Detection and risk assessment of adenoviruses in swimming pool water

AIMS: The role of swimming pool water as a source of human adenovirus (HAd) infection has previously been demonstrated. In this study, the risk of infection of HAds detected in a survey of swimming pool water from two indoor and one outdoor swimming pools over a period of 1 year was assessed. METHODS AND RESULTS: The HAds were concentrated from 1 l grab samples of swimming pool water using a silicon dioxide-based method. The extracted HAd DNA was amplified by means of a nested PCR method. Adenoviruses were detected in four of 26 samples (15.4%) from the indoor swimming pool A, eight of 38 samples (21.1%) from the indoor swimming pool B and three of 28 samples (10.7%) from the outdoor swimming pool C. Application of these results in an exponential risk assessment model indicated a daily risk of infection of 2.61 x 10(-3) for swimming pool A, 3.69 x 10(-3) for swimming pool B and 1.92 x 10(-3) for swimming pool C assuming a daily consumption of 30 ml of swimming pool water. CONCLUSIONS: No acceptable (tolerable) risk of infection has yet been recommended for swimming pool water. However, the quality of swimming pool water is generally expected to be similar to that of drinking water. One infection per 10 000 consumers per year has been recommended for drinking water. The risk of HAd infections calculated for the swimming pool water under investigation exceeded this acceptable risk. SIGNIFICANCE AND IMPACT OF THE STUDY: The finding that swimming pool water which conforms to generally accepted specifications for treatment, disinfection and indicator organisms constituted a risk of HAd infection, has implications for the swimming pool industry. The formulation of acceptable (tolerable) risks of infection for swimming pool water may be essential. Specifications will, therefore, have to be formulated to ensure that swimming pool water conforms to the acceptable risk of infection.     

Incidence of bacteremia in stressed and unstressed populations of the blue crab, Callinectes sapidus

The incidence of bacteremia in the blue crab, Callinectes sapidus, is reported to be in excess of 80%. Because these results have been controversial, a field study was initiated to determine the effect of commercial capture and handling stresses on the incidence and levels of infection in blue crabs. The majority (75%) of "unstressed" crabs which were captured individually and bled immediately upon removal from the water were bacteremic, with a geometric mean level of infection of 14 CFU/ml of hemolymph. Crabs collected by crab pot, confined within these pots for as long as 24 h, and sampled immediately after removal from the water had a similar mean level of infection. Crabs subjected to the stresses of commercial capture, handling, and transport showed a higher incidence of infection (91%) and a mean infection level of 46 CFU/ml. Injuries sustained by crabs during commercial handling are thought to be associated with the higher incidence of infection. Vibrio spp. were primarily responsible for progressive infections in commercially stressed crabs and were the predominant bacterial type in heavily infected crabs. Our results indicated that uninjured healthy crabs do not have sterile hemolymph but instead harbor low-level bacterial infections.     

Incidence of bacteremia in stressed and unstressed populations of the blue crab, Callinectes sapidus.

The incidence of bacteremia in the blue crab, Callinectes sapidus, is reported to be in excess of 80%. Because these results have been controversial, a field study was initiated to determine the effect of commercial capture and handling stresses on the incidence and levels of infection in blue crabs. The majority (75%) of "unstressed" crabs which were captured individually and bled immediately upon removal from the water were bacteremic, with a geometric mean level of infection of 14 CFU/ml of hemolymph. Crabs collected by crab pot, confined within these pots for as long as 24 h, and sampled immediately after removal from the water had a similar mean level of infection. Crabs subjected to the stresses of commercial capture, handling, and transport showed a higher incidence of infection (91%) and a mean infection level of 46 CFU/ml. Injuries sustained by crabs during commercial handling are thought to be associated with the higher incidence of infection. Vibrio spp. were primarily responsible for progressive infections in commercially stressed crabs and were the predominant bacterial type in heavily infected crabs. Our results indicated that uninjured healthy crabs do not have sterile hemolymph but instead harbor low-level bacterial infections.     

Risk assessment of adenoviruses detected in treated drinking water and recreational water

AIMS: Human adenoviruses (HAds) have previously been detected in sewage and polluted river and dam water, as well as treated drinking water. The 51 serotypes of HAds cause a wide range of infections with clinical manifestations associated with the gastrointestinal, respiratory and urinary tracts, and the eyes. Water may play a meaningful role in the transmission of many of these HAd serotypes, specifically the enteric HAds which are transmitted via the faecal-oral route. The presence of these viruses in water used for drinking and recreational purposes is considered to constitute a potential health risk. In this study, the risk of infection by the group of HAds previously detected over a period of 1 year in selected drinking water supplies, as well as river and dam water used for recreational purposes, was assessed. METHODS AND RESULTS: Adenoviruses were previously detected in nine of 204 (4.41%) samples of two drinking water supplies (A and B) treated and disinfected according to international specifications, in four of 51 (7.8%) samples of river water and nine of 51 (17.7%) samples of dam water. Application of these previously published results in an exponential risk assessment model indicated an annual risk of infection of 1.01 x 10(-1) and 1.7 x 10(-1) for drinking water supplies A and B, respectively, assuming a daily consumption of 2 l day(-1). The daily risk of infection constituted by HAds in the river water was calculated as 1.71 x 10(-4), and in the dam water as 3.12 x 10(-5), assuming a consumption of 30 ml of water per day. CONCLUSIONS: The risk of infection exceeded the tolerable risk of one infection per 10 000 consumers per year proposed for drinking water. However, the results for river and dam water used for recreational purposes were within the tolerable risk of one infection per 1000 bathers per day proposed for environmental waters used for recreational purposes. SIGNIFICANCE AND IMPACT OF THE STUDY: The study showed that the risk of HAd infection calculated for the drinking water supplies and the recreational water may overestimate the actual risk of infection, as conservative values were assumed for some of the variables. For a more accurate assessment of the potential risk of infection research should at least include a thorough investigation of the water consumption of individuals in South Africa, and the efficiency of recovery of the glass wool adsorption-elution method.     

Mitigation of Tomato Spotted Wilt Virus Infection and Symptom Expression by Water Stress

The effects of water stress on the infection and systemic movement of tomato spotted wilt virus (TSWV) in vegetative tomato plants are examined. Two groups of plants: water stressed (water potential ?1.31 MPa) and well-watered ones (water potential ?0.65 MPa), were mechanically inoculated on the third expanded leaf from the top. Each group was then divided into a well-watered and a water stressed one. TSWV inoculation was effective both on water-stressed and well-watered plants. In all groups, infection was first detected in roots, then in shoot apices and subsequently in the rest of the leaves. Systemic virus infection was somewhat slower in the water stressed plants. Post inoculation stress treatments had a significant effect in reducing infection as well as in attenuating TSWV symptoms. Results from the present work suggest that water stress may be effective in controlling virus concentration and systemic infection symptoms in tomato plants and could thus be used as a tool to study the regulation of development of TSWV infection in this system.     

Dynamics of tuberculosis in a naturally infected Badger population

The dynamics of bovine tuberculosis infection in a wild Badger population have been studied since 1981. Faeces are sampled regularly to determine the distribution of infection in the study area. Badgers in known infected social groups, plus the neighbouring groups, are periodically caught and subjected to full clinical and bacteriological examination to monitor the progression of infection within and between groups of Badgers. The interim results of this study are presented and discussed. During 5 years (1981–1985), the temporal spread of infection between social groups in the study area was slow and restricted. Some clinically affected Badgers have survived for long periods. Evidence of infection in individual Badgers indicates that pseudo-vertical (mother to cub) transmission may be important in the maintenance of infection in the Badger population. There is also evidence of horizontal transmission by bite wounding and aerosol infection. Badger mortality due to Mycobacterium bovis infection was low, and the prevalence of infection in male Badgers is greater than in females. The data accumulated so far show no apparent relationship between Badger population density and the prevalence of M. bovis infection, nor is there any evidence to support the hypothesis that M. bovis infection acts to depress Badger population density significantly below disease-free levels. Cattle and infected Badgers can co-exist for long periods without the disease necessarily being transmitted to cattle.     

Ecological characteristics of scale-bearing Chrysophyceae from The Netherlands

In 1979 50 sampling localities in the south-eastern part of The Netherlands have been sampled once, twice or thrice. The scale-bearing Chrysophyceae have been identified and their abundancies have been estimated. Determinations of pH, alkalinity and water temperature have been done, and the relation between the occurrence of the taxa and these environmental factors was studied. Cluster analysis (TWINSPAN) was used to detect groups of co-occurring taxa. The ordination of these taxa (DECORANA) showed that the first principal component was the pH and the second principal component was the water temperature. Although very few environmental data could be used, these results indicate the importance of the pH (and alkalinity) of fresh waters in the occurrence of the scale-bearing Chrysophyceae, whereas their quantitative development is greatly dependent on the water temperature. Experimental data are needed to test this hypothesis.     

Molecular dynamics simulations of urea and thermal-induced denaturation of S-peptide analogue

Molecular dynamics simulations of the S-peptide analogue AETAAAKFLREHMDS in water at 278 and 358 K, and in 8 M urea at 278 K were performed. The results show agreement with experiments. The helix is stable at low temperature (278 K), while at 358 K, unfolding is observed. The effects of urea on protein stability have been studied. The data support a model in which urea denatures proteins by: (1) diminishing the hydrophobic effect by displacing water molecules from the solvent shell around nonpolar groups; and (2) binding directly to amide units (NH and CO groups) via hydrogen bonds. The results of cluster analysis and essential dynamics analysis suggest that the mechanism of urea and thermal-induced denaturation may not be the same.     

Immunoregulatory role of intestinal surfactant-like particles during Salmonella typhimurium infection

Surfactants like particles (SLP) are secreted by Intestinal epithelium. These particles have the ability to lower surface tension of intestinal epithelial cells and contain small amounts of surfactant specific proteins A, B, and D. In the intestinal lumen they are known to function as lubricants and/or as a vehicle to deliver digestive enzymes to the luminal fluid. These particles have been found to have the ability in binding of uropathogenic E.coli. But their immunological function is not known. The present study was designed to assess the role of the SLP in the regulation of immune response during Salmonella (S) typhimurium infection using a rat an enteric model. The animals were divided in four different groups including control (PBS), rats fed fat diet (corn oil), rats fed fat diet followed with S. typhimurium infection and rats with S. typhimurium infection alone. The Peyer's patches (PP), intraepithelial (IE) and lamina propria (LP) mononuclear cells were isolated from the above-mentioned groups. These mononuclear cells were then incubated in presence of S. typhimurium lysate alone, SLP alone and S. typhimurium lysate and SLP together. T cell markers CD4 and CD8, cytokines mainly pro-inflammatory ones including IFN-gamma, TNF-alpha, IL-12 etc were studied under such conditions. In addition histological studies were also carried out under these conditions. We report in this study that SLP plays an important role in modulating the cytokine level during infection. The pro-inflammatory cytokines were found significantly reduced in SLP induced diet along with the infection group compared to the infection group alone. Histopathological studies revealed the breakdown of duodenal villi after infection while only broadening of villi was observed in rats given corn oil induced SLP along with infection. These results suggested an important immuno-modulatory role for SLP during Salmonella infection.     

Comparison of most probable number-PCR and most probable number-foci detection method for quantifying infectious Cryptosporidium parvum oocysts in environmental samples

Microbial contamination of public water supplies is of significant concern, as numerous outbreaks, including Cryptosporidium, have been reported worldwide. Detection and enumeration of Cryptosporidium parvum oocysts in water supplies is important for the prevention of future cryptosporidiosis outbreaks. In addition to not identifying the oocyst species, the U.S. EPA Method 1622 does not provide information on oocyst viability or infectivity. As such, current detection strategies have been coupled with in vitro culture methods to assess oocyst infectivity. In this study, a most probable number (MPN) method was coupled with PCR (MPN-PCR) to quantify the number of infectious oocysts recovered from seeded raw water concentrates. The frequency of positive MPN-PCR results decreased as the oocyst numbers decreased. Similar results were observed when MPN was coupled to the foci detection method (MPN-FDM), which was done for comparison. For both methods, infectious oocysts were not detected below 10(3) seeded oocysts and the MPN-PCR and MPN-FDM estimates for each seed dose were generally within one-log unit of directly enumerated foci of infection. MPN-PCR estimates were 0.25, 0.54, 0 and 0.66 log(10) units higher than MPN-FDM estimates for the positive control, 10(5), 10(4) and 10(3) seed doses, respectively. The results show the MPN-PCR was the better method for the detection of infectious C. parvum oocysts in environmental water samples.     

Transmission of white sturgeon iridovirus in Kootenai River white sturgeon Acipenser transmontanus

It is thought that white sturgeon iridovirus (WSIV) is transmitted vertically from adult white sturgeon Acipenser transmontanus to progeny, and that wild adults are carriers of this virus. Based on this assumption, egg disinfection trials were initiated using wild Kootenai River white sturgeon. Over 2 consecutive years, post-fertilized eggs were disinfected with iodine at concentrations ranging from 0 to 400 ppm. Eggs were incubated and progeny were reared on either de-chlorinated municipal or Kootenai River water. Juvenile sturgeon (mean weight 3.0 g) from these treatment groups were then subjected to a density stress (15 or 20 g(-1)) to manifest WSIV disease in individuals harboring the virus. In Year 1, mortality in all groups ranged from 6 to 37% and the use of municipal water was shown to significantly improve survival. However, WSIV infection was not detected in fish from any of the treatment groups or controls, and therefore did not contribute to the observed mortality. In Year 2, all treatment and control groups reared on Kootenai River water tested positive for WSIV infection and exhibited mortality ranging from 59 to 94%, but fish from groups reared on municipal water did not test positive for WSIV infection. This shows that that vertical transmission did not occur in this study. Horizontal transmission played a significant role in WSIV infection, but the lack of infection in Year 1 suggests a cyclic occurrence of the virus in the Kootenai River system. Although survival tended to be better in iodine-treated groups, the effects of iodine treatment in relation to WSIV transmission remain unknown. An important finding is that not all wild white sturgeon broodstock yield WSIV-positive progeny.     

Structure and dynamics of the dilauroylphosphatidylethanolamine lipid bilayer.

A 200-ps molecular dynamics (MD) simulation trajectory of a model dilauroylphosphatidylethanolamine (DLPE) bilayer in water at 315 K has been generated. Segmental order parameters, electron density profiles, and water pair distribution functions have been calculated. Comparison to experiment is made where possible. The dynamics of the system has been studied by analyzing the velocity autocorrelation functions (VAF) of both water and lipid atoms. Furthermore, the diffusive properties of water have been analyzed by computing the mean square displacement (MSD) and orientational correlation function (OCF) of water in two regions around the bilayer. The calculated order parameters show a behavior similar to the liquid crystalline phase of other bilayers, but the region around C1-C3 does not show the expected behavior. The electron density profile shows features that are characteristic of the liquid crystalline phase. The radial distribution functions suggest ordering of water near the charged head groups, which results in about 15 water molecules solvating each lipid molecule. We find from the VAF, MSD, and OCF calculation that the water molecules near the head groups of the lipid bilayer move more slowly than those further away. The VAF of the hydrocarbon chains have features of low-frequency motions that are probably cooperative nature in addition to the high-frequency motions associated with bond angle and torsional motions.     

Determination of Taxonomic Status of Pathogenic and Nonpathogenic Entamoeba histolytica Zymodemes Using Isoenzyme Analysis

ABSTRACT Entamoeba histolytica infection results in either asymptomatic colonization or invasion of host tissues leading generally to clinical symptoms. Zymodeme studies have demonstrated a correlation between isoenzyme profiles and clinical presentation. Thus, strains have been attributed to pathogenic or nonpathogenic groups according to their zymodeme. To determine the taxonomic relationship of these two groups, the isoenzyme profiles of 14 loci of 38 E. histolytica strains (pathogenic and nonpathogenic) and seven strains of other species of the same genus were analyzed. Genetic distance analysis clearly demonstrates the existence of two separate groups within the species E. histolytica .     

Determination of taxonomic status of pathogenic and nonpathogenic Entamoeba histolytica zymodemes using isoenzyme analysis.

Entamoeba histolytica infection results in either asymptomatic colonization or invasion of host tissues leading generally to clinical symptoms. Zymodemes studies have demonstrated a correlation between isoenzyme profiles and clinical presentation. Thus, strains have been attributed to pathogenic or nonpathogenic groups according to their zymodeme. To determine the taxonomic relationship of these two groups, the isoenzyme profiles of 14 loci of 38 E. histolytica strains (pathogenic and nonpathogenic) and seven strains of other species of the same genus were analyzed. Genetic distance analysis clearly demonstrates the existence of two separate groups within the species E. histolytica.     

Homogeneity of the morphological groups of bacteriophages infecting Bacteroides fragilis strain HSP40 and strain RYC2056

Bacteriophages infecting Bacteroides fragilis strains RYC2056 and HSP40 have been proposed as indicators of water quality. To accomplish this function, homogeneity of the group of phages detected by these strains is necessary to ensure that the final results are not due to the different kinetics of inactivation of the phages. To evaluate homogeneity, we observed by electron microscopy bacteriophages isolated from sewage with two Bacteroides fragilis strains (HSP40 and RYC2056). A predominant group of phages was observed, Siphoviridae with slightly curved tails. Detection of other minority groups, which could be present in the sample, was done with neutralization experiments by using antiserum against the majority group and with host mutants resistant to infection with the predominant phage. Although two other minority groups were observed, results showed that bacteriophages infecting B. fragilis strain HSP40 and strain RYC2056 form a homogeneous group, Siphoviridae with slightly curved tails being the most predominant in sewage.     

Conformations of model peptides in membrane-mimetic environments.

The influence of a membrane environment on the conformational energetics of a polypeptide chain has been investigated through studies of model peptides in a variety of membrane-mimetic media. Nuclear magnetic resonance (NMR) and circular dichroism (CD) data have been obtained for the peptides in bulk hydrophobic solvents, normal micelles, and reversed micelles. Several hydrophobic peptides which are sparingly soluble in water have been solubilized in aqueous sodium dodecyl sulfate (SDS) solution. NMR and CD data indicate that the micelle-solubilized peptides experience an environment with the conformational impact of bulk methanol, and have decreased conformational freedom. The site of residence of the peptides interacting with the micelles appears to be near the surfactant head groups, in a region permeated by water, and not in the micelle core. Strongly hydrophilic peptides have been solubilized in nonpolar solvents by reversed micelles. These peptides are located in small water pools in close association with the head groups of the surfactant. NMR and CD data show that there is a conformational impact of this interfacial water region on peptide solubilizates distinct from that of bulk water.     

The epidemiology of rotavirus gastroenteritis

The materials, comprehensively characterizing the water outbreaks of gastroenteritis with its etiology established and its clinico-epidemiological interpretation made, are presented. Along with other data, the water nature of the outbreak was verified by the detection of rotavirus antigen, made in the enzyme immunoassay, not only in patients, but also in water samples from the surface water source and from sewage. To isolate the infective agent from environmental objects, bentonite, a mineral sorbent, has been used for the first time. The authors believe that the water route plays the main (primary) role in the spread of rotavirus gastroenteritis and the adult population is the main reservoir of this infection. The fact the disease is registered mainly among younger age groups of the child population reflects only the maximum manifestation of rotavirus infection among these age groups.     

Trypanosoma cruzi: effect of benznidazole therapy combined with the iron chelator desferrioxamine in infected mice

Iron chelators have been employed in various studies aimed at evaluating the relationship between the iron status of the host and the development of infection. In the present study, the effects of benznidazole (BZ) therapy in combination with the iron chelator desferrioxamine (DFO) on the development of infection in mice inoculated with Trypanosoma cruzi Y strain have been investigated. Infected mice treated with DFO presented lower levels of parasitemia compared with infected untreated animals. Therapy with BZ for 21 days, with or without DFO, led to decreased parasitemia and reduced mortality, but BZ in combination with DFO treatment for 35 days (BZ/DFO-35) gave 0% mortality. All infected groups presented lower levels of iron in the liver, but serum iron concentrations were greater in DFO-35 and BZ/DFO-35, whereas hemoglobin levels were higher in BZ/DFO-35 and lower in DFO-35 compared with other treated groups. The percentage cure, determined from negative hemoculture and PCR results in animals that had survived for 60 days post-infection, was 18% for BZ and BZ/DFO-35, 42% for BZ combined with DFO for 21 days, and 67% for DFO-35. The results demonstrate that modification in iron stores increases BZ efficacy.     

Isolation and Characterization of Temperature-Sensitive Mutants of Vesicular Stomatitis Virus, New Jersey Serotype

Forty-eight temperature-sensitive (ts) mutants have been isolated from a wild-type strain of the New Jersey serotype of vesicular stomatitis virus (VSV) after exposure to the base analogue mutagen 5-fluorouracil. Of these mutants, 47 have been classified into 6 nonoverlapping complementation groups containing 21, 17, 4, 3, 2, and 1 mutant, respectively (1 mutant remaining unallocated). The ribonucleic acid (RNA) phenotype of 23 of these mutants has been established. Four of the six groups contain one or more mutants unable to synthesize detectable amounts of viral RNA under restrictive conditions (39 C). No complementation was observed in mixed infection with ts mutants from the five established complementation groups of the Indiana serotype of VSV.     

Characteristics of the meningococcal carrier state in organized collectives and its role in the development of generalized forms of meningococcal infection

Complex (epidemiological and bacteriological) investigations of the level and structure of meningococcal carriership among the members of organized collective bodies differing in the epidemiological situation with respect to meningococcal infection have been carried out. The absence of differences between the total level of meningococcal carriership and the morbidity rate with respect to the generalized forms of meningococcal infection has been shown. The presence of cases of meningococcal meningitis in the groups under study has been found to depend on the intensity of the circulation of certain meningococcal serogroups. The possibility of the ecological reservation of the causative agents of meningococcal infection as polyagglutinable forms has been suggested.