Cryptosporidiosis remains a significant health concern, particularly in immunocompromised hosts, for whom available treatments may have limited efficacy. This study evaluated the therapeutic effects of geraniol (GR), alone and in combination with nitazoxanide (NZ), in immunosuppressed mice experimentally infected with Cryptosporidium parvum. The mice were treated in groups with varying doses of GR (25 and 50 mg/kg), NZ (100 mg/kg), and their combination for 7 days. Treatment effects were assessed based on fecal oocyst shedding, intestinal antioxidant enzyme activities, Th1-associated cytokine gene expression, and selected hepatic and renal biochemical markers. GR significantly reduced oocyst shedding and improved antioxidant and immune-related parameters compared with untreated infected/immunosuppressed mice (p<0.01). The combination of GR with NZ produced a greater reduction in oocyst shedding than either treatment alone, with the GR 50 mg/kg + NZ 100 mg/kg regimen producing the greatest overall effect (p<0.001). The combined treatment was also associated with restoration of CAT and SOD activities and increased expression of IL-12 and IFN-γ (p<0.001). No evident treatment-related hepatic or renal toxicity was observed based on the evaluated biochemical markers. Overall, these findings provide preliminary experimental evidence that GR, particularly in combination with NZ, may warrant further investigation as an adjunctive approach for cryptosporidiosis. However, validation in additional animal models and ultimately well-designed clinical studies is required.

[This corrects the article DOI: 10.3389/fphar.2024.1491249.].

Giardiasis remains a significant global health issue, and its treatment is increasingly hampered by the emergence of drug resistance and cross-resistance between nitroimidazoles and nitazoxanide. In this study, we evaluated the in vitro antigiardial activity, cytotoxicity, and potential mechanisms of action of seven structural analogs (CLB 1-7) derived from a hybrid structure combining nitazoxanide and clofibric acid. Among them, compounds CLB-3 and CLB-5 emerged as the most potent candidates, demonstrating exceptional efficacy against a nitazoxanide-resistant (NTZr) strain of Giardia lamblia, with potencies up to 43 and 60 times higher than that of the reference drug, respectively. Furthermore, both compounds demonstrated high selectivity for the parasite, exhibiting low cytotoxicity in human Caco-2 cells. To investigate the mechanism of action, functional biochemical assays revealed that treatment with these derivatives significantly reduced the intracellular ATP levels and altered the ADP/ATP ratio, indicating a direct impact on the parasite's energy metabolism. These functional findings correlate closely with RT-qPCR transcriptional analyses and in silico molecular dynamics simulations; collectively, the data suggest a multi-target mechanism of action that may involve the disruption of energy homeostasis and the possible inhibition of key enzymes in the glycolytic pathway (PFOR, G6PD::6PGL, PK, and PPDK). In summary, these results identify compounds CLB-3 and CLB-5 as highly promising chemotherapeutic candidates for combating refractory giardiasis.

Advanced oxidation processes can efficiently degrade organic micropollutants in wastewater. However, chemical assessment alone does not ensure that treated effluents are biologically compatible for reuse, as residual contaminants and transformation products may have toxic effects. Thus, this study evaluated the efficiency of the previously optimized H2O2/S2O8[2‒]/UVC process for mitigating the ecotoxicological effects of municipal wastewater enriched with colchicine, nitazoxanide, and sulfamethoxazole, considering its potential application for agricultural reuse. The approach was based on multiple ecotoxicological parameters at different biological levels: oxidative stress in Eisenia andrei, phytotoxicity in Lactuca sativa, cytotoxicity and genotoxicity in Allium cepa, and growth inhibition of environmentally relevant microorganisms (Azospirillum brasilense and Saccharomyces cerevisiae). The microcontaminant-enriched effluent exhibited substantial genotoxicity (21%) and severe phytotoxicity, highlighting the risks of reusing effluent without advanced treatment. The H2O2/S2O8[2‒]/UVC process significantly reduced residual toxicity, lowering genotoxicity to 6% and increasing the plant growth index to values above 80%. This process did not induce cytotoxic effects or adversely affect beneficial microorganisms. These results suggest that the process can reduce the toxicity and biological impacts of microcontaminants, which could mitigate the ecotoxicological risks of wastewater intended for non-potable reuse, including in agriculture.

Cryptosporidiosis is a global health concern that causes mortality and morbidity in children and immunocompromised patients. The currently available anticryptosporidial agent, nitazoxanide (NTZ), has limited efficacy in cases of low immunity. This study aimed to evaluate the curative and protective effects of fenugreek (FNK) extract-loaded zinc oxide nanoparticles (ZnO NPs) against lung damage and intestinal dysplasia induced by experimental cryptosporidiosis. Eighty mice were divided into eight groups (ten mice per subgroup): non-infected non-treated, infected non-treated, infected receiving ZnO NPs, infected receiving NTZ, infected receiving FNK extract, infected receiving FNK- loaded ZnO NPs, infected receiving NTZ-loaded ZnO NPs, and infected receiving both NTZ and FNK extract-loaded ZnO NPs. Assessment parameters included oocyst shedding counts, histopathological and cyclin D1 immunohistochemical analysis in intestinal and lung tissues, evaluation levels of serum IL-17 levels and tissue oxidative stress markers (MDA, NO, SOD, and CAT), and expression of apoptotic gene markers (BCL-2 and caspase-3). The results demonstrated that treatment with ZnO NPs loaded NTZ and FNK extract yielded the most significant reduction in oocyst shedding, along with marked improvement in lung damage and intestinal dysplasia. Furthermore, this treatment was associated with negative cyclin D1 expression, near-normal levels of IL-17 and oxidative stress markers, and normalized BCL-2 and caspase-3 expression. Thus, the curative and protective activity of NTZ in cryptosporidiosis was enhanced when combined with FNK extract and loaded onto ZnO NPs.

The increasing prevalence of multidrug-resistant (MDR) and extensively drug-resistant (XDR)Mycobacterium tuberculosis has emerged as a major global health concern. Advanced molecular diagnostics, including rapid assays, have improved resistance detection; however, resource-limited settings remain underserved. Nanotechnology-based drug delivery systems, including polymeric carriers, solid lipid nanoparticles, and silver nanoparticles, can provide targeted delivery, prolonged release, and improved bioavailability. Drug repurposing strategies using existing drugs like linezolid, celecoxib, and nitazoxanide also show a promising option in combating drug-resistant TB while reducing treatment duration. Host-directed therapies (HDTs) and natural products offer complementary strategies by enhancing host immune responses and providing multitarget antimycobacterial effects. This review emphasizes the drug resistance mechanisms and advancements in novel theranostics. Despite significant progress, challenges related to cost-effective scalability, accessibility of advanced diagnostics, and optimization of therapeutic systems continue to limit clinical translation. Future research must prioritize multidisciplinary collaborations to accelerate drug development, enhance diagnostic capabilities, and expand nanotechnology-based solutions to address the TB epidemic effectively.

Cryptosporidiosis is a significant zoonotic parasite disease caused by Cryptosporidium parvum, impacting both humans and animals, with little therapeutic alternatives available. This work examined the phytochemical composition, antioxidant capacity, in silico antiparasitic potential, and in vitro oocysticidal activity of methanolic Pleurotus ostreatus extract (POE). Fruiting bodies extract was analyzed for phytochemical constituents. The total amounts of phenolics, flavonoids, tannins, and saponins were quantified spectrophotometrically, while antioxidant activity was evaluated by the reducing power test. Gas chromatography-mass spectrometry (GC-MS) chemical profiling revealed 18 identified compounds. Molecular docking was performed on three validated targets of C. parvum: N-myristoyltransferase (NMT), calcium-dependent protein kinase 1 (CDPK1), and lysyl-tRNA synthetase (KRS) to assess possible antiparasitic interactions. Several sterol derivatives produced favorable predicted binding scores, including Ergost-7-en-3-ol (3β) and ergosterol toward NMT (- 9.87 and - 9.28 kcal/mol, respectively), compared with nitazoxanide under the docking conditions. Interaction profiling identified possible ligand-target interactions with proteins involved in protein lipidation, Ca[2+]-dependent signaling, and translation pathways, but these mechanisms remain computational hypotheses pending biochemical validation. ADMET predictions suggested minimal systemic absorption for sterols, which may favor gut-localized activity and prolonged intestinal exposure against C. parvum while minimizing systemic toxicity. In vitro studies revealed a concentration- and time-dependent decrease in C. parvum oocyst numbers and viability. At a concentration of 1000 µg/mL, POE diminished oocyst counts by nearly 71% after 48 h and lowered viability to 37.41%, demonstrating greater reduction of oocyst count and viability than nitazoxanide under comparable conditions with LC50 of 460 µg/mL. These findings collectively underscored P. ostreatus as a potential natural antiparasitic agent, indicating that its sterol-rich extract may demonstrate oocysticidal activity via multi-target molecular interactions and direct impacts on oocyst integrity.