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.     

镧-铅复合污染下AM真菌对玉米生长和镧、铅吸收的影响

采用温室盆栽试验的方法,模拟不同程度的镧-铅复合污染土壤(50、200、800 mg·kg~(-1)),研究接种丛枝菌根(arbuscular mycorrhizal,AM)、真菌Claroideoglomus etunicatum(CE)和Rhizophagus intraradices(RI)对玉米(Zea mays L.)菌根侵染率、生物量、矿质营养元素吸收、C∶N∶P生态化学计量比、稀土镧(La)和重金属铅(Pb)吸收、转运的影响,旨在为稀土-重金属复合污染土壤的治理和修复提供科学依据.结果表明,AM真菌CE和RI均与玉米建立了共生关系,平均菌根侵染率为26.7%~95.8%;随着La-Pb复合污染含量的增加,玉米植株菌根侵染率、地上部和根部生物量以及N、P、K、Ca、Mg这5种矿质营养元素含量显著降低,而玉米植株C∶P和N∶P以及地上部和根部La、Pb含量显著增加.接种2种AM真菌使玉米植株生物量显著提高了17.8%~158.9%,地上部和根部P含量显著提高了24.5%~153.8%,降低了C∶P和N∶P,符合生长速率假设.在3种程度La-Pb复合污染含量土壤上,AM真菌使玉米植株根部Pb含量显著增加了51.3%~67.7%,地上部Pb含量显著降低了16.0%~67.7%,Pb从玉米根部向地上部的转运率降低了31.5%~54.7%;同时,接种AM真菌显著增加了轻度LaPb复合污染土壤上玉米植株的La含量,在中度La-Pb复合污染土壤上却显著减少了玉米地上部的La含量,增加了玉米根部的La含量,抑制了La从根部向地上部的转运,重度La-Pb复合污染土壤上均没有显著影响.试验结果初步证明,AM真菌具有促进稀土-重金属复合污染土壤植物修复的潜力,对于稀土-重金属复合污染土壤生态系统的植被恢复具有潜在应用价值.     

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.     

Hunting behaviour and breeding performance of northern goshawks Accipiter gentilis, in relation to resource availability, sex, age and morphology

Animal territories that differ in the availability of food resources will require (all other things being equal) different levels of effort for successful reproduction. As a consequence, breeding performance may become most strongly dependent on factors that affect individual foraging where resources are poor. We investigated potential links between foraging behaviour, reproductive performance and morphology in a goshawk Accipiter gentilis population, which experienced markedly different resource levels in two different parts of the study area (rabbit-rich vs. rabbit-poor areas). Our analyses revealed (1) that rabbit abundance positively affected male reproductive output; (2) that age, size and rabbit abundance (during winter) positively affected different components of female reproductive output; (3) that foraging movements were inversely affected by rabbit abundance for both sexes (for females, this may mainly have reflected poor provisioning by males in the rabbit-poor area); (4) that younger breeders (both in males and females) tended to move over larger distances than older individuals (which may have reflected both a lack of hunting experience and mate searching); and (5) that male body size (wing length) showed some covariation with resource conditions (suggesting possible adaptations to hunting agile avian prey in the rabbit-poor area). Although we are unable to establish firm causal relationships with our observational data set, our results provide an example of how territory quality (here, food abundance) and individual features (here, age and morphology) may combine to shape a predator's foraging behaviour and, ultimately, its breeding performance.