رسائل دكتوراة
English
2025
preparation and Characterization of Phyto-Copper Nanoparticles Using Phragmites australis and Evaluation of Their Biological and Veterinary Applications
Samir Derouiche , Frahtia Ahlem, Janetta Niemann
كلية علوم الطبيعة والحياة-جامعة الشهيد حمة لخضر - الوادي · الجزائر
This study aims to evaluate the synthesis, characterization, and biological activities—including antioxidant, anti-inflammatory, antibacterial, and antitumor properties—of coppernanoparticles (CuNPs) biosynthesized using aqueous extracts from the leaves and rhizomes ofPhragmitesaustralis. Furthermore, the study assesses the in vivo safety, toxicity, and potentialbiovaccination effect of these CuNPs in a broiler chicken model, with a focus on growthperformance, survival rate, and physiological biomarkers. The plant extracts have previouslyundergone both quantitative and qualitative phytochemical analysis. To confirm their size, shape,and stability CuNPs were characterized using various analytical techniques, such as UV-Visspectroscopy, FTIR, SEM, TEM, XRD, and Zeta potential. In vitro assessments were conductedto evaluate the antioxidant and anti-inflammatory properties of both plant extracts and plant-basedNPs using standard protocols. The investigation of antibacterial efficacy was performed on bothGram-positive and Gram-negative bacterial strains utilizing the disk diffusion method, whereinthe copper nanoparticles synthesized from P. australis exhibited intermediate effectiveness. Theanti-tumor activity of the plant-based CuNPs was assessed using the MTT assay. Following the invitro studies, in vivo experiments were performed in two phases: the first aimed to determine theacute toxicity of the NPs on 15 broiler chickens (Arbor Acres) divided into 5 groups 3 in each:control, 25 mg/Kg BW (Leaf-based CuNPs), 25 mg/Kg BW (Rhizome-based CuNPs), 50 mg/KgBW (Leaf-based CuNPs), and 50 mg/Kg BW (Rhizome-based CuNPs), while the second phaseevaluated the biovaccination effect of the NPs on 100 one-day-old broiler chicks that wererandomly divided into five groups (n=20). The first group was injected into the breast muscle, onday 4, with 5mg/Kg BW of leaf-based CuNPs, the second group with 5mg/Kg BW of rhizomebasedCuNPs,thethirdgroupwithamixtureofbothsynthesizedCuNPs(Leafandrhizome-basedCuNPs),thefourthGroupwasleftwithoutanyinjectionandthefifthgroupreceivedthestandardvaccinationandmedication.The three first groups were re-injected after 10 days with the samedose of the plant-based CuNPs, and after 10 days with a double dose. Four birds from each groupwere sacrificed 4 days after the last injection. Various hematological, biochemical, and oxidativestress parameters were assessed in both study phases. Liver histology was studied as well. Resultsof the phytochemical analysis of the plant extracts demonstrate their richness of different activecompounds: phenols, flavonoids, and terpenoids which may explain the important anti-inflammatory activity revealed according to hemolysis assay results. Regarding thecharacterization of P. australis-based Cu NPs, the UV-Vis absorption spectra indicated prominentpeaks at 312 nm and 300 nm for copper nanoparticles derived from leaf and rhizome sources,respectively. The crystalline structure of the phyto-synthesized Cu NPs was substantiated throughX-ray diffraction (XRD) analysis, revealing an estimated grain size of 18.06 nm for leaf-based CuNPs and 18.24 nm for rhizome-based counterparts. Furthermore, the average diameter of Cu NPswas calculated to be 13.22 nm for those derived from leaf sources and 19.76 nm for thoseoriginating from rhizome sources. Additionally, the synthesized nanoparticles demonstratedsignificant antioxidant properties, as evidenced by IC50values of 4.62 µg/ml and 78.99 µg/ml inDPPH and FRAP assays, respectively, for leaf-derived Cu NPs.The IC50values associated withanti-inflammatory activity in both assessments were notably pronounced, underscoring theconsiderable potential of green-synthesized Cu NPs. The cytotoxic effects of PhragmitesaustralismediatedCu NPs against MCF-7 cell lines exhibited a dose-dependent response at minimalconcentrations: 3.53 µg/ml for leaf-derived Cu NPs and 2.28 µg/ml for those derived fromrhizomes. Moreover, the acute toxicity findings show some changes in biochemical,hematological, and oxidative stress markers without signaling death. On the other hand, theoutcomes of the second phase of the in vivo study indicate that the injections of Phragmitesaustralis-based Cu NPs had no detrimental impact on the growth and survival of broiler chicks.Their weight gain and overall performance remained comparable to those of chickens that receivedvaccination, medication, and supplemental nutrition. Notably, the mortality rate in thebiovaccinated groups was approximately 8%, whereas it exceeded 50% in the untreated group.This suggests that these nanoparticles may not only support healthy development but alsosignificantly enhance survival rates in broiler chickens. Regarding the final weight at the end ofthe rearing period, the chickens in the group that received the green nanoparticles treatmentexhibited a weight comparable to those in the conventional treatment group. More notably, thegroup that received a mixture of P. australis-based copper nanoparticles surpassed the weight ofthe conventional group, reaching an average of 3,2 kg, compared to 3,09 kg in the conventionallytreated group. These findings highlight the potential of P. australis-based copper nanoparticlestreatments, especially when combined, to positively influence growth and weight gain in broilerchickens. This study points out the valuable advantages of copper nanoparticles (Cu NPs) as aharmless, cost-effective, and efficient product in broiler chicken farming. Additional research isimperative to examine the fundamental mechanisms and long-term consequences of integratingCu NPs into broiler farming, to optimize production outcomes and enhance the performance of thebirds.
