For the first time, Boliwong, an indigenous community in the Philippines, was surveyed for the prevalence of Cryptosporidium from April to December 2017. Cryptosporidium oocysts were detected in samples from the river, creek, and water pumps via immunomagnetic separation techniques, and from human and animal concentrated faecal samples using the modified Ziehl-Neelsen technique. Seven of the 24 water samples (29.2%) were positive for Cryptosporidium, with the highest concentration (0.8 oocyst/L) detected in the creek. Of 35 fecal samples from different animal groups, 8 (21.6%) were positive for Cryptosporidium oocysts. The highest intensity of oocyst shedding was detected in dogs (χ2 =8.00). Of the 137 human fecal samples, 39 (28.5%) were infected with Cryptosporidium. In this study, 3 risk factors were found to be associated with infection: (1) location (crude odds ratio [cOR], 16.39; 95% confidence interval [CI], 2.11 to 127.41; p=0.008), (2) drinking water from the natural spring (cOR, 0.29; 95% CI, 0.11 to 0.82; p<0.05), and (3) using an open pit as a sanitary toilet facility (cOR, 2.44; 95% CI, 1.14 to 5.20; p<0.05). When the cOR was adjusted, using an open pit as a sanitary toilet facility remained a significant risk factor of infection (adjusted OR, 0.41; 95% CI, 0.19 to 0.90; p<0.05). There is a potentially emerging Cryptosporidium zoonosis in Boliwong, Lagawe, Philippines. It is recommended that the toilet facilities and the water system in the community be rehabilitated to avoid any possible disease outbreak. Health education is also needed in the community to maintain proper hygiene and sanitation practices.

Leptospirosis remains a public health concern in the Philippines. Conventional diagnostic methods, including the microscopic agglutination test (MAT) and qPCR, are routinely used for outbreak response and surveillance. However, these methods often yield discordant results due to cross-reactivity, limited sensitivity, or lack of species-level resolution. To address these diagnostic gaps, this study optimized the Boonsilp 16S rRNA PCR assay and applied Sanger sequencing for accurate species identification of Leptospira in 92 archived DNA samples collected between 2018 and 2020. The sensitivity and specificity of the optimized assay were compared with those of MAT and qPCR. Species-level identification was confirmed via sequencing, and a phylogenetic tree was constructed. Among the 92 samples, 46 (50.0%) tested positive by qPCR, 39 (42.4%) by MAT, and 67 (72.8%) by at least one of the two methods. The optimized Boonsilp assay detected Leptospira in 23 samples (25.0%), of which 22 were also qPCR positive. Twenty-one samples were confirmed as L. interrogans, one as L. borgpetersenii, and one as an unclassified Leptospira species. One sample undetected by both MAT and qPCR tested positive using the optimized assay. Compared to the composite reference, the Boonsilp assay showed 32.8% sensitivity and 96.0% specificity. Phylogenetic analysis revealed multiple L. interrogans strains, including those closely related to reference sequences of Copenhageni, Manilae, and Canicola. While the optimized Boonsilp PCR assay demonstrates diagnostic value as an adjunct molecular tool to qPCR and MAT supporting species-level identification during outbreak surveillance, this warrants further validation in freshly isolated DNA samples.

Background and Aim: Acanthamoeba spp. is free-living protozoa capable of causing severe infections, notably Acanthamoeba keratitis, which is difficult to manage due to cyst resistance and the cytotoxicity of current treatments. Plant-derived com­pounds represent a promising alternative strategy. This study investigated the amoebicidal, anti-adhesive, and cytotoxic properties of Mangifera indica L. (mango) leaf extract against ocularly relevant Acanthamoeba spp. Materials and Methods: Crude ethanolic leaf extract of M. indica was prepared and evaluated against Acanthamoeba polyphaga American Type Culture Collection (ATCC) 30461 and Acanthamoeba castellanii ATCC 50739. Minimum inhibitory concentration (MIC) and minimum parasiticidal concentration were determined for trophozoites and cysts. Morphological changes were analyzed by scanning electron microscopy (SEM). Anti-adhesion assays were conducted using polystyrene surfaces, with a commercial multipurpose contact lens (CL) solution as a control. Cytotoxicity was tested in Vero cells using the 3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide assay to establish the minimum cytotoxic concentration. Results: The extract inhibited trophozoite growth at 2 mg/mL and demonstrated cysticidal activity at 4 mg/mL for A. polyph­aga and 32 mg/mL for A. castellanii. SEM revealed disruption of trophozoite morphology, loss of acanthopodia, and sur­face perforations in cysts. At MIC levels, adhesion was reduced by >70%, and even at 1/8 MIC, inhibition remained above 50%, comparable to a commercial multipurpose solution. Cytotoxicity assessment showed >80% Vero cell viability at 0.125 mg/mL, indicating a favorable therapeutic window. Conclusion: This is the first report demonstrating amoebicidal and anti-adhesive effects of M. indica L. leaf extract against ocular Acanthamoeba species. The dual trophozoiticidal and anti-adhesive actions, combined with low cytotoxicity, high­light its potential for development as a plant-based therapeutic agent, particularly in ocular formulations or CL disinfectants. Future work should focus on phytochemical isolation, mechanistic studies, and novel delivery systems to enhance efficacy and safety. Keywords: Acanthamoeba, amoebicidal activity, anti-adhesion, cytotoxicity, Mangifera indica L., ocular infection.