Protein post-translational modifications (PTMs) refer to the covalent attachment of chemical groups to polypeptide chains after protein synthesis, which expands protein chemical diversity and enhances functional versatility. Common PTM types include phosphorylation, glycosylation, and lysine acetylation (Kac), a major, reversible PTM regulated by lysine acetyltransferases (KATs) and lysine deacetylases (KDACs). Kac cross-regulates with other modifications to govern chromatin transcription and epigenetic programs, and occurs in histones, nucleoproteins, and cytoplasmic regulatory factors. In this study, we employed proteomic approaches to investigate lysine acetylation in proteins derived from snake-borne spargana for the first time. A total of 390 acetylated proteins and 655 Kac sites were identified. Subcellular localization analysis revealed these acetylated proteins were predominantly localized in the cytoplasm, followed by the mitochondria, nucleus, extracellular matrix, and plasma membrane. This study aims to expand our understanding of protein acetylation in Spirometra mansoni plerocercoid larvae and provide valuable information for designing effective drugs or vaccines to control sparganosis.

Anopheles aquasalis mosquito, a key Plasmodium vivax vector in coastal Latin America, can be sensitive to environmental shifts. This study assessed the impact of IPCC AR5-projected climate scenarios on mosquito survival, infectivity, and immunity. Four scenarios were simulated in digitally controlled climate rooms: a forest-based environmental control and three IPCC projections - (i) Scenario RCP 4.5 (light, +1.0°C and +250 ppm CO2 on current scenario); RCP Scenario 6.0 (intermediate, +2.5°C and +400 ppm CO2 over current scenario) and RCP Scenario 8.5 (extreme, +4.5°C and +850 ppm CO2 over current scenario). Female An. aquasalis mosquitoes were infected with P. vivax via artificial membrane feeding, and survival, infection, and expression of immune genes (Attacin, Cecropin, and Defensin) evaluated. All conditions supported parasite development, with no significant differences in infection rates (54 - 68%) or oocyst intensity. However, mosquito survival declined significantly under RCPs 6.0 and 8.5, with probabilities of 18.6% and 6.5% by day 7 post-infection respectively, versus 40% in the non-blood-fed controls (p<0.001). Attacin expression differed significantly between the experimental control and RCP 4.5 (P = 0.0379), and RCP 8.5 (P = 0.0016); RCPs 6.0 and 8.5 (P = 0.0210). Defensin also varied between RCP 4.5 and 6.0 (P = 0.0359); RCP 6.0 and experimental control (P = 0.0464), and RCP 8.5 and experimental control (P = 0.0274), suggesting decreased immune activation under higher heat and CO₂ stress. Cecropin showed no significant variations. While An. aquasalis retained infectivity under all simulated conditions, the increased mortality and altered immune gene expression observed in RCPs 6.0 and 8.5 suggest that future climatic stress may compromise vector fitness.

Ruta chalepensis L. is a medicinal species widely used in ethnomedicine for gastrointestinal disorders and parasitic diseases. Among its bioactive compounds, the furanocoumarins chalepensin and graveoline have shown antiparasitic activity. This study aimed to evaluate the nematocidal potential of graveoline and chalepensin, isolated from R. chalepensis, against Trichinella spiralis in both in vitro cultures and an experimental in vivo model. The compounds were obtained from leaves and stems and first tested against first-stage larvae (L1) in culture. Selectivity indices (SI) were calculated based on cytotoxicity in Vero cells. Based on in vitro efficacy, chalepensin was selected for evaluation in a murine model of trichinellosis at 50 mg/kg/day administered over three consecutive days against three different infection stages: intestinal (days 0-2), migrating (days 13-15), and encysted (days 34-36). Larvae per gram of muscle were quantified on day 43. Histological sections were analyzed for capsule morphology and inflammation. Additionally, molecular docking was performed to explore potential parasite targets. Both compounds exhibited superior activity compared to the R. chalepensis extract (LC₅₀ = 28.2 µg/mL; SI = 22.4). Chalepensin exhibited strong in vitro activity (LC₅₀ = 0.1 µg/mL; SI = 8561), superior to graveoline (LC₅₀ = 1.1 µg/mL; SI = 162). In vivo, chalepensin reduced larval burden by 90.7% (intestinal), 37.5% (migrating), and 37.0% (encysted). Histology revealed reduced capsule thickness and pericystic inflammation. Docking predicted high affinity for thymidylate synthase (ΔG = -7.175 kcal/mol), suggesting interference with DNA synthesis. Chalepensin demonstrates potent nematocidal activity against T. spiralis, supporting its potential utility as a phytochemical-based therapeutic candidate for the management of trichinellosis.

Myxozoa are a highly diverse group of cnidarian parasites, with approximately 3,070 described species. This study describes a new species, Myxobolus mirandensis n. sp., based on morphological, molecular, and histopathological analyses. The parasite was found in Salminus brasiliensis (dourado), one of the largest and most economically and recreationally important freshwater fish species in South America. Specimens of S. brasiliensis were collected from the Miranda River in the Brazilian Pantanal, with a prevalence of the infection of 63.6% (7/11). White and elongated plasmodia were observed in the gill filaments. Pear-shaped myxospores measured 10.1 ± 0.3 µm in length, 6.6 ± 0.3 µm in width, and 5.1 ± 0.1 µm in thickness. Nematocysts were elongated and occupied more than half of the spore body, measuring 4.9 ± 0.3 µm in length and 1.9 ± 0.2 µm in width. Histopathological analysis revealed plasmodia developing in the epithelial tissue at the distal ends of the gill filaments, resulting in compression of adjacent tissues and structures. A thin connective tissue capsule surrounded the plasmodia, and numerous granulocytic cells were observed in the tissue adjacent to the plasmodia. Sequencing of the small subunit ribosomal DNA (SSU rDNA) yielded a 1,954 bp fragment. Phylogenetic analysis showed M. mirandensis grouping with Myxobolus oliveirai and Myxobolus filamentum, both reported in Bryconidae hosts. This represents the seventh record of a myxozoan infecting S. brasiliensis.

Extreme climate variables are increasingly important for understanding the distribution of Biomphalaria snails, the intermediate hosts of Schistosoma mansoni. While many studies focus on long-term averages, our approach emphasizes the role of climatic extreme events and their seasonal timing in shaping habitat persistence. We applied spatially explicit Random Forest (RF) species distribution models with spatial cross-validation and bootstrap aggregation to produce robust and transferable predictions of snail habitat suitability, and then ranked the influence of extreme climate variables using SHapley Additive exPlanations (SHAP) values and partial dependence plots. The models performed well, with average area under curve (AUC) values of 0.75 across species and partial AUC ratios greater than one, confirming the robustness of the predictions. Precipitation seasonality, multi-month drought indices, surface absorbed solar radiation extremes, and diurnal air temperature range were the highest-ranked drivers, though their influence was not uniform across snail species. B. glabrata was most responsive to seasonal recharge, while B. straminea showed resilience to variability and often persisted in man-made habitats. B. tenagophila was more constrained by drought and radiation stress. Spatial comparisons between 1995 and 2020 indicated expansions and contractions in various states, with new hotspots emerging in southeastern and central Brazil, while habitat suitability declined in drought-prone regions, such as in the State of Pernambuco. These results demonstrate that climate extreme events, in addition to long-term baseline changes, drive the spatially heterogeneous redistribution of Biomphalaria habitats. Also, our findings highlight the need for species-specific monitoring, integration of water infrastructure management, and forward-looking surveillance strategies that address both climate variability and landscapes modified by humans.

Visceral leishmaniasis (VL) is a climate-driven disease with a complex epidemiological pattern and is difficult to predict, especially in regions like eastern Sudan, where disease patterns vary substantially from season to season. In this study, our objective was to forecast the monthly incidence of VL in Gedaref State based on key climatic drivers-precipitation, temperature, and humidity. To capture the complex dynamics of disease transmission, we propose a new hybrid model based on wavelet transforms and state-of-the-art deep learning models to decompose multi-scale patterns and learn linear and nonlinear relationships.. We developed and tested hybrid models: Wavelet-Gaussian Process Regression (Wavelet-GPR), Wavelet-Spatiotemporal Graph Neural Network (Wavelet-StemGNN), and Wavelet-Temporal Convolutional Network with Bidirectional LSTM (Wavelet-TCN-BiLSTM). These were compared against a traditional Vector Autoregressive model (VAR) as a baseline. We employed a fixed temporal split (2000-2018 for training; 2019-2022 for testing). All models were implemented using Python and their performance was evaluated using key forecasting metrics: RMSE, MAE, R, and MAPE. Our wavelet-GPR model outperformed both baseline and other deep learning methods in terms of prediction accuracy (RMSE = 43.93, MAE = 32.85, R = 0.93, MAPE = 14.6%), with the smallest prediction errors and well-calibrated 95% predictive intervals. The results of our study underscore the possibility of improving the disease surveillance and early warning systems by means of hybrid wavelet-based artificial intelligence models, particularly in resource constraints situations. The projection may be employed for monthly early warning bulletins and the strategic pre-positioning of diagnostics and medicines in Gedaref. The method is straightforward to modify to different VL locations in Sudan and East Africa, and it requires much re-calibration. Future projects will include more layers of environmental and socio-economic data to make predictions and planning operations easier.

Visceral leishmaniasis is a zoonosis of high epidemiological relevance, caused by protozoan parasites of the Leishmaniinae subfamily, mainly of the Leishmania genus and transmitted by hematophagous phlebotomine sand flies. Vector-borne disease control faces significant challenges, and innovative strategies towards the vector - such as the use of the endosymbiont bacteria Wolbachia - have gained prominence for their ability to manipulate the reproduction of their hosts and modulate their immunity, reducing pathogen transmission. However, little is known about natural Wolbachia infection in the sand fly population. This study aimed to assess the circulation of Wolbachia in sand flies from Montes Claros, a visceral leishmaniasis-endemic area in Minas Gerais, Brazil. A total of 1.191 females Lutzomyia longipalpis were analyzed, and Wolbachia DNA was detected in 30 samples (2.5%), with a homogeneous presence among the points sampled. DNA sequences revealed a single strain, wPup, that has not been previously described in sand flies. The positive Wolbachia samples were also tested for Leishmania spp, however, no DNA was detected.

The primary transmission of Trypanosoma cruzi, causative agent of Chagas disease, to humans and animals is through the feces and urine of Triatominae. Therefore, knowledge of their ecology and distribution is crucial for establishing effective surveillance programs with citizen participation. This is coupled with geographic distribution models to predict the risk of transmission in the areas most affected by Chagas disease. Therefore, this study aims to: a) document and validate citizen participation on the iNaturalist in the collection and observation of triatomines, as well as exploring risk factors; b) detect natural T. cruzi infection in triatomines using molecular and parasitological methods; c) to review the historical and recent literature of Triatoma gerstaeckeri (Stål 1859) in Mexico, and d) develop current and potential distribution models of T. gerstaeckeri to infer risk zones for parasite transmission in Mexico. Data of T. gerstaeckeri were collected from three sources: iNaturalist (n = 87), fieldwork data (n = 68), and scientific articles and theses (n = 882). iNaturalist observations were validated from photographs, specimen capture, and distribution overlap. The prevalence of natural infection by T. cruzi was 21.73 % (5/23) in T. gerstaeckeri from Nuevo Leon, with DTU strains TcI and TcIII, the first reports of DTU in the state. Citizen participation data like iNaturalist can potentially document the presence of triatomines at broad spatial scales. Still, they also represent an opportunity to engage and educate the public in the surveillance and control of these insect vectors associated with Chagas disease.

Chagas disease is a globally widespread parasitic infection caused by the flagellated protozoan Trypanosoma cruzi. It is classified as a neglected tropical disease, primarily affecting impoverished and rural regions where access to diagnosis and treatment is limited. Although treatment for this disease is effective, it is restricted to the acute phase, during which diagnosis is more challenging, reducing cure rates. Consequently, prevention remains the most effective control method, and recombinant proteins offer a promising strategy for vaccine development. Specifically, the proteins ASP-2 and TC24 have demonstrated immunoprotective activity in various experimental models. This study aimed to characterize the immune response elicited by the combined use of recombinant ASP-2 (from the intracellular stage) and TC24 (from the bloodstream stage), seeking to promote a synergistic protective effect compared to the response generated by the proteins used individually. To evaluate this, recombinant proteins rASP-2 and rTC24 were used as vaccine formulations to immunize female BALB/c mice as follows: Group 1: Saline solution; Group 2: 25 µg of rTC24 + aluminum hydroxide; Group 3: 25 µg of rASP-2 + aluminum hydroxide; Group 4: 12.5 µg of rTC24 + 12.5 µg of rASP-2 + aluminum hydroxide. The humoral immune response assessed IgG antibody levels by indirect ELISA of animal sera collected on days 0, 21, and 42 of the experiment, while the cellular response was evaluated by collecting and culturing splenocytes, assessing cytokines IFN-γ, TNF-α, interleukins 1β, 4, 6, 12, 17, and Toll-like receptor 4, quantified by real-time PCR. The results indicated a significant antibody production in the group where the proteins were combined (G4) compared to the control group (G1) on days 21 and 42. A significant increase in antibody production was also observed in group G4 on day 42 when compared to both groups using the isolated proteins (G2 and G3). Conversely, the cellular response showed an increase in IFN-γ and interleukins 1β and 17 in Group D, while the isolated ASP-2 protein induced the expression of TNF-α, interleukins 4 and 12, and Toll-like receptor 4. Western blotting using T. cruzi lysate and pooled serum confirmed the ability of the antibodies to recognize native parasite proteins. In conclusion, the combined use of proteins from different parasite life stages proved advantageous, indicating the induction of a mixed cellular immune response, predominantly of the Th1 and Th17 profiles.

Plasmodium falciparum remains a major public health challenge in the Jazan region, Saudi Arabia. This study investigated the influence of haemoglobin variants and ABO blood groups on malaria susceptibility, severity, and immune response among Saudi children. A total of 587 children were classified into malaria-free controls (MFC, n = 196), uncomplicated malaria (UM, n = 215), and severe malaria (SM, n = 176). Blood samples were analysed for P. falciparum infection, haemoglobin genotypes, ABO blood groups, and IgG levels using PCR, ELISA. The median age was 4 years for MFC and UM groups, and 5 years for SM. Blood group O predominated in MFC (49.5%), while blood group A was more frequent in UM (44.7%) and SM (41.5%). Carriers of the HbS allele had reduced odds ratio of SM versus MFC (OR=0.40, 95% CI: 0.22-0.72, P = 0.002). HbC allele carriers showed even stronger protective associations against SM versus MFC and UM in both unadjusted and adjusted models (P < 0.01). Blood group A was associated with higher susceptibility but did not reach statistical significance. Antibody analysis revealed higher total IgG and IgG subclass (IgG1-IgG4) levels in UM cases compared to MFC and SM (P < 0.001), suggesting a more effective immune response in UM. Hb S and C variants offer significant protection against severe malaria and high parasitaemia. Blood group A may be linked to increased susceptibility. These findings underscore the role of host genetic factors in malaria severity and may guide future prevention strategies in endemic areas.

The Sterile Insect Technique (SIT), which uses ionizing radiation, is a promising approach for suppressing Aedes aegypti, a major vector of arboviruses. However, limited knowledge of the transgenerational effects of gamma radiation may compromise the long-term effectiveness of SIT programs. This study investigated the impact of gamma irradiation on male Ae. Aegypti, and its consequences for fecundity, fertility, and genotoxicity across three generations (F0, F1, and F2). Male pupae were irradiated with 20, 30, 40, and 50 Gy and then mated with non-irradiated females. Fecundity and fertility were quantified for the parental and filial generations, and genotoxicity was assessed using the micronucleus assay in larval neuroblasts, adult male Malpighian tubules, and adult female oocytes. Radiation induced a dose-dependent reduction in fertility, reaching near-complete sterility (97.1%) at 50 Gy. Micronucleus formation confirmed genotoxic effects, particularly at 30 and 40 Gy, in both somatic and germline tissues. Heritable damage was evident in F1 and F2 generations, with sex-specific patterns suggesting differences in DNA damage repair and retention, providing new evidence that gamma radiation induces inheritable genomic instability in Ae. aegypti and confirming 50 Gy as an effective sterilizing threshold for SIT applications. The micronucleus assay proved to be a reliable and sensitive biomarker for detecting inherited genomic alterations, supporting its use in quality control and cytogenetic monitoring. These findings inform SIT dose optimization and long-term biosafety protocols.

Crimean-Congo haemorrhagic fever (CCHF) is a major public health concern in Türkiye, which has the highest global incidence. This study analysed national surveillance data from 2011 to 2024 to describe the epidemiology of CCHF and identify predictors of fatality through a retrospective cohort analysis using the CCHF Information System. All laboratory-confirmed cases reported between 2011 and 2024 were included. Descriptive statistics summarized demographic, clinical, and laboratory variables. Logistic regression was applied to determine fatality predictors, and adjusted odds ratios (ORadj) with 95% confidence intervals (CI) were calculated. Among 27,269 samples tested, 11,811 cases were laboratory-confirmed, of which 552 died (case fatality rate: 4.7%). Males comprised 60.5% of cases, and the mean age was 46.8 ± 18.5 years. Tick-bite or tick contact (62.2%) and close contact with animal blood or tissues (69.6%) were common exposure histories. Fatality was independently associated with increasing age (ORadj: 1.034; 95% CI: 1.029-1.040), male sex (ORadj: 1.3; 95% CI: 1.1-1.6), splenomegaly (ORadj: 2.0; 95% CI: 1.5-2.7), haemorrhagic findings (ORadj: 2.4; 95% CI: 2.0-3.0), platelet count.

Cryptosporidiosis remains a major health concern, particularly in immunocompromised individuals, due to limited effective treatment options. Nitazoxanide (NTZ) is currently the only FDA-approved drug for cryptosporidiosis, yet its efficacy is significantly reduced in immunosuppressed hosts. This study evaluated the therapeutic potential of disulfiram, an FDA-approved drug for alcoholism, alone and in combination with NTZ, against Cryptosporidium infection in immunocompromised mice. Forty immunosuppressed Swiss albino mice were divided into five equal groups: uninfected controls, infected untreated, NTZ-treated, disulfiram-treated, and combination-treated (NTZ + disulfiram). All infected mice were orally inoculated with ∼10⁴ Cryptosporidium oocysts and were treated with NTZ (250 mg/kg/day) and/or disulfiram (25 mg/kg/day) for 10 consecutive days. Efficacy was assessed through parasitological, histopathological, and ultrastructural analyses. The combination therapy achieved the highest fecal oocyst reduction: 34.3% after one week and 88.3% after two weeks. In comparison, NTZ and disulfiram monotherapies achieved 24% and 76% reductions, respectively, at two-weeks mark. In intestinal contents, the combination therapy resulted in 62.6% oocyst reduction versus 11.4% for NTZ and 36.2% for disulfiram at week two post treatment. Histopathological analysis revealed near-complete mucosal restoration in the combination group, whereas monotherapies showed limited or moderate recovery. Transmission electron microscopy confirmed full epithelial regeneration only in the dual therapy group, with intact microvilli, normal mitochondria, and restored cellular junctions. In conclusion, disulfiram, particularly when combined with NTZ, demonstrated enhanced anti-cryptosporidial efficacy and may serve as a promising adjunct therapy, mostly for immunocompromised patients.

Trachoma, caused by Chlamydia trachomatis, remains the leading infectious cause of preventable blindness globally, with children aged 1-9 years bearing the highest burden. Ethiopia continues to report a high prevalence despite ongoing control efforts. The Health Extension Program (HEP) aims to improve household hygiene and sanitation. This is the first study to quantify the direct association between household-level HEP adherence and active trachoma prevalence.

This study analyzed the time of dengue-related deaths in two regions of Peru during the atmospheric phenomena of El Niño Costero and Cyclone Yaku, which caused the largest dengue epidemic ever registered in Peru at that time.

Incarcerated populations are disproportionately affected by the hepatitis C virus (HCV) due to overlapping social, structural, and behavioral vulnerabilities. However, epidemiological data from some regions in Brazil, a continental country, remain scarce. Therefore, this study estimated the HCV exposure prevalence and identified associated factors among inmates in Goiás, a state in the Central-West region of Brazil. A cross-sectional study was conducted with 1083 inmates from prison units in Goiás. All participants were interviewed, and their serum samples were tested for anti-HCV using ELISA and confirmed by a line immunoassay. The prevalence of anti-HCV was 2.2 % (95 % CI: 1.42-3.28). A multivariate analysis showed that age > 32 years (adjusted odds ratio [aOR] = 4.821; 95 % confidence interval [CI]: 1.492-15.574; p = 0.009), history of injectable drugs use (aOR = 4.536; 95 % CI: 1.685-12.212; p = 0.003), history of sexually transmitted infections (STIs, aOR = 3.313; 95 % CI: 1.213-9.049; p = 0.019), and HBV infection (aOR = 3.147; 95 % CI: 1.052-9.411; p = 0.040) were associated with HCV exposure. These findings revealed that the anti-HCV prevalence is higher among incarcerated individuals in Goiás than in the general population in Brazil, and the use of injectable drugs as the factor with the strongest association with HCV exposure emphasized the importance of HCV transmission through the parenteral route. Additionally, the association with HBV infection and previous STIs highlights the need for integrated screening of multiple infections and ongoing surveillance for HCV within the prison environment.

Trypanosoma bubalisi is a newly identified mammalian trypanosome isolated from a freshwater leech (Hirudinaria manillensis) and the water buffalo (Bubalus bubalis) is considered its host. Despite the detailed morphological description of in vitro cultivated forms of this trypanosome, little is known about its ultrastructure. In this study, a detailed ultrastructure of T. bubalisi was characterized using scanning electron microscopy, transmission electron microscopy and focused ion beam-scanning electron microscopy. Two membrane-bound organelles were identified, multivesicular body-like vesicles (ve1) and large lipid-rich vesicles resembling reservosomes (ve2), alongside an extracellular 'beads-on-a-string' structure. These all appear to be associated with endocytosis or secretion. Three-dimensional reconstructions confirmed the organization and distribution of the two membrane-bound organelles. Functional assays using Tomato lectin and Lyso-Tracker demonstrated that ve2 is involved in endocytic uptake and may act as a terminal storage compartment. Compared to the pathogen causing Nagana, Trypanosoma brucei, T. bubalisi showed higher endocytic activity under both 27 °C and 4 °C conditions. These findings suggest that T. bubalisi possesses an active endocytic system and may share conserved mechanisms of material transport and storage with the pathogen causing Chagas' disease, Tryponosoma cruzi.

Crimean-Congo hemorrhagic fever virus (CCHFV) is endemic in >30 countries across Asia, Europe, the Middle East, and Africa. CCHFV causes Crimean-Congo hemorrhagic fever (CCHF), a disease characterized by bleeding and haemoptysis. Since the first diagnosis in 1944, 10,000-15,000 CCHF cases are reported worldwide annually. Around three billion people are at risk of CCHFV infection. The World Health Organization (WHO) announced CCHFV as a priority pathogen in December 2015 due to its widespread and significant healthcare threat. In this review, we track the outbreaks, discuss their origin, and analyze the geographical spread of the CCHFV. We will also discuss reservoirs, the modes of transmission, and the factors influencing CCHFV infection. Preventive measures will be highlighted with emphasis on the potential of artificial intelligence in outbreak prediction and disease surveillance.

Rodents and didelphids are highly diverse and abundant groups that can be found in different ecological niches, including anthropized environments. This can favor a high diversity of zoonotic agents, constituting a complex network of ecological interactions. Nematode parasites can have adaptative flexibility to emerge as new diseases, considering its ecology and diversity, so we aimed to evaluate the functional role of a network interaction between small mammals and nematodes parasites. We used a literature search to construct our network. A total of 52 articles were selected for analysis, from which 80 records of nematodes parasitizing small mammals were retrieved, being 24 species associated with Didelphimorphia and 56 with Rodentia. No nematode species were found parasitizing hosts from both orders. The network exhibited significant modularity (Q = 0.62, P = 0.01), but did not show significant overall specialization (H2 = 0.48, P = 0.23), nor nestedness (WNODF metric = 0.03, P = 0.1), but there is a nestedness patterns both within modules (WNODA = 0.17, P = 0.02) and across the entire network (WNODA = 0.03, P = 0.01). We observed 18 modules all related to the host family. All the hosts roles were classified as either ultraperipheral (81.58 %) or peripheral (18.42 %); and the nematodes were 90 % classified as ultraperipheral, 7.5 % classified as peripheral. Only Aspidodera raillieti and Guerrerostrongylus zetta were classified as provincial hubs. This could highlight the importance of niche sharing by hosts in the cycle transmission of those parasites and how the loss of these key species could result in module fragmentation and compromise system stability, emphasizing their critical role in network architecture and in hotspots of biodiversity and threaten biomes such as Atlantic Forest and Pantanal.

Numerous studies have explored the effects of nanoparticles on Giardia lamblia under in vitro and in vivo conditions, yet the findings remain inconsistent and incomplete due to the limited use of animal models. Green-synthesized copper nanoparticles (GSCNP) have shown broad-spectrum antimicrobial properties against bacteria, fungi, viruses, and parasites, including Leishmania, Echinococcus, and Toxoplasma gondii. This study investigated the anti-giardial efficacy of GSCNP biosynthesized using Rumex vesicarius extract, alone and in combination with metronidazole (MZ), in a murine model of giardiasis. GSCNP were synthesized via a hydroalcoholic extract of R. vesicarius acting as a reducing and capping agent. Treatments with GSCNP (5, 10, and 20 mg/kg), alone or combined with MZ (7.5 or 15 mg/kg), were assessed for cyst load, enzyme and electrolyte profiles, cytokine responses (IL-17, IFN-γ), secretory IgA, and apoptosis-related gene expression. Co-administration of GSCNP (20 mg/kg) with MNZ (7.5 mg/kg) significantly reduced cyst excretion (p < 0.05), achieving complete cyst elimination compared to MZ monotherapies (15 mg/kg = 12.3 cysts; 7.5 mg/kg = 39.6 cysts). This combination normalized serum Na (148.9 mEq/L), K (5.6 mEq/L), Cl (112.7 mEq/L), α-amylase (271.7 mU/mL), and lipase (5.8 mU/mL), while markedly elevating IL-17 (323.6 pg/mL) and IFN-γ (445.7 pg/mL) levels (p < 0.001). Gene expression analysis revealed upregulation of BCL-2 (5.97-fold) and moderate increases in caspase-3 (1.09-fold) and BAX (1.04-fold). These findings demonstrate that GSCNP, particularly in combination with subtherapeutic MZ, enhances anti-giardial efficacy through immunomodulatory and anti-apoptotic mechanisms, supporting their potential as a novel adjunctive therapy for giardiasis.

Protection against malaria induced by the sickle cell trait has been a major example of genetic selection for over half a century. It has been demonstrated that human populations in Africa have acquired a high frequency of haemoglobin S (HbS) allele because the heterozygous genotype (sickle cell trait) provides protection against the severe symptoms of Plasmodium falciparum infection leading to lower parasite numbers and increased clearance of infected cells by the body. However, there is scarcity of data from Chad. This study was conducted to determine the HbS allele frequency, and also to assess the possible protection, in subjects with uncomplicated malaria from N'Djamena, Chad. Dry blood spot samples were collected from patients with uncomplicated Plasmodium falciparum malaria, whose infection was identified by microscopy. To assess HbS allele frequency, human DNA was analyzed by PCR-RFLP. We collected samples from 320 (2/3 females) uncomplicated falciparum malaria cases, among which the allele frequency of the HbS variant allele was 6.72 %. Not significant association with the parasite density was found among the different HbS genotypes. This result confirms a non-negligible frequency of the HbS allele in the study population, providing for the first time data from Chad. The present report provides some information on the sickle prevalence in the study population but can not be interpreted as being representative of the wider population, since the subjects were recruited in a malaria survey.