Origin of objectionable odorous components in piggery wastes and the possibility of applying indicator components for studying odour development

Storage of swine manure is associated with the microbiological production of a variety of odorous chemicals including ammonia, organic acids and alcohols, and sulphides. Although largely the product of microbiological activity, little is known about the microorganisms present in swine manure. In order to gain a better understanding of the types and activities of the microorganisms present, representative strains of microorganisms were isolated from faeces and stored manure slurry, identified, and physiologically characterized. For swine manure slurry samples, total anaerobe colony counts were greatest when a non-selective, habitat simulating medium containing clarified swine manure slurry was used whereas the highest counts for faecal anaerobes were obtained on rumen fluid containing medium. Faecal and slurry samples were also plated onto the appropriate medium containing the antibiotics tetracycline, erythromycin and tylosin (10 micro g ml-1, individually) and the proportional counts of organisms capable of growing in the presence of these antibiotics determined. Randomly selected isolates from the highest dilutions were identified by 16 s rDNA sequence analysis, and selected physiological characteristics were determined. The results of these examinations indicate that the predominant culturable microorganisms from these environments are obligately anaerobic, low mol percentage G + C Gram positive bacteria (Firmicutes) who are members of Clostridial, Eubacterial, and Lactobacillus/Streptococcus phylogenetic groups. Isolates similar to Sporomusa and Flexibacter/Cytophaga/Bacteroides (CFB or Bacteroidetes) groups were also obtained. Although similar overall, faecal and slurry samples differed in bacterial composition. Manure slurry samples were dominated by organisms similar to Clostridium coccoides and Enterococcus species whereas the distribution of species present in faeces appeared much broader. Whereas most of the pure cultures could be assigned to known phylogenetic groupings, few could be identified as known species. Examination of some growth and physiological characteristics of faecal and slurry isolates showed these to be primarily carbohydrate fermenters, although some were able to ferment lactate and amino acids. When the ability of manure and faecal isolates to ferment protein, peptides and amino acids was examined, a relatively small percentage of these were able to do so and most of these fermented carbohydrates in addition to the amino acid sources provided. The predominant amino acid fermenters were most closely related to C. coccoides and C. botulinum, but representatives of the Bacteroides, Staphylococcus, Enterococcus and other phylogenetic groups were also found. The results reported here are compared with those obtained from clone libraries prepared from the same environmental samples.

Isolation, characterization and comparison of bacteria from swine faeces and manure storage pits

Despite intensive studies of microbial-community diversity, the questions of which kinds of microbial populations are associated with changes in community diversity have not yet been fully solved by molecular approaches. In this study, to investigate the impact of livestock wastewater on changes in the bacterial communities in groundwater, bacterial communities in subsurface aquifers were analyzed by characterizing their 16S rDNA sequences. The similarity coefficients of restriction fragment length polymorphism (RFLP) patterns of the cloned 16S ribosomal DNAs showed that the bacterial communities in livestock wastewater samples were more closely related to those in contaminated aquifer samples. In addition, calculations of community diversity clearly showed that bacterial communities in the livestock wastewater and the contaminated aquifer were much more diverse than those in the uncontaminated aquifer. Thus, the increase in bacterial-community diversity in the contaminated aquifer was assumed to be due to the infiltration of livestock wastewater, containing high concentrations of diverse microbial flora, into the aquifer. Phylogenetic analysis of the sequences from a subset of the RFLP patterns showed that the Cytophaga-Flexibacter-Bacteroides and low-G+C gram-positive groups originating from livestock wastewater were responsible for the change in the bacterial community in groundwater. This was evidenced by the occurrence of rumen-related sequences not only in the livestock wastewater samples but also in the contaminated-groundwater samples. Rumen-related sequences, therefore, can be used as indicator sequences for fecal contamination of groundwater, particularly from livestock.

Increase in bacterial community diversity in subsurface aquifers receiving livestock wastewater input.

Bacterial community dynamics in liquid swine manure during storage: molecular analysis using DGGE/PCR of 16S rDNA

Mineralization of organic matter in aquatic ecosystems with shallow waters occurs to a large extent in their sediments under anoxic conditions This is achieved by a community of bacteria, which are the catalysts in a sequence of processes Of the two possible terminal processes, methanogenesis and sulphate reduction, the first usually dominates in freshwater systems, whereas in estuarine and marine sediments electrons are mainly channelled to sulphate Interactions between sulphate-reducing and methanogenic bacteria are described Sulphate-reducing bacteria also show interactions with fermentative bacteria After a brief description of properties of sulphate-reducing and fermentative bacteria occurring in sediments, examples are given of interactions between them This is followed by the presentation of some results obtained from studies on competition for L-lactate between organisms belonging to both groups It is shown that sulphate-reducing bacteria could successfully compete for L-lactate when this was available in growth-limiting amounts with sufficient sulphate and iron Finally, a brief discussion is given of ecological niches of sulphide-oxidizing bacteria thriving in the upper sediment layers

Microbial Interactions in Sediment Communities

Media for enumeration of the microbiota of anaerobically stored piggery wastes were tested. Highest colony counts were obtained with 80 to 100% farm slurry supernatant included in the anaerobic roll tube media. Colony counts with these media numbered 2 X 10(9) to 12 X 10(9)/g (wet weight), which represents about 20% of the microscopic counts. Lower percentages of slurry supernatant in the media gave lower colony counts. Addition of glucose, cellobiose, and starch or of Trypticase to media with 20% slurry supernatant did not increase colony counts. Higher values were obtained when hemicellulose preparations were added to these media. Incubation at 25 degrees C gave the highest numbers. Incubation at 15 to 37 degrees C gave counts of about 70 and 10%, respectively, of those at 25 degrees C. Of the colonies picked for isolation, about 20% were obtained in pure culture. The isolates apparently belonged to the genera Peptococcus, Ruminococcus, Peptococcus, Ruminococcus, Pepostreptococcus, and Bacteroides.

Enumeration and isolation of anaerobic microbiota of piggery wastes.

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Enumeration and isolation of anaerobic microbiota of piggery wastes.