A rapid monitoring assay for the detection of Salmonella spp. and Salmonella Senftenberg strain W775 in composts

AIMS: The composting process needs to be validated for its hygienic status in order to ensure that it is free of pathogens. Generally, this is evaluated through temperature monitoring, or additionally through active inoculation and monitoring of indicator organisms. We aimed to develop a monitoring method for the heat-resistant indicator organism Salmonella enterica ssp. enterica serovar Senftenberg strain W775 for detection in composting biowastes. METHODS AND RESULTS: The method development is comprised of: (i) optimization of molecular detection of bacteria belonging to the genus Salmonella; (ii) identification of a DNA marker for Salmonella strain W775; and (iii) development of a multiplex polymerase chain reaction (PCR)-based on both DNA markers. Subsequently, Salmonella strain W775 was inoculated and monitored during composting of biowastes in an industrial composting facility. CONCLUSIONS: A highly sensitive and specific detection of viable cells was obtained by enriching the compost sample prior to multiplex PCR analysis. Complete inactivation of Salmonella strain W775 was obtained within 4 days in an industrial composting facility at temperatures ranging between 41 and 57 degrees C. SIGNIFICANCE AND IMPACT OF THE STUDY: We describe a monitoring method for the simultaneous detection of naturally occurring Salmonella strains and artificially introduced Salmonella strain W775 in composting biowastes that can be applied in routine analysis.     

Humic substances change during the co-composting process of municipal solid wastes and sewage sludge

The change of the degree of stability of compost during the composting process was a kind of guideline for our study. This stability was estimated by monitoring the chemical fractionation (extraction of humic and fulvic acids, and humin) during two cycles of composting. Change of humin (H), humic-like acid carbon (CHA) and fulvic-like acid carbon (CFA) fractions during the composting process of municipal solid wastes were investigated using two windrows W1 (100% of municipal solid wastes) and W2 (60% of municipal solid wastes and 40% of dried sewage sludge). Humin and fulvic acid fractions in the two windrows decreased since the start of composting process and tend to stabilize. At the end of composting process, humic acid fraction is more important in the windrow without sludge (W1) than the one with sludge (W2). The humification indexes used in this study showed that the humic-like acid carbon fraction production takes place largely during the phase of temperature increase (thermophilic phase), and it appeared very active in the windrow W2. At the end of composting process, the E₄/E₆ ratio value indicated that the compost of W1 is more mature than the compost of W2. The humification ratio (HR) allowed a correct estimation of compost organic matter stabilization level.     

Variation in microbial population during composting of agro-industrial waste

Two compost piles were prepared, using two ventilation systems: forced ventilation and ventilation through mechanical turning. The material to compost was a mixture of orange waste, olive pomace, and grass clippings (2:1:1 v/v). During the composting period (375 days), samples were periodically taken from both piles, and the enumeration of fungi, actinomycetes, and heterotrophic bacteria was carried out. All studied microorganisms were incubated at 25 and 55 °C after inoculation in appropriate growth media. Fungi were dominant in the early stages of both composting processes; heterotrophic bacteria proliferated mainly during the thermophilic stage, and actinomycetes were more abundant in the final stage of the composting process. Our results showed that the physical and chemical parameters: temperature, pH, moisture, and aeration influenced the variation of the microbial population along the composting process. This study demonstrated that composting of these types of wastes, despite the prolonged mesophilic stage, provided an expected microbial variation.     

Microbiological parameters as indicators of compost maturity

AIMS: The objectives of this study were to determine the changes of microbial properties of pig manure collected from pens with different management strategies and composted using different turning and moisture regimes; relate their association with humification parameters and compost temperature; and identify the most suitable microbial indicators of compost maturity. METHODS AND RESULTS: Six different microbial parameters, including total bacterial count, oxygen consumption rate, ATP content, dehydrogenase activity, and microbial biomass C and N, along with humification parameters [humic acid (HA), fulvic acid (FA) and HA : FA ratio] and compost temperature were monitored during composting. Significant positive correlations were found between temperature and microbial properties, including O2 consumption rate, ATP content, dehydrogenase activity, and microbial biomass N. The humification parameters also showed significant correlations with microbial properties of the manure compost. For instance, HA contents of pig manures was positively correlated with total aerobic heterotrophs, and microbial biomass N and C; and negatively correlated with O2 consumption rate, ATP content, and dehydrogenase activity. Among the six microbial parameters examined, dehydrogenase activity was the most important factor affecting compost temperature and humification parameters. Composting strategies employed in this study affected the speed of composting and time of maturation. If the moisture content is maintained weekly at 60% with a 4-day turning frequency, the pig manure will reach maturity in 56 days. CONCLUSIONS: The composting process went through predictable changes in temperature, microbial properties and chemical components despite differences in the initial pig manure and composting strategies used. Among the six microbial parameters used, dehydrogenase activity is the most suitable indicator of compost maturity. Compared with respiration rate, ATP content and microbial biomass procedures, dehydrogenase activity is the simplest, quickest, and cheapest method that can be used to monitor the stability and maturity of composts. SIGNIFICANCE AND IMPACT OF THE STUDY: The results presented here show that microbial parameters can be used in revealing differences between composts and compost maturity. The statistical relationship established between humification parameters and microbial parameters, particularly dehydrogenase activity, demonstrates that it is possible to monitor the composting process more easily and rapidly by avoiding longer and more expensive analytical procedures.     

Determining thermal inactivation of Escherichia coli O157:H7 in fresh compost by simulating early phases of the composting process

A three-strain mixture of Escherichia coli O157:H7 was inoculated into fresh dairy compost (ca. 10(7) CFU/g) with 40 or 50% moisture and was placed in an environmental chamber (ca. 70% humidity) that was programmed to ramp from room temperature to selected composting temperatures in 2 and 5 days to simulate the early composting phase. The surviving E. coli O157:H7 population was analyzed by direct plating and enrichment. Optimal and suboptimal compost mixes, with carbon/nitrogen (C/N) ratios of 25:1 and 16:1, respectively, were compared in this study. In the optimal compost mix, E. coli O157:H7 survived for 72, 48, and 24 h in compost with 40% moisture and for 72, 24, and 24 h with 50% moisture at 50, 55, and 60°C, respectively, following 2 days of come-up time (rate of heating up). However, in the suboptimal compost mix, the pathogen survived for 288, 72, and 48 h in compost with 40% moisture and for 240, 72, 24 h in compost with 50% moisture at the same temperatures, respectively. Pathogen survival was longer, with 5 days of come-up time compared with 2 days of come-up. Overall, E. coli O157:H7 was inactivated faster in the compost with 50% moisture than in the compost with 40% at 55 and 60°C. Both moisture and come-up time were significant factors affecting Weibull model parameters. Our results suggest that slow come-up time at the beginning of composting can extend pathogen survival during composting. Additionally, both the C/N ratio and the initial moisture level in the compost mix affect the rate of pathogen inactivation as well.     

Heat inactivation of Salmonella spp. in fresh poultry compost by simulating early phase of composting process

Aims: The purpose of this study was to determine the effect of moisture on thermal inactivation of Salmonella spp. in poultry litter under optimal composting conditions. Methods and Results: Thermal inactivation of Salmonella was studied in fresh poultry compost by simulating early phase of composting process. A mixture of three Salmonella serotypes grown in Tryptic soy broth with rifampin (TSB‐R) was inoculated in fresh compost with 40 or 50% moisture at a final concentration of c. 7 log CFU g^?1. The inoculated compost was kept in an environmental chamber which was programmed to rise from room temperature to target composting temperatures in 2 days. In poultry compost with optimal moisture content (50%), Salmonella spp. survived for 96, 72 and 24 h at 50, 55 and 60°C, respectively, as compared with 264, 144 and 72 h at 50, 55 and 60°C, respectively, in compost with suboptimal moisture (40%). Pathogen decline was faster during the come‐up time owing to higher ammonia volatilization. Conclusions: Our results demonstrated that Salmonella spp. survived longer in fresh poultry compost with suboptimal moisture of 40% than in compost with optimal moisture of 50% during thermophilic composting. High nitrogen content of the poultry compost is an additional factor contributing to Salmonella inactivation through ammonia volatilization during thermal exposure. Significance and Impact of the Study: This research validated the effectiveness of the current composting guidelines on Salmonella inactivation in fresh poultry compost. Both initial moisture level and ammonia volatilization are important factors affecting microbiological safety and quality of compost product.     

湿度对堆肥理化性质的影响

水分是堆肥微生物生命活动的基础 ,也是堆肥中重要的工艺控制参数。弄清湿度对堆肥微生物及理化性质的影响 ,对于优化堆肥工艺参数、提高堆肥效率、降低投资和运行成本具有重要意义。综述了堆肥湿度研究的动态 ,指出了当前研究中存在的问题 ,并提出了未来的研究方向。大量的研究表明 ,湿度低于 4 5 %或高于 6 5 %都不利于堆肥处理。湿度太高会导致堆料的压实度增加、FAS减少、透气性能降低 ,从而导致堆体内氧气供应不足、堆肥升温困难、有机物降解速率降低、堆肥周期延长。湿度过低 ,水分会限制堆肥微生物的新陈代谢 ,导致微生物活性下降、堆肥腐熟困难。由于鼓风、散热、水蒸发等会使堆体内存在湿度的空间变异 ,也会降低堆肥效率和堆肥产品的质量。另外 ,堆肥湿度还影响堆肥的保肥能力。由各文献得出结论 ,堆肥的最佳湿度范围一般为 5 0 %～ 6 0 %左右     

Determination of aeration rate and kinetics of composting some agricultural wastes

This study aimed to determine the aeration rate and its kinetics in aerobic composting of agricultural wastes. For this aim compost materials were prepared by mixing grass trimmings, tomato, pepper, and eggplant wastes. Four vertical forced aeration type reactors and one vertical natural convection type reactor were manufactured to apply four different aeration rates. CO2 rate and temperature changes were recorded in three different places in the reactors. Moisture content, pH and organic material rate were recorded each day. While process-monitoring parameters (CO2, temperature, pH, moisture content) were used for interpretation of the process, organic material degradation was used for interpretation of the process success. The seven different kinetic models were applied for modeling decomposition rate to the experimental values. According to the results, four of these models were found applicable to this study. These models were analyzed with some statistical methods as root mean square error (RMSE), chi-square (chi2), and modeling efficiency (EF). According to the statistical results of these models, the best model was found as: [Formula: see text] where kT is the rate of decomposition (g VS/g VS day); T the process temperature (degrees C); Mc the daily moisture content (%wb); C the daily CO2 rate in composting reactor (%) and a, b, c, d are constants. According to the results, the highest organic matter degradation and temperature value were obtained at the aeration rate of 0.4 l air min(-1)kg(om)(-1). Thus, it could be applied to this mixed materials composting process.     

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.     

Characteristics of dairy manure composting with rice straw

The aim of this work was to investigate the effects of aeration rate, aeration method, moisture content, and manure age on the characteristics of dairy manure composting with rice straw in terms of composting temperature, oxygen consumption rate, emission of odorous gases, and final compost property. It was found that the aeration rate of 0.25 L/min-kg VS was capable of achieving the highest composting temperature, longest retention time of high temperature, and less emission of odor gases. Except for the composting temperature reached, there was no significant difference between bottom-forced and top-diffusion aerations in terms of final compost property. The higher initial moisture content (65%) was more favorable for its higher temperature, longer retention time of high temperature, and more stable end compost obtained. Fresh manure showed better composting performance than the aged manure for its higher temperature reached in less time and less ammonia emission. Oxygen consumption rates were basically similar to those of temperatures. Most emissions of the odorous gases occurred during the first week of composting, therefore, special attention should be paid to this period of time for effective odor control.     

Microbiological analysis of composts produced on South Carolina poultry farms

Aims: The purpose of this study was to determine whether the methods used in compost operations of small and medium‐sized poultry forms resulted in the production of an amendment free of foodborne pathogens. Methods and Results: Nine compost heaps on five South Carolina poultry farms were surveyed at different stages of the composting process. Compost samples were analysed for coliforms and enriched for Escherichia coli O157:H7, Salmonella spp. and Listeria monocytogenes. The waste materials and composting practices differed among the surveyed farms. On two farms, new materials were added to heaps that had previously completed the active composting phase. Five compost heaps did not reach an internal temperature of 55°C, and c. 62% of all internal samples in the first composting phase contained moisture contents <40%. Escherichia coli was detected in 63% of the surface samples (n = 38) and 9·8% of the internal samples (n = 82) from the first composting phase, as compared with 16·7% of the surface samples (n = 12) and 0% internal samples (n = 24) from the second composting phase. Salmonella was detected in 26 and 6·1% of all surface and internal samples collected from heaps in the first composting phase, respectively, but was absent in all compost samples undergoing a second composting phase. The predominant Salmonella serotypes were Thompson, Montevideo and Anatum. Neither E. coli O157:H7 nor L. monocytogenes was detected in any of the samples. Conclusions: Our results indicate that the conditions at the compost surface are suitable for pathogen survival, and the complete composting process can result in the elimination of pathogens in poultry wastes. Significance and Impact of the Study: This research provides information regarding the effectiveness of the composting practices and microbiological quality of poultry compost produced by small‐ and medium‐sized farms. Ensuring the safety of compost that may be applied to soils should be an integral part of preharvest food safety programme.     

High-rate composting of barley dregs with sewage sludge in a pilot scale bioreactor

The feasibility of high-rate composting of barley dregs and sewage sludge was examined using a pilot scale bioreactor. A central composite design (CCD) was used to optimize the mix ratio of barley dregs/sewage sludge and moisture content. The performance of the bioreactor was monitored as a function of carbon decomposition rate (CDR) and total volatile solids (TVS) loss rate. The optimum range of mix ratio and moisture content was found to be 35-40% and 55-60%, respectively. High CO2 evolution rate (CER) and TVS loss rate were observed after 3 days of the composting and the compost was matured/stable after 7 days. Cardinal temperature model with inflection (CTMI) was used to analyze the compost stability with respect to CER as a parameter of composting efficiency. After examining the phytotoxicity, the compost can be promoted for land application.     

The fate of a genetically modified Pseudomonas strain and its transgene during the composting of poultry manure

The fate of the genetically modified (GM) Pseudomonas chlororaphis strain 3732 RN-L11 and its transgene (lacZ insert) during composting of chicken manure was studied using plate count and nested polymerase chain reaction (PCR) methods. The detection sensitivity of the nested PCR method was 165 copies of the modified gene per gram of moist compost or soil. Compost microcosms consisted of a 100-g mixture of chicken manure and peat, whereas soil microcosms were 100-g samples of sandy clay loam. Each microcosm was inoculated with 4 x 1010 CFU of P. chlororaphis RN-L11. In controlled temperature studies, neither P. chlororaphis RN-L11 nor its transgene could be detected in compost microcosms after incubation temperature was elevated to 45 degrees C or above for one or more days. In contrast, in the compost microcosms incubated at 23 degrees C, the target organism was not detected by the plate count method after 6 days, but its transgene was detectable for at least 45 days. In compost bins, the target organism was not recovered from compost microcosms or soil microcosms at different levels in the bins for 29 days. However, the transgene was detected in 8 of the 9 soil microcosms and in only 1 of the 9 compost microcosms. The compost microcosm in which transgene was detected was at the lower level of the bin where temperatures remained below 45 degrees C. The findings indicated that composting of organic wastes could be used to reduce or degrade heat sensitive GM microorganisms and their transgenes.     

Microbiological aspects of a municipal waste composting system

Summary The municipal solid waste of Rome, processed by the Dano plant, has been studied in the stabilization phase. During the different composting stages the following were determined: pile temperature, pH, moisture content, microbiological counts on compost extract and on other media, and mycological investigations. Twenty four days in January were necessary for the compost stabilization.     

Olive mill waste composting: A review

Olive mill wastes exacerbate environmental problems in Mediterranean countries. These wastes are highly phytotoxic and contain phenolic compounds, lipids and organic acids. They also contain high percentages of organic matter and a vast range of plant nutrients that could be reused as fertilizers for sustainable agricultural practices. In this paper, recent research on composting wastes of 2-phase and 3-phase olive mills is reviewed, concentrating on factors affecting composting such as bulking agents, aeration strategy, physicochemical characteristics (duration of the thermophilic phase, moisture content, organic matter, volatile solids, total organic carbon, water soluble carbon, total nitrogen, total phosphorus, potassium, C/N ratio, phenols and the humification process), and phytotoxicity. The review highlights the effects of composting operational factors (bulking agent, additives, and aeration strategy) on the physicochemical characteristics of the final compost, and the production of a good quality soil amender or fertilizer.     

生物垃圾堆肥的过程研究及腐熟度评价

目的：研究静态强制通风发酵工艺对合肥市农贸市场的生物废弃物进行好氧堆肥处理过程的物质变化和腐熟效果。方法：分析发酵过程中温度、含水率、挥发分含量、纤维素、半纤维素和木质素含量等物理指标和化学指标。结果：一次发酵中堆肥的温度在55℃以上的有3d,经过一次发酵和二次发酵,堆肥中挥发分含量由85.4%降低到51.2%。纤维素的含量由20.90%减少到15.48%,半纤维素的含量由7.20%减少到1.36%,,纤维素、半纤维素在二次发酵中降解率比一次发酵的小,木质素虽在一次发酵中基本没有降解。但在二次发酵中得到有效降解,木质素含量由8.30%减少到5.38%。堆肥后物料在70d堆肥后,基本上达到腐熟程度。结论：温度、含水率、挥发性物质、淀粉实验、生物可降解度实验等指标检测可以评价静态强制通风发酵工艺对合肥市农贸市场的生物废弃物进行好氧堆肥处理过程的物质变化和腐熟效果。     

环境条件对猪粪好氧堆肥过程的影响

禽畜粪便的大量排放给环境造成了巨大污染,其中猪场粪便和污水是最主要的污染源。为了实现猪粪的无害化、减量化和资源化,好氧堆肥是目前普遍应用和效果良好的实用技术。为了探明堆肥过程的生物学和化学过程,论本文主要从几个影响因素如温度、调理剂、含水率、通风方式、微生物菌剂等对猪粪好氧堆肥技术方面进行了综述。     

Co-composting of oil exhausted olive-cake, poultry manure and industrial residues of agro-food activity for soil amendment

The co-composting of exhausted olive-cake with poultry manure and sesame shells was investigated. These organic solid wastes were watered by the confectionary wastewater which is characterized by its high content of residual sugars raising its COD. Four aerated windrows were performed to establish the effects of confectionary by-products on the compost process. Different mixtures of the agro-industrial wastes were used. During the composting process, physico-chemical parameters (temperature, moisture, pH, electrical conductivity, total carbon and total nitrogen) were studied. The stability of the biological system was noticed after 70 days. The final products were characterized by their relatively high organic matter content, and low C/N ratio of 14-17. The humidification of the windrows with the wastewater seemed to have accelerated the composting process in comparison to a windrow humidified with water. In addition, the organic matter degradation was enhanced to reach 55-70%. The application of the obtained composts to soil appeared to significantly improve the soil fertility. Indeed, field experiments showed an increase in potato yield; the production was 30.5-37.5 tons ha(-1), compared to 30.5 tons ha(-1) with farm manure.     

Changes in physical,chemical and microbial parameters during the composting of municipal sewage sludge

Changes in physical, chemical and microbial parameters were investigated during the composting of municipal sewage sludge. Raw sewage sludge (30% dry matter) was mixed with compost from sewage sludge (85% dry matter) in 3:1 ratio (v/v). The mixture was divided into 4 windrows which were composted under the same conditions except the turning factor. The turning was every 7, 10, 15 days and according to the temperature which must be (55–65°C) for windrow 1 (W1), windrow 2 (W2), windrow 3 (W3) and windrow 4 (W4), respectively. Water was added to adjust the moisture content (40–60%). The composting process consisted of 2 periods; fermentation (12 weeks) and maturation (4 weeks). The results showed that the temperature reached the maximum after 12 weeks for W1 and 11 weeks for W2, W3 and W4 and then decreased. The final compost was nearly odourless and black, especially in case of W4. The general trend indicates a decrease in organic matter, organic carbon and nitrogen (N), whereas ash, potassium (K) and phosphorus (P) increased and consequently C/K and C/P ratios decreased. There was a slight increase in C/N ratio. The pH increased and then decreased to near neutrality at the end. The mesophilic bacteria increased during the fermentation period and decreased after that, whereas the thermophilic ones increased with increasing of temperature, decreased after 2 weeks and increased again during the fermentation period and then decreased. The mesophilic and thermophilic fungi were present during the first week and disappeared after that. The final compost was pathogens-free as indicated by the counts of coliforms and Salmonella.