Arcobacter butzleri in Sheep Ricotta Cheese at Retail and Related Sources of Contamination in an Industrial Dairy Plant

This study aimed to evaluate Arcobacter species contamination of industrial sheep ricotta cheese purchased at retail and to establish if the dairy plant environment may represent a source of contamination. A total of 32 sheep ricotta cheeses (1.5 kg/pack) packed in a modified atmosphere were purchased at retail, and 30 samples were collected in two sampling sessions performed in the cheese factory from surfaces in contact with food and from surfaces not in contact with food. Seven out of 32 samples (21.9%) of ricotta cheese collected at retail tested positive for Arcobacter butzleri at cultural examination; all positive samples were collected during the same sampling and belonged to the same batch. Ten surface samples (33.3%) collected in the dairy plant were positive for A. butzleri. Cluster analysis identified 32 pulsed-field gel electrophoresis (PFGE) patterns. The same PFGE pattern was isolated from more than one ricotta cheese sample, indicating a common source of contamination, while more PFGE patterns could be isolated in single samples, indicating different sources of contamination. The results of the environmental sampling showed that A. butzleri may be commonly isolated from the dairy processing plant investigated and may survive over time, as confirmed by the isolation of the same PFGE pattern in different industrial plant surface samples. Floor contamination may represent a source of A. butzleri spread to different areas of the dairy plant, as demonstrated by isolation of the same PFGE pattern in different production areas. Isolation of the same PFGE pattern from surface samples in the dairy plant and from ricotta cheese purchased at retail showed that plant surfaces may represent a source of A. butzleri postprocessing contamination in cheeses produced in industrial dairy plants.     

Prevalence and fingerprinting of Listeria monocytogenes strains isolated from raw whole milk in farm bulk tanks and in dairy plant receiving tanks

The incidence of Listeria species in raw whole milk from farm bulk tanks and from raw milk in storage at a Swedish dairy plant was studied. Listeria monocytogenes was found in 1.0% and Listeria innocua was found in 2.3% of the 294 farm bulk tank (farm tank) milk specimens. One farm tank specimen contained 60 CFU of L. monocytogenes ml(-1). L. monocytogenes was detected in 19.6% and L. innocua was detected in 8.5% of the milk specimens from the silo receiving tanks at the dairy (dairy silos). More dairy silo specimens were positive for both Listeria species during winter than during summer. Restriction enzyme analysis and pulsed-field gel electrophoresis were applied to 65 isolates of L. monocytogenes, resulting in 16 different clonal types. Two clonal types were shared by the farm tank milk and the dairy silo milk. All except one clonal type belonged to serovar 1/2a. In the dairy silo milk five clonal types were found more frequently and for a longer period than the others. No Listeria species were found in any other samples from the plant.     

Remote engineering for a cheese whey biorefinery: an Internet-based application for process design,economic analysis,monitoring, and control of multiple plant sites

The proteolysis of cheese whey with the aid of immobilized enzymes is an attractive alternative for this by-product of the dairy industry. Among some possible applications for whey protein hydrolysates, one may cite their use as protein source for individuals with reduced capacity of digestion, or with genetic metabolic disorders (phenylketonuria patients, for instance). The multipurpose plant that processes whey is named here as a cheese whey biorefinery. This work presents the remote control and monitoring of the whey biorefineries using the Internet. In an integrated environment, the web application also enables simulation and economic analyses of the process. This technology might allow small companies to access a remote “engineering centre”, with know-how on plant design and advanced control techniques. The idea can also be extended to large dairy companies, providing the remote control of geographically spread sites of production.     

Prevalence and Fingerprinting of Listeria monocytogenes Strains Isolated from Raw Whole Milk in Farm Bulk Tanks and in Dairy Plant Receiving Tanks

The incidence of Listeria species in raw whole milk from farm bulk tanks and from raw milk in storage at a Swedish dairy plant was studied. Listeria monocytogenes was found in 1.0% and Listeria innocua was found in 2.3% of the 294 farm bulk tank (farm tank) milk specimens. One farm tank specimen contained 60 CFU of L. monocytogenes ml⁻¹. L. monocytogenes was detected in 19.6% and L. innocua was detected in 8.5% of the milk specimens from the silo receiving tanks at the dairy (dairy silos). More dairy silo specimens were positive for both Listeria species during winter than during summer. Restriction enzyme analysis and pulsed-field gel electrophoresis were applied to 65 isolates of L. monocytogenes, resulting in 16 different clonal types. Two clonal types were shared by the farm tank milk and the dairy silo milk. All except one clonal type belonged to serovar 1/2a. In the dairy silo milk five clonal types were found more frequently and for a longer period than the others. No Listeria species were found in any other samples from the plant.     

Assessing and analysing contamination of a dairy products processing plant by Staphylococcus aureus using antibiotic resistance and PFGE

A dairy product processing plant was studied for 2.5 years to examine contamination with Staphylococcus aureus and try to correlate the source of contamination. Cultures were submitted to an antibiotic susceptibility test (AST) and characterised by Pulsed-field Gel Electrophoresis (PFGE) analysis. Results showed that 35.2% (19/51) of food handlers were asymptomatic carriers of S. aureus, and that 90.4% (19/21) of raw milk sampled was contaminated. Staphylococcus aureus was isolated from only 10 samples among more than 3200 investigated dairy products. No S. aureus contamination was found on machinery. The AST analysis demonstrated sensitivity of tested S. aureus to oxacillin, cephalothin, vancomycin, gentamicin, and sulfamethoxazole/trimethoprim. AST analysis generated eight different phenotypic profiles, but did not allow us to identify the source of contamination in seven of ten final products. PFGE analysis proved to be a sensitive method as it generated 42 different DNA banding profiles among the 48 S. aureus investigated, demonstrating a lack of predominance of endemic strains in the plant, contrary to suggestions raised by antibiotic resistance typing. Based on PFGE genotyping, S. aureus strains isolated from four contaminated final products were similar to four S. aureus isolated from raw milk. Five final products contained S. aureus different from all other strains collected, and one showed similarity to a strain isolated from a food handler. These results suggest contamination by raw milk as the main source of contamination of the final dairy products.     

Recycling of manure nutrients: use of algal biomass from dairy manure treatment as a slow release fertilizer

An alternative to land spreading of manure is to grow crops of algae on the N and P present in the manure and convert manure N and P into algal biomass. The objective of this study was to evaluate the fertilizer value of dried algal biomass that had been grown using anaerobically digested dairy manure. Results from a flask study using two soils amended with algal biomass showed that 3% of total algal nitrogen (N) was present as plant available N at day 0. Approximately 33% of algal N was converted to plant available N within 21 days at 25 degrees C in both soils. Levels of Mehlich-3 extractable phosphorus (P) in the two soils rose with increasing levels of algal amendment but were also influenced by existing soil P levels. Results from plant growth experiments showed that 20-day old cucumber and corn seedlings grown in algae-amended potting mix contained 15-20% of applied N, 46-60% of available N, and 38-60% of the applied P. Seedlings grown in algae-amended potting mixes were equivalent to those grown with comparable levels of fertilizer amended potting mixes with respect to seedling dry weight and nutrient content. These results suggest that dried algal biomass produced from treatment of anaerobically digested dairy manure can substitute for commercial fertilizers used for potting systems.     

Relation between substrate feeding pattern and development of filamentous bacteria in activated sludge processes: Part III. applications with industrial waste waters

Summary Laboratory scale activated sludge systems were operated under regimes of continuous or intermittent feeding of the waste water. Industrial waste waters from breweries, a dairy plant and a petro-chemical plant were investigated. The systems were started up with sludge from a municipal waste water plant or more often with sludges obtained from the corresponding industrial waste water treatment plants. It was found that intermittently fed systems produced sludges with better settleability characteristics than systems that were continuously supplemented with waste water. Our previous hypothesis that in intermittent systems floc forming bacteria become dominant as a result of higher substrate uptake rates was confirmed and may thus be extended to waste waters containing readily available substrates such as carbohydrates (brewery-and dairy waste water) or acids (petro-chemical waste water). Supplementation of brewery waste water with urea had a negative influence on sludge settleability, especially in continuously operated systems.     

Studies on the survival of enterohemorrhagic and environmental Escherichia coli strains in wastewater and in activated sludges from dairy sewage treatment plants

Survival of Escherichia coli O157:H7 strain isolated from milk in Poland and an environmental E. coli strain in wastewater from Garwolin and ?owicz dairies and in activated sludges from dairy sewage treatment plants as well as in dairy wastewater with activated sludges was examined. Environmental materials were contaminated with about 10(8) of target bacteria/ml of sample. The experiments were performed under temperature conditions typical of autumn-winter (6 degrees) and spring-summer (24 degrees C) seasons. It was found that the non-pathogenic E. coli strain survived longer in all media than the enterohemorrhagic serotype. E. coli O157:H7 bacteria were not detected (in direct plating method) in activated sludges after 21-28 days; in dairy wastewater as well as in wastewater with activated sludges after 21-25 days. These periods for environmental E. coli strain were 35-42 days (activated sludges), 25-28 days (wastewater with activated sludges). At higher temperature environmental E. coli were not detected in wastewater from ?owicz dairy sewage treatment plant after 25 days, but the bacteria were still present in wastewater from Garwolin dairy sewage tratment plant after 34 days. The obtained results show that the lack of environmental E. coli bacteria (as a indicator bacteria of fecal contamination) in dairy wastewater and in dairy wastewater with activated sludges could indicate the absence of pathogenic E. coli bacteria. Prolonged existence of the enterohemorrhagic serotype in activated sludges shows the need to treat activated sludges prior to the utilization of these materials as fertilizer.     

Characterization of micro-organisms isolated from dairy industry after cleaning and fogging disinfection with alkyl amine and peracetic acid

AIMS: To characterize micro-organisms isolated from Norwegian dairy production plants after cleaning and fogging disinfection with alkyl amine/peracetic acid and to indicate reasons for survival. METHODS AND RESULTS: Microbial samples were collected from five dairy plants after cleaning and fogging disinfection. Isolates from two of these production plants, which used fogging with alkylamino acetate (plant A), and peracetic acid (plant B), were chosen for further characterization. The sequence of the 16S ribosomal DNA, fatty acid analysis and biochemical characteristics were used to identify isolates. Three isolates identified as Rhodococcus erythropolis, Methylobacterium rhodesianum and Rhodotorula mucilaginosa were isolated from plant A and one Sphingomonas sp. and two M. extorquens from plant B. Different patterns of resistance to seven disinfectants in a bactericidal suspension test and variable degree of attachment to stainless steel were found. The strains with higher disinfectant resistance showed lower degree of attachment than susceptible strains. CONCLUSIONS: The study identifies and characterizes micro-organisms present after cleaning and fogging disinfection. Both surface attachment and resistance were shown as possible reasons for the presence of the isolates after cleaning and disinfection. SIGNIFICANCE AND IMPACT OF THE STUDY: These results contribute to the awareness of disinfectant resistance as well as attachment as mechanisms of survival in dairy industry. It also strengthens the argument of frequent alternation of disinfectants in the food processing industry to avoid the establishment of resistant house strains.     

Cup plant,an alternative to conventional silage from a LCA perspective

Purpose

The growing awareness of the importance of biodiversity in agroecosystems in increasing and ensuring the supply of biomass has led to heightened interest from governments and farmers in alternative crops. This article assesses one such alternative crop, cup plant (Silphium perfoliatum L.), in terms of the environmental aspects of cultivation for forage production. Many studies have previously focused on cup plant, but so far, this plant has not been assessed using the life cycle assessment (LCA) method.

Materials and methods

This study compares the environmental load of cup plant with the most commonly grown silage crops in Central European conditions—maize—and with another common forage crop—lucerne using LCA. The system boundaries include all the processes from cradle to farm gate and both mass-based (1 ton of dry matter) and area-based (1 ha of monoculture) functional units were chosen for the purposes of this study. The results cover the impact categories related to the agricultural LCAs, and the ReCiPe Midpoint (H) characterization model was used for the data expression, by using SimaPro 9.0.0.40 software.

Results

This study compares the cultivation of cup plant with the most commonly grown silage crop in Central European conditions—maize—and with another common forage crop—lucerne. The paper shows the potential of cup plant to replace conventional silage (maize and lucerne silage mix) with certain environmental savings in selected impact categories, and importantly, while still maintaining the same performance levels in dairy farming as with conventional silage, as already reported in previous publications. For the Czech Republic alone, this would, in practice, mean replacing up to 50,000 ha of silage maize and reducing the environmental load by about tens of percent or more within the various impact categories and years of cultivation.

Conclusion

Cup plant can replace the yield and quality of silage maize, represents a lower environmental load per unit of production and unit of area and generally carries many other benefits. Thus, cup plant is a recommendable option for dairy farming. Given the recent experience and knowledge of the issue, the cup plant can be considered an effective alternative to conventional silage.

     

Bioaccumulation and phyto-translocation of arsenic,chromium and zinc by Jatropha curcas L.: Impact of dairy sludge and biofertilizer

The present study was planned to remediate the metalloid and metal contaminated soil by using non-edible and economic plant species Jatropha curcas L. The experiment was conducted on pots to improve the survival rate, metal tolerance and growth response of the plant on soil; having different concentrations of arsenic, chromium and zinc. The soil was amended with dairy sludge and bacterial inoculum (Azotobacter chroococcum) as biofertilizer. The results of the study showed that the bioaccumulation potential was increased with increase in metalloid and metal concentration in soil system. Application of dairy sludge significantly reduces the DTPA-extractable As, Cr and Zn concentration in soil. The application of organic amendment stabilizes the As, Cr and Zn and reduced their uptake in plant tissues.     

Fermentation of plant-based dairy alternatives by lactic acid bacteria

Ethical, environmental and health concerns around dairy products are driving a fast-growing industry for plant-based dairy alternatives, but undesirable flavours and textures in available products are limiting their uptake into the mainstream. The molecular processes initiated during fermentation by lactic acid bacteria in dairy products is well understood, such as proteolysis of caseins into peptides and amino acids, and the utilisation of carbohydrates to form lactic acid and exopolysaccharides. These processes are fundamental to developing the flavour and texture of fermented dairy products like cheese and yoghurt, yet how these processes work in plant-based alternatives is poorly understood. With this knowledge, bespoke fermentative processes could be engineered for specific food qualities in plant-based foods. This review will provide an overview of recent research that reveals how fermentation occurs in plant-based milk, with a focus on how differences in plant proteins and carbohydrate structure affect how they undergo the fermentation process. The practical aspects of how this knowledge has been used to develop plant-based cheeses and yoghurts is also discussed.     

Genome-Scale Genotype-Phenotype Matching of Two Lactococcus lactis Isolates from Plants Identifies Mechanisms of Adaptation to the Plant Niche

Lactococcus lactis is a primary constituent of many starter cultures used for the manufacturing of fermented dairy products, but the species also occurs in various nondairy niches such as (fermented) plant material. Three genome sequences of L. lactis dairy strains (IL-1403, SK11, and MG1363) are publicly available. An extensive molecular and phenotypic diversity analysis was now performed on two L. lactis plant isolates. Diagnostic sequencing of their genomes resulted in over 2.5 Mb of sequence for each strain. A high synteny was found with the genome of L. lactis IL-1403, which was used as a template for contig mapping and locating deletions and insertions in the plant L. lactis genomes. Numerous genes were identified that do not have homologs in the published genome sequences of dairy L. lactis strains. Adaptation to growth on substrates derived from plant cell walls is evident from the presence of gene sets for the degradation of complex plant polymers such as xylan, arabinan, glucans, and fructans but also for the uptake and conversion of typical plant cell wall degradation products such as α-galactosides, β-glucosides, arabinose, xylose, galacturonate, glucuronate, and gluconate. Further niche-specific differences are found in genes for defense (nisin biosynthesis), stress response (nonribosomal peptide synthesis and various transporters), and exopolysaccharide biosynthesis, as well as the expected differences in various mobile elements such as prophages, plasmids, restriction-modification systems, and insertion sequence elements. Many of these genes were identified for the first time in Lactococcus lactis. In most cases good correspondence was found with the phenotypic characteristics of these two strains.     

Genetic diversity and spoilage potentials among Pseudomonas spp. isolated from fluid milk products and dairy processing plants

Degradation of milk components through various enzymatic activities associated with the contamination of dairy products by Pseudomonas spp. can reduce the shelf life of processed milk. Reliable methods for differentiating among Pseudomonas spp. strains are necessary to identify and eliminate specific sources of bacterial contamination from dairy processing systems. To that end, we assessed the genetic diversity and dairy product spoilage potentials among a total of 338 Pseudomonas spp. isolates from raw and pasteurized milk and from environmental samples collected from four dairy processing plants. The majority of isolates were identified as P. fluorescens and P. putida by API 20 NE. A total of 42 different ribotype patterns were identified among a subset of 81 isolates. The presence of many different ribotypes within this collection indicates high genetic diversity among the isolates and suggests multiple origins of contamination within the processing plant and in dairy products. The extracellular enzyme activity patterns among Pseudomonas isolates appeared to be associated with ribotypes. Isolates with the same ribotype frequently had the same extracellular protease, lecithinase, and lipase activities. For example, isolates grouped in ribotype 55-S-6 had the highest extracellular protease activity, while those in ribotypes 50-S-8 and 72-S-3 had the highest extracellular lipase activities. We conclude that ribotyping provides a reliable method for differentiating Pseudomonas strains with dairy food spoilage potential.     

The growth of Bacillus stearothermophilus on stainless steel

AIMS: To determine the potential for Bacillus stearothermophilus cells to form biofilms of significance in dairy manufacture. METHODS AND RESULTS: The ability of isolates of B. stearothermophilus from dairy manufacturing plants to attach to stainless steel surfaces was demonstrated by exposing stainless steel samples to suspensions of spores or vegetative cells and determining the numbers attaching using impedance microbiology. Spores attached more readily than vegetative cells. The attachment of cells to stainless steel was increased 10-100-fold by the presence of milk fouling the stainless steel. The growth of B. stearothermophilus as a biofilm on stainless steel surfaces was determined using a continuously flowing experimental reactor. Vegetative cells were released in greater numbers than spores from biofilms of most strains studied. Biofilms of one strain (B11) were studied in detail. Biofilms of > 106 cells cm-2 formed in the reactor and released approximately 106 cells ml-1 into milk passing over the biofilm. A doubling time of 25 min was calculated for this organism grown as a biofilm. CONCLUSION: The formation of biofilms of thermophilic Bacillus species within the plant appears to be a likely cause of contamination of manufactured dairy products. Methods to control the formation of biofilms in dairy manufacturing plants are required to reduce the contamination of dairy products with thermophilic bacilli. SIGNIFICANCE AND IMPACT OF THE STUDY: Biofilms of B. stearothermophilus growing in dairy manufacturing plants can explain the contamination of dairy products with these bacteria.     

Diversity among Lactobacillus paracasei phages isolated from a probiotic dairy product plant

Aims:  To evaluate the phage diversity in the environment of a dairy industry which manufactures a product fermented with a probiotic strain of Lactobacillus paracasei .
Methods and Results:  Twenty-two Lact. paracasei phages were isolated from an industrial plant that manufactures a probiotic dairy product. Among them, six phages were selected based on restriction profiles, and two phages because of their notable thermal resistance during sample processing. Their morphology, host range, calcium dependency and thermal resistance were investigated. All phages belonged to the Siphoviridae family (B1 morphotype), were specific for Lact. casei and paracasei strains showing identical host spectrum, and only one phage was independent of calcium for completing its lytic cycle. Some of the phages showed an extraordinary thermal resistance and were protected by a commercial medium and milk.
Conclusions:  Phage diversity in a probiotic product manufacture was generated to a similar or greater extent than during traditional yogurt or cheese making.
Significance and Impact of the Study:  This work emphasizes probiotic phage infections as a new ecological situation beyond yogurt or cheese manufactures, where the balanced coexistence between phages and strains should be directed toward a favourable state, thus achieving a successful fermentation.     

Phosphate solubilization and growth promotion by <Emphasis Type="Italic">Pseudomonas fragi</Emphasis> CS11RH1 (MTCC 8984), a psychrotolerant bacterium isolated from a high altitude Himalayan rhizosphere

Phosphate solubilization and growth promotion by Pseudomonas fragi CS11RH1 (MTCC 8984), a psychrotolerant bacterium isolated from a high altitude garlic rhizosphere from the Indian Himalayas, are reported here. The identity of the isolate was arrived on the basis of its biochemical features and sequencing of the 16S rRNA gene. The isolate grew and solubilized phosphate at temperatures ranging from 4 to 30°C. Besides solubilizing P it produced indole acetic acid (IAA) and hydrogen cyanide (HCN). Seed bacterization with the isolate significantly increased the percent germination, rate of germination, plant biomass and nutrient uptake of wheat seedlings. While Pseudomonas fragi is normally associated with the spoilage of dairy products stored at cold temperatures, this is an early report on the plant growth promoting ability of the bacterium.     

Growth of Jatropha curcas on heavy metal contaminated soil amended with industrial wastes and Azotobacter. A greenhouse study

The aims of the study were to evaluate the effect of organic wastes (biosludge and dairy sludge) and biofertilizer (Azotobacter chroococcum) on the planting conditions of Jatropha curcas in metal contaminated soils. Results showed that the plants survival rate in heavy metal contaminated soil increased with addition of amendments. Treatment T6 (heavy metal contaminated soils+dairy sludge+biofertilizer) observed to be the best treatment for growth (height and biomass) as compared with the treatment T5 (heavy metal contaminated soils+biosludge+biofertilizer). In addition, organic amendments provided nutrients such as carbon, N, P and K to support plant growth and reduced the metal toxicity to plant. The present study showed that metal contaminated lands/soils could be suitably remediated by adapting appropriate measures.     

The effects of pruning,fertilizers, and organic amendments on lowbush blueberry production

A four-year field study was conducted on a Hebert gravelly sandy loam (pH 4.5) in Nova Scotia to assess the effects of pruning management and seven fertility amendments on lowbush blueberry (Vaccinium augustifolium Ait) production (yield, above ground and root tissue composition) and soil fertility. Pruning by oil burning produced higher fruit yields than flail mowing but burning had the opposite effect on the plant N content (with a lesser influence on above ground Mn and Zn). None of the fertility treatments (chicken manure, dairy manure, swine manure, urea, sawdust, NPK, NPK+S+Lime+Micronutrients) produced fruit yields significantly greater than the control. Treatments provided the equivalent of 50kg total N/ha/2-yr cycle. Treatments influenced tissue N, P, K, Ca, Mg, B, Mn, Cu, Zn and Mo levels. In general, the three manure treatments produced the highest levels of plant macronutrients; the urea treatment produced the lowest levels of plant nutrients. In most cases, extractable levels of soil P, K, Ca and Mg were highly correlated with the plant tissue content of these elements. Overall, the dairy manure treated soils were the highest in soil fertility.