An in vivo evaluation of the effects of feed restriction regimens on wooden breast using ultrasound images as a predictive tool.

1. Gradual feed restriction was applied to broilers in order to reduce growth rate and, as a consequence, gradually impacts wooden breast myopathy occurrence. Ultrasound (US) images of breast muscle in live birds were correlated with breast fillets presenting wooden breast characteristics (WB). 2. A total of 1800 Cobb × Cobb 500 slow-feathering male chicks were fed one of the six feed restriction treatments with 12 replicates of 25 birds each, in a completely randomised design. Birds were fed ad libitum or were pair-fed to 50%, 60%, 70%, 80% or 90% of normal ad libitum intakes from 8 to 49 d to provide a gradual reduction in growth rate. Ultrasound images were obtained weekly from all birds and, in parallel, one bird per pen was weekly slaughtered and the major breast muscle was weighed and WB graded as 0 (normal), 1 (mild hardening in the upper), 2 (moderate hardening in the upper and/or lower), 3 (severe hardening) and 4 (severe hardening with haemorrhagic lesions and yellow fluid). Blood was taken for analysis of enzymes related to muscle cell breakdown. 3. Feed restriction applied at 50%, 60%, 70%, 80% and 90% of the ad libitum feed intake (FI) resulted in decreased body weight gain (BWG; P ≤ 0.05). 4. From 21 to 49 d, the increasing feed restriction led to linear increases (P ≤ 0.05) in WB scores, fibre density as well as breast depth and breast echogenicity. Creatine kinase, lactate dehydrogenase and aspartate aminotransferase concentration decreased linearly when broilers were feed restricted (P ≤ 0.05). 5. Wooden breast was positively correlated with echogenicity at 21 d (r = 0.510), 28 (r = 0.531), 35 (r = 0.470), 42 (r = 0.430) and 49 d (r = 0.548) (P ≤ 0.001). The use of breast echogenicity can be an additional tool to early detect alterations related to wooden breast.

Strychnos pseudoquina A. St. Hil.: a Brazilian medicinal plant with promising in vitro antiherpes activity.

AIMS: To investigate the anti-HSV and anti-inflammatory effects of a standardized ethyl acetate extract (SEAE) prepared with the stem bark of Strychnos pseudoquina, along with two isolated compounds: quercetin 3-O-methyl ether (3MQ) and strychnobiflavone (SBF). METHODS AND RESULTS: The mechanisms of action were evaluated by different methodological strategies. SEAE and SBF affected the early stages of viral infection and reduced HSV-1 protein expression. Both flavonoids elicited a concentration-dependent inhibition of monocyte chemoattractant protein-1 (MCP-1), whereas 3MQ reduced the chemokine release more significantly than SBF. Conversely, both compounds stimulated the production of the cytokines TNF-α and IL-1-ß in LPS-stimulated cells, especially at the intermediate and the highest tested concentrations. CONCLUSIONS: SEAE and SBF interfered with various steps of HSV replication cycle, mainly adsorption, postadsorption and penetration, as well as with ß and γ viral proteins expression; moreover, a direct inactivation of viral particles was observed. Besides, both flavonoids inhibited MCP-1 selectively, a feature that may be beneficial for the development of new anti-HSV agents. SIGNIFICANCE AND IMPACT OF THE STUDY: The results indicated that the samples present anti-HSV and anti-inflammatory activities, at different levels, which is an interesting feature since cold and genital sores are accompanied by an inflammation process.

Comparing the quality of life of adults with and without intellectual disability.

BACKGROUND: Quality of life (QOL) is an important issue for all citizens, comprising equity, equality, inclusion and participation of each person in society. The research purposes driving this investigation were (1) to compare QOL among people with and without intellectual disability (ID) and (2) to examine the predictors of QOL in both groups of participants. METHODS: Data were collected from 1929 adults, including 1264 individuals with an ID and the remainder (n = 665) were participants without a disability. QOL was assessed by the Portuguese version of the Personal Outcomes Scale. RESULTS: The findings supported significantly higher QOL for adults without ID. Furthermore, QOL is influenced by individual and environmental variables, stressing that the health status was the highest mutual predictor of QOL among those with and without ID. CONCLUSION: The gaps among both groups of participants should be addressed in service provision and policies. Taking into account the research findings, the personalised supports should enable the full inclusion and participation of adults with ID in community-based settings.

Antiherpes evaluation of soybean isoflavonoids.

The antiviral effects of soybean isoflavonoids have been investigated recently, especially those of genistein. It has been reported that this isoflavone is able to inhibit herpes simplex virus (HSV) replication, which is associated with skin and epithelial mucosa infections. The treatment of these infections with antiherpes drugs has resulted in the emergence of resistant viral strains. Based on this evidence, the aim of this study was to investigate the anti-HSV effects of soybean isoflavonoids: daidzein, genistein, glycitein, and coumestrol. Genistein and coumestrol inhibited HSV-1 (KOS and 29R strains, which are acyclovir sensitive and acyclovir resistant, respectively) and HSV-2 (333 strain) replication, whereas no antiviral effects were detected for daidzein and glycitein. The mechanisms of action were evaluated by different methodological strategies. Coumestrol affected the early stages of viral infection, and both compounds were able to reduce HSV-1 protein expression, as well as HSV-2 cell-to-cell spread.

Impact of a very low-energy diet on the fecal microbiota of obese individuals.

PURPOSE: Study how the dietary intake affects the fecal microbiota of a group of obese individuals after a 6-week very low-energy diet (VLED) and thereafter during a follow-up period of 5, 8, and 12 months. Additionally, we compared two different methods, fluorescent in situ hybridization (FISH) and real-time PCR (qPCR), for the quantification of fecal samples. METHODS: Sixteen subjects participated in a 12-month dietary intervention which consisted of a VLED high in protein and low in carbohydrates followed by a personalized diet plan, combined with exercise and lifestyle counseling. Fecal samples were analyzed using qPCR, FISH, and denaturing gradient gel electrophoresis. RESULTS: The VLED affected the fecal microbiota, in particular bifidobacteria that decreased approximately two logs compared with the baseline numbers. The change in numbers of the bacterial groups studied followed the dietary intake and not the weight variations during the 12-month intervention. Methanogens were detected in 56% of the participants at every sampling point, regardless of the dietary intake. Moreover, although absolute numbers of comparable bacterial groups were similar between FISH and qPCR measurements, relative proportions were higher according to FISH results. CONCLUSIONS: Changes in the fecal microbial numbers of obese individuals were primarily affected by the dietary intake rather than weight changes.

Simultaneous quantification of flavonoids from Achyrocline satureioides by a polar-reversed phase LC method--application to skin permeation/retention studies.

A selective and sensitive polar-reversed phase LC method was validated for simultaneous quantification of the main Achyrocline satureioides flavonoids (quercetin, luteolin, and 3-O-methylquercetin) in skin samples after permeation/retention studies from topical nanoemulsions. The method was linear in a range of 0.25 to 10.0 microg/mL exhibiting a coefficient of determination higher than 0.999 for all flavonoids. No interference of the nanoemulsion excipients or skin components was observed in the retention times of all flavonoids. The R.S.D. values for intra- and inter-day precision experiments were lower than 6.73%. Flavonoids recovery from nanoemulsions and skin matrices was between 90.05 and 109.88%. In a permeation/retention study with porcine ear high amount of 3-O-methylquercetin was found in the skin sample (0.92 +/- 0.22 microg/g) after two hours. The proposed method was suitable to quantify the main flavonoids of A. satureioides in skin permeation/retention studies from topical nanoemulsions.

In vivo anti-herpes simplex virus activity of a sulfated derivative of Agaricus brasiliensis mycelial polysaccharide.

Agaricus brasiliensis (syn. A. subrufescens), a basidiomycete fungus native to the Atlantic forest in Brazil, contains cell walls rich in glucomannan polysaccharides. The ß-(1 → 2)-gluco-ß-(1 → 3)-mannan was isolated from A. brasiliensis mycelium, chemically modified by sulfation, and named MI-S. MI-S has multiple mechanisms of action, including inhibition of herpes simplex virus (HSV) attachment, entry, and cell-to-cell spread (F. T. G. S. Cardozo, C. M. Camelini, A. Mascarello, M. J. Rossi, R. J. Nunes, C. R. Barardi, M. M. de Mendonça, and C. M. O. Simões, Antiviral Res. 92:108-114, 2011). The antiherpetic efficacy of MI-S was assessed in murine ocular, cutaneous, and genital infection models of HSV. Groups of 10 mice were infected with HSV-1 (strain KOS) or HSV-2 (strain 333). MI-S was given either topically or by oral gavage under various pre- and posttreatment regimens, and the severity of disease and viral titers in ocular and vaginal samples were determined. No toxicity was observed in the uninfected groups treated with MI-S. The topical and oral treatments with MI-S were not effective in reducing ocular disease. Topical application of MI-S on skin lesions was also not effective, but cutaneously infected mice treated orally with MI-S had significantly reduced disease scores (P < 0.05) after day 9, suggesting that healing was accelerated. Vaginal administration of MI-S 20 min before viral challenge reduced the mean disease scores on days 5 to 9 (P < 0.05), viral titers on day 1 (P < 0.05), and mortality (P < 0.0001) in comparison to the control groups (untreated and vehicle treated). These results show that MI-S may be useful as an oral agent to reduce the severity of HSV cutaneous and mucosal lesions and, more importantly, as a microbicide to block sexual transmission of HSV-2 genital infections.

Production of polysaccharide from Agaricus subrufescens Peck on solid-state fermentation.

The interest upon products obtained from fungi has increased during the recent years. Among the most noticeable, nutraceuticals, enzymes, and natural drugs occupy a privileged position. Fungal biomass for the obtainment of those products can be produced either by solid-state fermentation (SSF) or submersed fermentation. SSF has been employed for the production of spawn on pretreated wheat grains with the objective of increasing the fungal polysaccharide (glucomannans) contents. Among the important factors for the production of spawn, time of cooking, time of resting after grain cooking, consequently grain moisture, substrate pH, temperature of incubation, and initial inoculum amount are among the most significant. For wheat grains, cooking time of 21 min followed by a 24-min resting time has been shown as optimal for the production of glucomannans by the fungus Agaricus subrufescens (=Agaricus brasiliensis). Amendments of CaSO(4) (up to 3 %) and CaCO(3) (up to 1 %) had an important influence on the substrate pH. In general, better results for glucomannan production were obtained when no supplement was added or when up to 0.25 % CaCO(3) (pH 6.6) has been added to the mix. Our results demonstrate that the inoculum amount necessary for the best polysaccharide levels is around 10.3 %, while the best temperature is around 27.2 °C. Besides using the spawn for its main purpose, it could potentially and alternatively be used as nutraceutical due to the high levels of glucomannan observed (6.89 %), a compound technically proven to be a potent immunostimulatory and antitumoral agent.

Aneurysmal bone cyst of the mobile spine: the therapeutic role of embolization.

PURPOSE: Our aim is to define the role of embolization in the treatment of aneurysmal bone cyst of the spine in order to include this option in the decision making process. METHODS: From April 2004 to November 2009, seven patients with primary aneurysmal bone cyst of the mobile spine treated by embolization have been prospectively followed-up. All clinical presentations and imagings were recorded. There are many options of embolic agent and techniques used, but all aim to devascularize the tumor. The therapeutic protocol includes: embolization repeated every 8 weeks until the appearance of radiographic signs of healing. Complications, rate of healing and clinical outcome were analyzed. RESULTS: The number of embolizations varied from one to a maximum of seven without related intra- or post-operative complications. One patient, after four selective arterial embolizations, underwent direct percutaneous injection of embolic agents into the cyst. A clinical and radiographical response was achieved in all patients who were found alive and completely free of disease at mean follow-up of 46 months after last treatment and nobody crossed to surgical option. CONCLUSION: Embolization seems to be the first option for spinal aneurysmal bone cyst treatment because of the best cost-to-benefit ratio. It is indicated in intact aneurysmal bone cyst, when diagnosis is certain, when technically feasible and safe and when no pathologic fracture or neurologic involvements are found. If embolization fails, other options for treatment would still be available.

Staging and treatment of osteoblastoma in the mobile spine: a review of 51 cases.

PURPOSE: To define the role of Enneking staging system and of the consequent different treatment options on the outcome of osteoblastoma (OBL) of the spine. METHODS: A retrospective review of 51 patients with OBL of the mobile spine conducted to compare the outcomes among the different types of treatments at long term follow-up (25-229 months, av.90). These 51 patients were previously staged according to Enneking staging system and treatment selected accordingly. 10 stage two (st.2) OBLs were treated with intralesional excision and 41 stage three (st.3) OBLs were treated either by intralesional excision or en bloc resection. The intralesional excision group was divided considering the use or not of radiation therapy after surgery. The recurrence rate was compared among these groups and also considering previous open surgery ("non intact" vs. "intact"). The statistical significance was defined using the Fisher Exact test. RESULTS: No local recurrence occurred in the st.2 patients treated by intralesional excision. Considering the st.3 patients, 2 local recurrences out of 13 patients occurred in the en bloc resection (15.4 %) group. All occurred in "non intact" cases (67 %). In the intralesional group, 5 local recurrences out of 27 patients occurred (18 %) being none in the group that received radiation therapy after surgery. Two occurred in the "intact" (7 %) and three in the "non intact" group (75 %). Considering all patients, the difference between the recurrence rate between "intact" and "non intact" groups was statistically significant (p < 0.002). CONCLUSIONS: Intralesional excision proved to be effective in st.2 lesions and en bloc resection in st.3. Radiotherapy seems to be an effective adjuvant treatment when en bloc resection is not feasible or requires unacceptable functional sacrifices. The first treatment significantly affects the prognosis as previously treated patients have worse prognosis.

Genetic and molecular mechanisms of aluminum tolerance in plants.

Aluminum (Al) toxicity restricts root growth and agricultural yield in acid soils, which constitute approximately 40% of the potentially arable lands worldwide. The two main mechanisms of Al tolerance in plants are internal detoxification of Al and its exclusion from root cells. Genes encoding membrane transporters and accessory transcription factors, as well as cis-elements that enhance gene expression, are involved in Al tolerance in plants; thus studies of these genes and accessory factors should be the focus of molecular breeding efforts aimed at improving Al tolerance in crops. In this review, we describe the main genetic and molecular studies that led to the identification and cloning of genes associated with Al tolerance in plants. We include recent findings on the regulation of genes associated with Al tolerance. Understanding the genetic, molecular, and physiological aspects of Al tolerance in plants is important for generating cultivars adapted to acid soils, thereby contributing to food security worldwide.

Viability of human periodontal ligament fibroblasts in milk, Hank's balanced salt solution and coconut water as storage media.

AIM: To evaluate the effectiveness of various storage media at 5 °C for maintaining the viability of human periodontal ligament fibroblasts (PDLF). METHODOLOGY: Plates with PDLF were soaked in recently prepared Hank's balanced salt solution (HBSS), skimmed milk, whole milk, Save-A-Tooth(®) system's HBSS (Save), natural coconut water, industrialized coconut water or tap water (negative control) at 5 °C for 3, 6, 24, 48, 72, 96 and 120 h. Minimum essential medium (MEM) at 37 °C served as the positive control. PDL cell viability was determined by MTT assay. Data were statistically analysed by Kruskal-Wallis test complemented by the Scheffé test (α=5%). RESULTS: The greatest number of viable cells was observed for MEM. Skimmed and whole milk, followed by natural coconut water and HBSS, were the most effective media in maintaining cell viability (P<0.05). From 24 to 120 h, Save, industrialized coconut water and tap water were the worst storage media. CONCLUSIONS: Skimmed and whole milk had the greatest capacity to maintain PDLF viability when compared with natural coconut water, HBSS, Save, industrialized coconut water and tap water.

Progressive in vivo detection of wooden breast in broilers as affected by dietary energy and protein.

Wooden breast (WB) myopathy was investigated in broilers fed varying energy and protein at early ages. Correlation analyses were conducted between echogenicity of ultrasound images (US) of breast muscle from live birds and WB after slaughter. A total of 1,000 Cobb 500 one-day-old male chicks were fed on five dietary programs with eight replicates of 25 birds each, in a completely randomized design. Control feeds (commercially used ME and ideally balanced amino acids) or low-density feeds (low EP, with reductions of 50 kcal/kg ME and 0.20% dig. Lys compared to the control) were formulated. Feeds were provided in different periods: 1 to 7 d, 8 to 14 d, 15 to 21 d or 22 to 28 d. All broilers were fed a common basal diet thereafter until 49 d. Images using US were obtained once a week from all individuals and WB scored from one slaughtered bird per replication (0, normal; 1, mild hardening in the upper breast muscle; 2; moderate hardening in the upper and/or lower breast muscle; 3, severe hardening; 4, severe hardening with hemorrhagic lesions and yellow fluid). Blood was collected for enzyme investigation from the weekly slaughtered bird. Broilers had lower BWG and higher FCR when fed low EP feeds, regardless of the period fed when compared to the control (P < 0.001). Growth compensation, however, occurred afterwards such that all birds presented similar performance at the end. At 14, 21, and 28 d, broilers previously fed low EP feeds had lower WB scores (P < 0.001) compared to birds fed the control; however, both groups presented increased WB scores after 28 d. Wooden breast was positively correlated with breast echogenicity at 21 d (r = 0.31), 28 d (r = 0.43), 35 d (r = 0.21) and 42 d (r = 0.39). In conclusion, dietary energy and protein affected the development of WB scores in broilers and breast US images can be used as an early predictor of WB.

Assessment of adenovirus, hepatitis A virus and rotavirus presence in environmental samples in Florianopolis, South Brazil.

AIMS: To assess the presence of human adenovirus (HAdV), hepatitis A (HAV) virus and rotavirus A (RV-A) in environmental samples from the Southern region of Brazil and to provide viral contamination data for further epidemiological studies and governmental actions. METHODS AND RESULTS: Water samples from various sources (seawater, lagoon brackish water, urban wastewater, drinking water sources-with and without chlorination and water derived from a polluted creek) and oysters of two growing areas were analysed by enzymatic amplification (nested PCR and RT-PCR), quantification of HAdV genome (qPCR) and viral viability assay by integrated cell culture-PCR (ICC-PCR). From June 2007 to May 2008 in a total of 84 water samples, 54 (64·2%) were positive for HAdV, 16 (19%) for RV-A and 7 (8·3%) for HAV. Viability assays showed nonpositive samples for HAV; though, infectious viruses were confirmed for RV-A (12·5%) and HAdV (88·8%). Oyster samples by PCR were positive for HAdV (87·5%) and RV-A (8·3%), but none for HAV. Quantitative PCR in oysters showed means loads in genomic copies (gc) of 9·1 × 10(4) gc g(-1) (oyster farm south) and 1·5 × 10(5) gc g(-1) (oyster farm north) and in waters ranging from 2·16 × 10(6) (lagoon water) to 1·33 × 10(7) gc l(-1) (untreated drinking water). CONCLUSIONS: This study has shown a widespread distribution of the analysed viruses in this particular region with high loads of HAdV in the environment which suggests the relevance of evaluating these viruses as positive indicators of viral contamination of water. SIGNIFICANCE AND IMPACT OF THE STUDY: The environmental approach in this study provides data concerning the prevalence, viability and quantification of enteric viruses in environmental waters and oysters in the South region of Brazil and has indicated that their presence might pose a risk to population in contact with the environmental samples searched.

In vitro antiherpes effects of a C-glycosylflavonoid-enriched fraction of Cecropia glaziovii Sneth.

AIMS: To investigate the in vitro antiherpes effects of the crude aqueous extract obtained from Cecropia glaziovii leaves and their related fractions, the n-butanol fraction (n-BuOH) and the C-glycosylflavonoid-enriched fraction (MeOH(AMB)), and to determine the viral multiplication step(s) upon which this C-glycosylflavonoid-enriched fraction acts. METHODS AND RESULTS: The antiviral activity was evaluated against human herpes virus types 1 and 2 (HHV-1, HHV-2) by plaque reduction assay. The mode of action of the most active fraction was investigated by a set of assays, and the results demonstrated that MeOH(AMB) fraction exerts anti-herpes action by the reduction of viral infectivity (only against HHV-2); by the inhibition of virus entry into cells; by the inhibition of cell-to-cell virus spread as well as by the impaired levels of envelope proteins of HHV-1. The high-performance liquid chromatography (HPLC)-photo-diode array (PDA) analysis showed that the C-glycosylflavonoids are the major constituents of this fraction. CONCLUSIONS: These data showed that the MeOH(AMB) fraction has an antiviral activity against HHV types 1 and 2. The C-glycosylflavonoids are the major constituents of this fraction, which suggests that they could be one of the compounds responsible for the detected anti-herpes activity. SIGNIFICANCE AND IMPACT OF THE STUDY: The MeOH(AMB) fraction can be regarded as a phytopharmaceutical candidate for the treatment of herpetic infections.

Detection of enteric viruses in sewage sludge and treated wastewater effluent.

Sewage sludge and treated wastewater when contaminated with enteric virus and discharged into the environment, could pose a human health risk. The aim of study was to verify the presence and viability of enteric viruses in sewage sludge and treated wastewater at a local sewage plant in Florianopolis city, Brazil. Sewage sludge was concentrated by organic flocculation and polyethylene glycol precipitation and wastewater by electronegative membrane filtration and ultrafiltration by Centriprep Concentrator. Adenovirus (AdV), hepatitis A virus (HAV), and Rotavirus (RV) were examined for all samples for 12 months and Poliovirus (PV) was also tested for in sewage sludge samples. AdV was the most prevalent in both kind of samples, followed by RV, PV (in sludge) and HAV. Viral viability by cell culture (ICC-PCR) was: AdV: 100%, HAV: 16.7%, PV: 91.7%, RV: 25% in sludge and AdV: 66.6%, HAV: 66.6% and RV: 0% in wastewater. IFA for AdV in sludge ranged from 70 to 300 FFU/ml. QPCR for AdV ranged from 4.6 x 10(4) to 1.2 x 10(6) and from 50 to 1.3 x 10(4) gc/ml in sludge and wastewater, respectively. HAV quantification in sludge ranged from 3.1 x 10(2) to 5.4 x 10(2) gc/ml. In conclusion, it was possible to correlate presence and viability of enteric viruses in the environmental samples analyzed.

[Personal and social factors associated with the perception of health and the perception of happiness in a nonclinical adolescent population]. / Facteurs personnels et facteurs sociaux associés à la perception de santé et à la perception de bonheur dans une population adolescente non clinique.

INTRODUCTION: A theoretical conceptualization of the relationship between health and happiness and associated factors was studied using a structural equation model. METHOD: Data from the Portuguese part of a World Health Organization collaborative study, health behavior in school-aged children [Currie C, Smith R, Boyce W, et al. HBSC, a WHO cross national study: Research protocol for the 2001/2002 survey. Copenhagen: WHO; 2001], were used. A representative national sample (N=6131) of 11-, 13- and 15-year-old pupils was studied [Matos M.G., Equipa do Projecto Aventura Social. A saúde dos adolescentes portugueses: Quatro anos depois (La santé des adolescents portugais : quatre ans après). Lisboa: Edições FMH; 2003] and data were analyzed for the purpose of this paper. Results showed a significant positive correlation between perception of health and perception of happiness. A group of personal and social factors was associated with this correlation. RESULTS: Results demonstrated that personal factors had more impact on health perception, and social factors had more impact on happiness perception. Adjustment fit indexes were good. Socioeconomic status (SES) was significantly associated with a positive health perception, and SES was also significantly but modestly associated with the perception of happiness. DISCUSSION: Results highlighted the importance of a holistic and positive approach to adolescent health. This study confirmed the importance of family, peer relationship and positive evaluation of the school on adolescents' health and well-being. Results may suggest that preventive interventions in school settings should target both personal factors (e.g., promoting social competence) and social factors (e.g., promoting social support and acceptance). Interventions should also target physical and mental health in order to help adolescents to cope with the daily challenges they meet.

Biological activities of marine invertebrates extracts from the northeast brazilian coast.

This paper reports the in vitro antiproliferative effects, antiprotozoal, anti-herpes and antimicrobial activities of 32 organic extracts of 14 marine sponges and 14 corals collected in northeast Brazilian coast. The ethanolic extracts of the sponges Amphimedon compressa and Tedania ignis, and the acetone extract of Dysidea sp. showed relevant results concerning the antiproliferative effects against A549, HCT-8, and PC-3 cell lines by sulforhodamine B assay, but also low specificity. Concerning the antiprotozoal screening, the ethanolic extract of Amphimedon compressa and the acetone and ethanolic extracts of Dysidea sp. were the most active against Leishmania amazonensis and Trypanosoma cruzi expressing ß-galactosidase in THP-1 cells. In the preliminary anti-HSV-1 (KOS strain) screening, the ethanolic extracts of the sponges Amphimedon compressa, Haliclona sp. and Chondrosia collectrix inhibited viral replication by more than 50%. The most promising anti-herpes results were observed for the ethanolic extract of Haliclona sp. showing high selective indices against HSV-1, KOS and 29R strains (SI> 50 and >79, respectively), and HSV-2, 333 strain (IS>108). The results of the antibacterial screening indicated that only the ethanolic extract of Amphimedon compressa exhibited a weak activity against Enterococcus faecalis, Pseudomonas aeruginosa and Escherichia coli by the disk diffusion method. In view of these results, the extracts of Amphimedon compressa, Tedania ignis and Dysidea sp. were selected for further studies aiming the isolation and identification of the bioactive compounds with antiproliferative and/or antiprotozoal activities. The relevant anti-herpes activity of the ethanolic extract of Haliclona sp. also deserves special attention, and will be further investigated.

Comparative study of two extraction methods for enteric virus recovery from sewage sludge by molecular methods.

The aim of this study was to compare two nucleic acid extraction methods for the recovery of enteric viruses from activated sludge. Test samples were inoculated with human adenovirus (AdV), hepatitis A virus (HAV), poliovirus (PV) and rotavirus (RV) and were then processed by an adsorption-elution-precipitation method. Two extraction methods were used: an organic solvent-based method and a silica method. The organic-based method was able to recoup 20% of the AdV, 90% of the RV and 100% of both the PV and HAV from seeded samples. The silica method was able to recoup 1.8% of the AdV and 90% of the RV. These results indicate that the organic-based method is more suitable for detecting viruses in sewage sludge.

Evaluation of HA negatively charged membranes in the recovery of human adenoviruses and hepatitis A virus in different water matrices.

Human adenoviruses (HAdV) and hepatitis A virus (HAV) are shed in the faeces and consequently may be present in environmental waters, resulting in an increase in pathogen concentration that can affect water quality and human health. The aim of this study was to evaluate an adsorption-elution method which utilizes negatively charged membrane HA to determine the efficient recovery of HAdV and HAV from different water matrices and to combine this procedure with a qualitative molecular method (nested RT-PCR and nested PCR). The best efficiency recovery was achieved in distilled water and treated wastewater effluent (100%) for both viruses and in recreational lagoon water for HAV (100%). The efficiency recovery was 10% for HAdV and HAV in seawater and 10% for HAdV in lagoon water. The viral detection limit by nested PCR for HAV in water samples ranged between 20-0.2 FFU/mL and 250 and 25 TCID50/mL for HAdV. In conclusion, these results suggest that the HA negatively charged membranes vary their efficiency for recovery of viral concentration depending upon the types of both enteric viruses and water matrices.