Whole-mount in situ TUNEL method revealed ectopic pattern of apoptosis in cadmium treated naupliar larvae of barnacle (Balanus amphitrite Darwin)

The effect of cadmium on stage II naupliar larvae of barnacle (Balanus amphitrite Darwin) was investigated. Barnacle larvae were exposed to 0-15 microM CdCl(2) for 24 h. Apoptotic cells were stained by the terminal deoxyribonucleotidyl transferase mediated dUTP nick end labelling (TUNEL) method. Incidence of apoptosis, as measured by numbers of animals with ectopic pattern of apoptosis as well as numbers of apoptotic cells per animal, was assessed using confocal microscopy. An increase in incidence of apoptosis was observed in the experimental animals with an increase in cadmium concentration. Mortality increased, and motility decreased, when barnacle larvae were exposed to an increasing concentration of cadmium. The relationship between the occurrence of apoptosis and swimming behaviour was investigated in larvae exposed to 10 microM CdCl(2). Significant increases in apoptosis were detected in the non-motile and dead nauplii. This study suggested that whole-mount in situ TUNEL method may be used to study increased occurrence of apoptotic cells in crustacean larvae.     

Studies with natural and anthropogenic iodine isotopes: iodine distribution and cycling in the global environment

Iodine is an important trace element in geological and biological processes. We summarize here recent results and new data of experiments and observations carried out to improve the understanding of concentration levels and behavior of natural and anthropogenic iodine nuclides in the global environment. The distribution of stable iodine in the Earth's crust was estimated using concentration data in a suite of representative samples and the influence of subduction on the marine iodine cycle was investigated using (129)I systematics on iodine-rich brines from Japan. The importance of microorganisms for the natural iodine cycle is shown in recent studies of iodine sorption on soil and of iodine volatilization from terrestrial and marine environments. Levels of anthropogenic (129)I were measured in samples collected around a spent fuel reprocessing plant in Japan.     

Chlorophyll and suspended sediment mapping to the Caribbean Sea from rivers in the capital city of the Dominican Republic using ALOS AVNIR-2 data

This study aims to study the distribution of contaminants in rivers that flow into the Caribbean Sea using chlorophyll-a (Chl-a) and suspended sediment (SS) as markers and ALOS AVNIR-2 satellite sensor data. The Haina River (HN) and Ozama and Isabela Rivers (OZ-IS) that flow through the city of Santo Domingo, the capital of the Dominican Republic, were chosen. First, in situ spectral reflectance/Chl-a and SS datasets obtained from these rivers were acquired in March 2011 (case A: with no rain influence) and June 2011 (case B: with rain influence), and the estimation algorithm of Chl-a and SS using AVNIR-2 data was developed from the datasets. Moreover, the developed algorithm was applied to AVNIR-2 data in November 2010 for case A and August 2010 for case B. Results revealed that for Chl-a and SS estimations under cases A and B conditions, the reflectance ratio of AVNIR-2 band 4 and band 3 (AV4/AV3) and the reflectance of AVNIR-2 band 4 (AV4) were effective. The Chl-a and SS mapping results obtained using AVNIR-2 data corresponded with the field survey results. Finally, an outline of the distribution of contaminants at the mouth of the river that flows into the Caribbean Sea was obtained for both rivers in cases A and B.     

Bayesian Modeling of Enteric Virus Density in Wastewater Using Left-Censored Data

Stochastic models are used to express pathogen density in environmental samples for performing microbial risk assessment with quantitative uncertainty. However, enteric virus density in water often falls below the quantification limit (non-detect) of the analytical methods employed, and it is always difficult to apply stochastic models to a dataset with a substantially high number of non-detects, i.e., left-censored data. We applied a Bayesian model that is able to model both the detected data (detects) and non-detects to simulated left-censored datasets of enteric virus density in wastewater. One hundred paired datasets were generated for each of the 39 combinations of a sample size and the number of detects, in which three sample sizes (12, 24, and 48) and the number of detects from 1 to 12, 24 and 48 were employed. The simulated observation data were assigned to one of two groups, i.e., detects and non-detects, by setting values on the limit of quantification to obtain the assumed number of detects for creating censored datasets. Then, the Bayesian model was applied to the censored datasets, and the estimated mean and standard deviation were compared to the true values by root mean square deviation. The difference between the true distribution and posterior predictive distribution was evaluated by Kullback–Leibler (KL) divergence, and it was found that the estimation accuracy was strongly affected by the number of detects. It is difficult to describe universal criteria to decide which level of accuracy is enough, but eight or more detects are required to accurately estimate the posterior predictive distributions when the sample size is 12, 24, or 48. The posterior predictive distribution of virus removal efficiency with a wastewater treatment unit process was obtained as the log ratio posterior distributions between the posterior predictive distributions of enteric viruses in untreated wastewater and treated wastewater. The KL divergence between the true distribution and posterior predictive distribution of virus removal efficiency also depends on the number of detects, and eight or more detects in a dataset of treated wastewater are required for its accurate estimation.     

Viral Interference as a Factor of False-Negative in the Infectious Adenovirus Detection Using Integrated Cell Culture-PCR with a BGM Cell Line

Food and Environmental Virology - This study investigated the influence of viral interference on the detection of enteric viruses using the integrated cell culture (ICC)-PCR with a BGM cell line....     

Detection of Rotavirus Strains in Freshwater Clams in Japan

Bivalve molluscan shellfish like clams and oysters, etc., are capable to bioaccumulate surrounding contaminants from waters into their digestive systems and posing serious threats of food poisoning. Detection of rotaviruses (RVs) in shellfish is of particular importance because RVs are prone to genome reassortment resulting in the emergence of new RV variants that may compromise vaccine safety. Herein, we have detected the wild-type RVs and Rotarix/RotaTeq vaccine strains in freshwater clams collected on the riverside, Kawasaki city, from July 2019 to January 2020 and correlated the detected genotypes with that of gastroenteritis cases of nearby clinics to understand the transmission of RVs in the environment. The wild-type RVs were detected in 62 (64.6%) out of 96 freshwater clams in every study month: July, September, November, and January that are considered as off-season for RV infections. The most frequent genotypes were G2 (42.9%), G8 (28.6%), G3 (14.3%), G1 (7.1%), and G10 (7.1%), which remained comparable with genotypic distribution found in the clinical samples over the last few years indicating that these RVs may accumulate in clams since a long time. However, G10 genotype was detected in clam but not in clinical samples suggesting the presence of asymptomatic infection or RVs could be carried out from a long distance. Importantly, vaccine strains, RotaTeq (1%) but not Rotarix (0%), were also detected in a clam. Attention must be paid to monitoring the potential transmission of wild-type and vaccine RV strains in the environment to prevent the emergence of new variants generated from genome reassortment with vaccine strains.

     

Genetic population structure in a commercial marine invertebrate with long-lived lecithotrophic larvae: <Emphasis Type="Italic">Cucumaria</Emphasis><Emphasis Type="Italic">frondosa</Emphasis> (Echinodermata: Holothuroidea)

The patterns of genetic diversity and connectivity were investigated in Cucumaria frondosa, the most abundant sea cucumber in the North Atlantic, to assist in the management and conservation of this ecologically important marine invertebrate, which is the target of an emerging fishery. Mitochondrial DNA COI sequences of 334 C. frondosa were obtained and analyzed, mainly from its western North Atlantic range, where the commercial fishery is being developed, with complementary sampling in the mid- and eastern North Atlantic. Analysis of molecular variance showed no significant (P > 0.05) differences among subpopulations in the western region suggesting that it constitutes one panmictic population. The same analysis showed low, but significant differences between eastern and western Atlantic populations. Coalescent analyses using isolation with migration models and a Bayesian skyline plot indicated historical divergence and a general increase in population size prior to the last glacial maximum and highly asymmetric gene flow (nearly 100 times lower from west to east) between sea cucumbers from North America and Norway. Results suggest that subpopulations of C. frondosa within the western North Atlantic have been highly connected. We propose that aided by the high-connectivity local subpopulations can recover rapidly from natural (i.e., ice ages) or anthropogenic (i.e., overfishing) population declines through recruitment from deep refugia.     

Validation and quantification of extractable age pigments for determining the age of Antarctic krill (<Emphasis Type="Italic">Euphausia superba</Emphasis>)

Antarctic krill, Euphausia superba, hatched from eggs and maintained for four years, were sampled periodically for age-pigment analysis. Extractable pigments from the eye and eyestalk ganglia were quantified using fluorescence intensity and standardised against protein. Three peak fluorescence intensities were detected at wavelengths of excitation 280 nm, emission 625 nm (pigment 1); excitation 355 nm, emission 510 nm (pigment 2); and excitation 463 nm, emission 620 nm (pigment 3). There was a positive correlation between the quantity of pigments 1 and 3 and the age of Antarctic krill. A model was developed to predict age from pigment 3 and to compare it with other age proxies (carapace length and eyeball diameter). The quantity of pigment 3 was the best predictor of age. The pigment method can discriminate between similar sized krill aged 12 and 36 months. Age pigments provide an improved tool for age estimation in Antarctic krill, particularly if used in conjunction with other demographic information.