Tick-borne diseases (TBD) in dogs have been an emerging issue worldwide. In the Philippines, most reports are in the northern areas, which include Babesia, Ehrlichia, Anaplasma, and Hepatozoon spp. The Mycoplasma spp., a suspected TBD pathogen, has not been reported in Philippine dogs while Hepatozoon spp. has not been previously documented in the southern Philippines. The present study aimed to evaluate the presence of Mycoplasma and Hepatozoon spp. in dogs in Cebu, Philippines. A total of 100 dogs from four veterinary clinics and hospitals were tested for Hepatozoon and Mycoplasma spp. using PCR. The inclusion criteria used were presence or history of ticks, anemia and/or thrombocytopenia. Clinical signs of the dogs were also obtained. There were four and two dogs found positive for Hepatozoon and Mycoplasma spp., respectively using PCR, and two dogs were found positive for Hepatozoon spp. using PBSE. Clinical signs such as lethargy, inappetence, fever, weight loss, and paleness were observed in most of the subjects. This study is the first of Mycoplasma spp. detection in Philippine dogs, and the initial report of Hepatozoon spp. in the southern Philippines.

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The available drugs against piroplasmosis are insufficient and faced with several challenges, such as drug-resistant parasites and toxicity to treated animals. Therefore, the discovery of new drug compounds is necessary for the effective control of babesiosis and theileriosis. Methanolic Peganum harmala seed extract (MPHSE) and ethanolic Artemisia absinthium leaf extract (EAALE) have several medicinal properties. In the present study, the growth-inhibition effects of MPHSE and EAALE were evaluated in vitro and in vivo. The half-maximal inhibitory concentration (IC₅₀) values for MPHSE against Babesia bovis, B. bigemina, B. divergens, B. caballi, and Theileria equi were 24.9 ± 1.2, 77.1 ± 2.3, 61.1 ± 2.9, 80.8 ± 4.1, and 11.3 ± 2.1 μg/mL, respectively. EAALE exhibited IC₅₀ values of 43.3 ± 3.1, 39.2 ± 2.7, 38.5 ±3.7, 50.3 ± 2.1, and 28.2 ± 2.1 μg/mL against B. bovis, B. bigemina, B. divergens, B. caballi, and T. equi, respectively. The toxicity assay on Madin–Darby bovine kidney (MDBK), mouse embryonic fibroblast (NIH/3T3), and human foreskin fibroblast (HFF) cell lines showed that MPHSE affected the viability of MDBK, NIH/3T3, and HFF cell lines with half maximum effective concentration (EC₅₀) values of 611.7± 10.9, 870 ± 22, and ˃1500 μg/mL, respectively, while EAALE exhibited EC₅₀ values of 340.7 ± 8.5, 736.7 ± 9.3, and 1371.5 ± 17.3 μg/mL against MDBK, NIH/3T3, and HFF cell lines, respectively. In the in vivo experiment, MPHSE and EAALE oral treatments at 150 mg/kg inhibited the growth of Babesia microti in mice by 60% and 55.1%, respectively. These findings suggest that MPHSE and EAALE have the potential to be alternative remedies for treating piroplasmosis.

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Drug susceptibility of the Babesia parasite in vitro has been used to assess the anti-babesial activities of new candidates. Recently, the in vitro drug susceptibility assay has been performed by SYBR Green I (SG I) -based fluorescence assay. Herein, we investigated the validity of a fluorescence assay for the shipping process under various culture conditions. The highest fluorescence signal was observed in freeze-thaw of Babesia and Theileria parasites cultures incubated with lysis buffer containing SG I for a half hour. This result indicates the ability of this additional step to lysis of infected RBCs completely with releasing of parasite nucleic acid. Such that, the nucleic acid can interact with SG I stain. All tested drugs showed potential anti-babesial activity and slightly high selective index against the in vitro growth of the Babesia and Theileria parasites. The IC₅₀ values generated using the fluorescence assay with additional freeze-thaw step is comparable with those of IC₅₀ values obtained from the unmodified fluorescence assay and microscopy method.

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Pregnancy-associated malaria is a major health concern in Bududa district. To assess the prevalence of malaria infection in pregnancy in Bududa District; a hospital-based, crosssectional study was carried out. Two hundred fifty two participants were selected using consecutive simple non probability sampling on consenting. Capillary blood samples were collected. Thick and thin smears were made, stained using 3% Giemsa staining technique and examined under the light microscope for malaria parasites and species typing. Data was cleaned and entered using Epi information version 3.5.3, later converted by Microsoft access, excel and word and descriptive data analysed using SPSS. Prevalence of malaria was 8.73 % (22/252) (95% CI 3.8-13.7). Only P. falciparum species was identified and malaria infections were high in primigravid with 16.28% (14/86) than in multigravid (P-value < 0.001). Malaria infection still remains a health challenge among pregnant women with a prevalence of 8.73% (22/252); P. falciparum was the only species identified, primigravid pregnant women are most affected. Routine malaria screening in symptomatic and asymptomatic malaria infections in pregnancy for early diagnoses and prevent transmission is vital. It is crucial to intensify and sensitize mothers and communities about malaria control programmes.

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The resting cyst formation (encystment) is a survival strategy against environmental stressors, and is found in many species of single-celled eukaryotic organisms. The process incorporates cell differentiation accompanied by drastic morphological changes. This study presents observation via fluorescence microscopy on the encystation process of Colpoda cucullus. We confirmed that C. cucullus, which was identified by 18S rRNA analysis, contains lepidosomes, which are cyst-specific cell structures. The existence of these structures in Colpoda species is controversial at present, so we present background information concerning the controversy. Moreover, we reveal that lepidosomes acquire autofluorescence after formation, that is, encysting cells contain lepidosomes lacking autofluorescence while mature cysts contain lepidosomes exhibiting autofluorescence. In addition, we describe the process of formation of nuclear surrounding particles (NSPs), which are cyst-specific cell structures.

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