Ticks are key vectors of some important diseases of humans and animals. Although they are carriers of disease agents, the viability and development of ticks are not harmed by the infectious agents due to their innate immunity. Antimicrobial peptides directly protect hosts against pathogenic agents such as viruses, bacteria, and parasites. Among the identified and characterized antimicrobial peptides, defensins have been considerably well studied. Defensins, which contain intramolecular disulfide bridges between cysteine residues, are commonly found among fungi, plants, invertebrates, and vertebrates. The sequence of the tick hemolymph defensin (HEdefensin) gene from the hard tick Haemaphysalis longicornis was analyzed after identification and cloning from a cDNA library. HEdefensin has a predicted molecular mass of 8.15 kDa and a theoretical isoelectric point of 9.48. Six cysteine residues were also identified in the amino acids. The synthetic HEdefensin peptide only showed antibacterial activity against Gram-positive bacteria such as Micrococcus luteus. A fluorescence propidium iodide exclusion assay also showed that HEdefensin increased the membrane permeability of M. luteus. Additionally, an indirect fluorescent antibody test showed that HEdefensin binds to M. luteus. These results suggested that HEdefensin strongly affects the innate immunity of ticks against Gram-positive bacteria.

Rickettsia and Coxiella burnetii are zoonotic tick-borne pathogens that can cause febrile illnesses with or without other symptoms in humans but may cause subclinical infections in animals. There are only a few reports on the occurrence of these pathogens in cattle and water buffaloes in Southeast Asia, including the Philippines. In this study, molecular detection of Rickettsia spp. and C. burnetii in the blood and Rhipicephalus (Boophilus) microplus ticks of cattle and water buffaloes from five provinces in Luzon Island of the Philippines was done. A total of 620 blood samples of cattle and water buffaloes and 206 tick samples were collected and subjected to DNA extraction. After successful amplification of control genes, nested PCR was performed to detect gltA of Rickettsia and com1 of C. burnetii. No samples were positive for Rickettsia while 10 (cattle – 7, water buffaloes - 3) or 1.6% of blood and 5 or 1.8% of tick samples were C. burnetii-positive. Sequence analysis of the positive amplicons showed 99-100% similarity to reported C. burnetii isolates. This molecular evidence on the occurrence of C. burnetii in Philippine ruminants and cattle ticks and its zoonotic nature should prompt further investigation and surveillance to facilitate its effective control.