Nano magnetic-based ELISA and nano magnetic-based latex agglutination test for diagnosis of experimental trichinellosis.

Human trichinellosis is a worldwide foodborne public health threat. Detecting circulating antigens of Trichinella spiralis "T. spiralis" allows for an early diagnosis before larval encystation develops in skeletal muscles. For the first time, the present study aimed to formulate an effective nanomagnetic beads based-ELISA and -latex agglutination test (NMB-ELISA and NMB-LAT) to recognize T. spiralis adult worm crude extract antigen (AWCEA) in sera of experimentally infected mice. The study included thirty-eight mice classified into 3 groups; T. spiralis-infected group (GI) which was euthanized 6, 8, 10, 12, 14 days post-infection (dpi), other parasitic infections group (GII) and healthy control group (GIII). Rabbit anti-T. spiralis polyclonal antibodies (pAbs) were utilized to detect AWCEA in serum samples by sandwich ELISA, NMB-ELISA, and NMB-LAT. Using NMB-ELISA, AWCEA was detected in sera collected at 6 and 8 dpi, with a sensitivity of 50% and 75%, respectively, and a specificity of 100%. Whereas, sandwich ELISA and NMB-LAT couldn't detect the antigen at the same time intervals. Both ELISA formats were able to detect the antigen in samples collected at 10, 12, and 14 dpi with a sensitivity of 100% for NMB-ELISA and 25%, 75%, and 100% respectively, for sandwich-ELISA. Yet, NMB-LAT couldn't detect AWCEA until 12 dpi with a sensitivity of 50% and specificity of 75%. In conclusion, NMB-ELISA is a promising sensitive tool for early and specific diagnosis of acute trichinellosis. The use of NMB-LAT could be a helpful screening procedure in field surveys.

The effect of Nigella sativa oil- and wheat germ oil-loaded metal organic frameworks on chronic murine toxoplasmosis.

Treatment of chronic toxoplasmosis is challenging as the available drugs are effective only in the acute stage. Therefore, the current study aimed to investigate Nigella sativa oil (NSO) and wheat germ oil (WGO) loaded on copper-benzene tricarboxylic acid metal organic framework (Cu-BTC MOF) for treating chronic toxoplasmosis in a murine model. Eighty mice were divided into 8 groups (G); uninfected untreated negative control (GI), infected untreated positive control (GII), infected and treated with: Spiramycin (GIII), Spiramycin@Cu-BTC (GIV), Cu-BTC (GV), WGO@Cu-BTC (GVI), NSO@Cu-BTC (GVII) and combined WGO+NSO@Cu-BTC (GVIII). The infected groups were orally inoculated with 10 Toxoplasma gondii Me49 strain cysts/mouse. All drugs were orally administered for 14 consecutive days starting 8 weeks post-infection (wpi). The therapeutic efficacy was evaluated by parasitological (survival rate of mice and brain cyst burden) and histopathological (brain, liver, kidney, eye) parameters. At the end of 2-weeks therapy, the highest therapeutic outcome was achieved with GVII and GVIII exhibiting 100% survival, 64.3% and 51.4% reduction of brain cysts, and an apparent amendment of pathological insults. In the next place was GVI with 90% survival, 49.5% reduction of cysts and marked amelioration of pathological lesions. Meanwhile, GIII and GIV showed 80% survival, 42.4% and 41.8% reduction of cysts as well as minimal to moderate alleviation of tissue damage. The lowest effect was obtained with GV resulting in 70% survival and 24.4% reduction of cysts. The current results support the assertion that the new metal-based nanocomposites can be promising remedies of chronic toxoplasmosis particularly if conjugated with natural herbal extracts as NSO and WGO.

Efficient therapeutic effect of Nigella sativa aqueous extract and chitosan nanoparticles against experimentally induced Acanthamoeba keratitis.

Acanthamoeba keratitis (AK) is a devastating, painful corneal infection, which may lead to loss of vision. The development of resistance and failure of the currently used drugs represent a therapeutic predicament. Thus, novel therapies with lethal effects on resistant Acanthamoeba are necessary to combat AK. In the present study, the curative effect of Nigella sativa aqueous extract (N. sativa) and chitosan nanoparticles (nCs) and both agents combined were assessed in experimentally induced AK. All inoculated corneas developed varying grades of AK. The study medications were applied on the 5th day postinoculation and were evaluated by clinical examination of the cornea and cultivation of corneal scraps. On the 10th day posttreatment, a 100% cure of AK was obtained with nCs (100 µg/ml) in grades 1 and 2 of corneal opacity as well as with N. sativa 60 mg/ml-nCs 100 µg/ml in grades 1, 2, and 3 of corneal opacity, highlighting a possible synergistic effect. On the 15th day posttreatment, a 100% cure was reached with N. sativa aqueous extract (60 mg/ml). Moreover, on the 20th day posttreatment, N. sativa (30 mg/ml) provided a cure rate of 87.5%, while nCs (50 µg/ml) as well as N. sativa 30 mg/ml-nCs 50 µg/ml yielded a cure rate of 75%; the lowest percentage of cure (25%) was obtained with chlorhexidine (0.02%), showing a non-significant difference compared to the parasite control. The clinical outcomes were in agreement with the results of corneal scrap cultivation. The results of the present study demonstrate the effectiveness of N. sativa aqueous extract and nCs (singly or combined) when used against AK, and these agents show potential for the development of new, effective, and safe therapeutic alternatives.