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

BACKGROUND: 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. METHOD: 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. RESULTS: 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). CONCLUSION: 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).

First record of a possible trypanotolerant cattle breed in Latin America: Parasitological, serological, and clinical aspects.

Trypanosoma vivax infections are endemic in Africa, where they provoke trypanosomosis against which some local taurine breeds are tolerant and are thus named trypanotolerant. In Latin America, T. vivax was imported in 1919, since when it has been responsible for periodic outbreaks of the disease. This study assessed whether a South American taurine breed resilient to several parasitic and infectious diseases (Curraleiro Pé-Duro-CPD) can meet trypanotolerant criteria (control parasite proliferation, prevent anemia, survive without treatment, and maintain productivity). Three groups were established, each consisting of six animals (Group 1: CPD-infected; Group 2: Holstein/Gyr-infected; Group 3: Holstein/Gyr-uninfected, negative control). Groups 1 and 2 were infected with T. vivax on Day 0 and evaluated until day 532. Throughout the experimental period, parasitological (Woo and Brener), molecular (cPCR), serological (enzyme-linked immunosorbent assay - ELISA, indirect fluorescent antibody test - IFAT, immunochromatographic assay - IA), and clinical (hemogram, fever, weight loss) aspects were evaluated. During the acute phase of the disease, T. vivax was initially detected in Holstein/Gyr. Notably, the CPD animals restored their packed cell volume (PCV) values to the normal range 74 days after inoculations. In the chronic phase, two of the six CPD animals were positive by cPCR until D + 522 following immunosuppression with dexamethasone. Regarding serological aspects, the two CPD animals had positive tests until D + 532. The absence of T. vivax in blood during the chronic phase did not correspond to "self-cure". Holstein/Gyr animals exhibited fever on more evaluation days than CPD animals. Both breeds experienced weight loss, with Holstein/Gyr animals losing significantly more weight. On D + 25, the Holstein/Gyr group required treatment. During the 532 days, none of the CPD animals required treatment, even after being sensitized with dexamethasone. Animals from Group 3 tested negative for T. vivax throughout the experiment. This study demonstrated that CPD cattle fulfill the mentioned trypanotolerant criteria.

Effect of neem leaf extract (Azadirachta indica) in reducing the degree of parasitemia and apoptosis in C57BL mice with cerebral malaria.

Background and Aim: Brain malaria, which results from Plasmodium falciparum infection, is responsible for substantial fatalities and health issues. These processes, including cytoadherence, rosetting, and sequestration, induce an immune response, hypoxia, brain microvascular obstruction, disruption of the blood-brain barrier, and cell death. Parasitemia level can reveal the presence of infection and its association with apoptosis-related genes. Neem (Azadirachta indica) leaves with antimalarial properties could replace ineffective Indonesian malaria medications. This study was designed to evaluate the impact of neem leaf extract on cerebral malaria-induced parasitemia and neuron cell apoptosis in mice through an in vivo approach. Materials and Methods: 13-16 weeks old C57BL mice received infection by Plasmodium berghei strain ANKA. Parasitemia was estimated daily from the mice's tail blood. 8 mg, 12 mg, and 16 mg of a 96% ethanolic neem leaf extract were orally given for 6 days. Healthy, positive, and negative controls were included for treatment comparisons. On the 7th day, brain tissue was analyzed for (p > 0.05) gene expression. Through immunohistochemistry, both cell apoptosis in neurons expressing caspase-3 within a brain sample and the degree of parasitemia in a blood smear were assessed. The Pearson correlation test and one-way analysis of variance were employed to analyze the data. Results: Neem leaf extract reduces parasitemia and neuron cell apoptosis at multiple dosages (p < 0.000). Apoptosis in brain neurons and parasitemia show a strong positive correlation (r = +0.939). Neem leaf extract at doses of 12 and 16 mg was the most effective in reducing parasitemia levels and causing cell death. Conclusions: Neem leaf therapy significantly reduced the degree of parasitemia and cell apoptosis in C57BL mice compared with the control group without treatment (p = 0.05). This shows that neem leaves have the potential to be a candidate drug for malaria.

Asplenia-Associated Babesiosis: A Quagmire Traversed by Exchange Transfusion.

Babesiosis is a potentially life-threatening tick-borne parasitic infection. Severe disease in splenectomized individuals may require exchange transfusion. A 58-year-old male with a history of splenectomy presented with 2 weeks of subjective fever, weakness, and abdominal pain. He denied any rashes, tick bites, or recent travel. He had a motor vehicle accident a few years ago and had undergone an emergency splenectomy. On examination, the patient was febrile (39.3 °C), tachycardic (106/min), and jaundiced. Labs revealed anemia and thrombocytopenia. Computed tomography (CT) abdomen revealed asplenia. As it was summer, there was concern for a tick-borne illness. A peripheral smear showed schistocytes, and labs revealed hyperbilirubinemia, high lactate dehydrogenase (LDH), low haptoglobin, and reticulocytosis (13%), consistent with hemolysis. Testing for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), Ehrlichia, Borrelia, Anaplasma, and viral hepatitis was negative. Antibody testing for Babesia microti was positive. A blood parasite smear confirmed Babesia microti with a parasitemia of 9.5%. The patient received intravenous azithromycin and atovaquone for severe babesiosis. On day 2 of hospitalization, parasitemia increased to 14.7%. Hemoglobin and platelets dropped further on day 3. His parasite load remained consistently above 10% despite medical treatment. A decision was made for a red blood cell (RBC) exchange transfusion for severe disease, which was performed on the fourth day of hospitalization. Clinical improvement was seen after one session of exchange RBC transfusion. Hemoglobin remained stable, and thrombocytopenia improved 1 day after RBC exchange transfusion. Parasitemia dropped to 1.2% after 4 days of exchange transfusion, and azithromycin was switched to oral. He received 9 days of inpatient azithromycin and atovaquone. He was discharged with a plan to continue the oral antimicrobials for 3 more weeks. Asplenia and parasitemia > 10% are associated with severe babesiosis. Asplenia, in particular, is associated with severe infection, hospitalization, and prolonged duration of therapy. Exchange transfusion in severe babesiosis can be lifesaving.

Nonsteroidal Anti-Inflammatory Drugs and Experimental Chagas Disease: An Unsolved Question.

Chagas disease is a parasitic disease caused by the protozoan Trypanosoma cruzi with an acute, detectable blood parasites phase and a chronic phase, in which the parasitemia is not observable, but cardiac and gastrointestinal consequences are possible. Mice are the principal host used in experimental Chagas disease but reproduce the human infection depending on the animal and parasite strain, besides dose and route of administration. Lipidic mediators are tremendously involved in the pathogenesis of T. cruzi infection, meaning the prostaglandins and thromboxane, which participate in the immunosuppression characteristic of the acute phase. Thus, the eicosanoids inhibition caused by the nonsteroidal anti-inflammatory drugs (NSAIDs) alters the dynamic of the disease in the experimental models, both in vitro and in vivo, which can explain the participation of the different mediators in infection. However, marked differences are founded in the various NSAIDs existing because of the varied routes blocked by the drugs. So, knowing the results in the experimental models of Chagas disease with or without the NSAIDs helps comprehend the pathogenesis of this infection, which still needs a better understanding.

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.

Blood parasites of water frogs (Pelophylax esculentus complex) from the Danube Delta, Romania.

Water frogs of the genus Pelophylax host a variety of parasites, from protozoa to helminths. Among the blood parasites, representatives of Apicomplexa, Trypanosoma and Nematoda show the highest prevalence. In this study, we focused on blood parasites of water frogs living in the Danube Delta, Romania. In total, 74 individuals of P. ridibundus and eight individuals of P. esculentus from six localities were examined. Blood parasites were detected microscopically and using a molecular marker (18S rDNA). 89.77% of frogs from all investigated localities were found to be infected with at least one parasitic group, specifically with haemogregarines (84.09%), nematodes (1.14%), and trypanosomes (63.64%). The parasitemia of haemogregarines and trypanosomes differed significantly among the studied locations. There was no statistically significant difference in parasitemia between male and female hosts. However, adults were found to have a significantly higher parasitemia in comparison with subadults infected with haemogregarines. Correlation between parasitemia and the body length of frogs infected with haemogregarines was also significant (r = 0.226). By comparing the 18S rDNA sequences with the corresponding GenBank sequences, Hepatozoon species identified in water frogs showed a close similarity (98.1-99.8%) to Hepatozoon magna. Trypanosomes showed the highest sequence similarity to Trypanosoma sp. isolate R10 clone L2-3, Trypanosoma ranarum, and Trypanosoma cobitis.

Efectos de la infección del Trypanosoma cruzi en ratones de las cepas Balb/c y NIH / Effects of Trypanosoma cruzi infection in Balb/c and NIH mouse strains

La enfermedad de Chagas es una infección causada por el parásito Trypanosoma cruzi y transmitida por el vector Triatoma dimidiata, conocido en El Salvador como «chinche picuda¼. Esta enfermedad siempre ha sido de interés científico en modelos animales. Objetivo. Identificar el efecto de la infección por Trypanosoma cruzi en ratones de diferentes cepas (BALB/c y NIH) y sexo. Metodología. Se establecieron ocho grupos: cuatro infectados con Trypanosoma cruzi y cuatro grupos no infectados, distribuidos por cepa y sexo, con cinco ratones por grupo. Durante seis semanas se registró el peso corporal de los ratones. Además, se prepararon muestras de sangre de los grupos infectados en láminas para realizar los conteos de parasitemia. Al final del estudio, se extrajeron el bazo y el corazón de ambos grupos para los análisis estadísticos. Resultados. Los grupos infectados mostraron un incremento de peso en comparación a sus grupos controles. En la cepa NIH, las hembras presentaron una mayor parasitemia, mientras que en la cepa BALB/c fueron los machos los de mayor parasitemia. Los órganos de los grupos infectados fueron significativamente más grandes comparados a los de los grupos de control, excepto en el corazón de la cepa BALB/c. Respecto al peso de los órganos, se observaron diferencias significativas únicamente en el corazón de los machos de la cepa BALB/c, mientras que en el bazo ocurrió lo contrario. Conclusión. Los machos de la cepa BALB/c son más susceptibles al Trypanosoma cruzi, presentando niveles de parasitemia más altos entre los grupos estudiados

Chagas disease is an infection caused by the parasite Trypanosoma cruzi and transmitted by the vector Triatoma dimidiata, known in El Salvador as "chinche picuda". This disease has always been of scientific interest in animal models. Objective. Identify the effect of Trypanozoma cruzi infection in mice of different strains (BALB/c and NIH) and sex. Methodology. Eight groups were established: four infected with Trypanosoma cruzi and four uninfected groups, distributed by strain and sex, with five mice per group. The body weight of the mice was recorded for six weeks. In addition, blood samples from the infected groups were prepared on slides for parasitemia counts. At the end of the study, the spleen and heart were extracted from both groups for statistical analyses. Results. The infected groups showed an increase in weight compared to their control groups. In the NIH strain, females had higher parasitemia, whereas in the BALB/c strain, males had higher parasitemia. The organs of the infected groups were significantly larger compared to those of the control groups, except in the heart of the BALB/c strain. Regarding organ weight, significant differences were observed only in the heart of the male BALB/c strain, while the opposite was true for the spleen. Conclusion. Males of the BALB/c strain are more susceptible to Trypanosoma cruzi, presenting higher levels of parasitemia among the groups studied.

Late Parasitological Failure and Subsequent Isolated Gametocytemia of Uncomplicated Plasmodium falciparum Malaria in a Returned Traveler From Ghana, 2023.

Herein, we report a case of uncomplicated falciparum malaria with late parasitological failure in a 45-year-old businessman returning from Ghana. The patient visited the emergency department with high fever, headache, and dizziness. He traveled without antimalarial chemoprophylaxis. Laboratory tests led to the diagnosis of uncomplicated falciparum malaria with an initial density of 37,669 parasites per µL of blood (p/µL). The patient was treated with intravenous artesunate followed by atovaquone/proguanil. He was discharged with improved condition and decreased parasite density of 887 p/µL. However, at follow-up, parasite density increased to 7,630 p/µL despite the absence of any symptoms. Suspecting treatment failure, the patient was administered intravenous artesunate and doxycycline for seven days and then artemether/lumefantrine for three days. Blood smear was negative for asexual parasitemia after re-treatment but positive for gametocytemia until day 101 from the initial diagnosis. Overall, this case highlights the risk of late parasitological failure in patients with imported uncomplicated falciparum malaria.

Malaria in under-five children: prevalence and multi-factor analysis of high-risk African countries.

BACKGROUND: Malaria remains a significant public health challenge in Sub-Saharan Africa (SSA), particularly affecting under-five (UN5) children. Despite global efforts to control the disease, its prevalence in high-risk African countries continues to be alarming, with records of substantial morbidity and mortality rates. Understanding the association of multiple childhood, maternal, and household factors with malaria prevalence, especially among vulnerable young populations, is crucial for effective intervention strategies. OBJECTIVE: This study examines the prevalence of malaria among UN5 children in selected high-risk SSA countries and analyzes its association with various childhood, maternal, and household factors. METHODS: Data from the Malaria Indicator Surveys (MIS) spanning from 2010 to 2023 were analyzed. A weighted sample of 35,624 UN5 children from seven countries in sub-Saharan Africa (SSA) known for high malaria prevalence was considered in the analyses. Descriptive statistics and modified Poisson regression analysis were used to assess the association of multiple factors with malaria prevalence. Stata version 15 software was used in analyzing the data and statistical significance was set at a 5% significance level. RESULTS: The overall pooled prevalence of malaria among the studied population was 26.2%, with substantial country-specific variations observed. In terms of child factors, a child's age was significantly associated with malaria prevalence (APR = 1.010, 95% CI: 1.007-1.012). Children of mothers with higher education levels (APR for higher education = 0.586, 95% CI: 0.425-0.806) and Fansidar uptake during pregnancy (APR = 0.731, 95% CI: 0.666-0.802) were associated with lower malaria risk. Children from middle-wealth (APR = 0.783, 95% CI: 0.706-0.869) and rich (APR = 0.499, 95% CI: 0.426-0.584) households had considerably lower malaria prevalence compared to those from poor households. Additionally, rural residency was associated with a higher risk of malaria compared to urban residency (APR = 1.545, 95% CI: 1.255-1.903). CONCLUSION: The study highlights a notable malaria prevalence among under-five (UN5) children in high-risk SSA countries, influenced significantly by factors such as maternal education, Fansidar uptake during pregnancy, socioeconomic status, and residency. These findings underscore the importance of targeted malaria prevention strategies that address these key determinants to effectively reduce the malaria burden in this vulnerable population.

Parasitic Effects on the Congenital Transmission of Trypanosoma cruzi in Mother-Newborn Pairs.

Maternal parasitemia and placental parasite load were examined in mother-newborn pairs to determine their effect on the congenital transmission of Trypanosoma cruzi. Parasitemia was qualitatively assessed in mothers and newborns by the microhematocrit test; parasite load was determined in the placental tissues of transmitting and non-transmitting mothers by the detection of T. cruzi DNA and by histology. Compared to transmitter mothers, the frequency and prevalence of parasitemia were found to be increased in non-transmitter mothers; however, the frequency and prevalence of parasite load were higher among the transmitter mothers than among their non-transmitter counterparts. Additionally, serum levels of interferon (IFN)-γ were measured by an enzyme-linked immunosorbent assay (ELISA) in peripheral, placental, and cord blood samples. Median values of IFN-γ were significantly increased in the cord blood of uninfected newborns. The median IFN-γ values of transmitter and non-transmitter mothers were not significantly different; however, non-transmitter mothers had the highest total IFN-γ production among the group of mothers. Collectively, the results of this study suggest that the anti-T. cruzi immune response occurring in the placenta and cord is under the influence of the cytokines from the mother's blood and results in the control of parasitemia in uninfected newborns.

Antimalarial evaluation of alkyl-linked bis-thiadiazine derivatives in murine model infected with two Plasmodium strains.

Background and Purpose: Plasmodium falciparum and P. vivax are responsible for most malaria cases in humans in the African Region and the Americas; these parasites have developed resistance to classic antimalarial drugs. On the other hand, previous investigations of the alkyl-linked bis tetrahydro-(2H)-1,3,5-thiadiazine-2-thione (bis-THTT) derivatives compounds show satisfactory results against protozoan parasites such as Trypanosoma cruzi, Trypanosoma vaginalis, Trypanosoma brucei rhodesiense and Leishmania donovani. Therefore, it is possible to see some effect of bis-THTT derivatives on other protozoan parasites, such as Plasmodium. Experimental Approach: This study aimed to perform an in vivo biological evaluation of bis-THTT (JH1 to JH6) derivatives compounds as possible anti-malaria drugs in BALB/c mice infected with Plasmodium berghei ANKA and Plasmodium yoelii 17XL strains. In this work, we evaluated the compounds as potential antimalarial drugs in BALB/c mice infected with Plasmodium strains. Key Results: For each compound, we assess the percentages of parasitemia by smears from tail blood and the humoral response by indirect ELISA test using each compound as an antigen. We also evaluated the B lymphocyte response and the cytotoxicity of the bis-THTT derivatives compounds with MTT cell proliferation assays. Conclusions: Our results show that the bis-THTT derivatives JH2 and JH4 presented effective parasitemia control in mice infected with P. berghei; JH5 and JH6 compounds have similar infection control results as chloroquine in mice infected P. yoelii strain. The evaluation of bis-THTT derivatives compounds in a model of BALB/c mice infected with P. berghei and P. yoelii allowed us to conclude that some of them have an antimalarial effect; however, none of the tested compounds exceeded the efficiency of chloroquine.

Effect of moringa extract on parasitemia, monocyte activation and organomegaly among Mus musculus infected by Plasmodium berghei ANKA.

In Indonesia, malaria remains a problem, with 94,610 active cases in 2021 and its current therapy includes chloroquine and artemisinin; however, resistance has been commonly reported. To overcome this problem, studies about potential medicinal plants that can be used as antimalaria, such as moringa (Moringa oleifera) started to receive more attention. The aim of this study was to investigate the effects of moringa in parasitemia, monocyte activation, and organomegaly on animal model malaria. This experimental study used male Mus musculus, infected by Plasmodium berghei ANKA, as an animal malaria model. The extract was made by maceration of dry moringa leaves, which were then divided into three concentrations: 25%, 50%, and 75%. Dihydroartemisinin-piperazine was used as a positive control treatment, and distilled water as a negative control treatment. The animals were observed for six days to assess the parasitemia count and the number of monocyte activation. On day 7, the animals were terminated, and the liver, spleen, and kidney were weighed. The results showed that the effective concentrations in reducing parasitemia and inducing monocyte activation were 50% and 25% of moringa leaf extract, respectively. The smallest liver and spleen enlargement was observed among animals within the group treated with a 50% concentration of M. oleifera extract. In contrast, the smallest kidney enlargement was observed in the group treated with 25% of M. oleifera extract. Further analysis is recommended to isolate compounds with antimalarial properties in moringa leaves.

Malaria Parasitemia and Severe Health Complications in Children Under Five Years of Age in Nigeria: A Study Using the Demographic and Health Survey (DHS) Malaria Indicator Survey (MIS) 2021.

BACKGROUND: In Nigeria, 97% of the population is at risk of contracting malaria. It is transmitted by female Anopheles mosquitoes carrying the Plasmodium parasite and can be lethal. An estimated 55 million illnesses and 80,000 deaths per year result from it. Children under five are more likely to contract malaria. Efforts to control malaria in Nigeria include indoor residual spraying, insecticide-treated bed nets, and quick detection and treatment of confirmed cases with effective antimalarial medications. These attempts have been impeded by limited healthcare access, poor financing, and drug-resistant parasites. Thus, the study of the relationship between malaria complications and housing for children under five is essential. METHODS: The Demographic and Health Survey (DHS) Malaria Indicator Survey (MIS) 2021, a nationally representative data set from developing countries on population and health, was used for this study. A sample size of 13,727 was employed (n=13,727). Logistic regression analyses were conducted to test the association between the type of place of residence and malaria complications (outcome). RESULTS: Overall, 4.2% (n=570, weight HV005) of participants in the sample reported malaria complications. The results of the logistic regression revealed that children residing in urban settlements (aOR 0.37, 95% CI 0.37-0.37, p-value <0.001), children from the poorest class families (aOR 11.63, 95% CI 1.62-1.63, p-value 0.004), children from poorer class families (aOR 7.56, 95% CI 7.55-7.57, p-value <0.001), children from middle-class families (aOR 4.05, 95% CI 4.03-9.06, p-value <0.001), children from richer class families (aOR 1.22, 95% CI 2.21-2.23, p-value <0.001), children of mothers with primary education (aOR 0.42, 95% CI 2.32-4.112, p-value 0.001), children of mothers with secondary education (aOR 0.24, 95% CI 3.21-3.22, p-value <0.001), children of mothers with higher education (aOR 0.08, 95% CI 0.72-0.80, p-value <0.001), and children of the female gender (aOR 0.65, 95% CI 0.65-0.66, p-value <0.001) are all associated with severe malaria complications. CONCLUSIONS: In conclusion, the study examined malaria complications in Nigerian children under five by residency. The findings imply that rural children are more likely to have serious malaria complications than urban children. This emphasizes the necessity for targeted malaria therapies in rural areas with limited healthcare access.

Correlations between the degree of infection by wild strain of Toxoplasma gondii in vitro and porcine hematological parameters.

The apicomplexa Toxoplasma gondii is capable of actively proliferating in numerous types of nucleated cells, and therefore has a high potential for dissemination and resistance. Thus, the present work aimed to correlate the inoculum concentrations and amount of post-infection parasites with porcine hematological parameters (including biochemistry) through in vitro culture. Porcine blood was incubated with different concentrations of parasites (1.2 × 107, 6/3/1.5 × 106 cells/mL), then the concentrations of red blood cells (RBC) and their morphology, total and differential leukocytes, and free peptides were evaluated. In addition, eight different blood samples analyzed before inoculation, where subsequent multivariate analysis was applied to correlate different variables with trophozoite concentration. The results showed no significant variation (p < 0.05) in the relative levels of free peptides, or the relative percentage of RBC at all the parasite concentrations tested. However, the normalized percentages of leukocytes and neutrophils showed a significant reduction, while those of lymphocytes, eosinophils and monocytes showed the opposite behavior. Semi-automatic processing of images exhibited significant microcytosis and hypochromia. The multivariate analysis revealed a positive correlation between the amount number of protozoa (AP) and the variables: "Red cells" and "Neutrophils", an indifference between the AP and the content of free peptides, and the concentration of monocytes in the samples; and a negative correlation for AP and the percentages of lymphocytes and eosinophils. Our results suggest that specific changes in hematological parameters may be associated with different degrees of parasitemia, demanding a thorough diagnostic process and adequate treatment.

Molecular and haematological investigations on Anaplasma ovis infection in goats in Ahvaz, Iran: Insights into infection rate, haemolytic effect and breed influences.

BACKGROUND: Anaplasma ovis (A. ovis) is the predominant causative agent of anaplasmosis in goats and sheep in most tropical and subtropical regions of the world. However, there is considerable variation in reported infection rates, breed susceptibility, and controversial findings regarding the haemolytic effects of A. ovis infection in goats. OBJECTIVES: Thus, we investigated the molecular and haematological aspects of A. ovis infection in goats from Ahvaz city. METHODS: One hundred and fifty apparently healthy goats (74 blacks and 76 Najdi goats) were randomly sampled from six flocks in the Ahvaz suburb during ticks' activity season. Haematological evaluation, smear microscopic (SM) examination and PCR assay were performed to assess A. ovis infection. Additionally, the percentage of parasitemia was determined from blood smears. RESULTS: SM examination revealed that 25.7% of the goats displayed erythrocyte Anaplasma-like inclusion bodies. PCR analysis indicated that 54% of the goats were positive for A. ovis infection (44.6% of blacks and 63.2% of Najdi goats). No significant difference in haematological values was observed between healthy and infected goats based on PCR testing. However, a significant difference in haematological indices was observed between the group with parasitemia level of 0.01-0.02% (SM and PCR positive) compared to the healthy goats (SM and PCR negative), particularly concerning Hb, PCV and RBC count (p < 0.01). CONCLUSIONS: When the parasitemia exceeds 0.01%, A. ovis infection may disrupt haematological parameters in infected goats. The high prevalence of A. ovis infection (54%) among the studied goats underscores the importance of giving special attention to implementing necessary measures for disease control in the Ahvaz suburb.

Parasite Burden of Trypanosoma cruzi in Whole Blood and Buffy Coat Determined by Real-Time PCR in Individuals with Chronic Chagas Disease.

The objective of this study was to compare, by qPCR, the circulating blood parasite load of Trypanosoma cruzi in the buffy coat, and in whole blood mixed with boiled and unboiled guanidine hydrochloride-EDTA buffer, of individuals with chronic ChD. The concentration and purity of DNA were evaluated in a Nanodrop Denovix DS-11FX Series Spectrophotometer (DeNovix Inc., Wilmington, NC, USA). The parasite load was determined with the Taqman® qPCR system using a Stratagene Mx3000P thermocycler (Agilent Technologies, Santa Clara, CA, USA) with Cruzi 1 and Cruzi 2 satellite primers. Student's t-test with Bonferroni correction, Chi-squared (χ2) tests and Spearman's correlation coefficient were applied. The concentration and purity of DNA were higher in the buffy coat. Parasite DNA was detected and quantifiable in the three types of samples in seven patients, without statistically significant differences in the parasite load obtained. Higher correlations were found between the total DNA concentrations and the parasite loads obtained in the samples of the buffy coat.

Multiple myeloma and Chagas disease: qPCR as a marker for preemptive antiparasitic therapy: a case reports series and review

ABSTRACT Multiple myeloma (MM) associated with Chagas disease is rarely described. This disease and its therapy suppress T cell and macrophage functions and increase regulatory T cell function, allowing the increase of parasitemia and the risk of Chagas Disease Reactivation (CDR). We aimed to analyze the role of conventional (cPCR) and quantitative Polymerase Chain Reaction (qPCR) for prospective monitoring of T. cruzi parasitemia, searching for markers of preemptive antiparasitic therapy in MM patients with Chagas disease. Moreover, we investigated the incidence and management of hematological diseases and CDR both inside and outside the transplant setting in the MEDLINE database. We found 293 studies and included 31 of them. Around 1.9-2.0% of patients with Chagas disease were reported in patients undergoing Stem Cell Transplantation. One case of CDR was described in eight cases of MM and Chagas disease. We monitored nine MM and Chagas disease patients, seven under Autologous Stem Cell Transplantation (ASCT), during 44.56±32.10 months (mean±SD) using parasitological methods, cPCR, and qPCR. From these patients, three had parasitemia. In the first, up to 256 par Eq/mL were detected, starting from 28 months after ASCT. The second patient dropped out and died soon after the detection of 161.0 par Eq/mL. The third patient had a positive blood culture. Benznidazole induced fast negativity in two cases; followed by notably lower levels in one of them. Increased T. cruzi parasitemia was related to the severity of the underlying disease. We recommend parasitemia monitoring by qPCR for early introduction of preemptive antiparasitic therapy to avoid CDR.

Cytokine profile levels and their relationship with parasitemia and cardiomyopathy in people with Chagas disease in Spain. A prospective observational study.

Immunoregulatory networks may have a role in controlling parasitemia in the chronic phase of human Chagas disease. The aim was to describe the serum cytokine profile of Trypanosoma cruzi in chronically infected patients and to evaluate its relationship with parasitemia and Chagas cardiomyopathy.This prospective observational study included adult patients with chronic Chagas disease. Demographic and clinical data were collected, and peripheral blood samples were used to perform T. cruzi real-time polymerase chain reaction (RT-PCR) and determine the serum cytokine profile.Fifty-eight patients were included; 17 (29.3%) had positive RT-PCR results. This group had a higher median concentration of TNF-α (p = 0.003), IL-6 (p = 0.021), IL-4 (p = 0.031), IL-1ß (p = 0.036), and IL-17A (p = 0.043) than those with a negative RT-PCR. Patients with cardiac involvement had a higher median concentration of IL-5 (p = 0.016) than those without.These results reinforce the key role that cytokines play in Chagas disease patients with parasitemia and cardiac involvement.

A Non-Coding Fc Gamma Receptor Cis-Regulatory Variant within the 1q23 Gene Cluster Is Associated with Plasmodium falciparum Infection in Children Residing in Burkina Faso.

Antibodies play a crucial role in activating protective immunity against malaria by interacting with Fc-gamma receptors (FcγRs). Genetic variations in genes encoding FcγRs can affect immune cell responses to the parasite. In this study, our aim was to investigate whether non-coding variants that regulate FcγR expression could influence the prevalence of Plasmodium falciparum infection. Through bioinformatics approaches, we selected expression quantitative trait loci (eQTL) for FCGR2A, FCGR2B, FCGR2C, FCGR3A, and FCGR3B genes encoding FcγRs (FCGR), in whole blood. We prioritized two regulatory variants, rs2099684 and rs1771575, located in open genomic regions. These variants were identified using RegVar, ImmuNexUT, and transcription factor annotations specific to immune cells. In addition to these, we genotyped the coding variants FCGR2A/rs1801274 and FCGR2B/rs1050501 in 234 individuals from a malaria-endemic area in Burkina Faso. We conducted age and family-based analyses to evaluate associations with the prevalence of malarial infection in both children and adults. The analysis revealed that the regulatory rs1771575-CC genotype was predicted to influence FCGR2B/FCGR2C/FCGR3A transcripts in immune cells and was the sole variant associated with a higher prevalence of malarial infection in children. In conclusion, this study identifies the rs1771575 cis-regulatory variant affecting several FcγRs in myeloid and neutrophil cells and associates it with the inter-individual capacity of children living in Burkina Faso to control malarial infection.