Isolation and molecular identification of the etiological agents of streptococcosis in Nile tilapia (Oreochromis niloticus) cultured in net cages in Lake Sentani, Papua, Indonesia

Infections with Streptococcus spp. were observed in Nile tilapia cultured in net cages in Lake Sentani, Papua, Indonesia. Clinical signs included exophthalmia, erratic swimming, ascites in abdominal cavity, and external hemorrhages. Four types of bacterial colonies (SK, K10, P20, and M12) were isolated from the brain, kidney, and eyes. Based on phenotypic and genetic (16S rDNA sequencing) characteristics, the isolates were identified as Streptococcus iniae (SK), Streptococcus agalactiae (K10 and P20) and Lactococcus garvieae (M12). The latter species has not been previously isolated or reported from fish streptococcosis in Indonesia. Intraperitoneal injection of healthy tilapia with the bacterial species caused significant morbidity (70%) within 3 days and 100% mortality at 6 days post injection. Experimental infections and reisolation of the bacteria from morbid and dead fish suggest they are the causative agents of streptococcosis, which rendered high mortality among cage cultured Nile tilapia in Lake Sentani. Our results suggest the need for developing diagnostic tools for accurate identification of the agents of streptococcosis. As tilapia aquaculture continues to expand as a means of food production and livelihood in Indonesia, it becomes crucial to ensure that fish resources are monitored and protected from the adverse effects of infectious diseases

Identification of harmful cyanobacteria in the Sacramento-San Joaquin Delta and Clear Lake, California by DNA barcoding.

Accurate identification of cyanobacteria using traditional morphological taxonomy is challenging due to the magnitude of phenotypic plasticity among natural algal assemblages. In this study, molecular approach was utilized to facilitate the accurate identification of cyanobacteria in the Sacramento-San Joaquin Delta and in Clear Lake in Northern California where recurring blooms have been observed over the past decades. Algal samples were collected from both water bodies in 2011 and the samples containing diverse cyanobacteria as identified by morphological taxonomy were chosen for the molecular analysis. The 16S ribosomal RNA genes (16S rDNA) and the adjacent internal transcribed spacer (ITS) regions were amplified by PCR from the mixed algal samples using cyanobacteria generic primers. The obtained sequences were analyzed by similarity search (BLASTN) and phylogenetic analysis (16S rDNA) to differentiate species sharing significantly similar sequences. A total of 185 plasmid clones were obtained of which 77 were successfully identified to the species level: Aphanizomenon flos-aquae, Dolichospermum lemmermannii (taxonomic synonym: Anabaena lemmermannii), Limnoraphis robusta (taxonomic synonym: Lyngbya hieronymusii f. robusta) and Microcystis aeruginosa. To date, Dolichospermum and Limnoraphis found in Clear Lake have only been identified to the genus lavel by microscopy. During the course of this study, morphological identification and DNA barcoding confirmed A. flos-aquae as the predominant cyanobacterium in the Sacramento-San Joaquin Delta indicating a shift from M. aeruginosa that have dominated the blooms in the past decade. Lastly, the species-specific identification of Limnoraphis robusta in Clear Lake is another significant finding as this cyanobacterium has, thus far, only been reported in Lake Atitlan blooms in Guatemala

Henneguya sp. in yellowfin goby Acanthogobius flavimanus from the San Francisco Estuary.

Myxozoan spores were observed in yellowfin goby Acanthogobius flavimanus collected from Suisun Marsh, San Francisco Estuary (SFE). Although histopathological changes associated with the parasite were not observed, the spores formed plasmodia that partially blocked the gastric and intestinal mucosa and gut lumen and may affect the perfomance and survival of the yellowfin goby. Morphological features of the spores resembled Henneguya sp. and molecular analysis of the 18S ribosomal DNA (Domain III) confirmed close similarity to H. rhinogobii and H. pseudorhinogobii isolated from the Japanese freshwater goby. The yellowfin goby myxozoan however, is likely an undescribed species based on phylogenetic analysis and morphologic features. Detailed description of vegetative and spore stages are currently lacking for proposal to a new species of Henneguya. A specific PCR test was developed, which confirmed a 100% prevalence of the parasite among randomly collected gobies in group 1 (N = 30) and group 2 (N = 15) at termination of the study at one month in captivity. The myxozoan was also detected from 18 gobies (12%) that died in the first group within two weeks in captivity. Apparently healthy gobies that served as controls did not reveal the presence of the myxozoan by PCR. This study documents the occurrence of a potentially new species of myxozoan in the yellowfin goby and underscores the detection of a parasitic infection in an introduced fish in the SFE. Although the pathogenesis of the myxozoan was not assessed and the prevalence as reported here is restricted to a comparatively small collection site in Suisun slough, the reemergence, identification, and ecological relevance of the parasite on goby populations in the SFE may be investigated in the future using the specific diagnostic tool developed in this study

In situ hybridization: A detection tool for fish pathogens and its application on recent advances on whirling disease research

A non-radioactive in situ hybridization (ISH) protocol was used as a diagnostic tool for several fish pathogens including Candidatus Xenohaliotis californiensis, the etiologic agent of withering syndrome, which is a new disease in wild and cultured abalone Haliotis spp., the microsporidian Nucleospora salmonis and the myxozoan parasite Tetracapsula bryosalmonae (previously referred to as PKX) that causes proliferative kidney disease in salmonids. Most applications of the ISH protocol in our -laboratory were used in pathogenesis studies of whirling disease caused by the myxosporean Myxobolus cerebralis

Anthropogenic debris in seafood: Plastic debris and fibers from textiles in fish and bivalves sold for human consumption.

The ubiquity of anthropogenic debris in hundreds of species of wildlife and the toxicity of chemicals associated with it has begun to raise concerns regarding the presence of anthropogenic debris in seafood. We assessed the presence of anthropogenic debris in fishes and shellfish on sale for human consumption. We sampled from markets in Makassar, Indonesia, and from California, USA. All fish and shellfish were identified to species where possible. Anthropogenic debris was extracted from the digestive tracts of fish and whole shellfish using a 10% KOH solution and quantified under a dissecting microscope. In Indonesia, anthropogenic debris was found in 28% of individual fish and in 55% of all species. Similarly, in the USA, anthropogenic debris was found in 25% of individual fish and in 67% of all species. Anthropogenic debris was also found in 33% of individual shellfish sampled. All of the anthropogenic debris recovered from fish in Indonesia was plastic, whereas anthropogenic debris recovered from fish in the USA was primarily fibers. Variations in debris types likely reflect different sources and waste management strategies between countries. We report some of the first findings of plastic debris in fishes directly sold for human consumption raising concerns regarding human health