Toxoplasmosis, a parasitic disease resulting from Toxoplasma gondii infection, remains prevalent worldwide, and causes great harm to immunodepressed patients, pregnant women and newborns. Although various molecular approaches to detect T. gondii infection are available, they are either costly or technically complex. This study aimed at developing a rapid visual detection assay using recombinase-aided amplification (RAA) and lateral flow dipstick (LFD) coupled with CRISPR-Cas13a fluorescence (RAA-Cas13a-LFD) to detect T. gondii. The RAA-Cas13a-LFD assay was performed in an incubator block at 37 °C within 2 h, and the amplification results were visualized and determined through LFD by the naked eye. The detection limit was 1 × 10⁻⁶ ng/μL by our developed RAA-Cas13a-LFD protocol, 100-fold higher than that by qPCR assay (1 × 10⁻⁸ ng/μL). No cross-reaction occurred either with the DNA of human blood or Ascaris lumbricoides, Digramma interrupta, Entamoeba coli, Fasciola gigantica, Plasmodium vivax, Schistosoma japonicum, Taenia solium, and Trichinella spiralis, and the positive rate by RAA-Cas13a-LFD assay was identical to that by qPCR assay (1.50% vs. 1.50%) in detecting T. gondii infection in the unknown blood samples obtained from clinical settings. Our findings demonstrate that this RAA-Cas13a-LFD assay is not only rapid, sensitive, and specific and allows direct visualization by the naked eye, but also eliminates sophisticated and costly equipment. More importantly, this technique can be applied to on-site surveillance of T. gondii.

Dust in the home environment is thought to be a potential trigger for increasing allergic diseases, such as allergic rash, rhinitis, asthma, and other conditions, associated with dust mites. To verify the status of dust mite prevalence in indoor surroundings, we collected 189 dust samples from the air conditioner filters (n = 75) and floors (n = 114) of households, schools, and hotels in the Anhui area, China. All samples were measured for dust mite breeding rate and breeding density under light microscopy and analyzed for dust mite species Dermatophagoides farinae 1 (Der f 1) and Dermatophagoides pteronyssinus 1 (Der p 1) allergen using enzyme-linked immunosorbent assay (ELISA). The dust mite breeding rates were 34.67% (26/75) and 20.18% (23/114), respectively, in the dust samples from the floor and air conditioning filters. The breeding density was the highest in households (10/g), followed by schools (9/g) and hotels (4/g). ELISA indicated that the allergen threshold (2.0 µg/g dust) of Der f 1 was exceeded in only two samples and Der p 1 in one sample. Additionally, a questionnaire was used to investigate the health knowledge on allergic diseases involved in indoor facilities, finding that most allergy sufferers were aware that indoor dust might be responsible for their conditions. The findings suggest that regular maintenance of indoor hygiene and cleaning of air-conditioning filters should reduce the risks of exposure to indoor allergens.

Toxoplasma gondii is an obligate parasitic protozoon that transmits to animals and humans via ingested food. Cats that act as T. gondii’s final hosts play a critical role in T. gondii transmission by shedding millions of oocysts. Timely diagnosis of infected cats is essential for preventing toxoplasmosis because oocysts are a putative T. gondii source in epidemiology. We developed a new visual LAMP assay targeting the B1 gene to analyze single oocysts in cat feces in this study. The amplification result could be visually estimated based on the color change. LAMP assay analytical sensitivity was 10¹ copies/µL for the B1 gene plasmid, which was tenfold better than the PCR reaction. There were no cross-reactions with other parasites. The LAMP assay can detect a single T. gondii oocyst in 200 mg of cat feces. The LAMP assay detected a single oocyst in 200 mg cat feces at a higher rate than the PCR assay (83.3% vs. 50.0%).

Moutan Cortex essential oil (MCEO) is considered to be a promising botanical insecticide. However, like most oils, MECO has several limitations, including instability and poor solubility. Nanoencapsulation technology is an excellent strategy for stabilizing essential oils because of its controlled release, enhanced efficacy, and strengthened biological activity. The present study investigated the acaricidal efficacy of pure MCEO and its encapsulated nanoemulsion (NE) and mesoporous silica nanoparticles (MSNs) against the house dust mite Dermatophagoides farinae using contact bioassays, fumigant bioassays, repellent bioassays, and the observation of toxic symptoms. MCEO-MSNs obtained in the study successfully encapsulated MCEO with an encapsulation efficiency of 63.83%. The acaricidal mortality experiments revealed that MCEO-NE and MCEO-MSN showed more significant toxicity against D. farinae than did pure MCEO. The nanomaterials showed better larvicidal and nymphicidal activities than pure MCEO at a high concentration (12-h LC₉₀). Notably, the repellent effect experiment showed that MCEO-NE and MCEO-MSN had long-term and stable repellent effects on D. farinae, indicating the sustained release and persistence of the nanomaterials. More toxicity symptoms were observed in the IM-type group than in the KD-type group, suggesting that the MCEO nanoparticles have adverse effects on the respiratory system. Nanomaterials and MCEO promoted superoxide dismutase (SOD) activity and inhibited acetylcholinesterase (AChE) activity in D. farinae. In addition, the binding sites of paeonol to SOD and AChE were found through molecular docking. These findings demonstrate the potential of MCEO as a biological acaricide, which merits further investigation.