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This study assess the quality of wastewater through the detection and quantification of important viruses causing gastroenteritis at different stages of the wastewater treatment process in an activated-sludge wastewater treatment plant with ultraviolet disinfection. Ten sampling events were carried out in a campaign along a period of 18 months collecting wastewater samples from the influent, after the activated-sludge treatment, and after the final disinfection with UV radiation. Samples were concentrated through ultracentrifugation and analysed using retro-transcription, PCR and real time quantitative PCR protocols, for detection and quantification of Group A Rotavirus (RVA), Human Astrovirus (HAstV), Norovirus Genogroup II (NoV GII) and Human Adenovirus (HAdV). HAdV (100%), NoV GII (90%), RVA (70%) and HAstV (60%) were detected in influent samples with concentration from 1·4 (NoV GII) to 8·0 (RVA) log 10  gc l -1 . Activated-sludge treatment reached well quality effluents with low organic material concentration, although nonstatistical significant differences were registered among influent and postactivated sludge treatment samples, regarding the presence and concentration for most viruses. All post-UV samples were negative for NoV GII and HAstV, although RVA and HAdV were detected in 38% and 63% of those samples respectively, with concentration ranging from 2·2 to 5·5 and 3·1 to 3·4 log 10  gc l -1 ., Significance and Impact of the Study: This study demonstrates that an activated-sludge wastewater treatment plant with UV disinfection reduces to levels below the detection limit those single-stranded RNA viruses as noroviruses and astroviruses and reach significant lower levels of rotaviruses and adenoviruses after the complete treatment process., (© 2017 The Society for Applied Microbiology.)

Aim: To analyse group A rotavirus (RVA) environmental contamination in waters used for calves' consumption and to assess viral viability in dairy farm water sources., Methods and Results: We analysed 202 samples of water used for calves' consumption and RVA was detected by RT-qPCR in 35·1% (95% CI: 28·9-42·0%). A marked pattern of seasonality was observed with higher frequency of detection in colder than warmer months (P = 0·002). There was no association between viral load and season or between the number of milking cows in the herd and the detection of RVA in the farm. The viability of the RVA particles detected was confirmed by isolation of RVA in cell culture from 5 of 10 water samples. Furthermore, an RVA waterborne outbreak of neonatal calf diarrhoea was described., Conclusions: We demonstrate that RVA is frequent in dairy farm waters, and that the virus is infectious and capable of generating a diarrhoea outbreak., Significance and Impact of the Study: Neonatal diarrhoea syndrome leads to economic losses to the livestock industry worldwide. To determine transmission routes is essential to take action in this regard and reduce the impact that this syndrome has for the livestock production. The results obtained in this work alert the dairy industry and highlight that mitigation strategies are crucial to improve the microbiological quality of this water., (© 2018 The Society for Applied Microbiology.)

Aims: To determine the prevalence and molecular epidemiology of norovirus (NoV) genogroup I (GI) and GII in Uruguay., Methods and Results: One hundred and sixteen sewage samples were collected in six cities (Bella Unión, Salto, Paysandú, Fray Bentos, Melo and Treinta y Tres) from March 2011 to April 2013, viruses were concentrated by ultracentrifugation and NoV studies were performed by semi-nested RT-PCR (partial capsid region). NoV were detected in samples from all the cities and detected in 72% (84/116) of the samples with nine of them belonging to GI, 48 to GII and 27 to both genogroups. Remarkably, a high genetic diversity was identified: GII.2 (n = 13), GII.4 (n = 13), GI.1 (n = 5), GI.4 (n = 5), GI.8 (n = 4), GII.13 (n = 4), GII.1 (n = 3), GII.6 (n = 3), GI.3 (n = 1), GI.5 (n = 1), GI.6 (n = 1), GII.3 (n = 1), GII.17 (n = 1). Interestingly, a complete replacement of GII.4 New Orleans 2009 by GII.4 Sydney 2012 variants during 2012 was evidenced., Conclusion: This study reveals a high circulation of different NoV GI and GII genotypes in sewage evidencing a replacement of GII.4 variants., Significance and Impact of Study: This approach can be used as an indicator of the presence of a new GII.4 variant which can originate an increase in acute gastroenteritis outbreaks worldwide., (© 2016 The Society for Applied Microbiology.)

Aims: The aim of this study was to investigate the presence of the recently identified human astrovirus (HAstV) and to increase the knowledge of the molecular epidemiology of classical HAstV detected in Uruguay., Methods and Results: Recently identified and classical HAstV genotypes were investigated by RT-PCR targeting the ORF1b and ORF2 genome regions in 20 samples obtained between September 2011 and April 2013 in two cities of the eastern region of Uruguay. Four of 20 samples (20%) were identified as MLB-1 genotype and it was found a new MLB-1 classification through the segregation of the worldwide reported MLB-1 strains in two genetic lineages proposed and named: MLB-1a and MLB-1b. Fourteen (70%) samples were positive for classical HAstV and 12 of them were successfully sequenced and genotyped as: HAstV-1 (n = 10), HAstV-2 and HAstV-5 (one sample each)., Conclusion: These results constitute the first report in the Latin American region concerning the molecular detection and characterization of MLB-1 HAstV strains in environmental samples., Significance and Impact of the Study: This study highlights the benefits of an environmental surveillance to study emerging enteric viruses circulating in human societies., (© 2015 The Society for Applied Microbiology.)

Prevention of malaria in pregnant women is an utmost priority because the disease can cause serious maternal and neonatal complications. Maternal complications include marked anemia, increased risk of severe disease, and mortality, while the fetus or neonate is at risk of prematurity, anemia, and low birthweight. Pregnant women living in malaria endemic areas may be semiimmune to a particular Plasmodium spp. but when traveling to other regions, sometimes within their same country, where malaria epidemiology is different, may develop severe malaria complications. Here, we describe our experience in northeastern Venezuela associated with unfavorable outcomes of imported malaria cases among pregnant women who traveled to other Venezuelan regions with different malaria epidemiology. Travel medicine practitioners should be aware and educate their pregnant patients regarding the risk of malaria even when living in malaria endemic areas and traveling to other endemic areas such as occurs in Venezuela.

Clinico-epidemiological features of pediatric patients with malaria due Plasmodium vivax that developed anemia and thrombocytopenia requiring hospitalization are herein reported. Over a 3-year period, 78 children with P. vivax infection were admitted to our Hospital in Sucre, Venezuela. Clinical manifestations at admission were 93.59 per cent fever, 41.03 per cent chills and 14.10 per cent headache, among others. On paraclinical evaluations 94.87 percent presented with anemia (10.26 per cent severe), 25.64 percent with malnutrition, and 10.26 percent had intestinal parasitosis. The mean hemoglobin levels on admission were 8.09 g/dl and mean platelet counts 127 402 cells/mm3. Among these patients 58.97 per cent developed thrombocytopenia (24.36 per cent severe) requiring transfusion in 25.64 per cent of patients. After antimalarial treatment with chloroquine and primaquine and supportive care all patients were successfully discharged. No deaths or further complications were seen, except for persistent mild thrombocytopenia in 17.95 per cent of the patients.

PRL at a physiological concentration (10(-8) M) produced a very rapid and transient increase in 45Ca efflux in freshly isolated hepatocytes, which reached the highest value within 5 min and returned to baseline level after 20 min. PRL-induced 45Ca2+ efflux resulted in a loss of 15% of total cell calcium, which was similar to that found in vasopressin-treated cells. However, in contrast with the PRL effect, 45Ca2+ efflux induced by vasopressin was sustained. We demonstrate by using two different approaches, glycogen phosphorylase-a activation and direct cytosolic calcium concentration [( Ca2+]i) measurements, that PRL elicits a [Ca2+]i increase. The treatment of hepatic cells with PRL caused a 4-fold stimulation in glycogen phosphorylase-alpha activity after 2 min of PRL addition. Direct [Ca2+]i determination in fluo-3-loaded hepatocytes showed a 11% increase after 5 min of PRL addition. Similar data were observed in hepatocytes stimulated either with vasopressin (10(-7) M) or calcium ionophore A23187 (200 nM). The increase in [Ca2+]i promoted by PRL was independent of extracellular calcium or voltage-operated calcium channels. The data demonstrate that calcium is involved in the intracellular signaling of PRL in liver cells and that PRL initiates its action by a Ca2+ mobilization from the intracellular stores.