Effect of Length of Time before Incorporation on Survival of Pathogenic Bacteria Present in Livestock Wastes Applied to Agricultural Soil

In response to reports that the contamination of food can occur during the on-farm primary phase of food production, we report data that describes a possible cost-effective intervention measure. The effect of time before soil incorporation of livestock wastes spread to land on the rate of decline of zoonotic agents present in the waste was investigated. Fresh livestock wastes were inoculated with laboratory-cultured Salmonella, Listeria, and Campylobacter spp. and Escherichia coli O157 before they were spead onto soil. Incorporation of the spread wastes was either immediate, delayed for 1 week, or did not occur at all. Bacterial decline was monitored over time and found to be significantly more rapid for all waste types when they were left on the soil surface. There were no significant differences in initial bacterial decline rates when wastes were spread in summer or winter. Our results indicate that not incorporating contaminated livestock wastes into soil is a potential intervention measure that may help to limit the spread of zoonotic agents further up the food chain. The implications of these findings are discussed in relation to current advice for livestock waste disposal.     

Decline of zoonotic agents in livestock waste and bedding heaps

AIMS: To measure the rates of decline of zoonotic agents introduced into heaps of spent bedding and faecal wastes generated by commercially farmed livestock and managed in a similar way to that of a working farm. METHODS AND RESULTS: Livestock isolates of Salmonella, pathogenic Listeria, Campylobacter and Escherichia coli O157 were laboratory cultured and used to inoculate 5 m3 heaps of cattle, sheep or pig wastes mixed with bedding materials. Decline of each of the infectious agents was monitored with time as was the temperature inside each heap. Temperatures of >50 degrees C were typically achieved at the core of the heaps. Pathogen decline was rapid, typically <3 days for a 1-log reduction in levels. The longest time that zoonotic agents were isolated from the heaps was 93 days. CONCLUSIONS: Movement of heaps of livestock bedding waste from animal pens to a secondary store, and storing them under conditions conducive for increased temperature is a simple and cost-effective treatment for rapidly lowering levels of zoonotic agents in solid farm wastes. SIGNIFICANCE AND IMPACT OF THE STUDY: This study demonstrates a simple and cheap treatment that can be used to help prevent the spread of zoonotic agents through agricultural environments.     

Fate of pathogens present in livestock wastes spread onto fescue plots

Fecal wastes from a variety of farmed livestock were inoculated with livestock isolates of Escherichia coli O157, Listeria monocytogenes, Salmonella, Campylobacter jejuni, and Cryptosporidium parvum oocysts at levels representative of the levels found in naturally contaminated wastes. The wastes were subsequently spread onto a grass pasture, and the decline of each of the zoonotic agents was monitored over time. There were no significant differences among the decimal reduction times for the bacterial pathogens. The mean bacterial decimal reduction time was 1.94 days. A range of times between 8 and 31 days for a 1-log reduction in C. parvum levels was obtained, demonstrating that the protozoans were significantly more hardy than the bacteria. Oocyst recovery was more efficient from wastes with lower dry matter contents. The levels of most of the zoonotic agents had declined to below detectable levels by 64 days. However, for some waste types, 128 days was required for the complete decline of L. monocytogenes levels. We were unable to find significant differences between the rates of pathogen decline in liquid (slurry) and solid (farmyard manure) wastes, although concerns have been raised that increased slurry generation as a consequence of more intensive farming practices could lead to increased survival of zoonotic agents in the environment.     

Fate of Pathogens Present in Livestock Wastes Spread onto Fescue Plots

Fecal wastes from a variety of farmed livestock were inoculated with livestock isolates of Escherichia coli O157, Listeria monocytogenes, Salmonella, Campylobacter jejuni, and Cryptosporidium parvum oocysts at levels representative of the levels found in naturally contaminated wastes. The wastes were subsequently spread onto a grass pasture, and the decline of each of the zoonotic agents was monitored over time. There were no significant differences among the decimal reduction times for the bacterial pathogens. The mean bacterial decimal reduction time was 1.94 days. A range of times between 8 and 31 days for a 1-log reduction in C. parvum levels was obtained, demonstrating that the protozoans were significantly more hardy than the bacteria. Oocyst recovery was more efficient from wastes with lower dry matter contents. The levels of most of the zoonotic agents had declined to below detectable levels by 64 days. However, for some waste types, 128 days was required for the complete decline of L. monocytogenes levels. We were unable to find significant differences between the rates of pathogen decline in liquid (slurry) and solid (farmyard manure) wastes, although concerns have been raised that increased slurry generation as a consequence of more intensive farming practices could lead to increased survival of zoonotic agents in the environment.     

The air-borne distribution of zoonotic agents from livestock waste spreading and microbiological risk to fresh produce from contaminated irrigation sources

Aims:  To assess the risks of zoonotic agents in dissemination of livestock wastes into the environment by airborne distribution. To subsequently assess the survival time of zoonotic agents, introduced in irrigation water, on the phylloplane of produce.
Methods and Results:  An Escherichia coli marker was introduced into pig slurry which was spread using a rain gun sprayer. Air sampling was undertaken to determine the distance reached by the marker. No recoveries were observed at a distance of 250 m. Borehole water, contaminated with zoonotic agents, was used to irrigate field plots sown with lettuce and spinach. Decline in bacterial numbers on the phylloplane was observed with time. After initial rapid decreases, we were unable to detect any pathogen from the phylloplane, 1 month after contamination.
Conclusions:  These preliminary results suggest that the risks to public health from the aerosolized spread of bacteria during slurry spreading by rain gun are low. Although, zoonotic agents on crop phylloplanes perish quickly, the risks of overhead irrigation of fresh produce 3 weeks before harvest should still be considered.
Significance and Impact of the Study:  These preliminary results improve our understanding on the fate of zoonotic agents in the environment. Spreading liquid livestock wastes by an airborne mechanism may not pose a significant public health risk. Detection of zoonotic agents 3 weeks after contamination of lettuce and spinach means that consideration should be given by the farmers until the time of harvest, when irrigating fresh produce with water that may have been directly or indirectly contaminated by livestock wastes.     

Declines of zoonotic agents in liquid livestock wastes stored in batches on-farm

AIM: To measure the decline rates of zoonotic agents introduced into liquid livestock wastes in on-farm storage tanks. METHODS AND RESULTS: Salmonella spp., Escherichia coli O157, Campylobacter jejuni, Listeria monocytogenes and Cryptosporidium parvum, propagated in laboratory-controlled conditions, were inoculated into 35,000-l volumes of fresh livestock wastes (pig slurries, cattle slurries and dirty waters). D-values for bacteria were six to 44 days, and for C. parvum were 133 to 345 days. Campylobacter jejuni declined significantly more rapidly than the other bacterial pathogens, while E. coli O157 declined significantly more slowly. On average, bacterial declines were not affected by the season of waste deposition and storage or by the dry matter content of the wastes, but were more rapid in dirty waters than in pig slurries. The physiciochemical composition of wastes in each category varied significantly. CONCLUSIONS: Zoonotic agents can survive for several months during storage of liquid livestock wastes. Livestock wastes should be batch-stored and not subjected to continuous additions. SIGNIFICANCE AND IMPACT OF THE STUDY: This study indicates that batches of liquid livestock waste, if contaminated with bacterial pathogens, should be stored for 6 months to reduce contamination levels. Alternative strategies for reducing C. parvum levels in liquid livestock wastes should be explored.     

Analyses of livestock production, waste storage, and pathogen levels and prevalences in farm manures

Survey results describing the levels and prevalences of zoonotic agents in 1,549 livestock waste samples were analyzed for significance with livestock husbandry and farm waste management practices. Statistical analyses of survey data showed that livestock groups containing calves of <3 months of age, piglets, or lambs had higher prevalences and levels of Campylobacter spp. and Escherichia coli O157 in their wastes. Younger calves that were still receiving milk, however, had significantly lower levels and prevalence of E. coli O157. Furthermore, when wastes contained any form of bedding, they had lowered prevalences and levels of both pathogenic Listeria spp. and Campylobacter spp. Livestock wastes generated by stock consuming a diet composed principally of grass were less likely to harbor E. coli O157 or Salmonella spp. Stocking density did not appear to influence either the levels or prevalences of bacterial pathogens. Significant seasonal differences in prevalences were detected in cattle wastes; Listeria spp. were more likely to be isolated in March to June, and E. coli O157 was more likely to be found in May and June. Factors such as livestock diet and age also had significant influence on the levels and prevalences of some zoonotic agents in livestock wastes. A number of the correlations identified could be used as the basis of a best-practice disposal document for farmers, thereby lowering the microbiological risks associated with applying manures of contaminated livestock to land.     

Analyses of Livestock Production, Waste Storage, and Pathogen Levels and Prevalences in Farm Manures

Survey results describing the levels and prevalences of zoonotic agents in 1,549 livestock waste samples were analyzed for significance with livestock husbandry and farm waste management practices. Statistical analyses of survey data showed that livestock groups containing calves of <3 months of age, piglets, or lambs had higher prevalences and levels of Campylobacter spp. and Escherichia coli O157 in their wastes. Younger calves that were still receiving milk, however, had significantly lower levels and prevalence of E. coli O157. Furthermore, when wastes contained any form of bedding, they had lowered prevalences and levels of both pathogenic Listeria spp. and Campylobacter spp. Livestock wastes generated by stock consuming a diet composed principally of grass were less likely to harbor E. coli O157 or Salmonella spp. Stocking density did not appear to influence either the levels or prevalences of bacterial pathogens. Significant seasonal differences in prevalences were detected in cattle wastes; Listeria spp. were more likely to be isolated in March to June, and E. coli O157 was more likely to be found in May and June. Factors such as livestock diet and age also had significant influence on the levels and prevalences of some zoonotic agents in livestock wastes. A number of the correlations identified could be used as the basis of a best-practice disposal document for farmers, thereby lowering the microbiological risks associated with applying manures of contaminated livestock to land.     

Efficacy of organic wastes amendment on the establishment and biomass yield of grass species on a degraded Alfisol

Abstract

Deficiencies of organic matter and essential nutrients are important features of derelict/degraded lands; these characteristics have profound effects on the establishment of functional soil-plant systems on such lands. This work reports on the impact of organic wastes amendments on the establishment, growth and biomass yield of grass mixtures grown on a degraded soil. The experimental site was a heavily compacted and eroded land due to excavation (scrapping) of topsoil layer for urban construction activities. Zero, 10 and 20cm layers each of substrates collected from an old (abandoned) municipal refuse and from site affected by long-term surface disposal of livestock litter and topsoil collected under fallow vegetation were applied on soil surface, before the establishment of a mixture of grass species. Substrates collected from old municipal refuse contained high amount of essential nutrients and metals and, on application, produced upward shift in soil pH and electrical conductivity. Nevertheless, these properties neither significantly retarded plant growth nor impaired biomass yield. Plant establishment and biomass yield were comparable under topsoil application and organic waste amended soil and produced significantly higher biomass yield when spread at 20cm layers were made of them. This experimental group also produced superior biomass yield over the unamended control. The agronomic benefits of organic waste incorporation stemmed from improved soil properties in surface horizon and contents of essential nutrients supportive of the establishment and development of plant cover. Nutrient supply is related to soil organic matter status, organic wastes are therefore important to the reconstruction of effective nutrient cycling and the eventual functional soil-plant system on this degraded ecosystem. The goal of these revegetation efforts is to improve soil and plant productivity, plant diversity, conservation of native grasslands and aesthetic.     

Livestock waste treatment systems for reducing environmental exposure to hazardous enteric pathogens: Some considerations

Intensive livestock production systems produce significant quantities of excreted material that must be managed to protect water, air, and crop quality. Many jurisdictions mandate how livestock wastes are managed to protect adjacent water quality from microbial and chemical contaminants that pose an environmental and human health challenge. Here, we consider innovative livestock waste treatment systems in the context of multi-barrier strategies for protecting water quality from agricultural contamination. Specifically, we consider some aspects of how enteric bacterial populations can evolve during manure storage, how their fate following land application of manure can vary according to manure composition, and finally the challenge of distinguishing enteric pathogens of agricultural provenance from those of other sources of fecal pollution at a policy-relevant watershed scale. The beneficial impacts of livestock waste treatment on risk to humans via exposure to manured land are illustrated using quantitative microbial risk assessment (QMRA) scenarios. Overall, innovative livestock treatment systems offer a crucially important strategy for making livestock wastes more benign before they are released into the broader environment.     

Survival of E. coli O157:H7 in organic wastes destined for land application

AIM: To determine the persistence of Escherichia coli O157 in contrasting organic wastes spread to land and to assess the potential environmental risk associated with the disposal of these wastes to land. METHODS AND RESULTS: Twenty-seven organic wastes originating from slaughterhouses, wastewater treatment plants (raw and treated sewage), creameries and farms (bovine slurry), were inoculated with E. coli O157:H7 and incubated at 10 degrees C. Although pathogen numbers gradually declined in all the wastes, albeit at different rates even in the same waste type, E. coli O157:H7 was still viable in 77% of organic wastes tested after 2 months. CONCLUSIONS: Long-term storage of organic wastes led to a significant and gradual decline in E. coli O157:H7 numbers. Consequently, storage may be a useful means of reducing the pathogen load of wastes destined for land application. However, in most cases, long-term storage cannot be expected to completely eliminate E. coli O157:H7 from waste. SIGNIFICANCE AND IMPACT OF THE STUDY: Our results indicate that current legislation may be insufficient to protect the environment from E. coli O157:H7 contamination from untreated wastes spread to land.     

Effect of Antimicrobial Agents on Livestock Waste Emissions

Various antimicrobial agents were evaluated with the purpose of reducing the microbial fermentation in stored cattle waste and the resulting odor emissions. Duplicate sealed 2-L flasks with 500 ml waste slurry, with and without antimicrobial inhibitors, were used to measure the production of short-chain volatile fatty acids, lactate, and total fermentation gas over 27–30 days. A combination of chlorhexidine diacetate (2 mM), iodoacetate (2 mM), and α-pinene (3.8 mM) reduced gas production 80% (1000 ml to 200 ml) and total volatile fatty acid production 50% (145 mM to 72 mM). Pinene had little antimicrobial effect; rather, it served as an effective masking agent, giving the waste a less offensive odor. A combination of chlorhexidine diacetate and the deaminase inhibitor, diphenyliodonium chloride (1.3 mM) had a similar effect in reducing short-chain volatile fatty acid production (145 mM to 80 mM). It is concluded that a combination of antimicrobial agents may be useful in controlling odor emissions and conserving organic matter in livestock wastes, therefore providing a potentially more useful byproduct waste when used as plant fertilizer. Received: 22 November 1999 / Accepted: 5 January 2000     

Awareness and Practices Relating to Zoonotic Diseases Among Smallholder Farmers in Nepal

Increasing livestock production to meet growing demands has resulted in greater interactions at the livestock–wildlife–human interface and more opportunities for zoonotic disease spread. Zoonoses impose enormous burdens on low-income countries like Nepal, where populations are largely dependent on livestock production and access to shared grazing lands, often near protected areas, due to population pressures. Several livestock-associated zoonoses have been reported in Nepal; however, little is known regarding Nepali farmers’ knowledge of zoonoses and opportunities for disease management. We conducted a cross-sectional study to investigate Nepali farmers’ awareness of zoonoses, assess current health challenges, and evaluate disease prevention and control practices. We found that awareness of zoonotic pathogens was limited, especially in informally educated and illiterate farmers; the majority of which were women. Further, farmers’ preventive herd health, food safety, and sanitation practices were not associated with their awareness. Several farmers reported high-risk practices despite being aware of zoonotic diseases, suggesting a disconnect between the farmers’ awareness and practice. Our study highlights the need for improving Nepali farmers’ knowledge of zoonoses and disease prevention measures. Closing these awareness-practice gaps will require an improved understanding of risk and effective drivers of behavior change, alongside engagement of farmers in development of zoonotic disease prevention programs that encourage participation of both male and female farmers across all levels of education.     

Distribution and decline of human pathogenic bacteria in soil after application in irrigation water and the potential for soil-splash-mediated dispersal onto fresh produce

Aims: To improve our understanding of the survival and splash‐mediated transfer of zoonotic agents and faecal indicator bacteria introduced into soils used for crop cultivation via contaminated irrigation waters. Methods and Results: Zoonotic agents and an Escherichia coli marker bacterium were inoculated into borehole water, which was applied to two different soil types in early‐, mid‐ and late summer. Decline of the zoonotic agents was influenced by soil type. Marker bacteria applied to columns of two soil types in irrigation water did not concentrate at the surface of the soils. Decline of zoonotic agents at the surface was influenced by soil type and environmental conditions. Typically, declines were rapid and bacteria were not detectable after 5 weeks. Selective agar strips were used to determine that the impact of water drops 24–87 μl could splash marker bacteria from soil surfaces horizontal distances of at least 25 cm and heights of 20 cm. Conclusions: Soil splash created by rain‐sized water droplets can transfer enteric bacteria from soil to ready‐to‐eat crops. Persistence of zoonotic agents was reduced at the hottest part of the growing season when irrigation is most likely. Significance and Impact of the Study: Soil splash can cause crop contamination. We report the penetration depths and seasonally influenced declines of bacteria applied in irrigation water into two soil types.     

Zoonotic agents in small ruminants kept on city farms in southern Germany

Sheep and goats are popular examples of livestock kept on city farms. In these settings, close contacts between humans and animals frequently occur. Although it is widely accepted that small ruminants can carry numerous zoonotic agents, it is unknown which of these agents actually occur in sheep and goats on city farms in Germany. We sampled feces and nasal liquid of 48 animals (28 goats, 20 sheep) distributed in 7 city farms and on one activity playground in southern Germany. We found that 100% of the sampled sheep and 89.3% of the goats carried Shiga toxin-producing Escherichia coli (STEC). The presence of Staphylococcus spp. in 75% of both sheep and goats could be demonstrated. Campylobacter spp. were detected in 25% and 14.3% of the sheep and goats, respectively. Neither Salmonella spp. nor Coxiella burnetii was found. On the basis of these data, we propose a reasonable hygiene scheme to prevent transmission of zoonotic agents during city farm visits.     

Investigation of a potential zoonotic transmission of orthoreovirus associated with acute influenza-like illness in an adult patient

Bats are increasingly being recognized as important reservoir hosts for a large number of viruses, some of them can be highly virulent when they infect human and livestock animals. Among the new bat zoonotic viruses discovered in recent years, several reoviruses (respiratory enteric orphan viruses) were found to be able to cause acute respiratory infections in humans, which included Melaka and Kampar viruses discovered in Malaysia, all of them belong to the genus Orthoreovirus, family Reoviridae. In this report, we describe the isolation of a highly related virus from an adult patient who suffered acute respiratory illness in Malaysia. Although there was no direct evidence of bat origin, epidemiological study indicated the potential exposure of the patient to bats before the onset of disease. The current study further demonstrates that spillover events of different strains of related orthoreoviruses from bats to humans are occurring on a regular basis, which calls for more intensive and systematic surveillances to fully assess the true public health impact of these newly discovered bat-borne zoonotic reoviruses.     

Dehydrogenase activity in Lumbricus terrestris casts and surrounding soil affected by addition of different organic wastes and Zn

A laboratory experiment was conducted to determine the effects of different organic wastes such as wheat straw (WS), tea production waste (TEW), tobacco production waste (TOW), cow manure (CM) and hazelnut husk (HH) on dehydrogenase activity (DHA) in casts of earthworm Lumbricus terrestris and surrounding soil using 5% (dry weight) application rates associated with increasing doses of Zn (0, 50, 100, 250, 500 and 1000 microg g(-1)). Twenty one days after treatment of Zn and organic wastes, the DHA analyses were carried out on collected casts and soil samples. In general, all organic waste treatments influenced the DHA, the contents of organic C, N and available Zn in earthworm L. terrestris casts and the surrounding soil in comparison with the control. DHA in casts exceeded that in the surrounding soil without Zn additions. After Zn application of 50 microg Zn g(-1) in all organic waste treatments and the control, the DHA level in casts and surrounding soil increased significantly. It decreased by application rates of 100, 250, 500 and 1000 microg Zn g(-1) consecutively in all organic waste applications. The addition of wastes with low C/N ratio and high Zn content (TEW, TOW, CM) inhibited the DHA in both cast and surrounding soil.     

Design and application of a pre-composting test step to determine the effect of high fat food wastes on an industrial scale in-vessel composting system

The high fat content in food wastes was suspected to inhibit an industrial in-vessel composting process from reaching the European Union Animal By-Product Regulation (composting temperature >70 °C for 1 h). The aim of this study was to design a test step to guide the mixing ratio of food waste to green waste to meet the regulation. A 15-compartment composting unit was designed to contain the compost mixes. Sausage and cheese wastes were mixed with green waste at 1:1; 1:2; 1:3 and 1:4 ratios by wet weight volume. Only the sausage waste mix ratio of 1:4 gave an average temperature of 70 °C for at least 1 h after 2 days of composting (fat content - 17%; C: N ratio - 8.6). All the cheese waste mixes did not reach 70 °C after 15 days of composting. This study demonstrated that using a simple pre-composting test step could reduce the chances of process failure during industrial composting. Although both sausage and cheese wastes are high in fat, they performed very differently in the composting process. Two linear equations were fitted to model the impact of these wastes on the maximum composting temperature.     

The significance of mycotoxins in the framework of assessing workplace related risks

Mycotoxins are fungal metabolite which may in some cases exhibit a high health hazard potential. Mycotoxins can show carcinogenic, mutagenic, toxic, teratogenic or immunotoxic effects. Mycotoxin exposure in the workplace may occur through inhalation and skin contact,e.g. during occupational handling of organic matter such as livestock feed, food products, or waste. Various studies suggest that both acute and chronic effects can occur, depending at least on the exposure level. The magnitude of the potential health risks associated with a respiratory or dermal intake of mycotoxins has largely remained unclear to date. However, according to the directive 2000/54/EC on biological agents and the corresponding German Biological Agents Ordinance, employers are also required to consider the potential hazards posed by toxic effects of biological agents when assessing workplace risks. The aim of this article, therefore, is to present some basis information that should facilitate an evaluation of the significance of mycotoxins in the context of assessing workplace risks. It also provides suggestions for occupational health and safety measures.     

A yield control approach to assess phytoavailability of Zn and Cu in irradiated,composted sewage sludges and composted manure in field experiments: I. Zinc

The threat of spreading diseases is a serious concern when organic wastes are applied to farmland. Irradiation and composting are effective methods to reduce pathogens. Field experiments were conducted to assess the influence of these pathogen-eliminating methods on plant availability of Zn in the wastes. Four organic wastes: digested and dewatered (DSS), irradiated (DISS), composted (DICSS) sewage sludge and composted livestock manure (CLM) were applied during two growing seasons at 10, 20, 30, and 40 t solids ha^–1 year^–1. Available N and K in the wastes were estimated and N and K fertilizers were added to the soil to equalize available N and K supply among treatments to avoid dilution of crop Zn concentration. A control treatment (CT) received fertilizers but no waste. Lettuce, snap beans and petunias were grown in 1990, and two cuts of lettuce were harvested in 1991. The influence of waste Zn application on crop Zn concentration was studied within approximately equal crop yields. Crop Zn concentration increased in all crops treated with DSS or DISS, and often reached a maximum at the 30 t ha^–1 rate of application, then slightly decreased at 40 t ha^–1. The response of crop Zn concentration to the amount of Zn applied in the wastes was best described by quadratic equations. Waste application also significantly increased soil Zn availability index, which was a function of DTPA (diethylenetriamine pentaacetic acid)-extractable soil Zn and soil pH. The index was highly correlated with crop Zn concentration. Although Zn concentration in DICSS was similar to those in DSS and DISS, Zn applied in DICSS did not increase crop Zn concentration or soil availability index. Composting reduced the availability of Zn. The similar concentrations of Zn in DSS and DISS in both years allowed the use of a paired t-test to determine the differences in crop Zn concentration caused by application of DSS and DISS. Zinc applied in DISS produced a higher Zn concentration in bean pods than Zn applied in DSS (t > T _0.05 at P < 0.02, df = 15), indicating that irradiation increased phytoavailability of Zn in the sludge. However, no similar effect was found in Zn concentrations in the two cuts of lettuce in 1991 or in soil Zn availability index.