**1. Introduction**

48 Management of Organic Waste

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and P and Enzyme Activities in Soil Incubated with the Earthworms *Metaphire guillelmi* or *Eisenia fetida*. *Soil Biology and Biochemistry*, vol. 32, No. 2, pp. 2055-2062, Animal production pose potential hazards of environmental contamination with pathogenic microorganisms. These are particularly related to a subsequent storage processing and utilization of animal organic wastes (manure, fertilizer, wastewater, sludges etc.). A major source of pathogenic microorganisms in the environment are excrements from clinically and subclinically infected farm animals.

Handling, storage, treatment and use of different forms of animals excrements entails two principle problems: epizootological or epidemiological and hygienical. Solid excrements contain high numbers of common intestinal microflora (*E. coli*, faecal streptococci, lactobacilli etc.), bacteria that are pathogenic also for man (salmonellae, mycobacteria, listeriae etc.), protozoa (*Isospora* spp., *Balantidium coli*) and eggs or larvae of enteronematodes (*Ascaris suum, Oesophagostomum* sp., *Trichuris suis* etc.) (Lauková et al., 2000; Krupicer et al., 2000).

The parasitic propagative stages, mainly endoparasitic protozoa and helminths develop mostly outside their host´s organism. *A. suum* eggs are hygienically the most problematic ones. They are amongst the helminth eggs most resistant to environmental factors and may survive in the nature for many years, therefore, they tend to accumulate in the environment (soil, water) and serve as an infectious entity for both man and animals (Papajová and Juriš, 2009). The cell wall of *A suum* egg is enveloped with an outer layer formed by acid polysaccharides and proteins, central layer consisting of proteins (25%) and lipids (75%, particularly alpha glycosides). Thus this resistant cell wall protects eggs against effects of chemicals and drying (Eckert, 1992).

Regarding the spread of helminthoses, domestic animals (dogs, cats) are also of great importance because they live in a close contact with man. Infection and way of transmission of the disease depends on the way of breeding and on the breeding environment where the animal occurs. An important factor of the risk of infection transmission is also possibility of animal to move outside its housing (yard, move in nature), or the use of a dog (hunting or social). The most frequent way of transmission of parasitic diseases is through the contact (free-living animals with a domestic ones), or through contaminated environment with

The Sanitation of Animal Waste Using Anaerobic Stabilization 51

monitor the physical-chemical changes in pig slurry treated by ecologically acceptable and energetically beneficial anaerobic stabilisation, changes in the properties of anaerobically digested slurry stored in ground lagoon and the effect of anaerobically digested slurry stores in ground lagoon on the survival of parasitic germs. The impact of lime (two types) on the survival of parasitic organisms in anaerobic stored manure and dog excrements

To determine helminth eggs count in slurry (input and output samples from bioreactor and in lagoon samples – supernatant and sediment), 50 ml from each of the 1 l sample was taken

*A. suum* eggs were isolated by dissection of a distal uterine part of female pig ascaris. The distal uterine ends were then removed to a glass homogenizer and processed. The water

We used the ''artificial contamination of lagoon and organic wastes'' approach to make sure

Model eggs were inoculated by a micropipette into polyurethane carriers, prepared according to Plachý and Juriš (1995), at a dose of 1 000 eggs per one carrier. A porous cellular plastic - soft expanded polyurethane, commercially known as a plastic foam, was used as a material for the carriers. It is an additive product of polyisocyanates and compounds with a high content of hydroxylic groups. It consists of a network of interconnected cells, resembling a honeycomb. Its polyurethane structure allows for a sufficient contact of helminth eggs with the environment, preventing them from release and consequently improving their recovery (Picture 1). For mechanical protection the carriers were placed to perforated plastic net (Picture 2) before introducing them into the organic wastes. Three samples were taken and analysed at each sampling interval. The eggs were re-isolated from the inoculated carriers as follows: the carriers were cut into small pieces and washed in a mortar with 3 · 5 ml portions of saline, thoroughly stirred and filtered through a sieve into test tubes. After centrifugation, sediments were transferred to

The viability of exposed unembryonated *A. suum* eggs was determined by incubation up to the embryonated stage (Picture 6) in a thermostat at 26°C for 21 days. Petri dishes with *A. suum* eggs were aerated daily with micropipette. The developmental ability of *A. suum* eggs was compared with that of the control eggs which were kept in distilled water under aerobic

The following changes in physical and chemical properties of the solid and liquid wastes were monitored: pH, dry mater (DM), inorganic (IM) and organic (OM) matter, ammonium

+), total nitrogen (Nt), chemical oxygen demand (COD), soluble and insoluble

and examined by a sedimentation-flotation mathod (Cherepanov, 1982).

that there is s sufficient number of positive samples in our observations.

suspension of eggs was stored in an Erlenmayer flask in a refrigerator at 4°C.

mixed with hay, was also studied.

**2. Materials and methods 2.1 Parasitological methods** 

Petri dishes.

conditions.

ions (NH4

substances and C:N ratio.

**2.2 Physical and chemical methods** 

developmental stages (oocysts, eggs, larvae). Through faeces of infected dogs and cats the germs of parasitozoonoses spread into the environment. It is especially the case of cysts of intestinal parasitic protozoa – *Entamoeba histolytica, Giardia intestinalis, Balantidium coli, Toxoplasma gondii*, the eggs of tapeworms *Dipylidium* sp., *Echinococcus* sp. and parasitic nematodes (Antolová et al., 2004; Matsuo and Nakashio, 2005; Miterpáková et al., 2006). Regarding public health helminthozoonoses caused by *Toxocara* sp. (in dogs) and *Ascaris* sp. (in pigs) are very significant, especially due to their zoonotic character connected with the syndrome *larva migrans*.

Humans became infected usually orally *(per os)* by ingestion of substrates (soil, vegetables, etc.) with embryonated *Toxocara* eggs*.* Many symptoms are associated with this infection, including changes in blood cell counts and affection of various organs, as the ascaris larvae can migrate throughout the body. The symptoms of infection are often non-specific and may be mistaken with those of other infectious agents (common viral diseases, diarrhoea) or we may not observe any clinical signs. Toxocariasis manifests itself in two distinct forms: visceral, *larva migrans visceralis,* and ocular, *larva migrans ocularis* (Despommier, 2003).

*A. suum* infects pigs and is of major economical significance due to production losses linked to reduced feed conversion efficiency and losses to the mean industry associated with the condemnation of "milk-spot" livers (Dubinský et al., 2000). *Ascaris* infects over a quarter of the world´s human population (1.47 billion people worldwide) and clinically affects ~335 million people (Crompton, 1999).

The above-mentioned helminthozoonoses are classified among epidemiologically "low-risk" parasitozoonoses, because the propagative stages develop in the outdoor environment into the infectious stage and potentially secondarily contaminate the food chain. Therefore, a direct contact with infected animal, but also contaminated environment, or contaminated food chain (water, vegetables) are considered as a potential risk factor.

Attention has receantly been paid also to the problem of hygienical hazards in terms of the treatment and use of animal excrements and their application to soil as valuable nutrients for cultivated plants. The hazards are mainly connected with the quantity of continually produced solid and liquid wastes. The results is ecological disbalance, mainly with respect to environmental load with pathogenic microorgamism and nitrous organic substances. Animal organic wastes are also sources of greate amounts of gases releases. The most dangerous of them are ammonia and methane. Ammonia released into the atmosphere is irritating and toxic to the biotic component of the environment. On the other hand, animal excrements can supply essential plant nutrients and improve the fertility of soil by adding organic matter.

Therefore, to prevent health risks (for human as well as for animals) and odour nuisance from animal wastes, different methods for a satisfactory utilisation and sanitation have been researched (Schwartzbrod et al., 1989, Tofant et al., 1999; Juriš et al., 2000; Sasáková et al., 2005; Papajová and Juriš, 2009). There are big variations in the treatment of animal wastes (aerobic and anaerobic stabilization, composting etc.).

For the above-mentioned reasons our studies concentrated on anaerobic stabilization of liquid (slurry) and solid (manure, excrements) animal waste. The aims of our study were to monitor the physical-chemical changes in pig slurry treated by ecologically acceptable and energetically beneficial anaerobic stabilisation, changes in the properties of anaerobically digested slurry stored in ground lagoon and the effect of anaerobically digested slurry stores in ground lagoon on the survival of parasitic germs. The impact of lime (two types) on the survival of parasitic organisms in anaerobic stored manure and dog excrements mixed with hay, was also studied.
