search.noResults

search.searching

dataCollection.invalidEmail
note.createNoteMessage

search.noResults

search.searching

orderForm.title

orderForm.productCode
orderForm.description
orderForm.quantity
orderForm.itemPrice
orderForm.price
orderForm.totalPrice
orderForm.deliveryDetails.billingAddress
orderForm.deliveryDetails.deliveryAddress
orderForm.noItems
PRESERVATIVES 29


Table 3: Comparison of Log reduction and % kill Initial (Ni)


5,000,000 5,000,000 5,000,000 5,000,000 5,000,000 5,000,000


500,000 50,000 5,000 500 50 10


Recovered (Nx) Killed (Ni-Nx) (log10


Log reduction Ni-log10


% kill Nx) [(Ni-Nx)/Ni]*100


4,500,000 190 4,950,000 299 4,995,000 4,999,500 4,999,950 4,999,990


3 4 5


5.6989


organisms. In contrast a population that has a narrow variation on MIC values will likely show slower initial action followed by a rapid and complete kill. The growth phase of the culture also


determines the tolerance to biocides. The growth curve shown in Figure 2 shows the 4 phases of growth of any population. Test organisms are usually prepared in either log (exponential growth) or stationary phase and determined by the age of the culture. Standard Test methods usually stipulate the period of incubation of a test culture to ensure it is in the appropriate growth phase. Cultured bacterial cells supplied with excess nutrients grow quickly during exponential growth phase. Once there is a shortage of nutrients or excess of waste products, the culture enters stationary and then decline phases. Bacteria respond to the starvation stress with growth rate reduction and induction of defense mechanisms.5,6


As a result, they


may become more tolerant to biocides. Cultures in exponential growth are actively metabolising and are therefore more susceptible to the effect of biocides due to increased free radical production from interference with normal metabolism. Bacteria usually become more resistant to environmental stress during slowing down of growth. This has been seen in both steady state and batch growth of cells and by comparing mid exponential with stationary phase growth, large differences in efficacy have been demonstrated. The most tolerant cells are those in decline phase. While only viable cells will be counted to determine the initial cell concentration, the population may contain up to ten times the number of dead cells than viable cells. The dead cells will undergo lysis and release cell debris and intact enzyme systems that may interfere with the efficacy of the biocide.6 Test cultures may be prepared by


growing them in a liquid medium or on a solid agar. Cells grown in liquid media have been shown to be hardier and less susceptible to antimicrobial attack than those washed from solid agar surfaces. Almost all test methods include a


April 2020


antimicrobial cutting boards while acceptable for antibacterial socks that just need to stop growth rather than kill any organisms.


99.9 99.99 99.999 99.9998


requirement for the number of cells to be included in the inoculum along with how they are prepared. Cells grown in liquid medium or washed from solid agar require diluting to the required concentration. The dilution of the cells may result in a carryover of nutrients into the inoculum, particularly from liquid cultures. This may be removed by centrifuging and resuspending the cells in a non-nutrient solution, such as isotonic saline. Other test methods require the cells to be diluted in nutrient broth7 nutrient broth.8


or even a 1:500 dilution of These variations result in


different supply of necessary nutrients to the cells to support growth and they may also inhibit the biocide under test. The number of cells included in the inoculum is critical to the test result and will be further discussed later in this paper.


Step 2: Add test organisms to the sample under test


The inoculum may vary in volume, placement and method of addition. The volume of inoculum added may affect the efficacy by diluting the test material if too much or repeated inoculations are conducted. The inoculum may be dripped in or onto a test samples, sprayed on and then mixed or left on the surface, all may affect the final outcome.


Step 3: Allow a desired contact period


The contact time of the microorganisms with the test material must be realistic and sensible. Disinfectant tests typically allow 5-8 minutes contact while preservative efficacy tests allow days and weeks to see an effect. These times reflect actual in use periods. Tests for antibacterial hand wash are typically conducted using 30 seconds and 5 minutes contact. Results for 5 minutes contact are often reported and are entirely unrealistic if the hand wash is to be rinsed off. However, if it is a leave on product, it may be a reasonable time. The standards for antibacterial surfaces and textiles utilises a 24 hour contact period5,7 which may be totally inappropriate, such as for medical staff uniforms or


Step 4: Neutralise the active All antimicrobial tests require that the effect of the antimicrobial agent must be neutralised or stopped following the required contact time so that surviving organisms can be recovered. If this does not occur, the killing effect on cells may be allowed to continue well past the required contact time and may even inhibit growth of surviving organisms resulting in incorrect test results. If available a neutraliser specific for the antimicrobial is used, such as sodium thiosulphate for chlorinated water.


Otherwise general neutralisers, such as Lecithin and Tweens are used in varying concentrations. A list of examples of neutralisers is included in Annex C of ISO 119320:2019.10


If it is not possible to


neutralise the biocide then dilution of the test solution may be used. It is usual to find the result where no organisms are recovered reported as <10cfu/mL due to at least a 1:10 dilution in neutraliser. If this cannot be validated, then a result of <100cfu/mL is often seen indicating further dilution to 1:100 was necessary. It is usually considered that if a 1:1000 dilution is required to validate the neutraliser system, then the test product is antimicrobial. All antimicrobial tests require that the neutraliser is validated to ensure a correct result has been achieved.


Step 5: Determine survivors The enumeration method of surviving organisms may also affect the test result. The diluent and agar type chosen may influence recovery as surviving organisms may have been sub lethally injured. Agars with lower nutrient value, such as plate count agar (PCA) or synthetic agars like R2A, has been found to give better recovery than a high nutrient agar such as Tryptone Soya Agar (TSA) and the choice of agar used may affect the results obtained.11 Counts conducted using the spread plate technique, spreading the inoculum over the surface of pre-poured agar plates, have been claimed to give a higher recovery than pour plates, where molten agar is poured onto the recovered organisms and mixed, due to less chance of heat stress on the surviving organisms. The counter argument is that pour plates give an additional dilution step thereby improving recovery of surviving organisms by reducing the concentration of any carried over preservative/biocide. Pour plates also have an increased sensitivity over spread plates due to a higher volume of inoculum into the plates.


PERSONAL CARE EUROPE


Page 1  |  Page 2  |  Page 3  |  Page 4  |  Page 5  |  Page 6  |  Page 7  |  Page 8  |  Page 9  |  Page 10  |  Page 11  |  Page 12  |  Page 13  |  Page 14  |  Page 15  |  Page 16  |  Page 17  |  Page 18  |  Page 19  |  Page 20  |  Page 21  |  Page 22  |  Page 23  |  Page 24  |  Page 25  |  Page 26  |  Page 27  |  Page 28  |  Page 29  |  Page 30  |  Page 31  |  Page 32  |  Page 33  |  Page 34  |  Page 35  |  Page 36  |  Page 37  |  Page 38  |  Page 39  |  Page 40  |  Page 41  |  Page 42  |  Page 43  |  Page 44  |  Page 45  |  Page 46  |  Page 47  |  Page 48  |  Page 49  |  Page 50  |  Page 51  |  Page 52  |  Page 53  |  Page 54  |  Page 55  |  Page 56  |  Page 57  |  Page 58  |  Page 59  |  Page 60  |  Page 61  |  Page 62  |  Page 63  |  Page 64  |  Page 65  |  Page 66  |  Page 67  |  Page 68  |  Page 69  |  Page 70  |  Page 71  |  Page 72  |  Page 73  |  Page 74  |  Page 75  |  Page 76  |  Page 77  |  Page 78  |  Page 79  |  Page 80  |  Page 81  |  Page 82  |  Page 83  |  Page 84  |  Page 85  |  Page 86  |  Page 87  |  Page 88  |  Page 89  |  Page 90  |  Page 91  |  Page 92  |  Page 93  |  Page 94  |  Page 95  |  Page 96  |  Page 97  |  Page 98  |  Page 99  |  Page 100  |  Page 101  |  Page 102  |  Page 103  |  Page 104  |  Page 105  |  Page 106  |  Page 107  |  Page 108  |  Page 109  |  Page 110  |  Page 111  |  Page 112  |  Page 113  |  Page 114  |  Page 115  |  Page 116  |  Page 117  |  Page 118  |  Page 119  |  Page 120  |  Page 121  |  Page 122  |  Page 123  |  Page 124  |  Page 125  |  Page 126  |  Page 127  |  Page 128  |  Page 129  |  Page 130  |  Page 131  |  Page 132  |  Page 133  |  Page 134  |  Page 135  |  Page 136  |  Page 137  |  Page 138  |  Page 139  |  Page 140  |  Page 141  |  Page 142  |  Page 143  |  Page 144  |  Page 145  |  Page 146  |  Page 147  |  Page 148  |  Page 149  |  Page 150  |  Page 151  |  Page 152  |  Page 153  |  Page 154  |  Page 155  |  Page 156  |  Page 157  |  Page 158  |  Page 159  |  Page 160  |  Page 161  |  Page 162  |  Page 163  |  Page 164  |  Page 165  |  Page 166  |  Page 167  |  Page 168  |  Page 169  |  Page 170  |  Page 171  |  Page 172  |  Page 173  |  Page 174  |  Page 175  |  Page 176  |  Page 177  |  Page 178  |  Page 179  |  Page 180  |  Page 181  |  Page 182  |  Page 183  |  Page 184  |  Page 185  |  Page 186  |  Page 187  |  Page 188  |  Page 189  |  Page 190  |  Page 191  |  Page 192  |  Page 193  |  Page 194  |  Page 195  |  Page 196