Objective: To identify centrifugation parameters that maximize platelet recovery while preserving function in canine PRP. Methods: Whole blood from 10 healthy dogs was processed by single-spin (10 min at 800, 1,200, 1,500, or 2,000 rpm) or dual-spin (two-time immediate sequential repetition of the same centrifugation speed for 5 min) using a fixed-angle rotor (radius = 15 cm; 1,500 rpm = 364 × g equivalent). Platelet recovery and white blood cell (WBC) contamination were quantified, and platelet aggregation and dense-granule secretion (2-MeSADP, thrombin), thromboxane (Tx) A2 generation (TxB 2 enzyme-linked immunosorbent assay [ELISA]), protein kinase B (AKT)/extracellular signal-regulated kinase (ERK) phosphorylation (immunoblot), and platelet-derived growth factor (PDGF)-BB release (ELISA) were measured. Results: Among single-spin conditions, PRP obtained at 1,500 rpm/10 min yielded the highest platelet recovery (≈25.1%) with < 1% WBC carryover. This PRP exhibited maximum 2-MeSADP- and thrombin-induced platelet aggregation and secretion, TxA2 generation, AKT/ ERK phosphorylation, and PDGF-BB release, suggesting that optimizing the centrifugal speed is crucial for achieving the optimal PRP collection with the utmost platelet function.Compared with the optimal single-spin condition, dual-spin at 1,500 rpm/5+5 min further increased platelet recovery (≈48.7%) and enhanced functional readouts. Conclusions and Relevance: A dual-spin at 1,500 rpm/5+5 min protocol produces leukocytepoor PRP with superior platelet recovery and function in dogs, supporting more effective and reproducible clinical and research applications.

Malaria elimination and control require prompt and accurate diagnosis for treatment plan. Since microscopy and rapid diagnostic test (RDT) are not sensitive particularly for diagnosing low parasitemia, highly sensitive diagnostic tools are required for accurate treatment. Molecular diagnosis of malaria is commonly carried out by nested polymerase chain reaction (PCR) targeting 18S rRNA gene, while this technique involves long turnaround time and multiple steps leading to false positive results. To overcome these drawbacks, we compared highly sensitive cytochrome oxidase gene-based single-step multiplex reaction with 18S rRNA nested PCR. Cytochrome oxidase (cox) genes of P. falciparum (cox-III) and P. vivax (cox-I) were compared with 18S rRNA gene nested PCR and microscopy. Cox gene multiplex PCR was found to be highly specific and sensitive, enhancing the detection limit of mixed infections. Cox gene multiplex PCR showed a sensitivity of 100% and a specificity of 97%. This approach can be used as an alternative diagnostic method as it offers higher diagnostic performance and is amenable to high throughput scaling up for a larger sample size at low cost.