infection rate of 18% was evident in tissues and blood vessels in various organs. Furthermore, the thyroid had masses classified as thyroid carcinomas. Immunohistochemistry with anti- Plasmodium falciparum HSP70 antibody revealed positive signals in erythrocytes and intravascular leucocytes. Further microscopy analysis using a Hemacolor-stained impression smear revealed a high parasitaemia with an asynchronous infection showing all erythrocytic stages. Molecular diagnosis by PCR identified Plasmodium relictum, lineage GRW11 as the aetiological agent. The bird presented died most likely due to an acute infection as evidenced by the high blood parasitaemia, leading to major erythrocyte destruction. Further analyses of feral pigeons (n = 22) did not reveal any additional cases of Plasmodium infections.ConclusionThis study reports the first mortality associated with P. relictum lineage GRW11. The study supports previous studies, suggesting that Plasmodium infections are not frequent in pigeons. Host conditions like immunosuppression due to the tumour may have influenced the infection outcome in this fatal case. Use of anti-P. falciparum HSP70 antibody for detection of P. relictum antigens for immune assays in blood and tissue samples will be a useful tool for future studies.

Inpatient and postdischarge mortality among children with anaemia and malaria parasitaemia in Kenya: a cohort study

BackgroundAnaemia and malaria are leading causes of paediatric hospitalisation and inpatient mortality in sub-Saharan Africa. However, there is limited empirical data on survival following hospital discharge. We aimed to estimate...

Design, synthesis, in vitro and in vivo trypanosomaticidal efficacy of novel 5-nitroindolylazines

Leishmaniasis and trypanosomiasis rank among lethal vector-borne parasitic diseases that are endemic in tropical and sub-tropical countries. There are currently no preventive vaccines against them, and once diagnosed, a handful of less effective drugs clinically accessible are the only therapeutic options offered to treat these ailments. And although curable, the eradication and elimination of these diseases are hampered by the emergence of multidrug-resistant strains of the causal pathogens. Hence, there is accrued necessity for the development of new, effective, and affordable drugs. In recent decades, several molecular scaffolds, including nitroaromatics, endoperoxides, etc., have been attempted as building blocks to generate new effective clinical antitrypanosomatid agents with low toxicity so far to no avail. In this regard, a series of nitroindolylazine derivatives was synthesised in a three-step process involving nucleophilic substitution (SN), hydrazination and Schiff base condensation reactions, and was evaluated against various Leishmania and Trypanosoma species and strains. Several promising hits portraying leishmanicidal and trypanocidal with in vitro submicromolar activities, and devoid of toxicity on mammalian cells were uncovered. Among these, nitrofurylazine 11 (Tc IC50: 0.08 ± 0.03 μM) and nitrothienylazine 13 (Tc IC50: 0.09 ± 0.01 μM) were evaluated in vivo against Trypanosoma congolense, the causative agent of nagana, which is livestock most virulent trypanosome species in mice-infected preliminary study. However, only partial efficacy was observed as all mice succumbed due to high parasitemia within 13 days post-infection during the treatment. The translational potential of antileishmanial and antichagasic hits, as well as further identification of their molecular targets, will be assessed in future research.

Basic coagulation parameters and platelet count among malaria patients attending at Addis Zemen Primary Hospital, Northwest Ethiopia.

Malaria is an intravascular parasitic-related blood disease that causes bleeding, coagulopathy, and thrombocytopenia. However, limited data shows the effect of Plasmodium species infection on basic coagulation parameters and platelet count. Thus, this study aimed to assess basic coagulation parameters and platelet count among malaria patients. A cross-sectional study was conducted among 240 study participants (120 cases and 120 controls) from June 1, 2021, to February 30, 2022. A convenient sampling technique was employed to select study participants. The blood sample was collected by a trained laboratory technologist for platelet counts, prothrombin time (PT), partial thromboplastin time (PTT), international normalization ratio (INR), blood film, and serological testing. The collected data were analyzed in SPSS version 23. Data were analyzed by the Mann-Whitney U test, Kruskal Wallis H, and Spearman's rank-order correlation tests. Descriptive findings were presented through median, tables, and chart. In all cases, a P-value < 0.05 was considered statistically significant. The percentage of mild, moderate, and high malaria parasitemia levels per microliter of blood was 21.7%, 20%, and 58.3%, respectively. The overall median malaria parasitemia was 10,304 per microliter of blood. Among malaria patients, 77.5%, 61.7%, and 51.7% had prolonged PT, INR, and APTT, respectively as compared to control. Moreover, 26.7% of Plasmodium-infected participants had mild thrombocytopenia as compared to the control group (P < 0.001). The value of PT, APTT, and INR were significantly elevated, whereas the level of platelet count was inversely reduced when the malaria parasitemia level increased as compared to controls (p < 0.001).

Do sexual differences in life strategies make male lizards more susceptible to parasite infection?

Female and male hosts may maximise their fitness by evolving different strategies to compensate for the costs of parasite infections. The resulting sexual dimorphism might be apparent in differential relationships between parasite load and body condition, potentially reflecting differences in energy allocation to anti-parasitic defences. For example, male lacertids with high body condition may produce many offspring while being intensely parasitised. In contrast, female lacertids may show a different outcome of the trade-offs between body condition and immunity, aiming to better protect themselves from the harm of parasites. We predicted that females would have fewer parasites than males and a lower body condition across parasitaemia levels because they would invest resources in parasite defence to mitigate the costs of infection. In contrast, the male strategy to maximise access to females would imply some level of parasite tolerance and, thus, higher parasitaemia. We analysed the relationship between the body condition of lizards and the parasitemias of Karyolysus and Schellackia, two genera of blood parasites with different phylogenetic origins, in 565 females and 899 males belonging to 10 species of the Lacertidae (Squamata). These lizards were sampled over a period of 12 years across 34 sampling sites in southwestern Europe. The results concerning the Karyolysus infections were consistent with the predictions, with males having similar body condition across parasitaemia levels even though they had higher infection intensities than females. On the other hand, females with higher levels of Karyolysus parasitaemia had lower body condition. This is consistent with the prediction that different life strategies of male and female lacertids can explain the infection patterns of Karyolysus. In contrast, the parasitaemia of Schellackia was consistently low in both male and female hosts, with no significant effect on the body condition of lizards. This suggests that lizards of both sexes maintain this parasite below a pathogenic threshold.

Molecular Surveillance of Artemisinin Combination Therapy Resistance Markers in Kelch13 Gene of Plasmodium falciparum Isolates Collected From Yobe State Nigeria

Background: Despite the discovery of artemisinin and artemisinin combination therapies (ACTs), resistance emerged due to single-nucleotide polymorphisms (SNPs) at the Kelch13 propeller domain of Plasmodium falciparum; the predominant parasite causing malaria. This research aimed at surveying the gene responsible for reduced parasite clearance of the first-line antimalarial ACT in Yobe State, Nigeria. Methods: This study analyzed the blood samples of 300 patients (18-50 years) from 3 different hospitals in Yobe State for malaria using a rapid diagnostic test (malarial strip test). The Giemsa microscopy protocol was performed on the samples that tested positive in order to increase specificity for high parasitemia. Positive samples from microscopy were confirmed by COXIII gene polymerase chain reaction amplification. The fragments of the P. falciparum kelch13 gene encompassing the propeller domains were amplified and sequenced to detect polymorphism. Results: The result of multiple sequence alignment of 16 successfully sequenced samples performed alongside the reference 3D7 strain revealed nine novel mutations that have not been reported elsewhere. Two were synonymous (S485S and A675A), and seven were non-synonymous (E461V, S485R, D648H, E668Q, S679L, H697Y, and W706M). The genetic diversity parameters obtained in this study indicated non-severe mutations with haplotype diversity [Hd=0.662, D (Tadjima)=-2.16692 and D* (Fu and Li) -3.08435]. Seven different haplotypes were discovered, and the result of the haplotype network demonstrated Hap_1 to be the predominant haplotype in the population. Conclusion: The absence of the WHO-validated markers may suggest that ACTs are still effective in Yobe.

Repeat controlled human Plasmodium falciparum infections delay bloodstream patency and reduce symptoms

Resistance to clinical malaria takes years to develop even in hyperendemic regions and sterilizing immunity has rarely been observed. To evaluate the maturation of the host response against controlled repeat exposures to P. falciparum (Pf) NF54 strain-infected mosquitoes, we systematically monitored malaria-naïve participants through an initial exposure to uninfected mosquitoes and 4 subsequent homologous exposures to Pf-infected mosquitoes over 21 months (n = 8 males) (ClinicalTrials.gov# NCT03014258). The primary outcome was to determine whether protective immunity against parasite infection develops following repeat CHMI and the secondary outcomes were to track the clinical signs and symptoms of malaria and anti-Pf antibody development following repeat CHMI. After two exposures, time to blood stage patency increases significantly and the number of reported symptoms decreases indicating the development of clinical tolerance. The time to patency correlates positively with both anti-Pf circumsporozoite protein (CSP) IgG and CD8 + CD69+ effector memory T cell levels consistent with partial pre-erythrocytic immunity. IFNγ levels decrease significantly during the participants’ second exposure to high blood stage parasitemia and could contribute to the decrease in symptoms. In contrast, CD4-CD8 + T cells expressing CXCR5 and the inhibitory receptor, PD-1, increase significantly after subsequent Pf exposures, possibly dampening the memory response and interfering with the generation of robust sterilizing immunity.

Prevalence and Characteristics of Plasmodium vivax Gametocytes in Duffy-Positive and Duffy-Negative Populations across Ethiopia.

Plasmodium parasites replicate asexually in human hosts. The proportion of infections that carries gametocytes is a proxy for human-to-mosquito transmissibility. It is unclear which proportion of Plasmodium vivax infections in Duffy-negative populations carries gametocytes. We determined the prevalence and characteristics of P. vivax gametocytes in Duffy-positive and -negative populations across broad regions of Ethiopia. Finger-prick blood samples were collected for microscopic and molecular screening of Plasmodium parasites and Duffy status of individuals. Molecular screening of Plasmodium species and Duffy blood group genotyping was done using SYBR green and the Taqman quantitative polymerase chain reaction method. Of the 447 febrile patients who were shown to be P. vivax smear positive, 414 (92.6%) were confirmed by molecular screening as P. vivax and 16 (3.9%) of them were from Duffy-negative individuals. Of these, 5 of 16 (31.3%) Duffy-negative P. vivax-infected samples were detected with gametocytes. Of the 398 Duffy-positive P. vivax-infected samples, 150 (37.7%) were detected with gametocytes, slightly greater than that in Duffy-negative samples. This study highlights the presence of P. vivax gametocytes in Duffy-negative infections, suggestive of human-to-mosquito transmissibility. Although P. vivax infections in Duffy-negative individuals were commonly associated with low parasitemia, some of these infections were shown to have relatively high parasitemia and may represent a prominent erythrocyte invasion capability of P. vivax, and hidden reservoirs that can contribute to transmission. A better understanding of P. vivax transmission biology and gametocyte function particularly in Duffy-negative populations would aid future treatment and management of P. vivax malaria in Africa.

Quantitative PCR as a marker for preemptive therapy and its role in therapeutic control in Trypanosoma cruzi/HIV coinfection.

Trypanosoma cruzi and HIV coinfection can evolve with depression of cellular immunity and increased parasitemia. We applied quantitative PCR (qPCR) as a marker for preemptive antiparasitic treatment to avoid fatal Chagas disease reactivation and analyzed the outcome of treated cases. This mixed cross-sectional and longitudinal study included 171 Chagas disease patients, 60 coinfected with HIV. Of these 60 patients, ten showed Chagas disease reactivation, confirmed by parasites identified in the blood, cerebrospinal fluid, or tissues, 12 exhibited high parasitemia without reactivation, and 38 had low parasitemia and no reactivation. We showed, for the first time, the success of the timely introduction of benznidazole in the non-reactivated group with high levels of parasitemia detected by qPCR and the absence of parasites in reactivated cases with at least 58 days of benznidazole. All HIV+ patients with or without reactivation had a 4.0-5.1 higher chance of having parasitemia than HIV seronegative cases. A positive correlation was found between parasites and viral loads. Remarkably, treated T. cruzi/HIV-coinfected patients had 77.3% conversion from positive to negative parasitemia compared to 19.1% of untreated patients. Additionally, untreated patients showed ~13.6 times higher Odds Ratio of having positive parasitemia in the follow-up period compared with treated patients. Treated and untreated patients showed no differences regarding the evolution of Chagas disease. The main factors associated with all-cause mortality were higher parasitemia, lower CD4 counts/μL, higher viral load, and absence of antiretroviral therapy. We recommend qPCR prospective monitoring of T. cruzi parasitemia in HIV+ coinfected patients and point out the value of pre-emptive therapy for those with high parasitemia. In parallel, early antiretroviral therapy introduction is advisable, aiming at viral load control, immune response restoration, and increasing survival. We also suggest an early antiparasitic treatment for all coinfected patients, followed by effectiveness analysis alongside antiretroviral therapy.

Oral infectivity through carnivorism in murine model of Trypanosoma cruzi infection.

Oral transmission of T. cruzi is probably the most frequent transmission mechanism in wild animals. This observation led to the hypothesis that consuming raw or undercooked meat from animals infected with T. cruzi may be responsible for transmitting the infection. Therefore, the general objective of this study was to investigate host-pathogen interactions between the parasite and gastric mucosa and the role of meat consumption from infected animals in the oral transmission of T. cruzi. Cell infectivity assays were performed on AGS cells in the presence or absence of mucin, and the roles of pepsin and acidic pH were determined. Moreover, groups of five female Balb/c mice were fed with muscle tissue obtained from mice in the acute phase of infection by the clone H510 C8C3hvir of T. cruzi, and the infection of the fed mice was monitored by a parasitemia curve. Similarly, we assessed the infective capacity of T. cruzi trypomastigotes and amastigotes by infecting groups of five mice Balb/c females, which were infected orally using a nasogastric probe, and the infection was monitored by a parasitemia curve. Finally, different trypomastigote and amastigote inoculums were used to determine their infective capacities. Adhesion assays of T. cruzi proteins to AGS stomach cells were performed, and the adhered proteins were detected by western blotting using monoclonal or polyclonal antibodies and by LC-MS/MS and bioinformatics analysis. Trypomastigote migration in the presence of mucin was reduced by approximately 30%, whereas in the presence of mucin and pepsin at pH 3.5, only a small proportion of parasites were able to migrate (∼6%). Similarly, the ability of TCTs to infect AGS cells in the presence of mucin is reduced by approximately 20%. In all cases, 60-100% of the animals were fed meat from mice infected in the acute phase or infected with trypomastigotes or amastigotes developed high parasitemia, and 80% died around day 40 post-infection. The adhesion assay showed that cruzipain is a molecule of trypomastigotes and amastigotes that binds to AGS cells. LC-MS/MS and bioinformatics analysis, also confirmed that transialidase, cysteine proteinases, and gp63 may be involved in TCTs attachment or invasion of human stomach cells because they can potentially interact with different proteins in the human stomach mucosa. In addition, several human gastric mucins have cysteine protease cleavage sites. Then, under our experimental conditions, consuming meat from infected animals in the acute phase allows the T. cruzi infection. Similarly, trypomastigotes and amastigotes could infect mice when administered orally, whereas cysteinyl proteinases and trans-sialidase appear to be relevant molecules in this infective process.

Classification and clinical significance of immunogenic cell death-related genes in Plasmodium falciparum infection determined by integrated bioinformatics analysis and machine learning

BackgroundImmunogenic cell death (ICD) is a type of regulated cell death that plays a crucial role in activating the immune system in response to various stressors, including cancer cells and pathogens. However, the involvement of ICD in the human immune response against malaria remains to be defined.MethodsIn this study, data from Plasmodium falciparum infection cohorts, derived from cross-sectional studies, were analysed to identify ICD subtypes and their correlation with parasitaemia and immune responses. Using consensus clustering, ICD subtypes were identified, and their association with the immune landscape was assessed by employing ssGSEA. Differentially expressed genes (DEGs) analysis, functional enrichment, protein-protein interaction networks, and machine learning (least absolute shrinkage and selection operator (LASSO) regression and random forest) were used to identify ICD-associated hub genes linked with high parasitaemia. A nomogram visualizing these genes' correlation with parasitaemia levels was developed, and its performance was evaluated using receiver operating characteristic (ROC) curves.ResultsIn the P. falciparum infection cohort, two ICD-associated subtypes were identified, with subtype 1 showing better adaptive immune responses and lower parasitaemia compared to subtype 2. DEGs analysis revealed upregulation of proliferative signalling pathways, T-cell receptor signalling pathways and T-cell activation and differentiation in subtype 1, while subtype 2 exhibited elevated cytokine signalling and inflammatory responses. PPI network construction and machine learning identified CD3E and FCGR1A as candidate hub genes. A constructed nomogram integrating these genes demonstrated significant classification performance of high parasitaemia, which was evidenced by AUC values ranging from 0.695 to 0.737 in the training set and 0.911 to 0.933 and 0.759 to 0.849 in two validation sets, respectively. Additionally, significant correlations between the expressions of these genes and the clinical manifestation of P. falciparum infection were observed.ConclusionThis study reveals the existence of two ICD subtypes in the human immune response against P. falciparum infection. Two ICD-associated candidate hub genes were identified, and a nomogram was constructed for the classification of high parasitaemia. This study can deepen the understanding of the human immune response to P. falciparum infection and provide new targets for the prevention and control of malaria.

Molecular surveillance of Kelch 13 polymorphisms in Plasmodium falciparum isolates from Kenya and Ethiopia

BackgroundTimely molecular surveillance of Plasmodium falciparum kelch 13 (k13) gene mutations is essential for monitoring the emergence and stemming the spread of artemisinin resistance. Widespread artemisinin resistance, as observed in Southeast Asia, would reverse significant gains that have been made against the malaria burden in Africa. The purpose of this study was to assess the prevalence of k13 polymorphisms in western Kenya and Ethiopia at sites representing varying transmission intensities between 2018 and 2022.MethodsDried blood spot samples collected through ongoing passive surveillance and malaria epidemiological studies, respectively, were investigated. The k13 gene was genotyped in P. falciparum isolates with high parasitaemia: 775 isolates from four sites in western Kenya (Homa Bay, Kakamega, Kisii, and Kombewa) and 319 isolates from five sites across Ethiopia (Arjo, Awash, Gambella, Dire Dawa, and Semera). DNA sequence variation and neutrality were analysed within each study site where mutant alleles were detected.ResultsSixteen Kelch13 haplotypes were detected in this study. Prevalence of nonsynonymous k13 mutations was low in both western Kenya (25/783, 3.19%) and Ethiopia (5/319, 1.57%) across the study period. Two WHO-validated mutations were detected: A675V in three isolates from Kenya and R622I in four isolates from Ethiopia. Seventeen samples from Kenya carried synonymous mutations (2.17%). No synonymous mutations were detected in Ethiopia. Genetic variation analyses and tests of neutrality further suggest an excess of low frequency polymorphisms in each study site. Fu and Li’s F test statistic in Semera was 0.48 (P > 0.05), suggesting potential population selection of R622I, which appeared at a relatively high frequency (3/22, 13.04%).ConclusionsThis study presents an updated report on the low frequency of k13 mutations in western Kenya and Ethiopia. The WHO-validated R622I mutation, which has previously only been reported along the north-west border of Ethiopia, appeared in four isolates collected from eastern Ethiopia. The rapid expansion of R622I across Ethiopia signals the need for enhanced monitoring of the spread of drug-resistant P. falciparum parasites in East Africa. Although ACT remains currently efficacious in the study areas, continued surveillance is necessary to detect early indicators of artemisinin partial resistance.

The Parasitemia has Contributed to the Severity of Cases of Visceral Leishmaniasis.

Visceral leishmaniasis (VL) occurs due to the evolution, virulence, and adaptation of Leishmania, vector biology, host immune system evasion, and reservoir hosts. Parasitemia can be involved as a warning regarding the clinical severity of VL The present study aims to evaluate the relationship between parasitemia and the prognosis of individuals with VL. Blood and bone marrow samples from individuals with VL were analyzed to identify parasite and quantify or measure parasite burden. Individuals were classified in the clinical score model of risk of death by disease proposed by Coura-Vital et al. (PLoS Negl Trop Dis 8(12): e33742014, 2014). 39/74 individuals presented a better prognosis, and 35/74 individuals presented a worse prognosis. HIV + VL co-infection was present in 32 individuals, of which 12 were considered severe. The group aged 51 to 64 was classified as severe, with a decrease in leukocytes (p-value 0.0295) and neutrophils (p-value 0.0476). L. infantum DNA was identified in blood and bone marrow, in 69 individuals, and not detected in 5 individuals. The quantification of the parasite showed greater parasitemia in bone marrow (P = 0.0003) with an average of 4.70 × 104 Leishmanias/mL about blood, with 0.29 × 104 Leishmanias/mL. Individuals in the age group aged 51 to 64 co-infected with HIV + VL had higher parasitemia (p-value 0.0150) with 2.44 × 104 Leishmanias/mL in blood and bone marrow than in the group aged 20 to 50. Parasitemia, measured by molecular biology in blood and bone marrow, was related to the worst clinical prognosis of VL in the age group aged 51 to 64.

Enhanced Antimalarial and Antisequestration Activity of Methoxybenzenesulfonate-Modified Biopolymers and Nanoparticles for Tackling Severe Malaria.

Severe malaria is a life-threatening condition that is associated with a high mortality. Severe Plasmodium falciparum infections are mediated primarily by high parasitemia and binding of infected red blood cells (iRBCs) to the blood vessel endothelial layer, a process known as sequestration. Here, we show that including the 5-amino-2-methoxybenzenesulfonate (AMBS) chemical modification in soluble biopolymers (polyglutamic acid and heparin) and poly(acrylic acid)-exposing nanoparticles serves as a universal tool to introduce a potent parasite invasion inhibitory function in these materials. Importantly, the modification did not add or eliminated (for heparin) undesired anticoagulation activity. The materials protected RBCs from invasion by various parasite strains, employing both major entry pathways. Two further P. falciparum strains, which either expose ligands for chondroitin sulfate A (CSA) or intercellular adhesion molecule 1 (ICAM-1) on iRBCs, were tested in antisequestration assays due to their relevance in placental and cerebral malaria, respectively. Antisequestration activity was found to be more efficacious with nanoparticles vs gold-standard soluble biopolymers (CSA and heparin) against both strains, when tested on receptor-coated dishes. The nanoparticles also efficiently inhibited and reversed the sequestration of iRBCs on endothelial cells. First, the materials described herein have the potential to reduce the parasite burden by acting at the key multiplication stage of reinvasion. Second, the antisequestration ability could help remove iRBCs from the blood vessel endothelium, which could otherwise cause vessel obstruction, which in turn can lead to multiple organ failure in severe malaria infections. This approach represents a further step toward creation of adjunctive therapies for this devastating condition to reduce morbidity and mortality.

In vitro delayed response to dihydroartemisinin of malaria parasites infecting sickle cell erythocytes

BackgroundDecreased efficacy of artemisinin-based combination therapy (ACT) for Plasmodium falciparum malaria has been previously reported in patients with sickle cell disease (SCD). The main purpose of this study was to investigate the in vitro susceptibility of isolates to dihydro-artemisinin (DHA) to provide a hypothesis to explain this treatment failure.MethodsIsolates were collected from patients attending health centres in Abidjan with uncomplicated P. falciparum malaria. The haemoglobin type has been identified and in vitro drug sensitivity tests were conducted with the ring stage assay and maturation inhibition assay.Results134 isolates were obtained. Parasitaemia and haemoglobin levels at inclusion were lower in patients with haemoglobin HbSS and HbSC than in patients with normal HbAA. After ex vivo RSA and drug inhibition assays, the lowest rate of parasitic growth was found with isolates from HbAS red cells. Conversely, a significantly higher survival rate of parasites ranging from 15 to 34% were observed in isolates from HbSS. Isolates with in vitro reduced DHA sensitivity correlate with lower RBC count and haematocrit and higher parasitaemia at inclusion compared to those with isolates with normal DHA sensitivity. However, this decrease of in vitro sensitivity to DHA was not associated with Kelch 13-Propeller gene polymorphism.ConclusionThis study highlights an in vitro decreased sensitivity to DHA, for isolates collected from HbSS patients, not related to the Pfkelch13 gene mutations. These results are in line with recent studies pointing out the role of the redox context in the efficacy of the drug. Indeed, SCD red cells harbour a highly different ionic and redox context in comparison with normal red cells. This study offers new insights into the understanding of artemisinin selective pressure on the malaria parasite in the context of haemoglobinopathies in Africa.

Seroepidemiology of Asymptomatic Plasmodium falciparum CoInfections with Hepatitis B And C Virus among Blood Donors in Modakeke Area Office, Ile Ife, Osun State, Nigeria

Despite the remarkable achievements in the reduction of malaria mortality, the disease had remained the most common cause of morbidity and mortality. Infection with hepatitis B virus (HBV) and hepatitis C virus (HCV) might associate with increased mortality rate as the infections may predisposes to the development of serious liver diseases such as liver cirrhosis, liver failure and hepatocellular carcinoma (HCC). The aim of the present study was to determine the Asymptomatic Plasmodium falciparum co-infections with hepatitis B and C viruses among blood donors in Modakeke Area Office, Ile Ife, Osun State, Nigeria. The study was conducted in Modakeke Area Office, Ile Ife, Osun State. The study was conducted in Modakeke health facility, Ile Ife, Osun State. Five (5) ml of venous blood was obtained from each of 400 blood donors into plain bottles and labeled accordingly. Thick and thin films of the donor’s blood samples were made on slides dried for five minutes at 37°C then stained using Giemsa stain. The slides were then examined microscopically for presence and densities of malaria parasites. Species identification and determination of the Hematological parameters were also conducted on the slides. Sera were separated from the remaining aliquots of the blood and screened for hepatitis-B and C surface antigens (HBsAg and anti-HCV). The overall mean parasitaemia of malarious blood donors was 786.20µl/blood while that of blood donors with hepatitis virus infection was 53 representing 13.3% of the total donor examined. Donors infected with HBV, HCV and co-infection of HBV and HCV had viraemia of 25 (6.3%), 21 (5.3%) and 7(1.8%) respectively. Among the age groups, high prevalence of Hepatitis viral infection rate of 57.9% was recorded among aged group 18-26 years. The infection rate of 15.8%, 13.6% and 10.5% of HBV, HCV was recorded from age group 18-26years respectively. The highest co-infection rate of HBV and HCV of 18.8% was recorded from age group 54-62years, while high mean malaria parasitaemia of 1,882.78 (µl of blood) was also recorded among donors aged 18-26 years. High hepatitis viral infection rate of 22.2% with 16.7% of HCV were recorded among females, while 6.3% of HBV and 1.9% of co-infection was observed among males and the mean malaria parasitaemia of 2,382.80 µl of blood was observed among females. High prevalence of Hepatitis infection rate of 17.1%, 8.6% of HCV, (6.3%) of HBV and 2.3% of co-infection were recorded among single donors. The mean malaria parasitaemia of 786.89 (µl of blood) was observed among married donors, there was significant difference between marital status and hepatitis infection (p= 0.0043) and between HCV (p=0.009). High prevalence of hepatitis viraemia of 21.4% and co-infection 5.6% was recorded among artisans, while 14.6% HBV infection rate was recorded among civil servants and 14.3% of HCV was recorded among donors who are farmers. The mean malaria parasitaemia of 1,150.15 µl/blood was recorded among donors who are traders. Donors that have acquired primary education have high prevalence of hepatitis infection rate of 38.5%, with 15.4% of HBV and (23.1%) co-infection (HBVand HCV). HCV (6.7%) was high among donors with secondary education; high mean malaria parasitaemia of 2,096.84 µl of blood was recorded among donors with primary education. Blood B+ve had high hepatitis viral infection rate of 21.7%, HCV 13.0% and co-infection of 5.8% while 7.8% of HBV was recorded among O+ve donors. The mean malaria parasitaemia of 1,378.50 (µl of blood) was recorded among B-ve donors. The total mean PCV in the study was 42.63, White blood cell count was 5,281.25, lymphocytes count was 42.88, eosinophils was 1.53 and neutrophil count was 55.72. The total percentage of elevated AST was 5.0%, ALT was 6.0% and ALB was 10.5%. There was significant difference between malaria and ALT (p=0.000) also, there was significant difference between HCV and ALB (p=0.000). There was significant difference between HBV and AST (p=0.001), ALT (p=0.028) and ALB (p=0.028).The hepatitis viral infection had significant difference with ALB (p=0.000). The logistic regression shows that there was an association between hepatitis viral infection and aged group 27-35years (p=0.022), female (p=0.006), trading (p=0.001), tertiary (p=0.003) A+ve (p=0.018) and B+ve (p=0.001). In conclusion, Although, the prevalence of malaria was low, while that of hepatitis viral infection was high in the local government area, critical screening of blood donors is of prime importance because they serve as an asymptomatic reservoirs and a potential source of transmission of these infections. The strengthening of creating awareness on the general public regarding Malaria, HBV and HCV transmission and prevention should be embarked on.

Malaria parasitaemia and its impact on biological parameters among children

Malaria remains a major public health problem in children in endemic areas. This study aimed to determine its prevalence, intensity, and assess how biological parameters like RBC count, haemoglobin, haematocrit, glycaemia, platelet count and WBC count vary with respect to parasitaemia in children <16 years attending the Nkwen District Hospital, northwest Cameroon. The study was a hospital-based cross-sectional study conducted between March-May 2023. Structured, closed-ended questionnaires were administered to obtain information. Patients' temperature was measured using an infrared forehead digital thermometer. Malaria was diagnosed by RDT and positive samples Giemsa-stained for parasitaemia. Full blood count was performed using a haemolyser and glycaemia measured using a glucometer. In total, 321 children were examined. Overall prevalence of malaria (all P. falciparum) was 22.7% (73/321), with 24.7% (18/73), 34.2% (25/73) and 41.1% (30/73) having low, moderate and high parasitaemias, respectively. Overall GMPD was 2.670.8±179.9/μL; children aged 6-10 years were hit hardest (5.377.7 ± 3.2/μL). Malaria-positive children had significantly lower RBC count, Hb concentration, Hct, blood sugar, WBC and platelet counts (p<0.05) compared to those that were negative. Among positive children, RBC count, Hct, Hb, lymphocyte and platelet count each showed a significant (p<0.05) decrease while total WBC and granulocyte count each showed a significant (p<0.05) increase with increasing levels of parasitaemia. Changes in biological parameters during malaria are sensitive but poor specific indicators of malaria because they may overlap with symptoms of other infections. More attention should be given to children aged 6-10 years during strategic planning and design of malaria control programmes.

Infecção natural por Anaplasma marginale durante as primeiras semanas de vida de bezerros de uma propriedade leiteira da Amazônia Oriental

ABSTRACT: This study detected Anaplasma marginale in calvesusing blood smears and nested PCR (nPCR) and to compare the results with the clinical signs presented by calves on a dairy farm in the municipality of Castanhal, located northeast of the state Pará (1°07’19.1”S and 47°53’53.0”W), eastern Amazon. To this end, 192 blood samples were collected from 24 animals at 1-20, 21-41 and 42-60 days of age. Blood smears and nPCR with primers for the msp5 gene were performed. The prevalence of A. marginale was 61.5% (118/192) for the blood smear technique and nPCR (msp5). The manifestation of clinical signs of anaplasmosis also increased significantly over the course of the study (P &lt; 0.0001), being lower in animals aged 1-20 days, but increasing among those aged 21-41 and 42-60 days. These signs were characterized by apathy, fever, weight loss, diarrhea, dehydration, and hypochromic mucous membranes. Regarding the evaluation of the diagnostic techniques, no significant difference was observed in the detection of A. marginale between the blood smear and nPCR (P = 0.995), but the agent’s rickets increased on Day 47 (P &lt; 0.01) in both tests, thereby demonstrating a near-linear pattern of increase in rickets over the 60 days, with a consequent decrease in globular volume. This shows that of the 24 animals studied, 21 were infected at some point during the study period. Additionally, there was no significant difference between blood smears and nPCR, probably due to medium and high parasitemia, which were directly related to the clinical signs and decrease in globular volume.