Norovirus GII.Pe Genotype: Tracking a Foodborne Outbreak on a Cruise Ship Through Molecular Epidemiology,Brazil, 2014

Norovirus (NoV) is recognized as the most common cause of foodborne outbreaks. In 2014, an outbreak of acute gastroenteritis occurred on a cruise ship in Brazil, and NoV became the suspected etiology. Here we present the molecular identification of the NoV strains and the use of sequence analysis to determine modes of virus transmission. Food (cream cheese, tuna salad, grilled fish, orange mousse, and vegetables soup) and clinical samples were analyzed by ELISA, conventional RT-PCR, qRT-PCR, and sequencing. Genogroup GII NoV was identified by ELISA and conventional RT-PCR in fecal samples from 5 of 12 patients tested (41.7%), and in the orange mousse food sample by conventional RT-PCR and qRT-PCR. Two fecal GII NoV samples and the orange mousse GII NoV sample were successfully genotyped as GII.Pe (ORF 1), revealed 98.0–98.8% identities among them, and shared phylogenetically distinct cluster. Establishing the source of a NoV outbreak can be a challenging task. In this report, the molecular analysis of the partial RdRp NoV gene provided a powerful tool for genotyping (GII.Pe) and tracking of outbreak-related samples. In addition, the same fast and simple extraction methods applied to clinical samples could be successfully used for complex food matrices, and have the potential to be introduced in routine laboratories for screening foods for presence of NoV.     

Use of Preservative Agents and Antibiotics for Increased Poliovirus Survival on Positively Charged Filters

Environmental surveillance of poliovirus (PV) and other non-enveloped viruses can help identify silent circulation and is necessary to certify eradication. The bag-mediated filtration system is an efficient method to filter large volumes of environmental waters at field sites for monitoring the presence of viruses. As filters may require long transit times to off-site laboratories for processing, viral inactivation or overgrowth of bacteria and fungi can interfere with virus detection and quantification (Miki and Jacquet in Aquatic Microb Ecol 51(2):195–208, 2008). To evaluate virus survival over time on ViroCap^? filters, the filters were seeded with PV type 1 (PV1) and/or MS2 and then dosed with preservatives or antibiotics prior to storage and elution. These filters were stored at various temperatures and time periods, and then eluted for PV1 and MS2 recovery quantification. Filters dosed with the preservative combination of 2% sodium benzoate and 0.2% calcium propionate had increased virus survival over time when stored at 25 °C, compared to samples stored at 25 °C with no preservatives. While elution within 24 h of filtration is recommended, if storage or shipping is required then this preservative mixture can help preserve sample integrity. Addition of an antibiotic cocktail containing cephapirin, gentamicin, and Proclin^? 300 increased recovery after storage at 4 and 25 °C, when compared to storage with no antibiotics. The antibiotic cocktail can aid sample preservation if access to appropriate antibiotics storage is available and sample cold chain is unreliable. This study demonstrated that the use of preservatives or antibiotics is a simple, cost-effective method to improve virus detection from ViroCap cartridge filters over time.     

DNA Heat Treatment for Improving qPCR Analysis of Human Adenovirus in Wastewater

PCR inhibitory substances in complex sample matrices can cause false negatives or under-estimation of target concentration. This study assessed DNA heat treatment for reducing inhibition during qPCR analysis of human adenovirus (HAdV) in wastewater samples. Inhibition was reduced by heat treating DNA, where mean HAdV concentration was increased by 0.71 log₁₀ GC/L (and up to 3.04 log₁₀ GC/L in one case), and replicate variability and false negatives were reduced. DNA heat treatment should be further investigated for improving reliability of HAdV concentration estimates in water, which can support more accurate assessment of health risks associated with viral pathogen exposure.     

Assessment and Evaluation of an Integrated Hybrid Anaerobic–Aerobic Sewage Treatment System for the Removal of Enteric Viruses

The capability of a cost-effective and a small size decentralized pilot wastewater treatment plant (WWTP) to remove enteric viruses such as rotavirus, norovirus genogroup I (GGI), norovirus genogroup II (GGII), Hepatitis E virus (HEV), and adenovirus was studied. This pilot plant is an integrated hybrid anaerobic/aerobic setup which consisted of anaerobic sludge blanket (UASB), biological aerated filter (BAF), and inclined plate settler (IPS). Both the UASB and BAF are packed with a non-woven polyester fabric (NWPF). Results indicated that the overall log₁₀ reductions of enteric viruses’ genome copies through the whole system were 3.1 ± 1, 3.3 ± 0.5, and 2.6 ± 0.9 log₁₀ for rotavirus, norovirus GGI, and adenovirus, respectively. Reduction efficiency for both norovirus GGII and HEV after the different treatment steps could not be calculated because there were no significant numbers of positive samples for both viruses. The overall reduction of rotavirus infectious units through the whole system was 2.2 ± 0.8 log₁₀ reduction which is very close to the overall log₁₀ reduction of adenovirus infectious units through the whole system which was 2.1 ± 0.8 log₁₀ reduction. There was no considerable difference in the removal efficiency for different rotavirus G and P types. Adenovirus 41 was the only type detected in the all positive samples. Although the pilot WWTP investigated is cost effective, has a small footprint, does not need a long distance network pipes, and easy to operate, its efficiency to remove enteric viruses is comparable with the conventional centralized WWTPs.     

The effect of flower-like and non-flower-like visual properties on choice of unrewarding patterns by bumblebees

How do distinct visual stimuli help bumblebees discover flowers before they have experienced any reward outside of their nest? Two visual floral properties, type of a pattern (concentric vs radial) and its position on unrewarding artificial flowers (central vs peripheral on corolla), were manipulated in two experiments. Both visual properties showed significant effects on floral choice. When pitted against each other, pattern was more important than position. Experiment 1 shows a significant effect of concentric pattern position, and experiment 2 shows a significant preference towards radial patterns regardless of their position. These results show that the presence of markings at the center of a flower are not so important as the presence of markings that will direct bees there.     

Effects of exotic fish farms on bird communities in lake and marine ecosystems

Salmon farming is a widespread activity around the world, also known to promote diverse environmental effects on aquatic ecosystems. However, information regarding the impact of salmon farming on bird assemblages is notably scarce. We hypothesize that salmon farming, by providing food subsidies and physical structures to birds, will change their local community structure. To test this hypothesis, we conducted a seasonal monitoring of bird richness, abundance, and composition at paired salmon pen and control plots in two marine and two lake sites in southern Chile, from fall 2002 to summer 2004. Overall, salmon farming had no significant effects on species richness, but bird abundance was significantly and noticeably higher in salmon pens than in controls. Such aggregation was mainly accounted for by the trophic guilds of omnivores, diving piscivores, carrion eaters, and perching piscivores, but not by invertebrate feeders, herbivores, and surface feeders. Species composition was also significantly and persistently different between salmon pens and controls within each lake or marine locality. The patterns described above remained consistent across environment types and seasons indicating that salmon farming is changing the community structure of birds in both lake and marine habitats by promoting functional and aggregation responses, particularly by favoring species with broader niches. Such local patterns may thus anticipate potential threats from the ongoing expansion of the salmon industry to neighboring areas in Chile, resulting in regional changes of bird communities, toward a less diverse one and dominated by opportunistic, common, and generalist species such as gulls, vultures, and cormorants.     

Seasonal changes in burrow geometry of the common mole rat (Rodentia: Bathyergidae)

Sociality in mole rats has been suggested to have evolved as a response to the widely dispersed food resources and the limited burrowing opportunities that result from sporadic rainfall events. In the most arid regions, individual foraging efficiency is reduced, and energetic constraints increase. In this study, we investigate seasonal differences in burrow architecture of the social Cryptomys hottentotus hottentotus in a mesic region. We describe burrow geometry in response to seasonal weather conditions for two seasons (wet and dry). Interactions occurred between seasons and colony size for the size of the burrow systems, but not the shape of the burrow systems. The fractal dimension values of the burrow systems did not differ between seasons. Thus, the burrow complexity was dependent upon the number of mole rats present in the social group.