Elucidating the surface macroplastic load, types and distribution in mangrove areas around Cebu Island, Philippines and its policy implications.

The Philippines is identified as one of the major marine plastic litter polluters in the world with a discharge of approximately 0.75 million tons of marine plastic debris per year. However, the extent of the plastic problem is yet to be defined systematically because of limited research. Thus, this study aims to quantify plastic litter occurrence in mangrove areas as they function as sinks for plastic litter due to their inherent nature of trapping plastics. To define the extent of marine plastic pollution on an island scale, mangrove areas in 14 municipalities around Cebu Island were sampled, with 3 to 9 transects in each site depending on the length of coastline covered by mangroves. Sampling and characterization of both plastics and the mangrove ecosystem was performed in three locations along the transect - landward, middle, and seaward. A total of 4501 plastic items were sampled throughout the study sites with an average of 1.29 ± 0.67 items/m2 (18.07 ± 8.79 g/m2). The average distribution of plastic loads were 2.68 ± 1.9 items/m2 (38.52 ± 25.35 g/m2), 0.27 ± 0.10 items/m2 (6.65 ± 4.67 g/m2), and 0.94 ± 0.61 items/m2 (9.04 ± 4.28 g/m2) for the landward, middle, and seaward locations, respectively. The most frequent plastic types found were i) packaging, ii) plastic bags and iii) plastic fragments. The plastic loads and types suggest that most plastic wastes trapped in mangroves come from the nearby communities. Fishing-related plastics originated from the sea and were transported across the mangrove breadth. The findings confirm that mangroves are major traps of plastic litter that might adversely affect the marine ecosystem. The study underscores the urgent need for waste mitigation measures, including education, community engagement, infrastructure, technological solutions and supporting policies.

Phenolic acids and flavonoids from Salvia plebeia and HPLC-UV profiling of four Salvia species.

We isolated and purified phenolic acids and flavonoids from the ethanolic extract of Salvia plebeia using silica gel and a Sephadex LH-20 column chromatography. Spectroscopy revealed the isolated compounds were caffeic acid, rosmarinic acid, hispidulin, luteolin, jaceosidin, nepitrin, homoplantaginin, 6-hydroxyluteolin 7-O-glucoside, 6-methoxynaringenin 7-O-glucoside, naasanone, and cosmosiin. Quantitative analyses, using high-performance liquid chromatography coupled with UV (HPLC-UV), revealed that the major flavonoid from S. plebeia was 6-hydroxyluteolin 7-O-glucoside (100.63 mg/g) and the most abundant phenolic acid was rosmarinic acid (47.73 mg/g). Furthermore, among four other Salvia species, S. officinalis contained the highest overall phenolic acid and flavonoid level but these were still lower than S. plebeia. These results can help assess the potential of phenolic acids and flavonoids as potent sources of pharmacological ingredients from different Salvia species extracts.

Molecular Detection and Characterization of Borrelia garinii (Spirochaetales: Borreliaceae) in Ixodes nipponensis (Ixodida: Ixodidae) Parasitizing a Dog in Korea.

The present study aimed to detect and characterize Borrelia spp. in ticks attached to dogs in Korea. Overall, 562 ticks (276 pools) attached to dogs were collected and tested for Borrelia infection by PCR targeting the 5S-23S rRNA intergenic spacer region (rrf-rrl). One tick larva (pool level, 0.4%; individual level, 0.2%) was confirmed by sequencing Borrelia garinii, a zoonotic pathogen. For molecular characterization, the outer surface protein A (ospA) and flagellin genes were analyzed. Phylogenetic ospA analysis distinguished B. garinii from B. bavariensis, which has been recently identified as a novel Borrelia species. On the other hand, phylogenetic analysis showed that single gene analysis involving rrf-rrl or flagellin was not sufficient to differentiate B. garinii from B. bavariensis. In addition, the B. garinii-infected tick was identified as Ixodes nipponensis by sequencing according to mitochondrial 16S rRNA and the second transcribed spacer region. To our knowledge, this is the first study to report the molecular detection of B. garinii in I. nipponensis parasitizing a dog in Korea. Continuous monitoring of tick-borne pathogens in ticks attached to animals is required to avoid disease distribution and possible transmission to humans.

Differential identification of Anaplasma in cattle and potential of cattle to serve as reservoirs of Anaplasma capra, an emerging tick-borne zoonotic pathogen.

Bovine anaplasmosis is a tick-borne, infectious, non-contagious disease caused by Anaplasma marginale, A. centrale, A. bovis, and zoonotic A. phagocytophilum. Recently, Anaplasma capra detected in goats was identified as a novel zoonotic pathogen. To determine whether A. capra can infect bovines, we used PCR to differentially diagnose Anaplasma spp. in 1219 South Korean cattle by performing multilocus gene typing and restriction fragment length polymorphism (RFLP). Nucleotide sequencing and phylogenetic analysis detected the 16S rRNA gene of A. bovis and four genes from A. capra in 12 (1.0%) and five (0.4%) cattle, respectively. Supplementary discrimination between A. bovis and A. capra was accomplished by RFLP. The 16S rRNA, msp4, groEL, and gltA genes of A. capra identified in this study had much lower degrees of identity to those in A. centrale and other Anaplasma spp. A. phagocytophilum was not detected in any of the tested cattle. Although the prevalence was low, this study suggests the potential of cattle to serve as reservoirs of A. capra. Thus, further studies are needed to clarify the pathogenesis of A. capra in cattle and its possible involvement in transmission to humans.

Eimeria species in cattle with diarrhoea in the Republic of Korea regarding age, season and nature of diarrhoea.

This study evaluated the prevalence of Eimeria species, particularly E bovis, E zuernii and E auburnensis that are pathogenic to cattle, in faecal samples collected from cattle with diarrhoea reared in the Republic of Korea by using microscopy and PCR. In addition, the prevalence of Eimeria species was analysed according to age, type of cattle, region, season and nature of diarrhoea. Overall, Eimeria species were identified in 279 of the 1261 (22.1 per cent) faecal samples through microscopy, and statistical analysis revealed a lower prevalence in calves aged than three weeks or less and higher prevalence in cattle with haemorrhagic diarrhoea. Of the 279 microscopy-positive samples, E bovis, E zuernii and E auburnensis were identified in 100 (7.9 per cent), 83 (6.6 per cent) and 27 (2.1 per cent) faecal samples, respectively, by using PCR. To the authors' knowledge, this study is the first to apply PCR for epizootiology of bovine coccidiosis.

The effects of contemporary selection and dispersal limitation on the community assembly of acidophilic microalgae.

Extremophilic microalgae are primary producers in acidic habitats, such as volcanic sites and acid mine drainages, and play a central role in biogeochemical cycles. Yet, basic knowledge about their species composition and community assembly is lacking. Here, we begin to fill this knowledge gap by performing the first large-scale survey of microalgal diversity in acidic geothermal sites across the West Pacific Island Chain. We collected 72 environmental samples in 12 geothermal sites, measured temperature and pH, and performed rbcL amplicon-based 454 pyrosequencing. Using these data, we estimated the diversity of microalgal species, and then examined the relative contribution of contemporary selection (i.e., local environmental variables) and dispersal limitation on the assembly of these communities. A species delimitation analysis uncovered seven major microalgae (four red, two green, and one diatom) and higher species diversity than previously appreciated. A distance-based redundancy analysis with variation partitioning revealed that dispersal limitation has a greater influence on the community assembly of microalgae than contemporary selection. Consistent with this finding, community similarity among the sampled sites decayed more quickly over geographical distance than differences in environmental factors. Our work paves the way for future studies to understand the ecology and biogeography of microalgae in extreme habitats.

Serological Detection of Antibodies against Anaplasma spp. in Cattle Reared in the Gyeongsangbuk-do, Korea.

Anaplasmosis is a tick-borne, non-contagious, zoonotic disease caused by Anaplasma spp., which include Anaplasma marginale, A. centrale, A. phagocytophilum, A. platys, A. ovis, and A. bovis. Recently, in Korea, the prevalence of Anaplasma spp. has been investigated in some animals, such as dogs, horses, goats, cats, and Korean water deer. In cattle, A. marginale is the most virulent species and regarded as the typical type of species. However, data on the seroprevalence of Anaplasma spp. in cattle in Korea during the last decade is limited. This study was designed to investigate the seroprevalence of bovine anaplasmosis in Korea. From 2010 to 2013, blood samples were collected from 568 cattle. Forty animals (7.0%) tested seropositive for Anaplasma spp. by cELISA. Despite that current bovine anaplasmosis seropositivity rate in the Gyeongsangbuk-do is lower than those in tropical countries, anaplasmosis needs to be regarded as a concerning disease. The identification of the specific Anaplasma species infecting cattle in this province requires additional molecular studies. Moreover, further monitoring and control programs for bovine anaplasmosis is required, and the information from this study will be beneficial to develop these programs.

DNA barcode assessment of Gracilaria salicornia (Gracilariaceae, Rhodophyta) from Southeast Asia.

BACKGROUND: DNA barcoding is becoming a widely applied tool for the quick and accurate identification of species. The evolution of the mitochondrial cytochrome c oxidase subunit I (COI) gene is sufficiently rapid to allow discrimination between closely related species and biogeographic subgroups within species. Gracilaria salicornia was originally described as being from Manila, the Philippines, and is distributed throughout Asia and the Indian Ocean. To more accurately define this species and its genetic diversity owing to the confusion of identification historically, DNA barcoding using the 5' end of the COI gene of the mitochondrial genome was applied to specimens collected from the Philippines, Malaysia, Thailand, China, and Japan, and they were compared to other gracilarian species. RESULTS: Within species, the COI marker yielded two clusters with nucleotide divergences of 0.0-1.3%. This divergence is slightly higher than the typical intraspecific variation for red algae. A total of eight COI haplotypes were found for G. salicornia, comprising the following groups: H1-H3 from the Philippines; H4 from Okinawa in Japan; H5-H7 from Malaysia, Thailand, and China; and H8 from Thailand. CONCLUSION: Although this work concentrated on a limited geographical region of a widespread taxon, the data shows intraspecific molecular divergences in G. salicornia and provides further evidence that DNA barcodes are useful tools for identifying species boundaries and examining biogeographical haplotypes for the genus Gracilaria.