Detection and enumeration of Listeria monocytogenes in a sewage treatment plant in Iraq

Listeria monocytogenes was isolated from a sewage treatment plant in Baghdad, Iraq, at all stages of treatment. The treatment processes did not yield a sewage sludge cake or a final discharge free of listerias. The agricultural practice of using such sewage products as fertilizers could become a route of spreading the organism in Iraq, particularly by infecting animals that consume vegetation in fields spread with such sewage. Dewatering of sewage reduced the number of L. monocytogenes but long periods of exposure to sun would be needed to obtain a 'safe' sewage sludge cake.     

Effects of sewage treatment on the removal of Listeria monocytogenes

Two sewage treatment plants in Baghdad, Iraq, were investigated to assess the effects of the different treatment stages on the removal of Listeria monocytogenes . The bacteria were severely affected after the activation and digestion stages at both plants. A dramatic decrease in numbers of listerias after each of these two stages was noticed during the cold months (September-January). The organisms were able to survive these treatments and were present in the final effluent and even in low numbers in the sewage sludge cake. Sufficient dewatering of sewage sludge is recommended to obtain sewage free of listerias. Improvements in the isolation procedure of L. monocytogenes from such heavily contaminated material is also discussed.     

Effects of sewage treatment on the removal of Listeria monocytogenes

Two sewage treatment plants in Baghdad, Iraq, were investigated to assess the effects of the different treatment stages on the removal of Listeria monocytogenes. The bacteria were severely affected after the activation and digestion stages at both plants. A dramatic decrease in numbers of listerias after each of these two stages was noticed during the cold months (September-January). The organisms were able to survive these treatments and were present in the final effluent and even in low numbers in the sewage sludge cake. Sufficient dewatering of sewage sludge is recommended to obtain sewage free of listerias. Improvements in the isolation procedure of L. monocytogenes from such heavily contaminated material is also discussed.     

Storage effects of sewage sludge cake on the survival of Listeria monocytogenes

Sewage sludge cake is widely used as an agricultural fertilizer in Iraq. Listeria monocytogenes was shown to be present in small numbers in this material despite sewage treatments. In an attempt to reduce the numbers of this pathogen in this sewage end product, the survival of L. monocytogenes was monitored in a heap of sewage sludge cake stored for over 23 weeks on farm land. The organisms were reduced in numbers and eliminated to undetectable limits during 8 weeks of storage under subtropical weathering and did not recover even 2 months after disappearance. Dewatering processes seem to have some affect on the survival of the bacteria. Therefore, solar dewatering by heaping the sewage sludge cake and exposing it to sun for no less than 8 weeks is recommended to obtain a listeria-free product.     

Storage effects of sewage sludge cake on the survival of Listeria monocytogenes

Sewage sludge cake is widely used as an agricultural fertilizer in Iraq. Listeria monocytogenes was shown to be present in small numbers in this material despite sewage treatments. In an attempt to reduce the numbers of this pathogen in this sewage end product, the survival of L. monocytogenes was monitored in a heap of sewage sludge cake stored for over 23 weeks on farm land. The organisms were reduced in numbers and eliminated to undetectable limits during 8 weeks of storage under subtropical weathering and did not recover even 2 months after disappearence. Dewatering processes seem to have some affect on the survival of the bacteria. Therefore, solar dewatering by heaping the sewage sludge cake and exposing it to sun for no less than 8 weeks is recommended to obtain a listeria-free product.     

Listeria monocytogenes contamination of crops grown on soil treated with sewage sludge cake

A l -G hazali , M.R. & A l -A zawi , S.K. 1990. Listeria monocytogenes contamination of crops grown on soil treated with sewage sludge cake. Journal of Applied Bacteriology 69 , 642–647.
Listeria monocytogenes was found in the sewage sludge cake which is commonly used as an agricultural fertilizer in Iraq. Soils treated with this material were contaminated with the organism. Pot and field experiment showed that crops grown on treated soil became contaminated with L. monocytogenes and when alfalfa plant was grown on farmland soil treated with sewage sludge cake, listerias were found on 10% of 50 plants sampled at harvest, but the organism was detected only in low numbers on these crops (≤ 5 cells/g). This could add to the risk to animals and man.     

Listeria monocytogenes contamination of crops grown on soil treated with sewage sludge cake

Listeria monocytogenes was found in the sewage sludge cake which is commonly used as an agricultural fertilizer in Iraq. Soils treated with this material were contaminated with the organism. Pot and field experiment showed that crops grown on treated soil became contaminated with L. monocytogenes and when alfalfa plant was grown on farmland soil treated with sewage sludge cake, listerias were found on 10% of 50 plants sampled at harvest, but the organism was detected only in low numbers on these crops (less than or equal to 5 cells/g). This could add to the risk to animals and man.     

Isolation and Enumeration of Listeria monocytogenes from Sewage, Sewage Sludge and River Water

Listeria monocytogenes in sewage, sewage sludge and river water was isolated by enrichment at 4°C with subculture and enrichment in thiocyanate, naladixic acid broth and plating on to Tryptose Agar. The results indicated that L. monocytogenes is present in sewage and sewage sludge in considerable numbers and that this organism survives longer than Salmonella spp. on land sprayed with sewage sludge.     

Comparison of a cultural method with ListerScreen plus Rapid'L.mono or PCR-ELISA methods for the enumeration of L. monocytogenes in naturally contaminated sewage sludge

Cultural methods used to count Listeria monocytogenes in sewage sludge are laborious and time consuming, and alternative methods are needed to reduce analysis time and improve detection limits. In this study, a survey of L. monocytogenes in sewage sludge is presented with a comparative study between a cultural method and immunomagnetic separation using a ListerScreen test followed by identification of L. monocytogenes with Rapid'L.mono agar or PCR-ELISA. These two alternative methods improved the detection of L. monocytogenes in different types of sludge, irrespective of their physical and chemical characteristics. The ListerScreen method coupled with detection of L. monocytogenes on Rapid'L.mono offers the advantage of being less sophisticated than the molecular method and allows isolation of the organism, which may be useful in epidemiological studies. However, ListerScreen coupled with PCR-ELISA proved best for high-sensitivity detection of L. monocytogenes in sewage samples.     

Occurrence of Listeria sp and L monocytogenes in sewage sludge used for land application: effect of dewatering,liming and storage in tank on survival of Listeria species

The application of sewage sludge to agricultural land is widely used in France. To determine the impact of sludge treatments, concentrations of Listeria sp., Listeria monocytogenes and faecal indicators were monitored in five types of sludge from three sewage treatment plants in Angers (France) and its suburbs over a 1-year period. On the whole, bacteria were reduced in numbers through sludge treatments. Apart from liming, which leads to reduced levels of bacteria below detection limits, other sludge treatments did not eliminate Listeria sp. and faecal indicators. Listeria sp. and L. monocytogenes were found respectively in 87% and 73% of dewatered sludges and in 96% and 80% of sludges stored in tanks. Concentrations of L. monocytogenes, ranging from 0.15 to 20 MPN g(-1) dry matter in dewatered sludge and from 1 to 240 MPN g(-1) dry matter in sludge stored in tanks, did not show seasonal variations. Spreading of sanitised sludge onto agricultural land results in the addition of 10(6)-10(8) L. monocytogenes per hectare per year, which may contribute to the increase in the dissemination of this pathogenic species in the environment.     

Heteroduplex mobility assay for the identification of Listeria sp and Listeria monocytogenes strains: application to characterisation of strains from sludge and food samples

One hundred and ten Listeria sp. isolates from sewage sludge were identified according to phenotypic and genotypic methods. The Listeria sp. strains isolated from five types of sludge from three sewage treatment plants in Angers (France) and the surrounding area included L. monocytogenes (55.5%), L. innocua (29.1%), L. seeligeri (13.6%) and L. welshimeri (1.8%). The majority of L. monocytogenes strains belonged to serotypes 4b, 1/2b and 1/2a. Moreover, a heteroduplex mobility assay based on the 16S rRNA sequences was tested for its ability to identify the six species of the genus Listeria. This study, performed on 283 Listeria sp. strains from human, food and sewage sludge samples, showed that all the species were distinguishable from one another. L. innocua and L. seeligeri showed respectively three and two distinct banding patterns. Within L. monocytogenes, four groups (I-IV) were defined. The majority of food and environmental isolates were clustered in group I and it is noteworthy that group IV clustered epidemiologic isolates and strains belonging to serotypes 4b, 1/2a and 1/2b.     

Oxygen diffusion and microbial activity in the composting of dehydrated sewage sludge cakes

The effective diffusivity for oxygen within sewage sludge cakes was measured, and in turn used for the study of microbial activity in the sludge cake composting. The diffusivity was found to be proportional to the one and a half power of the sludge cake porosity. Composting experiments were done using two types of sludge cakes, 5 mesh sieved and pelletized. It was found that the microbial activity in the inner core region of the pelletized sludge cake was significantly less than that in the outer region, even through the rate of oxygen diffusion was fast enough to support the microorganisms' activities. It appears, therefore, that microorganisms acting to degrade organic matter grow primarily on the macroscopic interface between solids and air.     

Human enteropathogen load in activated sewage sludge and corresponding sewage sludge end products

This study demonstrated a significant reduction in the concentrations of Cryptosporidium parvum and Cryptosporidium hominis oocysts, Giardia lamblia cysts, and spores of human-virulent microsporidia in dewatered and biologically stabilized sewage sludge cake end products compared to those of the respective pathogens in the corresponding samples collected during the sludge activation process.     

Fate of Cryptosporidium oocysts, Giardia cysts, and microbial indicators during wastewater treatment and anaerobic sludge digestion.

The extent of reduction in selected microorganisms was tested during both aerobic wastewater treatment and anaerobic digestion of sludge at the wastewater treatment plant in Ottawa to compare the removal of two encysted pathogenic protozoa with that of microbial indicators. Samples collected included the raw wastewater, the primary effluent, the treated wastewater, the mixed sludge, the decanted liquor, and the cake. All of the raw sewage samples were positive for Cryptosporidium oocysts and Giardia cysts, as well as for the other microorganisms tested. During aerobic wastewater treatment (excluding the anaerobic sludge digestion), Cryptosporidium and Giardia were reduced by 2.96 log10 and 1.40 log10, respectively. Clostridium perfringens spores, Clostridium perfringens total counts, somatic coliphages, and heterotrophic bacteria were reduced by approximately 0.89 log10, 0.96 log10, 1.58 log10, and 2.02 log10, respectively. All of the other microorganisms were reduced by at least 3.53 log10. Sludge samples from the plant were found to contain variable densities of microorganisms. Variability in microbial concentrations was sometimes great between samples, stressing the importance of collecting a large number of samples over a long period of time. In all cases, the bacterial concentrations in the cake (dewatered biosolids) samples were high even if reductions in numbers were observed with some bacteria. During anaerobic sludge digestion, no statistically significant reduction was observed for Clostridium perfringens, Enterococcus sp., Cryptosporidium oocysts, and Giardia cysts. A 1-2 log10 reduction was observed with fecal coliforms and heterotrophic bacteria. However, the method utilized to detect the protozoan parasites does not differentiate between viable and nonviable organisms. On the other hand, total coliforms and somatic coliphages were reduced by 0.35 log10 and 0.09 log10, respectively. These results demonstrate the relative persistence of the protozoa in sewage sludge during wastewater treatment.     

The potential of some soil amendments to improve tree growth on metalliferous mine wastes

Trees are commonly planted on metalliferous mine wastes as a means of reclaiming these contaminated substrates. Other land-uses are often limited. However tree establishment is often poor owing to hostile substrate properties such as acidity, low macronutrient concentrations, high trace element concentrations, poor substrate structure and poor water retention. Soil amendments are a potentially cheap amelioration option. A factorial experiment was set-up to investigate the effects of seven amendments on the substrate chemistry and the growth and leaf chemistries of alder and sycamore. The amendments included ammonium nitrate, mineral phosphate and sulphate of potash fertilizers, calcareous sand, a superexpanding polyacrylamide gel polymer, cake sewage sludge and diatomite. The last two amendments are used here in a novel capacity. The most successful amendments with regard to their growth and leaf chemistry effects were found to be cake sludge and diatomite for alder and mineral phosphate, calcareous sand and cake sludge for sycamore. The least successful amendments were ammonium nitrate, sulphate of potash and calcareous sand for alder and ammonium nitrate for sycamore. The results differed according to the autoecological properties of the two species, with alder being easily the most productive. The implications of these results for tree-planting schemes are discussed.     

Human Enteropathogen Load in Activated Sewage Sludge and Corresponding Sewage Sludge End Products

This study demonstrated a significant reduction in the concentrations of Cryptosporidium parvum and Cryptosporidium hominis oocysts, Giardia lamblia cysts, and spores of human-virulent microsporidia in dewatered and biologically stabilized sewage sludge cake end products compared to those of the respective pathogens in the corresponding samples collected during the sludge activation process.     

Comparison of sludge and clinical isolates of Listeria monocytogenes

AIMS: Listeria monocytogenes strains isolated in the same geographical area from sewage sludge and from patients presenting with listeriosis were compared. METHODS AND RESULTS: All isolates were typed by serotyping, phage typing and SmaI/ApaI pulsed-field gel electrophoresis (PFGE). Among the sludge isolates (n=32), 22 subtypes could be distinguished by the combination of all typing methods. The human isolates (n=11) were distributed into 10 subtypes which clearly differed from those observed among sludge isolates, except for one cluster formed by two related human isolates which showed high similarity in PFGE patterns (SmaI: 92%; ApaI: 89.5%) with one sludge isolate. CONCLUSION: These results suggest the existence of an epidemiological link between sludge and human isolates, but they may also be reflecting the distribution of L. monocytogenes types within the environment. SIGNIFICANCE AND IMPACT OF THE STUDY: Sludge and human L. monocytogenes may be related but further epidemiological studies are necessary to elucidate this point.     

Salmonella isolated in sewage sludge traced back to human cases of salmonellosis

AIM: The main aim of this study was to investigate a possible connection between the Salmonella content in sewage sludge and human cases of salmonellosis. An additional aim was to survey the antimicrobial resistance situation in Salmonella isolated from Swedish sewage sludge. METHODS AND RESULTS: The Salmonella strains were compared by restriction enzyme analysis combined with pulsed field gel electrophoresis and antimicrobial susceptibility testing. This study suggests a link between Salmonella isolated from sewage sludge and human Salmonella isolates. CONCLUSIONS: This study demonstrates that Salmonella spp. isolated in sewage treatment plants (STP) originate from infected humans and survive treatment at STP. It also highlights the risk of spreading resistant Salmonella strains from sewage sludge to the environment. SIGNIFICANCE AND IMPACT OF THE STUDY: As Salmonella spp. originating from infected humans can survive the treatment at STP, the risk of Salmonella spp. being spread with sewage sludge to the environment and then to people and animals is enhanced. The threat to society is even worse if the bacteria are resistant to antimicrobial agents.     

Demonstration of solids-associated virus in wastewater and sludge.

Data presented demonstrate the relatively high multiplicity of solids-associated virus in field samples, i.e., wastewater, sludge, and soils. Influent, effluent, and chlorinated effluent samples showed 16.1 to 100% of the total virus demonstrated in samples to be solids associated. Three techniques for freeing solids-associated virus are described and compared. Using sonication of solids and polyethylene glycol concentration, virus was demonstrated in fully digested sludge (60 days at 34 C), sand at the site of a sewer leak, and dried sludge cake and mud 900 m downstream from a sewage disposal site. These data emphasize the inadequacy of virus concentration techniques that do not include the processing of solids. In situ elution failed to free solids-associated virus.     

Demonstration of solids-associated virus in wastewater and sludge.

Data presented demonstrate the relatively high multiplicity of solids-associated virus in field samples, i.e., wastewater, sludge, and soils. Influent, effluent, and chlorinated effluent samples showed 16.1 to 100% of the total virus demonstrated in samples to be solids associated. Three techniques for freeing solids-associated virus are described and compared. Using sonication of solids and polyethylene glycol concentration, virus was demonstrated in fully digested sludge (60 days at 34 C), sand at the site of a sewer leak, and dried sludge cake and mud 900 m downstream from a sewage disposal site. These data emphasize the inadequacy of virus concentration techniques that do not include the processing of solids. In situ elution failed to free solids-associated virus.