Antibiotic resistance in coagulase negative staphylococci isolated from wastewater and drinking water

This study reports the antibiotic resistance patterns of coagulase negative staphylococci (CNS) isolated from a drinking water treatment plant (WTP), a drinking water distribution network, responsible for supplying water to the consumers (WDN), and a wastewater treatment plant (WWTP), responsible for receiving and treating domestic residual effluents. Genotyping and the 16S rRNA gene sequence analysis demonstrated a higher diversity of species both in the WTP (6 species/19 isolates) and WWTP (12 species/47 isolates) than in the WDN (6 species/172 isolates). Staphylococcus pasteuri and Staphylococcus epidermidis prevailed in the WTP and WDN and Staphylococcus saprophyticus in the WWTP. Staphylococci with reduced susceptibility (resistance or intermediary phenotype) to beta-lactams, tetracycline, clindamycin and erythromycin were observed in all types of water and belonged to the three major species groups. The highest resistance rate was found against erythromycin, presumably due to the presence of the efflux pump encoded by the determinant msrA, detected in the majority of the resistant isolates. This study demonstrates that antibiotic resistant CNS may colonize different types of water, namely drinking water fulfilling all the quality standards.     

Phylogenetic diversity of aqueous microorganisms separated after the advanced tertiary of tap water

The species composition of bacterial community of the drinking tap water and water samples taken at different purification stages of the water conditioning plant at an enterprise producing special drinks has been analyzed. This study involves the analysis of 16S rRNA gene sequences and the use of molecular biology methods based on the phylogenetic analysis. The following bacterial species were identified: Bacillus nanhaiensis, Brevibacterium frigoritolerans, and Lysinibacillus fusiformis. All the studied strains belong to phylum Firmicutes, class Bacilli representing Gram-positive microorganisms that are not pathogenic to human. The presence of the bacteria investigated in different habitats such as the drinking chlorinated water, in the presence of stress factors (arid climate, high or low temperatures, depleted soil, and the occurrence of disinfectants) indicates their ability to easily adapt to new living conditions expanding their habitat.     

Bottled mineral water as a potential source of antibiotic resistant bacteria

The antibiotic resistance phenotypes of the cultivable bacteria present in nine batches of two Portuguese and one French brands of commercially available mineral waters were examined. Most of the 238 isolates recovered on R2A, Pseudomonas Isolation agar or on these culture media supplemented with amoxicillin or ciprofloxacin, were identified (based on 16S rRNA gene sequence analysis) as Proteobacteria of the divisions Beta, Gamma and Alpha. Bacteria resistant to more than three distinct classes of antibiotics were detected in all the batches of the three water brands in counts up to 10² CFU/ml. In the whole set of isolates, it was observed resistance against all the 22 antimicrobials tested (ATB, bioMérieux and disc diffusion), with most of the bacteria showing resistance to three or more classes of antibiotics. Bacteria with the highest multi-resistance indices were members of the genera Variovorax, Bosea, Ralstonia, Curvibacter, Afipia and Pedobacter. Some of these bacteria are related with confirmed or suspected nosocomial agents. Presumable acquired resistance may be suggested by the observation of bacteria taxonomically related but isolated from different brands, exhibiting distinct antibiotic resistance profiles. Bottled mineral water was confirmed as a possible source of antibiotic resistant bacteria, with the potential to be transmitted to humans.     

Diversity and antibiotic resistance of Aeromonas spp. in drinking and waste water treatment plants

The taxonomic diversity and antibiotic resistance phenotypes of aeromonads were examined in samples from drinking and waste water treatment plants (surface, ground and disinfected water in a drinking water treatment plant, and raw and treated waste water) and tap water. Bacteria identification and intra-species variation were determined based on the analysis of the 16S rRNA, gyrB and cpn60 gene sequences. Resistance phenotypes were determined using the disc diffusion method.Aeromonas veronii prevailed in raw surface water, Aeromonas hydrophyla in ozonated water, and Aeromonas media and Aeromonas puntacta in waste water. No aeromonads were detected in ground water, after the chlorination tank or in tap water. Resistance to ceftazidime or meropenem was detected in isolates from the drinking water treatment plant and waste water isolates were intrinsically resistant to nalidixic acid. Most of the times, quinolone resistance was associated with the gyrA mutation in serine 83. The gene qnrS, but not the genes qnrA, B, C, D or qepA, was detected in both surface and waste water isolates. The gene aac(6’)-ib-cr was detected in different waste water strains isolated in the presence of ciprofloxacin. Both quinolone resistance genes were detected only in the species A. media. This is the first study tracking antimicrobial resistance in aeromonads in drinking, tap and waste water and the importance of these bacteria as vectors of resistance in aquatic environments is discussed.     

Occurrence and diversity of Arcobacter spp. along the Llobregat River catchment, at sewage effluents and in a drinking water treatment plant

The presence of Arcobacter species in faecally contaminated environmental waters has previously been studied. However, the ability to eliminate Arcobacter during the water treatment processes that produce drinking water has been little studied. We have investigated the prevalence and diversity of Arcobacter spp. throughout the year at 12 sampling points in the Llobregat River catchment (Catalonia, Spain) including 3 sites at a drinking water treatment plant. Positive samples for Arcobacter spp., came predominantly from the most faecally polluted sites. Recovery rates from all sites were greater in the spring (91.7%) and summer (83.3%) than in autumn and winter (75.0% in both cases), but this trend was not statistically evaluated due to the limited number of samples. Among the 339 colonies analyzed, the most prevalent species by multiplex PCR and 16S rDNA restriction fragment length polymorphism were Arcobacter butzleri (80.2%), followed by Arcobacter cryaerophilus (19.4%) and Arcobacter skirrowii (0.3%). Isolates showed a high genotype diversity as determined by the enterobacterial repetitive intergenic consensus PCR. In fact, 91.2% (309/339) of the colonies had different genotypes, i.e. 248 of them among the 275 isolates of A. butzleri and 60 among the 63 isolates of A. cryaerophilus and 1 genotype of A. skirrowii. Arcobacter was never detected or isolated from finished drinking water, demonstrating that water treatment is effective in removing Arcobacter species.     

Biodegradation of geosmin by a novel Gram-negative bacterium; isolation, phylogenetic characterisation and degradation rate determination

Biologically active sand filters within water treatment plants (WTPs) are now recognised as an effective barrier for the removal of geosmin. However, little is known regarding the actual microbiological processes occurring or the bacteria capable of degrading geosmin. This study reports the enrichment and isolation of a Gram-negative bacterium, Geo48, from the biofilm of a WTP sand filter where the isolate was shown to effectively degrade geosmin individually. Experiments revealed that Geo48 degraded geosmin in a planktonic state by a pseudo-first-order mechanism. Initial geosmin concentrations ranging from 100 to 1000 ng/l were shown to directly influence geosmin degradation in reservoir water by Geo48, with rate constants increasing from 0.010 h⁻¹ (R² = 0.93) to 0.029 h⁻¹ (R² = 0.97) respectively. Water temperature also influenced degradation of geosmin by Geo48 where temperatures of 11, 22 and 30 °C resulted in rate constants of 0.017 h⁻¹ (R² = 0.98), 0.023 h⁻¹ (R² = 0.91) and 0.019 h⁻¹ (R² = 0.85) respectively. Phylogenetic analysis using the 16S rRNA gene of Geo48 revealed it was a member of the Alphaproteobacteria and clustered with 99% bootstrap support with an isolate designated Geo24, a Sphingopyxis sp. previously described as degrading geosmin but only as a member of a bacterial consortium. Of the previously described bacteria, Geo48 was most similar to Sphingopyxis alaskensis (97.2% sequence similarity to a 1454 bp fragment of the 16S rRNA gene). To date, this is the only study to report the isolation and characterisation of a Gram-negative bacterium from a biologically active sand filter capable of the sole degradation of geosmin.     

Intensive exploitation of a karst aquifer leads to Cryptosporidium water supply contamination

Groundwater from karst aquifers is an important source of drinking water worldwide. Outbreaks of cryptosporidiosis linked to surface water and treated public water are regularly reported. Cryptosporidium oocysts are resistant to conventional drinking water disinfectants and are a major concern for the water industry. Here, we examined conditions associated with oocyst transport along a karstic hydrosystem, and the impact of intensive exploitation on Cryptosporidium oocyst contamination of the water supply. We studied a well-characterized karstic hydrosystem composed of a sinkhole, a spring and a wellbore. Thirty-six surface water and groundwater samples were analyzed for suspended particulate matter, turbidity, electrical conductivity, and Cryptosporidium and Giardia (oo)cyst concentrations. (Oo)cysts were identified and counted by means of solid-phase cytometry (ChemScan RDI^®), a highly sensitive method. Cryptosporidium oocysts were detected in 78% of both surface water and groundwater samples, while Giardia cysts were found in respectively 22% and 8% of surface water and groundwater samples. Mean Cryptosporidium oocyst concentrations were 29, 13 and 4/100 L at the sinkhole, spring and wellbore, respectively. Cryptosporidium oocysts were transported from the sinkhole to the spring and the wellbore, with respective release rates of 45% and 14%, suggesting that oocysts are subject to storage and remobilization in karst conduits. Principal components analysis showed that Cryptosporidium oocyst concentrations depended on variations in hydrological forcing factors. All water samples collected during intensive exploitation contained oocysts. Control of Cryptosporidium oocyst contamination during intensive exploitation is therefore necessary to ensure drinking water quality.     

Identification of bacterial populations in drinking water using 16S rRNA-based sequence analyses

Intracellular RNA is rapidly degraded in stressed cells and is more unstable outside of the cell than DNA. As a result, RNA-based methods have been suggested to study the active microbial fraction in environmental matrices. The aim of this study was to identify bacterial populations in drinking water by analyzing 16S rRNA-based clone libraries. Hollow-fiber ultrafiltration was used to concentrate bacterial communities from 40 l of tap water collected at 12 different times during three different summer months from a single point-of-use. Total RNA was extracted from the microbial concentrates and used to develop 16S rRNA-based clone libraries. Phylogenetic analyses of 1231 partial 16S rRNA gene sequences showed that difficult-to-classify bacterial sequences were the most predominant clones, representing 57.6% of the sequences analyzed. Within these unclassified clades, most sequences were closely related to sequences retrieved from previous DNA- and RNA-based drinking water studies. Other bacterial groups represented in this study included Proteobacteria, cyanobacteria, Actinobacteria, Bacteroidetes, and Planctomycetes. Overall, the results suggest that these bacterial groups are amongst potentially active bacteria in drinking water. Diversity analyses of clones generated show that while overall diversity is similar amongst the different months, membership changes with respect to time. The results from this study further improve our understanding of the molecular diversity and bacterial population dynamics of drinking water microbial communities. Moreover, these results provide the sequence foundation for the development of molecular assays that target active drinking water bacteria.     

Influence of Asellus aquaticus on Escherichia coli, Klebsiella pneumoniae, Campylobacter jejuni and naturally occurring heterotrophic bacteria in drinking water

Water lice, Asellus aquaticus (isopoda), frequently occur in drinking water distribution systems where they are a nuisance to consumers and water utilities. Whether they are solely an aesthetic problem or also affect the microbial water quality is a matter of interest. We studied the influence of A. aquaticus on microbial water quality in non-chlorinated drinking water in controlled laboratory experiments. Pure cultures of the indicator organisms Escherichia coli and Klebsiella pneumoniae and the pathogen Campylobacter jejuni as well as naturally occurring heterotrophic drinking water bacteria (measured as heterotrophic plate counts, HPC) were investigated in microcosms at 7 °C, containing non-sterilised drinking water, drinking water sediment and A. aquaticus collected from a non-chlorinated ground water based drinking water supply system. Concentrations of E. coli, K. pneumoniae and C. jejuni decreased over time, following a first order decay with half lives of 5.3, 18.4 and 1.3 days, respectively. A. aquaticus did not affect survival of indicators and pathogens substantially whereas HPC were influenced by presence of dead A. aquaticus. Growth rates increased with an average of 48% for bacteria grown on R-2A agar and an average of 83% for bacteria grown on yeast extract agar when dead A. aquaticus were present compared to no and living A. aquaticus present. A. aquaticus associated E. coli, K. pneumoniae and C. jejuni were measured (up to 25 per living and 500 per dead A. aquaticus) and so were A. aquaticus associated heterotrophic bacteria (>1.8*10⁴ CFU per living and >6*10⁴ CFU per dead A. aquaticus). A. aquaticus did not serve as an optimised habitat that increased survival of indicators and pathogens, since A. aquaticus associated E. coli, K. pneumoniae and C. jejuni were only measured as long as the bacteria were also present in the water and sediment.     

Microbial quality assessment of household greywater

A monitoring program was undertaken to assess the microbial quality of greywater collected from 93 typical households in Melbourne, Australia. A total of 185 samples, comprising 75 washing machine wash, 74 washing machine rinse and 36 bathroom samples were analysed for the faecal indicator Escherichia coli. Of these, 104 were also analysed for genetic markers of pathogenic E coli and 111 for norovirus (genogroups GI and GII), enterovirus and rotavirus using RT-PCR. Enteric viruses were detected in 20 out of the 111 (18%) samples comprising 16 washing machine wash water and 4 bathroom samples. Eight (7%) samples were positive for enterovirus, twelve (11%) for norovirus genogroup GI, one (1%) for norovirus genogroup GII and another (1%) for rotavirus. Two washing machine samples contained more than one virus. Typical pathogenic E. coli were detected in 3 out of 104 (3%) samples and atypical enteropathogenic E. coli in 11 (11%) of samples. Levels of indicator E. coli were highly variable and the presence of E. coli was not associated with the presence of human enteric viruses in greywater. There was also little correlation between reported gastrointestinal illness in households and detection of pathogens in greywater.     

Effect of GAC pre-treatment and disinfectant on microbial community structure and opportunistic pathogen occurrence

Opportunistic pathogens in potable water systems are an emerging health concern; however, the factors influencing their proliferation are poorly understood. Here we investigated the effects of prior granular activated carbon (GAC) biofiltration [GAC-filtered water, unfiltered water, and a blend (30% GAC filtered and 70% unfiltered water)] and disinfectant type (chlorine, chloramine) on opportunistic pathogen occurrence using five annular reactors (ARs) to simulate water distribution systems, particularly premise plumbing. GAC pre-treatment effectively reduced total organic carbon (TOC), resulting in three levels of influent TOC investigated. Quantitative polymerase chain reaction (q-PCR) provided molecular evidence of natural colonization of Legionella spp., Mycobacterium spp., Acanthamoeba spp., Hartmannella vermiformis and Mycobacterium avium on AR coupons. Cultivable mycobacteria and amoeba, including pathogenic species, were also found in bulk water and biofilm samples. While q-PCR tends to overestimate live cells, it provided a quantitative comparison of target organisms colonizing the AR biofilms in terms of gene copy numbers. In most cases, total bacteria and opportunistic pathogens were higher in the three undisinfected ARs, but the levels were not proportional to the level of GAC pre-treatment/TOC. Chlorine was more effective for controlling mycobacteria and Acanthamoeba, whereas chloramine was more effective for controlling Legionella. Both chlorine and chloramine effectively inhibited M. avium and H. vermiformis colonization. Pyrosequencing of 16S rRNA genes in coupon biofilms revealed a significant effect of GAC pre-treatment and disinfectant type on the microbial community structure. Overall, this study provides insights into the potential of different disinfectants and GAC biofilters at the treatment plant and in buildings to control downstream opportunistic pathogens and broader drinking water microbial communities.     

Assessment of phylogenetic diversity of bacterial microflora in drinking water using serial analysis of ribosomal sequence tags

We examined chlorinated drinking water samples from three different surface water treatment plants for bacterial 16S rDNA diversity using the serial analysis of V6 ribosomal sequence tag (SARST-V6) method. A considerable degree of diversity was observed in each sample, with an estimated richness ranging from 173 to 333 phylotypes. The community structure shows that there are differences in bacterial evenness between sampled sites. The taxonomic composition of the microbial communities was found to be dominated by members of the Proteobacteria (57.2-77.4%), broadly distributed among the classes Alphaproteobacteria, Betaproteobacteria, Gammaproteobacteria, and Deltaproteobacteria. Additionally, a large proportion of sequences (6.3-36.5%) were found to be distantly related to database sequences of unknown phylogenetic affiliation. Given the apparent significance of this bacterial group in drinking water, a 16S rDNA analysis was performed and confirmed their presence and phylogeny. Notwithstanding the potential under-representation of certain bacterial phyla using the SARST-V6 primer pairs, as revealed by a refined computer algorithm, our results suggest that 16S rDNA corresponding to a variety of eubacterial groups can be detected in finished drinking water, suggesting that this water may contain a higher level of bacterial diversity than previously observed.     

The genetic diversity of human noroviruses detected in river water in Korea

We studied the genetic diversity of human noroviruses in river waters by RT-nested PCR and phylogenetic analysis. During 2002-2003, water samples were collected from four rivers in Gyeonggi Province, South Korea. Among the 58 samples, 32 (55.2%) and 26 (44.8%) showed positive results with noroviruses belonging to genogroups I (GI) and II (GII), respectively. The phylogenetic analysis grouped 8 and 7 genotypes in GI and GII, respectively. The major types were GI/1, GI/13, and GII/15, and GI/1 and GI/3 were temporarily distributed. Most GI- and GII-grouped strains were closely related to the reference strains from neighboring countries, China and Japan, and GII/4-related strains had similar sequences to strains recognized as worldwide epidemic outbreaks. The strains circulating between countries are of particular concern to the outbreaks of noroviral diseases in Korea and must be periodically monitored in the natural environments.     

Characterization of bromate-reducing bacterial isolates and their potential for drinking water treatment

The objective of the current study was to isolate and characterize several bromate-reducing bacteria and to examine their potential for bioaugmentation to a drinking water treatment process. Fifteen bromate-reducing bacteria were isolated from three sources. According to 16S rRNA gene sequencing, the bromate-reducing bacteria are phylogenetically diverse, representing the Actinobacteria, Bacteroidetes, Firmicutes, and α-, β-, and γ-Proteobacteria. The broad diversity of bromate-reducing bacteria suggests the widespread capability for microbial bromate reduction. While the cometabolism of bromate via nitrate reductase and (per)chlorate reductase has been postulated, five of our bromate-reducing isolates were unable to reduce nitrate or perchlorate. This suggests that a bromate-specific reduction pathway might exist in some microorganisms. Bioaugmentation of activated carbon filters with eight of the bromate-reducing isolates did not significantly decrease start-up time or increase bromate removal as compared to control filters. To optimize bromate reduction in a biological drinking water treatment process, the predominant mechanism of bromate reduction (i.e., cometabolic or respiratory) needs to be assessed so that appropriate measures can be taken to improve bromate removal.     

Occurrence and fate of the angiotensin II receptor antagonist transformation product valsartan acid in the water cycle – A comparative study with selected β-blockers and the persistent anthropogenic wastewater indicators carbamazepine and acesulfame

The substantial transformation of the angiotensin II receptor antagonist valsartan to the transformation product 2′-(2H-tetrazol-5-yl)-[1,1′-biphenyl]-4-carboxylic acid (referred to as valsartan acid) during the activated sludge process was demonstrated in the literature and confirmed in the here presented study. However, there was a severe lack of knowledge regarding the occurrence and fate of this compound in surface water and its behavior during drinking water treatment. In this work a comparative study on the occurrence and persistency of valsartan acid, three frequently used β-blockers (metoprolol, atenolol, and sotalol), atenolol acid (one significant transformation product of atenolol and metoprolol), and the two widely distributed persistent anthropogenic wastewater indicators carbamazepine and acesulfame in raw sewage, treated wastewater, surface water, groundwater, and tap water is presented. Median concentrations of valsartan acid in the analyzed matrices were 101, 1,310, 69, <1.0, and 65 ng L⁻¹, respectively. Treated effluents from wastewater treatment plants were confirmed as significant source. Regarding concentration levels of pharmaceutical residues in surface waters valsartan acid was found just as relevant as the analyzed β-blockers and the anticonvulsant carbamazepine. Regarding its persistency in surface waters it was comparable to carbamazepine and acesulfame. Furthermore, removal of valsartan acid during bank filtration was poor, which demonstrated the relevance of this compound for drinking water suppliers. Regarding drinking water treatment (Muelheim Process) the compound was resistant to ozonation but effectively eliminated (≥90%) by subsequent activated carbon filtration. However, without applying activated carbon filtration the compound may enter the drinking water distribution system as it was demonstrated for Berlin tap water.     

One-year weekly survey of noroviruses and enteric adenoviruses in the Tone River water in Tokyo metropolitan area, Japan

To investigate the actual fluctuations in the concentrations of noroviruses (NoVs) GI and GII, and enteric adenoviruses (EAdVs) in river water and its relationship with the number of acute infectious gastroenteritis patients, one-year weekly quantitative monitoring of NoVs GI and GII and EAdVs was performed in the Tone River in Japan where the surface water is utilized for the main production of drinking water for the Tokyo Metropolitan Area from October 2009 to September 2010. Noroviruses GI and GII and EAdVs were detected in 28 (54%), 33 (63%), and 23 (44%) of the 52 samples (1 L each), respectively. The concentrations of NoVs GI and GII and EAdVs fluctuated strongly and were more abundant in winter and early spring. The concentration of NoVs GI was transiently greater than 10,000 copies/L. The number of acute infectious gastroenteritis patients in the upper river basin was highly correlated with all the viral concentrations, while general microbial indicator data such as turbidity and heterotrophic plate count were independent of viral concentration as suggested in previous studies. To the best of our knowledge, this is the first study that clearly shows the strong correlation of the number of gastroenteritis with virus contamination in lower river basin.     

Comparison of the microbiological and chemical characterization of harvested rainwater and reservoir water as alternative water resources

Rainwater harvesting (RWH) offers considerable potential as an alternative water supply. In this study, all of the harvested rainwater samples met the requirements for grey water but not for drinking water. In terms of microbiological parameters, total coliform (TC) and Escherichia coli (EC) were measured in 91.6% and 72%, respectively, of harvested rainwater samples at levels exceeding the guidelines for drinking water, consistent with rainfall events. In the case of the reservoir water samples, TC and EC were detected in 94.4% and 85.2%, respectively, of the samples at levels exceeding the guidelines for drinking water. Both indicators gradually increased in summer and fall. The highest median values of both TC and EC were detected during the fall. Chemical parameters such as common anions and major cations as well as metal ions in harvested rainwater were within the acceptable ranges for drinking water. By contrast, Al shows a notable increase to over 200 μg L^− 1 in the spring due to the intense periodic dust storms that can pass over the Gobi Desert in northern China. In terms of statistical analysis, the harvested rainwater quality showed that TC and EC exhibit high positive correlations with NO₃⁻ (ρ_TC = 0.786 and ρ_EC = 0.42) and PO₄⁻ (ρ_TC = 0.646 and ρ_EC = 0.653), which originally derive from catchment contamination, but strong negative correlations with Cl⁻ (ρ_TC = − 0.688 and ρ_EC = − 0.484) and Na⁺ (ρ_TC = − 0.469 and ρ_EC = − 0.418), which originate from seawater.     

The association of drinking water treatment and distribution network disturbances with Health Call Centre contacts for gastrointestinal illness symptoms

There are relatively few studies on the association between disturbances in drinking water services and symptoms of gastrointestinal (GI) illness. Health Call Centres data concerning GI illness may be a useful source of information. This study investigates if there is an increased frequency of contacts with the Health Call Centre (HCC) concerning gastrointestinal symptoms at times when there is a risk of impaired water quality due to disturbances at water works or the distribution network. The study was conducted in Gothenburg, a Swedish city with 0.5 million inhabitants with a surface water source of drinking water and two water works. All HCC contacts due to GI symptoms (diarrhoea, vomiting or abdominal pain) were recorded for a three-year period, including also sex, age, and geocoded location of residence. The number of contacts with the HCC in the affected geographical areas were recorded during eight periods of disturbances in the water works (e.g. short stops of chlorine dosing), six periods of large disturbances in the distribution network (e.g. pumping station failure or pipe breaks with major consequences), and 818 pipe break and leak repairs over a three-year period. For each period of disturbance the observed number of calls was compared with the number of calls during a control period without disturbances in the same geographical area. In total about 55, 000 calls to the HCC due to GI symptoms were recorded over the three-year period, 35 per 1000 inhabitants and year, but much higher (>200) for children <3 yrs of age. There was no statistically significant increase in calls due to GI illness during or after disturbances at the water works or in the distribution network. Our results indicate that GI symptoms due to disturbances in water works or the distribution network are rare. The number of serious failures was, however limited, and further studies are needed to be able to assess the risk of GI illness in such cases. The technique of using geocoded HCC data together with geocoded records of disturbances in the drinking water network was feasible.     

Natural and acquired resistance of mice to infection by airborne Klebsiella pneumoniae after subchronic intoxication by cadmium administered orally

Mice daily ingested about 22 mg of cadmium per kg of body weight in drinking water for 30 days. On the 30th day, the liver and kidneys of the mice contained about 18 μg of Cd²⁺ perg of fresh organ. A group of these mice was immunized against Klebsiella pneumoniae using two injections of vaccine, the first on the 7th day and the second on the 14th day of intoxication. On the 28th day, the non-immunized and the immunized mice were infected via a respiratory route by one lethal dose 50% of K. pneumoniae (the LD₅₀ for the immunized mice was 2.4 times higher than the LD₅₀ for the non-immunized mice). Comparison with the non-intoxicated control mice showed that the ingestion of Cd²⁺ did not significantly modify the natural resistance or the acquired resistance of the mice to the infection by airborne K. pneumoniae.     

Environmental impact assessment - A bibliography with abstracts: by B. B. Clark,R. Bisset and P. Wathern,Mansell Publishing, 3 Bloomsbury Place,London WC1A 2QA, 1980, 516 pp. Price: $23.50

The effect of fluoride administration on zinc levels in liver, kidney and bone, and on protein content (μg/mg tissue wt) in liver and kidney was investigated by subjecting albino mice for 16 weeks to different concentrations of fluoride (0 ppm — control; 10 ppm — group I; 25 ppm — group II; 50 ppm — group III; 100 ppm — group IV; and 200 ppm — group V) in drinking water. Zinc levels decreased significantly in liver and bone of groups III, IV and V; the decrease being 17%, 25% and 44% (p < 0.005) in the case of liver, and 14% (p < 0.01), 24% (p < 0.005), and 39% (p < 0.005) in the case of bone in groups III, IV and V respsectively. Zinc depletion in the kidney of the fluoridated animals was only statistically significant (p < 0.005) in groups IV and V. Similarly the protein content also decreased in liver and kidney (in liver significant at p < 0.005, p < 0.01 and p < 0.005; in kidney significant at p < 0.005, p < 0.05 and p < 0.005 in groups III, IV and V respectively). The experimental animals, especially in groups IV (100 ppm F) and V (250 ppm F), also exhibited retardation in growth.