A note on the use of Rappaport-Vassiliadis medium (RIO/RV) for the isolation of salmonellas from sewage and sewage sludges

The use of a modified Rappaport broth for the selective enrichment of salmonellas in sewage sludge is described. Comparative trials were carried out using Muller Kauffman—Tetrathionate (MKT) broth and Rappaport-Vassiliadis (R10/RV) medium for selective enrichment. Results have indicated that R10/RV broth is more selective and is to be preferred for routine monitoring.     

Comparison of Muller-Kauffmann tetrathionate broth with Rappaport-Vassiliadis (RV) medium for the isolation of salmonellas from sewage sludge

The Rappaport-Vassiliadis (RV, modification of Rappaport's) medium was superior to Muller-Kauffmann tetrathionate broth (MKT) for the enrichment and isolation of salmonellas from sewage sludge samples, either naturally contaminated or artificially inoculated with salmonella. Isolation rates for RV (42°C) were higher even when MKT (43°C) performance was improved by the use of brilliant green sulphamandelate agar. Kinetic results show that the suppression of the competing flora was poorer in MKT than in RV. The use of high inoculum to medium volume ratios (e.g. 1:5000) increased the isolation rates from RV and kinetic data explain why this was so.     

Comparison of Muller-Kauffmann tetrathionate broth with Rappaport-Vassiliadis (RV) medium for the isolation of salmonellas from sewage sludge

The Rappaport-Vassiliadis (RV, modification of Rappaport's) medium was superior to Muller-Kauffmann tetrathionate broth (MKT) for the enrichment and isolation of salmonellas from sewage sludge samples, either naturally contaminated or artificially inoculated with salmonella. Isolation rates for RV (42 degrees C) were higher even when MKT (43 degrees C) performance was improved by the use of brilliant green sul-phamandelate agar. Kinetic results show that the suppression of the competing flora was poorer in MKT than in RV. The use of high inoculum to medium volume ratios (e.g. 1:5000) increased the isolation rates from RV and kinetic data explain why this was so.     

A note on the performance of Rappaport's medium, compared with Rappaport-Vassiliadis broth, in the isolation of salmonellas from meat products, after pre-enrichment

Salmonellas were isolated from meat products using a slightly modified Rappaport's enrichment medium (R25), Rappaport-Vassiliadis procedure (Rappaport's broth containing 10 ml instead of 30 ml of Malachite Green solution and incubated at 43^oC instead of 37^oC), and Muller-Kauffmann's tetrathionate broth. From 255 samples, 89 were found positive with the Rappaport-Vassiliadis procedure, 83 with the R25 broth, whereas only 43 were positive with Muller-Kauffmann broth. It is concluded that the R25 medium may be used as an alternative to the more effective Rappaport-Vassiliadis broth when the only available incubation temperature is 37^oC.     

Dynamics of salmonella isolation with modified Rappaport's medium (R10)

Enhanced growth of salmonellas in Rappaport's medium as modified by Vassiliadis et al. (1976) after pre-enrichment in buffered peptone water during the first 6 h was obtained by replacement of tryptone by soya peptone. The competing bacteria, i.e. those which grow on brilliant green agar and which may interfere with the isolation of salmonellas when Rappaport's medium (R10) incubated at 43 degrees C is used for enrichment were inhibited or reduced in numbers when the normal amount of 5 g soya peptone/litre was used. When the amount was increased to 10 g/l, growth occurred, mainly of Enterobacter and Klebsiella species. The isolation of salmonellas was found to be largely dependent on the number and the ratio of their competitors. Every measure taken to reduce the number of competitive bacteria increases the possibility of isolating salmonellas. This explains the effect of improved selectivity of Rappaport's medium when small inocula are used. Rapid onset of growth of salmonellas by employing soya peptone introduces the possibility of using shorter incubation times 48 h.     

Dynamics of salmonella isolation with modified Rappaport's medium (R10)

Enhanced growth of salmonellas in Rappaport's medium as modified by Vassiliadis et al. (1976) after pre-enrichment in buffered peptone water during the first 6 h was obtained by replacement of tryptone by soya peptone. The competing bacteria, i.e. those which grow on brilliant green agar and which may interfere with the isolation of salmonellas when Rappaport's medium (R10) incubated at 43°C is used for enrichment were inhibited or reduced in numbers when the normal amount of 5 g soya peptone/litre was used. When the amount was increased to 10 g/l, growth occurred, mainly of Enterobacter and Klebsiella species. The isolation of salmonellas was found to be largely dependent on the number and the ratio of their competitors. Every measure taken to reduce the number of competitive bacteria increases the possibility of isolating salmonellas. This explains the effect of improved selectivity of Rappaport's medium when small inocula are used. Rapid onset of growth of salmonellas by employing soya peptone introduces the possibility of using shorter incubation times than 48 h.     

A note on the isolation of salmonellas from environmental samples using three formulations of Rappaport's broth

The efficiency of isolation of salmonellas from 412 seagull intestines and 247 polluted water samples was compared using three formulations of Rappaport's medium (RV, RV-soya and R25). A modification of RV medium (RV-soya) was shown to be the most efficient of the three media and it was shown that the duration of incubation of this medium could be restricted to 24 h. Inoculation ratios were compared for RV and RV-soya and 1:100 was shown to be significantly better than 1:20. It is concluded that RV-soya is at least as efficient as the standard RV medium and its use can therefore be recommended.     

A comparison of isolation procedures for salmonellas from polluted water using two forms of Rappaport's medium

The efficiency of Rappaport's broth ( RB10 ) and Rappaport's broth containing novobiocin ( NRB10 ) were compared for the isolation of salmonellas from polluted water, both as direct enrichment media and after pre-enrichment in buffered peptone water. Ninety samples were examined and 41 were found to contain salmonellas by at least one of the procedures used. Direct inoculation of the sample into RB10 resulted in the recovery of salmonellas from only 29.3% of the samples found to be positive. The use of NRB10 as a direct enrichment medium increased the percentage recovery to 78.0% of the total positive samples. Pre-enrichment in buffered peptone water allowed the recovery of salmonellas from a total of 41 samples whereas direct enrichment recovered them from only 32. No significant difference was demonstrated in the efficiencies of RB10 and NRB10 in recovering salmonellas after pre-enrichment in buffered peptone water. Three selective agars were used: no significant difference in their ability to recover salmonellae was demonstrated.     

A comparison of isolation procedures for salmonellas from polluted water using two forms of Rappaport's medium

F ricker , C.R. 1984. A comparison of isolation procedures for salmonellas from polluted water using two forms of Rappaport's medium. Journal of Applied Bacteriology 56 , 305–309.
The efficiency of Rappaport's broth (RB10) and Rappaport's broth containing novobiocin (NRB10) were compared for the isolation of salmonellas from polluted water, both as direct enrichment media and after pre-enrichment in buffered peptone water. Ninety samples were examined and 41 were found to contain salmonellas by at least one of the procedures used. Direct inoculation of the sample into RB10 resulted in the recovery of salmonellas from only 29.3% of the samples found to be positive. The use of NRB10 as a direct enrichment medium increased the percentage recovery to 78.0% of the total positive samples. Pre-enrichment in buffered peptone water allowed the recovery of salmonellas from a total of 41 samples whereas direct enrichment recovered them from only 32. No significant difference was demonstrated in the efficiencies of RB10 and NRB10 in recovering salmonellas after pre-enrichment in buffered peptone water. Three selective agars were used; no significant difference in their ability to recover salmonellae was demonstrated.     

A note on the preparation and use of a selective differential medium for the isolation of Acinetobacter spp. from clinical sources

A selective, differential medium is reported for the isolation of Acinetobacter spp. With this medium it was found that not one of 30 people carried Acinetobacter spp. in their faeces, but 2 of 40 individuals carried it on their skin.     

A note on a selective medium for the isolation of cutaneous propionibacteria

A selective medium containing 6 μg/ml furoxone was developed for routine isolation of cutaneous propionibacteria from mixed floras including staphylococci.     

A comparison of the effect of different sewage sludges on young barley

Summary Soil treated with three kinds of sewage sludge (activated, liquid digested, filtered digested sludge) was cropped three times with young barley.All three sludges produced a P-response, and a small N-response. For the first crop this was more marked with the liquid sludges than the filter-cake, from which the soluble nutrients had been removed in the filtrate. It was not possible to separate fully the effects of N and P in heavy sludge applications from possible harmful effects of heavy metals, but there was no certain harm from five times the recommended maximum application, and certainly no harm from lesser applications.Even after 12 months the fractions avilable/total Cu, Ni and Zn added in the sludges were greater (0.8, 0.2, 0.4) than those native in the soil, but their availabilities to young barley had substantially decreased during that period.By the end of 12 months there appeared to be no differences between the availabilities of these elements from different sludges, whatever may have been their original forms of combination. Relative to the total amount present, the concentration of Ni in water extracts of the soils was considerably greater than that of Zn and Cu. re]19760505     

A selective medium for the isolation of Arcobacter from meats

A method, including enrichment in Arcobacter Selective Broth (ASB) and isolation on semisolid Arcobacter Selective Medium (ASM) under aerobic conditions at 24°C, is described for the isolation of Arcobacter from retail meat products. Selective agents used in ASB and ASM were cefoperazone, trimethoprim, piperacillin and cycloheximide. Arcobacters were isolated from 53 (24.1%) of 220 poultry meat products and also, at lower incidence from samples of beef and pork. The isolates were identified as A. butzleri or A. butzleri -like and belonged to a wide variety of serotypes and biotypes.     

A note on the effect of the lactoperoxidase systems on salmonellas in vitro and in vivo

The lactoperoxidase system (LPS), a natural bactericidal system in milk, was investigated for its activity against salmonellas in vivo and in vitro. In acidified raw milk, in which the LPS was supplemented with an exogenous supply of H2O2, the numbers of salmonellas decreased rapidly. Different salmonella serotypes were affected to the same extent; rough strains, however, were more susceptible than smooth strains. When calves were fed on fresh milk, containing the LPS, and challenged with Salmonella typhimurium in doses of either 10(9) or 10(10), the clinical findings and salmonella excretion patterns were similar to those of control calves fed on heated milk. It was concluded that further studies, perhaps in the field, are necessary to evaluate LPS as a possible non-antibiotic system to control salmonellosis.