Coxsackievirus B4 as a Causative Agent of Diabetes Mellitus Type 1: Is There a Role of Inefficiently Treated Drinking Water and Sewage in Virus Spreading?

This study proposed to detect the enterovirus (EV) infection in children with type 1 diabetes mellitus (T1D) and to assess the role of insufficiently treated water and sewage as sources of viral spreading. Three hundred and eighty-two serum specimens of children with T1D, one hundred serum specimens of children who did not suffer from T1D as control, and forty-eight water and sewage samples were screened for EV RNA using nested RT-PCR. The number of genome copies and infectious units of EVs in raw and treated sewage and water samples were investigated using real-time (RT)-PCR and plaque assay, respectively. T1D markers [Fasting blood glucose (FBG), HbA1c, and C-peptide], in addition to anti-Coxsackie A & B viruses (CVs A & B) IgG, were measured in control, T1D-negative EV (T1D–EV^?), and T1D-positive EV (T1D–EV⁺) children specimens. The prevalence of EV genome was significantly higher in diabetic children (26.2%, 100 out of 382) than the control children (0%, 0 out of 100). FBG and HbA1c in T1D–EV^? and T1D–EV⁺ children specimens were significantly higher than those in the control group, while c-peptide in T1D–EV^? and T1D–EV⁺ children specimens was significantly lower than that in the control (n = 100; p < 0.001). Positivity of anti-CVs A & B IgG was 70.7, 6.7, and 22.9% in T1D–EV⁺, T1D–EV^?, and control children specimens, respectively. The prevalence of EV genome in drinking water and treated sewage samples was 25 and 33.3%, respectively. The prevalence of EV infectious units in drinking water and treated sewage samples was 8.5 and 25%, respectively. Quantification assays were performed to assess the capabilities of both wastewater treatment plants (WWTPs) and water treatment plants (WTPs) to remove EV. The reduction of EV genome in Zenin WWTP ranged from 2 to 4 log₁₀, while the reduction of EV infectious units ranged from 1 to 4 log₁₀. The reduction of EV genome in El-Giza WTP ranged from 1 to 3 log₁₀, while the reduction of EV infectious units ranged from 1 to 2 log₁₀. This capability of reduction did not prevent the appearance of infectious EV in treated sewage and drinking water. Plaque purification was performed for isolation of separate EV isolates from treated and untreated water and sewage samples. Characterization of the EV amplicons by RT-PCR followed by sequencing of these isolates revealed high homology (97%) with human coxsackievirus B4 (CV B4) in 60% of the isolates, while the rest of the isolates belonged to poliovirus type 1 and type 2 vaccine strains. On the other hand, characterization of the EV amplicons by RT-PCR followed by sequencing for T1D–EV⁺ children specimens indicated that all samples contained CV B4 with the same sequence characterized in the environmental samples. CV B4-contaminated drinking water or treated sewage may play a role as a causative agent of T1D in children.     

Assessment of the Virological Quality of Marine and Running Surface Waters in NW Greece: A Case Study

The virological quality of surface marine and running water samples collected from Igoumenitsa gulf and Kalamas river (NW Greece) was assessed from October 2012 to September 2013. Sampling sites were exposed to different land and/or anthropogenic effects. Seawater samples were collected monthly from five sampling stations (new harbor, old harbor, wastewater treatment plant outlet, protected Natura area, Drepano beach). Viral targets included human adenoviruses (hAdVs), as index human viruses, while noroviruses (NoVs) and hepatitis A virus (HAV) were also studied. Kalamas river samples were collected seasonally, from three sampling stations (Soulopoulo, Dam, Sagiada-estuaries), while viral targets included also porcine adenoviruses (pAdVs) and bovine polyoma viruses (bPyVs), as additional index viruses. All water samples were analyzed for standard bacterial indicators, as well. Physicochemical and meteorological data were also collected. Based on the standard bacterial indices, both sea and river water samples did not exceed the limits set according to Directive 2006/7/EU. However, positive samples for hAdVs were found occasionally in all sampling sites in Igoumenitsa gulf (23.3%, 14/60) showing fecal contamination of human origin. Moreover, HAV was detected once, in the sampling site of the old port (at 510 GC/L). Most of the Kalamas water samples were found positive for hAdVs (58.3%, 7/12), while human noroviruses GI (NoVGI) (8.3%, 1/12) and GII (NoVGII) (16.7%, 2/12) were also detected. HAV, pAdVs, and bovine polyomaviruses (bPyVs) were not detected in any of the analyzed samples. No statistically significant correlations were found between classic bacterial indicators and viral targets, nor between viruses and meteorological data. Overall, the present study contributed to the collection of useful data for the biomonitoring of the region, and the assessment of the overall impact of anthropogenic activities. It provided also valuable information for the evaluation of the risk of waterborne viral infections and the protection of public health. It was the first virological study in the area and one of the few in Greece.     

The Impact of Winter Relocation and Depuration on Norovirus Concentrations in Pacific Oysters Harvested from a Commercial Production Site

Oysters contaminated with norovirus present a significant public health risk when consumed raw. In this study, norovirus genome copy concentrations were determined in Pacific oysters (Magallana gigas) harvested from a sewage-impacted production site and then subjected to site-specific management procedures. These procedures consisted of relocation of oysters to an alternative production area during the norovirus high-risk winter periods (November to March) followed by an extended depuration (self-purification) under controlled temperature conditions. Significant differences in norovirus RNA concentrations were demonstrated at each point in the management process. Thirty-one percent of oyster samples from the main harvest area (Site 1) contained norovirus concentrations >?500 genome copies/g and 29% contained norovirus concentrations <?100 genome copies/g. By contrast, no oyster sample from the alternative harvest area (Site 2) or following depuration contained norovirus concentrations >?500 genome copies/g. In addition, 60 and 88% of oysters samples contained norovirus concentrations <?100 genome copies/g in oysters sampled from Site 2 and following depuration, respectively. These data demonstrate that site-specific management processes, supported by norovirus monitoring, can be an effective strategy to reduce, but not eliminate, consumer exposure to norovirus genome copies.     

Carbon, nitrogen, and phosphorus stoichiometry of plankton and the nutrient regime in Cabo Frio Bay, SE Brazil

This long-term study, performed during the years 2003–2005 and 2008–2009, investigated the carbon (C), nitrogen (N), and phosphorus (P) contents of the phyto- and zooplankton communities and the nutrient regime of Cabo Frio Bay, SE Brazil. The information intends to serve as baseline of the plankton C, N, and P stoichiometry for the calibration of biogeochemical and ecological models in support to future findings related to the local and regional phenomena of climatic change. Cabo Frio Bay is a small semienclosed system set adjacent to a region subject to sporadic coastal upwelling. Zooplankton exhibited average annual C, N, and P contents of 11.6?±?6.9 %, 2.8?±?1.8 %, and 0.18?±?0.08 %, and phytoplankton (>20 μm) 6.8?±?6.0 %, 1.6?±?1.5 %, and 0.09?±?0.08 %, respectively. The C/N/P ratios correspond to the lowest already found to date for a marine environment. The low C contents must have been brought about by a predominance of gelatinous zooplankton, like Doliolids/ Salps and also Pteropods. Average annual nutrient concentrations in the water were 0.21?±?0.1 μM for phosphate, 0.08?±?0.1 μM for nitrite, 0.74?±?1.6 μM for nitrate, and 1.27?±?1.1 μM for ammonium. N/P ratios were around 8:1 during the first study period and 12:1 during the second. The plankton C/N/P and N/P nutrient ratios and elemental concentrations suggest that the system was oligotrophic and nitrogen limited. The sporadic intrusions of upwelling waters during the first study period had no marked effect upon the systems metabolism, likely due to dilution effects and the short residence times of water of the bay.     

The reproduction of Enchytraeus sp.—technical improvement for the counting of juveniles

Soil-dwelling annelids of the genus Enchytraeus are used in ecotoxicology for testing of chemicals mixed in artificial soil or for testing of wastes and soils of unknown quality. ISO 16387 describes a method for determining the effects of substances or contaminated soils on survival and reproduction of Enchytraeus albidus or of the smaller species Enchytraeus buchholzi or Enchy-traeus crypticus. After the total test duration of 6 (or 4)?weeks, the juveniles hatched in the meantime are counted. There are several possible extraction techniques, which are always followed by counting the juveniles by hand, but none of them seems easy to handle. We proposed a new modification of the worm extraction method using flotation of fixed and stained juveniles followed by taking a photograph. The digital image of the juveniles is evaluated by computer processing. It makes the counting of juveniles much easier and less labor intensive.     

A comparative study on mesozooplankton abundance and diversity between a protected and an unprotected coastal area of Andaman Islands

The study was carried out to understand the variability in phytoplankton production (Chlorophyll a) and mesozooplankton diversity from two different shallow coastal regions of south Andaman viz. Port Blair Bay (PBB), the only real urban area among the islands and Mahatma Gandhi Marine National Park, a Marine Protected Area (MPA) at Wandoor. Seasonal sampling was carried out during the Northeast monsoon (NEM—November 2005), Intermonsoon (IM—April 2006), and Southwest monsoon (SWM—August 2006). Significant (P?<?0.05) seasonal variation was observed in the environmental variables at both the regions. Higher average chlorophyll a (Chl. a) and mesozooplankton standing stock were observed at PBB compared to MPA, but the seasonal variation observed was marginal at both the study areas. Chl. a showed a steep increasing gradient from outer to the inner regions of the PBB. The number of zooplankton taxa recorded at both areas was quite similar, but marked differences were noticed in their relative contribution to the total abundance. Eventhough the Copepoda dominated at both the areas, the non-copepod taxa differed significantly between the regions. Dominance of carnivores such as siphonophores and chaetognaths were noticed at PBB, while filter feeders such as appendicularians and decapod larvae were more abundant at MPA. A total of 20 and 21 copepod families was recorded from PBB and MPA, respectively. Eleven species of chaetognaths were observed as common at both areas. Larval decapods were found to be predominant at MPA with 20 families; whereas, at PBB, only 12 families were recorded. In the light of the recent reports on various changes occurring in the coastal waters of the Andaman Islands, it is suspected that the difference in Chl. a as well as the mesozooplankton standing stock and community structure observed between the two study areas may be related to the various anthropogenic events influencing the coastal waters.     

Impact of rapid urbanisation and industrialisation on river sediment metal contamination

This study aimed at evidencing contaminant inputs from a rapidly growing population and the accompanying anthropogenic activities to river sediments. The Fez metropolitan area and its impacts on the Sebou's sediments (the main Moroccan river) were chosen as a case study. The Fez agglomeration is surrounded by the river Fez, receiving the wastewaters of this developing city and then flowing into the Sebou. The sediment cores from the Fez and Sebou Rivers were extracted and analysed for major elements, butyltins and toxic metals. Normalised enrichment factors and geoaccumulation index were calculated. Toxicity risk was assessed by two sets of sediment quality guideline (SQG) indices. A moderate level of contamination by butyltins was observed, with monobutyltin being the dominant species across all sites and depths. The lowest level of metal pollution was identified in the Sebou's sediments in upstream of Fez city, whilst the Fez' sediments were heavily polluted and exhibited bottom-up accumulation trends, which is a clear signature of recent inputs from the untreated wastewaters of Fez city. Consequently, the sediments of Fez and Sebou at the downstream of the confluence were found to be potentially toxic, according to the SQG levels. This finding is concerned with aquatic organisms, as well as to the riverside population, which is certainly exposed to these pollutants through the daily use of water. This study suggests that although Morocco has adopted environmental regulations aiming at restricting pollutant discharges into the natural ecosystems, such regulations are neither well respected by the main polluters nor efficiently enforced by the authorities.