Source tracking of fecal contamination in Asian green mussels (Perna viridis) harvested in Manila Bay, Philippines by molecular detection and genotyping of Cryptosporidium spp.

Manila Bay, a multipurpose body of water located around Metro Manila, Philippines, is progressively deteriorating because of massive pollution. Reports have shown that the bay and its aquatic resources (i.e., seafood) are contaminated with fecal matter and enteric pathogens, posing a threat to public health and industry. This problem raises the need for a microbial source tracking methodology as a part of the rehabilitation efforts in the bay. Bivalve mollusks cultivated in water can serve as sentinel species to detect fecal pollution and can complement water monitoring. With the use of polymerase chain reaction and DNA sequence analysis, this study detected Cryptosporidium spp. in Asian green mussels (Perna viridis) cultivated and harvested in Manila Bay and sold in Bulungan Seafood Market, Parañaque, Philippines, from 2019 to 2021 with an overall occurrence of 8.77% (n = 57). The analysis of the 18S rDNA segment revealed three genotypes from Cryptosporidium-positive samples, namely, Cryptosporidium sp. rat genotype IV (60%), C. galli (20%), and C. meleagridis (20%). These findings suggest fecal pollution in bivalve cultivation sites coming from sewage, nonpoint, and agricultural sources. The presence of C. meleagridis, the third most common cause of human cryptosporidiosis, in mussels poses a threat to human health. Thus, there is a need to establish routine detection and source tracking of Cryptosporidium spp. in Manila Bay and to educate seafood consumers on food safety.

Attachment of potential cultivable primo-colonizing bacteria and its implications on the fate of low-density polyethylene (LDPE) plastics in the marine environment.

Plastics released in the environment become suitable matrices for microbial attachment and colonization. Plastics-associated microbial communities interact with each other and are metabolically distinct from the surrounding environment. However, pioneer colonizing species and their interaction with the plastic during initial colonization are less described. Marine sediment bacteria from sites in Manila Bay were isolated via a double selective enrichment method using sterilized low-density polyethylene (LDPE) sheets as the sole carbon source. Ten isolates were identified to belong to the genera Halomonas, Bacillus, Alteromonas, Photobacterium, and Aliishimia based on 16S rRNA gene phylogeny, and majority of the taxa found exhibit a surface-associated lifestyle. Isolates were then tested for their ability to colonize polyethylene (PE) through co-incubation with LDPE sheets for 60 days. Growth of colonies in crevices, formation of cell-shaped pits, and increased roughness of the surface indicate physical deterioration. Fourier-transform infrared (FT-IR) spectroscopy revealed significant changes in the functional groups and bond indices on LDPE sheets separately co-incubated with the isolates, demonstrating that different species potentially target different substrates of the photo-oxidized polymer backbone. Understanding the activity of primo-colonizing bacteria on the plastic surface can provide insights on the possible mechanisms used to make plastic more bioavailable for other species, and their implications on the fate of plastics in the marine environment.

Potential of sediment bacterial communities from Manila Bay (Philippines) to degrade low-density polyethylene (LDPE).

The persistence of plastics and its effects in different environments where they accumulate, particularly in coastal areas, is of serious concern. These plastics exhibit signs of degradation, possibly mediated by microorganisms. In this study, we investigated the potential of sediment microbial communities from Manila Bay, Philippines, which has a severe plastics problem, to degrade low-density polyethylene (LDPE). Plastics in selected sites were quantified and sediment samples from sites with the lowest and highest plastic accumulation were collected. These sediments were then introduced and incubated with LDPE in vitro for a period of 91 days. Fourier transform infrared spectroscopy detected the appearance of carbonyl and vinyl products on the plastic surface, indicating structural surface modifications attributed to polymer degradation. Communities attached to the plastics were profiled using high-throughput sequencing of the V4-V5 region of the 16S rRNA gene. Members of the phylum Proteobacteria dominated the plastic surface throughout the experiment. Several bacterial taxa associated with hydrocarbon degradation were also enriched, with some taxa positively correlating with the biodegradation indices, suggesting potential active roles in the partial biodegradation of plastics. Other taxa were also present, which might be consuming by-products or providing nourishment for other groups, indicating synergy in utilizing the plastic as the main carbon source and creation of a microenvironment within the plastics biofilm. This study showed that sediment microbes from Manila Bay may have naturally occurring microbial groups potentially capable of partially degrading plastics, supporting previous studies that the biodegradation potential for plastics is ubiquitously present in marine microbial assemblages.

Transport of floating litter within Manila Bay, Philippines.

The increasing amount of marine litter is a global issue that is also being experienced within the Manila Bay Philippines. To better understand the behavior of litter within the bay, particle tracking simulations of floating litter released from several sources were conducted. Forward-in-time (PTM-FIT) simulation of particle movements showed that during the southwest monsoon season, litter particles released by river sources tend to accumulate toward five hotspots located along the northeastern coastline of the bay. Backward-in-time (PTM-BIT) simulation of particles released from the identified hotspots showed coastal and foreign sources contributing to the litter collected within these areas. PTM-FIT simulations during the southwest monsoon season showed that particles tend to circulate locally within the bay before being transported toward the hotspots. By contrast, PTM-FIT simulations during the northwest monsoon season showed that litter particles tend to travel toward the mouth of the bay, an implication for global marine litter pollution.

Isolation, identification and antibiogram profiling of bacteria isolated from water, seafood, and macroplastic samples from Baseco Beach, Manila Bay

Background and Objective@#Manila Bay plays an important role both in economics and ecology because it serves as the major economic center of the Philippines and as it harbors different habitats and biodiversity. Unfortunately, it is threatened by various pollutions including the unregulated discharge of wastewater from industrial, agricultural, and household sectors and improper disposal of trash such as macroplastics among others. All these contributes to the current state of Manila Bay. This study identified bacteria isolated from water, seafood and floating macroplastic samples from Baseco Beach, Manila Bay and determined their antibiogram profiles. @*Methodology@#Bacterial isolates were obtained from water, seafoods and macroplastic samples from Baseco Beach, Manila Bay using conventional culture techniques. Identification of the isolates was done using Vitek-2 Automated System and antibiogram profiling was done using Kirby-Bauer Disk Diffusion Susceptibility Test. @*Results and Conclusions@#A total of 30 bacterial isolates were obtained from different samples from water, seafood and macroplastic samples from Baseco Beach, Manila Bay. These isolates were identified and found to belong to 13 different bacterial species with Bacillus spp. comprising 33.33% of the isolates (10 out of 30), and Vibrio alginolyticus comprising 23.33% of the isolates (7 out of 30) and the other species comprise the remaining 43.34% (Pseudomonas spp., Vibrio fluvialis, Klebsiella pneumoniae, Shewanella alga, Sphingomonas paucimobilis, Staphylococcus haemolyticus, Chryseobacterium indologenes, Myroides sp. and Aeromonas salmonicida). Of these, six out of 30 isolates (20%) showed susceptibility to all six representative antibiotics used (Cefazolin 30μg, Gentamicin 10 μg, Chloramphenicol 30 μg, ampicillin 10 μg, Cefuroxime 30 μg, Ceftazidime 30 μg) while 7 isolates (23.33%) were resistant to only one class of antibiotic. Moreover, 17 out of 30 isolates (56.66%) were resistant to two or more classes of antibiotic while only one isolate (3.33%) was found to be resistant to gentamicin. All 30 isolates (100%) were susceptible to chloramphenicol. Interestingly, three antibiotic resistant (AMR) bacteria were isolated from macroplastics namely Pseudomonas oleovorans (S2), Vibrio alginolyticus (S5), and Pseudomonas alcaligenes (S29) which were all resistant to ampicillin and cefazolin. This is the first study in the Philippines to isolate AMR bacteria from macroplastics from Manila Bay. The presence of AMR bacteria in macroplastics shows that these materials can be a reservoir for its dynamics and distribution. Lastly, with the emergence of antimicrobial resistant bacteria, the elucidation of the antibiogram profile of bacteria is necessary to determine its implication sand threats to public health. This study served as a baseline study of presence of AMR bacteria in macroplastic samples from Manila Bay.

Species check-list for Tintinnids of the Philippines Archipelago (Protozoa, Ciliophora).

Tintinnids are an essential link between nano- and macro- planktons in the food webs of the marine environment. It is also known that tintinnids are one of themajor components of marine planktonic ciliates and has a cosmopolitan character. In the Philippine archipelago, which is recognized as a center of marine biodiversity, tintinnids checklist has not been done or published. Therefore, a checklist is presented in this study based on a compilation of previous tintinnids studies conducted at the Philippines waters. As a result of the studies done since 1941 up to present, a total of 114 taxa belonging to 14 families and 37 genera were listed. The Philippines coastal waters record a total of 50 species while the open seas document 72 species to date.

Cryptosporidium species from common edible bivalves in Manila Bay, Philippines.

Manila Bay is one of the major propagation sites of edible bivalves in the Philippines. Studies have shown that bivalves might be contaminated with human pathogens like the protozoan parasite Cryptosporidium, one of the major causes of gastroenteritis in the world. In this study, Cryptosporidium from four species of edible bivalves were isolated using a combination of sucrose flotation and immunomagnetic separation. Using direct fluorescent antibody test, Cryptosporidium oocysts were found in 67 out of 144 samples collected. DNA sequence analysis of the 18S rRNA gene of the isolates detected C. parvum and C. hominis (major causes of human cryptosporidiosis) and C. meleagridis (causes infection in avian species). Analysis of the 60kDa glycoprotein gene further confirmed the genotypes of the Cryptosporidium isolates. This study is the first to provide baseline information on Cryptosporidium contamination of Manila Bay where bivalves are commonly cultured.

Spatiotemporal variability of hypoxia and eutrophication in Manila Bay, Philippines during the northeast and southwest monsoons.

Hypoxia in Manila Bay, Philippines was previously reported during the northeast monsoon (dry season) in February 2010. In this study, four more field surveys of the same 31 stations were conducted in July 2010, August 2011 and 2012 (wet season, southwest monsoon), and February 2011 (dry season, northeast monsoon). During the wet season, bottom hypoxia spread northward towards the coast with dissolved oxygen (DO) ranging from 0.12 to 9.22 mg/L and the bay-wide average reaching 2.10 mg/L. Nutrient levels were elevated, especially near the bottom where dissolved inorganic nitrogen reached 22.3 µM (July 2010) and phosphorus reached 5.61 µM (August 2011). High nutrient concentrations often coincided with low near-bottom DO content. Our work builds on the preliminary assessment of hypoxia in Manila Bay, the importance of repeated temporal studies, and shows hypoxia to prevail significantly during the southwest monsoon (wet season) when increased freshwater discharge caused strong water column stratification.