The effect of wiping and spray-wash temperature on bacterial retention on abraded domestic sink surfaces

The relative cleanability of artificially abraded stainless steel, enamelled steel, mineral resin and polycarbonate domestic sinks was assessed by examining bacterial retention after cleaning. Two cleaning regimes were used: the mechanical action of wiping combined with a spray-rinse, and spray-washing at a range of temperatures. After wiping, stainless steel retained 0.5–1 log order fewer bacteria than the enamel sinks which in turn were 0.5 log order cleaner than the mineral resin and polycarbonate sinks. After spray-washing, stainless steel retained 0.5 log order fewer bacteria than enamel which in turn was 0.5 log order cleaner than the polycarbonate and mineral resin. Extending the number of wipes or increasing spray-wash temperature enhanced bacterial removal but, in general, did not change the relative cleanability of the sink materials. As a cleaning technique, wiping was shown to be more effective than spray-washing in reducing bacterial numbers. SEM studies showed that bacteria were typically retained in surface imperfections, particularly pits and crevices such that surfaces which sustained the most extensive damage due to abrasion retained higher numbers of bacteria.     

Calcium content and distribution as a function of growth and transformation in the mouse 3T3 cell

Total Ca content and that fraction of Ca sensitive to removal by the chelator ethylene glycol-bis(β-aminoethyl ether)N,N,N',N'-tetraacetate (EGTA) have been investigated in the mouse 3T3 cell as a function of growth stage, transformation with SV40 virus, and serum levels of the media. Cells were allowed to grow through several doublings in media containing (45)Ca. The cellular content of (45)Ca was used to access total cell Ca. That fraction of (45)Ca removed by EGTA was presumed to represent primarily surface-localized Ca. The data are expressed on a per cell volume basis to compensate for size differences as a function of growth stage and transformation. During exponential growth phase, the 3T3 cell contains 525pmol Ca/μl cell volume. Of this, approx. 457 pmol/μl is not removable by EGTA and, presumably, is cytoplasmically located. This value is in close agreement with previous studies on the HeLa cell (470 pmol Ca/μl cell water after the removal of the surface Ca). The low level of EGTA- removable Ca present in the 3T3 cell during early exponential growth (68 pmol Ca/μl cell volume) increases progressively with increasing cell density, and upon quiescence it is sevenfold greater. In contrast, SV40- transformed 3T3 cells growing exponentially possess total levels of Ca which are approximately two-thirds the levels of the normal 3T3 cell. However, their EGTA-sensitive Ca is not significantly different from that of exponentially growing, normal 3T3 cells. As the transformed cells continue to grow at high density, their total ca and their sensitivity to EGTA do not change, in contrast to the normal 3T3 cell. Thus, an increase in Ca associated with the cell surface appears to be correlated with growth inhibition. This has been investigated further by regulating growth of the normal and transformed cell with alterations in the serum level of the media. In 4 percent calf serum the normal cell is stopped from continued proliferation. Growth stoppage under these conditions is characterized by a nearly fourfold increase in EGTA-removable Ca, similar to the increase observed upon quiescence in depleted 10 percent serum. Similar treatment of the transformed cell does not reduce its growth rate, nor does it significantly alter Ca distribution. However, at 0.5 percent medium serum levels, the SV40 3T3 growth rate is substantially reduced and, under these conditions, EGTA-removable Ca increases twofold.     

Recombinant adeno-associated virus type 2-mediated gene delivery into the <Emphasis Type="Italic">Rpe65</Emphasis><Superscript>-/-</Superscript> knockout mouse eye results in limited rescue

Background  

Leber's congenital amaurosis (LCA) is a severe form of retinal dystrophy. Mutations in the RPE65 gene, which is abundantly expressed in retinal pigment epithelial (RPE) cells, account for approximately 10–15% of LCA cases. In this study we used the high turnover, and rapid breeding and maturation time of the Rpe65 ^-/- knockout mice to assess the efficacy of using rAAV-mediated gene therapy to replace the disrupted RPE65 gene. The potential for rAAV-mediated gene treatment of LCA was then analyzed by determining the pattern of RPE65 expression, the physiological and histological effects that it produced, and any improvement in visual function.     

Importance of factors determining the effective lifetime of a mass, long-lasting, insecticidal net distribution: a sensitivity analysis

Background

Long-lasting insecticidal nets (LLINs) reduce malaria transmission by protecting individuals from infectious bites, and by reducing mosquito survival. In recent years, millions of LLINs have been distributed across sub-Saharan Africa (SSA). Over time, LLINs decay physically and chemically and are destroyed, making repeated interventions necessary to prevent a resurgence of malaria. Because its effects on transmission are important (more so than the effects of individual protection), estimates of the lifetime of mass distribution rounds should be based on the effective length of epidemiological protection.

Methods

Simulation models, parameterised using available field data, were used to analyse how the distribution's effective lifetime depends on the transmission setting and on LLIN characteristics. Factors considered were the pre-intervention transmission level, initial coverage, net attrition, and both physical and chemical decay. An ensemble of 14 stochastic individual-based model variants for malaria in humans was used, combined with a deterministic model for malaria in mosquitoes.

Results

The effective lifetime was most sensitive to the pre-intervention transmission level, with a lifetime of almost 10 years at an entomological inoculation rate of two infectious bites per adult per annum (ibpapa), but of little more than 2 years at 256 ibpapa. The LLIN attrition rate and the insecticide decay rate were the next most important parameters. The lifetime was surprisingly insensitive to physical decay parameters, but this could change as physical integrity gains importance with the emergence and spread of pyrethroid resistance.

Conclusions

The strong dependency of the effective lifetime on the pre-intervention transmission level indicated that the required distribution frequency may vary more with the local entomological situation than with LLIN quality or the characteristics of the distribution system. This highlights the need for malaria monitoring both before and during intervention programmes, particularly since there are likely to be strong variations between years and over short distances. The majority of SSA's population falls into exposure categories where the lifetime is relatively long, but because exposure estimates are highly uncertain, it is necessary to consider subsequent interventions before the end of the expected effective lifetime based on an imprecise transmission measure.     

First detection, isolation and molecular characterization of infectious salmon anaemia virus associated with clinical disease in farmed Atlantic salmon (Salmo salar) in Chile

Background  

Several forms of progressive retinal atrophy (PRA) segregate in more than 100 breeds of dog with each PRA segregating in one or a few breeds. This breed specificity may be accounted for by founder effects and genetic drift, which have reduced the genetic heterogeneity of each breed, thereby facilitating the identification of causal mutations. We report here a new form of PRA segregating in the Border Collie breed. The clinical signs, including the loss of night vision and a progressive loss of day vision, resulting in complete blindness, occur at the age of three to four years and may be detected earlier through systematic ocular fundus examination and electroretinography (ERG).     

Do European Standard Disinfectant tests truly simulate in‐use microbial and organic soiling conditions on food preparation surfaces?

Aims: The results from European standard disinfectant tests are used as one basis to approve the use of disinfectants in Europe. The design of these laboratory‐based tests should thus simulate as closely as possible the practical conditions and challenges that the disinfectants would encounter in use. No evidence is available that the organic and microbial loading in these tests simulates actual levels in the food service sector. Methods and Results: Total organic carbon (TOC) and total viable count (TVC) were determined on 17 visibly clean and 45 visibly dirty surfaces in two restaurants and the food preparation surfaces of a large retail store. These values were compared to reference values recovered from surfaces soiled with the organic and microbial loading, following the standard conditions of the European Surface Test for bactericidal efficacy, EN 13697. Conclusions: The TOC reference values for clean and dirty conditions were higher than the data from practice, but cannot be regarded as statistical outliers. This was considered as a conservative assessment; however, as additional nine TOC samples from visibly dirty surfaces were discarded from the analysis, as their loading made them impossible to process. Similarly, the recovery of test organisms from surfaces contaminated according to EN 13697 was higher than the TVC from visibly dirty surfaces in practice; though they could not be regarded as statistical outliers of the whole data field. No correlation was found between TVC and TOC in the sampled data, which re‐emphasizes the potential presence of micro‐organisms on visibly clean surfaces and thus the need for the same degree of disinfection as visibly dirty surfaces. Significance and Impact of the Study: The organic soil and the microbial burden used in EN disinfectant standards represent a realistic worst‐case scenario for disinfectants used in the food service and food‐processing areas.     

A new technique for the performance evaluation of clean-in-place disinfection of biofilms

A concentric cylinder reactor (CCR) is described that enables the steady-state kinetics of microbial biofilms to be evaluated under conditions of constant nutrient flow and variable shear-stress. The reactor has been used to evaluate the influence of fluid dynamic shear on the extent and mode of detachment of bacteria from biofilms. Using a food factory isolate of Pseudomonas aeruginosa, a general increase in the overall growth rate and detachment of the biofilms (cfu cm⁻² min⁻¹) with time was shown for each biofilm, regardless of the prevailing shear. As the shear rate was increased beyond 0.123 ms⁻¹, populations tended toward a pseudo steady-state. Sudden changes in shear force, however, caused dramatic changes in the productivity of steady-state populations. The CCR provides an effective means of testing disinfectant activity, particularly for clean-in-place situations, and allows for an examination of the residual effects of a cleansing programme on a treated surface for three different chemical classes of disinfectant. Utilisation of the CCR would, therefore, provide enhanced ability to determine the efficacy and efficiency of chemical products for use in sanitation protocols. Journal of Industrial Microbiology & Biotechnology (2000) 25, 235–241. Received 09 February 2000/ Accepted in revised form 27 June 2000     

Cleanability in relation to bacterial retention on unused and abraded domestic sink materials

The relative cleanability of stainless steel, enamelled steel, mineral resin and polycarbonate domestic sink materials was assessed by comparing the number of organisms remaining on surfaces after cleaning. In unused condition all materials, other than one enamelled steel, were equally cleanable. Stainless steel, abraded artificially or impact damaged to a similar degree as stainless steel subjected to domestic wear, retained approximately one log order less bacteria after cleaning than the other materials subjected to the same treatments. Little difference in cleanability was recorded between the abraded surfaces of the other materials although enamelled steel surfaces were less cleanable than mineral resin or polycarbonate after impact damage, because of the greater susceptibility of enamelled steel to damage by this treatment. When cleaning time was extended beyond 10 s for the abraded and impact damaged materials, their cleanability was not enhanced as compared with stainless steel. Changes in surface finish after abrasion were assessed by surface roughness measurement and scanning electron microscopy. Surfaces with poor cleanability before and after abrasion were characterized by pitting, crevices or jags. These surfaces are likely to retain more bacteria because of increased numbers of attachment sites, a larger bacterial/material surface contact area and topographical areas in which applied cleaning shear forces are reduced. Materials that resist surface changes, e.g. stainless steel, will remain more hygienic when subjected to natural wear than materials which become more readily damaged.     

Regulation of rat ovarian cell growth and steroid secretion

A cultured rat ovarian cell line (31 A-F(2)) was used to study the effect of growth factors (epidermal growth factor [EGF] and fibroblast growth factor [FGF]), a survival factor (ovarian growth factor [OGF]), a hormone (insulin), and an iron-binding protein (transferring) on cell proliferation and steroid production under defined culture conditions. EGF and insulin were shown to be mitogenic (half-maximal response at 0.12 nM and 0.11 muM, respectively) for 31A-F(2) cells incubated in serum-free medium. EGF induced up to three doublings in the cell population, whereas insulin induced an average of one cell population doubling. FGF, OGF, and transferrin were found not to have any prominent effect on cell division when incubated individually with 31A-F(2) cells in serum-free medium. However, a combination of EGF, OGF, insulin, and transferrin stimulated cell division to the same approximate extent as cells incubated in the presence of 5 percent fetal calf serum. EGF or insulin did not significantly affect total cell cholesterol levels (relative to cells incubated in serum-free medium) when incubated individually with 31A-F(2) cells. However, cell cholesterol levels were increased by the addition of OGF (250 percent), FGF (370 percent), or a combination of insulin and EGF (320 percent). Progesterone secretion from 31A-F(2) cells was enhanced by EGF (25 percent), FGF (80 percent), and insulin (115 percent). However, the addition of a mitogenic mixture of EGF, OGF, insulin, and transferrin suppressed progesterone secretion 150 percent) below that of control cultures. These studies have permitted us to determine that EGF and insulin are mitogenic factors that are required for the growth of 31A-F(2) cells and that OGF and transferrin are positive cofactors that enhance growth. Also, additional data suggest that cholesterol and progesterone production in 31A-F(2) cells can be regulated by peptide growth factors and the hormone insulin.