Source: SOUTH DAKOTA STATE UNIVERSITY submitted to
SAFETY AND QUALITY IMPROVEMENT OF MILK AND CHEESE.
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
TERMINATED
Funding Source
HATCH
Reporting Frequency
Annual
Accession No.
0230889
Grant No.
(N/A)
Project No.
SD00H424-12
Proposal No.
(N/A)
Multistate No.
(N/A)
Program Code
(N/A)
Project Start Date
Oct 1, 2012
Project End Date
Sep 30, 2017
Grant Year
(N/A)
Project Director
Anand, SA.
Recipient Organization
SOUTH DAKOTA STATE UNIVERSITY
PO BOX 2275A
BROOKINGS,SD 57007
Performing Department
Dairy Science
Non Technical Summary
Safety and quality enhancement have always been key words in the management of dairy production and manufacturing systems. The importance has further increased due to ever expanding supply chains and the complexity of operations. The recent signing of Food Safety Modernization Act 2011 (FSMA) puts further emphasis on the safety of the U.S. food supply by shifting the focus of federal regulators from response to prevention of contamination (FDA, 2011). Issues related to the cross contamination of milk at the production level with thermoduric spore formers still remain unresolved. These thermoduric spore-forming bacteria could not only limit the shelf life of milk but can also influence the quality of ripening of cheese. Some of these organisms such as Bacillus cereus are potential pathogens and thus can pose health risks. In addition, the risk posed by the incidence of potential pathogens in raw milk is not clearly understood. Our hypothesis is that it is possible to reduce the cross contamination of milk with bacteria including thermoduric spore formers. This can be accomplished by improving HACCP based management practices, better cleaning strategies especially to prevent bacterial biofilms on contact surfaces, and by combining some non-thermal treatment of milk such as ultrasonication to be able to enhance the effectiveness of pasteurization. It would also be of interest to study the competitive inhibition of thermoduric thermophiles during cheese ripening by the nonstarter lactic acid bacteria.
Animal Health Component
(N/A)
Research Effort Categories
Basic
(N/A)
Applied
(N/A)
Developmental
(N/A)
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
3083450110020%
3084010104010%
5013450110025%
5023430110025%
5024010110010%
5024010104010%
Knowledge Area
501 - New and Improved Food Processing Technologies; 308 - Improved Animal Products (Before Harvest); 502 - New and Improved Food Products;

Subject Of Investigation
3430 - Cheese; 4010 - Bacteria; 3450 - Milk;

Field Of Science
1040 - Molecular biology; 1100 - Bacteriology;
Goals / Objectives
Objective 1: To study the role of thermoduric bacteria in influencing shelf life of milk products such as cheese. Objective 2: Reducing the cross contamination in dairy processing environments by controlling bacterial biofilms.
Project Methods
Study 1: To study the role of thermoduric bacteria in influencing shelf life of milk products such as cheese. Studies will be conducted on incidence, growth and emergence of predominance in thermoduric bacterial counts during the Cheddar cheese making process. Bacterial community specific studies with regard to the spoilage microflora can help us determine and control undesirable biochemical changes occurring in cheese during ripening. During the initial phase of this study, thermoduric and thermophilic sporeformers will be screened at milk reception, holding, and processing steps in a cheese plant. We also plan to take cheese samples with defects and isolate the related spoilage bacteria from these samples. Representative bacterial isolates will be used to manufacture cheese at the lab scale to study the events under controlled conditions. We would also study their interactions with different types of starters that could help control these spoilage bacteria and prevent cheese defects. Studies will also be conducted to evaluate the biochemical activities of these bacteria leading to undesirable changes. Total Nitrogen, Water-soluble Nitrogen, and 12% TCA-soluble Nitrogen will be measured to track the proteolytic changes, while Total free AA levels in pH 4.6-soluble fraction of the cheeses will be determined by the Cd-ninhydrin method. Urea-PAGE of pH 4.6-insoluble fractions of the cheeses during ripening will be conducted to obtain degradation patterns. Free fatty acid analysis will be helpful in monitoring starter esterases, and lipoprotein lipase activities. Study 2: Reducing the cross contamination in dairy processing environments by controlling bacterial biofilms. Our previous studies have revealed the presence of multispecies biofilms on reverse osmosis membranes, many of which were found to be resistant to the current CIP protocols being followed by the industry. Frequent presence of biofilms in dairy processing areas reflects the inadequacy of the currently used CIP protocols. One of the important cleaning steps is the enzyme action that helps to break down the biofilm matrices for better cleaning. We hypothesize that it is possible to identify a cocktail of enzymes that would help clean the biofilms in a more effective manner. We plan to develop in vitro biofilms using resistant bacterial isolates that we have obtained from the membrane biofilm consortia of 2 to 14 months old membrane cartridges from active industrial systems. Such resistant biofilms will be tested against the existing and modified CIP protocols under both static and dynamic conditions in order to identify a better cleaning process. We propose to test enzyme modifications with surfactants and test them against resistant biofilms. Tests will be conducted against the lab grown biofilms that will be developed from the biofilm consortia isolates under both static and dynamic conditions using a biofilm reactor. The cross flow studies will help evaluate the efficacy of the modified process in a simulated environment.

Progress 10/01/12 to 09/30/17

Outputs
Target Audience:Food and dairy industry. Dairy producers. Researchers. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?List of M.S. and Ph.D. students, and Post Docs who participated in these projects, leading to completion of their respective degrees. 1. Dalia Khan, M.S. Dairy Manufacturing. 2017. Studying microflora of semi-hard cheese, and sporulation within contact surface biofilms. 2. Taghreed Almalki, M.S. Dairy Manufacturing. 2017. Effect of ultrasonication on biofilm forming ability of common dairy sporeformers. 3. Nuria Garcia-Fernandez, Post Doc. Dairy Manufacturing. 2017. Evaluating enzyme formulations for biofilm removal from dairy separation membranes. 4. Shivali Jindal, M.S. Dairy Manufacturing. 2017. Biofilm formation by common dairy sporeformers on native and modified stainless steel surfaces. 5. Dikshi Bawa, M.S. Dairy Manufacturing. 2016. Inactivation of thermoduric sporeformers and spores in milk by continuous ultrasonication and hydrodynamic cavitation in combination with thermal treatments. 6. Nuria Garcia-Fernandez, Ph.D. Dairy Manufacturing. 2016. Role of exopolymeric substances in biofilm formation on dairy separation membranes. 7. Somil Gupta, M.S. Dairy Manufacturing. 2015. Studies related to microbially induced corrosion of stainless steel 304 and 316. 8. Khilendra Bhanduriya, M.S. Dairy Manufacturing. 2015. Microbial aspects related to thermoduric bacteria during cheese manufacturing and ripening. 9. Sowmya Marka, M.S. Dairy Manufacturing. 2014. Modifications of a typical CIP protocol to control biofilms on dairy processing membranes. 10. Kimberly Buehner, M.S. Dairy Manufacturing. 2014. The incidence of spores and thermoduric bacteria in Midwest dairies and commercial NFDM samples and their reduction via ultrasonication. 11. Akimoto Ichinomiya, M.S.Dairy Manufacturing. 2014. Development and optimization of food-grade antimicrobial lactic acid bacteria isolated from raw milk. (Co-advised) 12. Joan Hegerfeld-Baker, Ph.D. Nutrition, Excercises and food Science. 2013. Influence of STEM education and technology in selecting food and agriculture careers. (Co-advised) 13. Som N. Khanal, M.S. Dairy Manufacturing. 2012. A study on effectiveness of ultrasonication in inactivating thermoduric aerobic sporeformers and spores in nonfat milk. 14. Diwakar Singh, M.S. 2012.Dairy Manufacturing. Evaluation of a typical CIP protocol to control bacterial biofilms on whey reverse osmosis membranes. 15. Robert John Jjuuko, M.S. Nutrition, Excercises and food Science. 2012. Antibiotic residues in feed (DDGS) and Food. Health and Nutritional Sciences. (Co-Advised). List of current M.S. and Ph.D. students, who completed a part of the above projects. 1. Steve Beckman, Ph.D.Dairy Manufacturing. (2014-18). 2. Nancy Awasthi, Ph.D.Dairy Manufacturing. (2016-18). 3. Neha, Ph.D.Dairy Manufacturing. (2016-18). 4. Shayanti Minj, Ph.D.Dairy Manufacturing. (2017-20). 5. Pratibha Chaudhary, M.S.Dairy Manufacturing. (2017-18). List of M.S. and Ph.D. students, who were awarded for the research accomplishments. 1. Neha, N, Dairy Recognition and Educational Foundation Scholarship, 2017. 2. Neha, N, ADSA 2017, 3 min thesis presentation award. 3. Neha, N, Sanford Research 2017, Graduate completion award. 4. Awasti, N, Dairy Recognition and Educational Foundation Scholarship, 2017. 5. Jindal, S, Distinguished Master's Thesis Award, 2017, South Dakota State University. List of undergrdauate researchers, who were trained under this project during 2012-2017. 1. April A Johnson, 2017, (Animal, Poultry, and Veterinary Science, Tuskegee University), 'Genomic forensics for improving food safety.' 2. Connor Bushman, 2017, 'An examination of environmental Listeria's antibiotic resistance and biofilm persistence.' 3. Shelby Riddle, 2017, 'To compare the population change in sporeformers and spores during storage of raw and pasteurized milk'. 4. Logan Johnson, 2016, 'Microbial changes during various stages of ice cream manufacture and storage.' 5. Jae Young Sim, 2016, 'Effect of cavitation on microbial quality of milk.' 6. Lauren A. Gonzalez, 2015, 'Influence of environment on Bacillus licheniformis Spore Formation.' 7. Jae Young Sim, 2015, 'Effect of feeding Lactic acid bacteria as 'Direct Fed Microbials' on the microbial quality of raw milk.' 8. Sara Sontag, 2014-2015, 'Effect of feeding Lactic acid bacteria as 'Direct Fed Microbials' on the microbial quality of raw milk.' 9. Melissa Young, 2014, 'Inactivation of thermoduric spore formers in milk concentrates by ultrasonication.' 10. Weina Liu, 2013, 'Interaction of starter cultures and gas producing thermoduric bacteria to control the blowing or slit defects of Cheddar cheese.' 11. Brittany Sipp, 2013, 'Inactivation of thermoduric spore formers and spores in milk concentrates by a combination of ultrasonication and pasteurization.' 12. Giovana Charite, 2013, 'Incidence and characterization of thermoduric spore formers and spores in nonfat dry milk.' 13. Allison Sewell, 2012, 'Screening of thermoduric bacterial cheese isolates for proteolysis, lipolysis, and gas production activities.' 14. Kaylee King, 2012, 'Comparing biofilms formed under different processing substrates.' 15. Megan Huegli, 2012-13, 'Role of thermoduric bacteria in split defects found in Swiss cheese.' 16. Eric Flynn, 2012, 'Spore and sporeforming bacterial counts in raw milk'. 17. Megan Huegli, 2012, 'Survival of sporeformers in whole milk by thermal treatment.' How have the results been disseminated to communities of interest?Peer-reviewed publications: 18 Students trained (postdoc, graduate students, undergraduate students): 37 Presentations (poster or oral): 34 Commercial collaborations: 3 What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? Objective 1: To study the role of thermoduric bacteria in influencing shelf life of milk products such as cheese. (100% accomplished) Our work successfully demonstrated the application of hydrodynamic cavitation as a potential non-thermal method to inactivate thermoduric sporeformers in milk. This technology could be used for applications such as manufacturing high quality dairy products, such as cheese. The process of hydrodynamic cavitation (60Hz frequency at 200L/h flow rate) was combined with HTST pasteurization to inactivate the thermally resistant sporeformers such as Bacillus coagulans, Geobacillus stearothermophilus, Bacillus stearothermophilus, and Bacillus licheniformis. A six-pass hydrodynamic cavitation process effectively reduced most of thermoduric vegetative cells by three logs, and reduced the viable endospores by one log. The outcome of this study showed promising results for application of hydrodynamic cavitation in reduction of spoilage bacteria in milk. The findings also led to acquiring an additional National Dairy Council grant to scale up the process at a pilot plant level. Those studies are now under progress. In another project we used response surface models to develop contour plots for predicting the population kinetics of sporeformers and spores in raw milk. In this project, we studied the changes in population of vegetative and spores of B. licheniformis during low temperatures storage in spiked raw milk samples. Regression models were developed to identify optimum time-temperature combinations where no significant change or a minimum shift in population was observed. Rate of population change of sporeformers was observed to be significantly higher at 10 and 12 °C from approx. 4.0 log cfu/mL to 4.99 ± 0.10 and 7.05 ± 0.27 cfu/mL, respectively. On the other hand, dissolved oxygen decreased from 20.4 to 17.7 %, with the increase in holding duration due to increased microbial activity. The pH changes were due to different growth behavior of the organism at different temperatures. The model suggested that raw milk can be held at 4 and 6° C up to 72 h and 8° C for 55 h without any significant increase in microbial populations. Objective 2: Reducing cross contamination in dairy processing environments by controlling bacterial biofilms. (100% accomplished) Our work related to cleaning of biofilms produced some very useful data that resulted in establishing a partnership with Ecolab to scale-up a new enzyme-based cleaning technology. This helped us demonstrate the pilot scale application of an enzyme based cleaner. The National Dairy Council funded this collaborative project. We studied various enzyme based cleaning protocols to successfully remove lab-grown membrane biofilms of single and mixed species of sporeformers. A combination of three enzymes brought out maximum reduction in membrane biofilms. The membrane material UF PVDF supported the most resilient biofilm formation. Enzyme formulation (EC1) was not effective on RO biofilms, while formulation (EC2) was more effective against UF PES biofilms compared to the UF PVDF. The formulated enzymes performed better than EC1 and EC2 for cleaning RO PA and UF PES membranes, respectively, although were not effective against UF-PVDF biofilms. The enzyme combination formulation (T5) was the most effective in cleaning biofilms. The average reductions were 1.0-2.0 logs for various treatments. This was also validated in a pilot scale trial involving two pre-fouled UF-PES membranes where the experimental CIP using T5 produced a significantly greater flux (155%), compared to standard CIP (73%) using EC2. Further studies are in progress for next steps towards the potential commercialization of the formulation. These enzymes have the potential to help reduce chemical concentrations and extend the life of the dairy filtration membranes, while improving product quality and profitability. We completed work on biofilm prevention by using modified plate heat exchangers in collaboration with Kansas State University and the University of Massachusetts. In this collaboration, the surface modified coupons were provided by the collaborating partners, while we were responsible to develop resilient biofilms on these coupons and then demonstrate the effectiveness of the cleaning protocols, both under lab and pilot scale operations. Ni-PTFE coating was found to be effective in reducing the biofilm formation (to 1.0 log cfu/cm2 or lower) both under lab conditions and pilot scale operations for a 17h pasteurizer run. We also investigated the effect of cell surface properties on the attachment behavior of different aerobic spore forming bacteria and their spores that are commonly encountered in dairy industry. The results indicated higher adhesion tendency, about a log higher for spores compared to vegetative cells of aerobic sporeforming bacteria. As the vegetative cells and spores of B. sporothermodurans and G. stearothermophilus demonstrated significantly greater attachment as compared to B. licheniformis, thus it can be interpreted that HHRS showed a greater attachment tendency as compared to thermo-tolerant sporeformers. Additional grant funds for 2016-17 were secured from National Dairy Council. Research in the area of control of Listeria during the manufacture of ice cream progressed further in collaboration with a major US ice cream manufacturer. The work under this grant is in progress. The early findings helped us to evaluate the importance of environmental cross contamination as a predictor of injured cell recovery in the ice cream mix. Only high levels of cross contamination (log 4.0 or more) offered any injured cells recovery, and some early attempts were made to develop Listeria risk prediction models based on response surface modeling. The current research in this area is focused at the recovery potential of injured cells. Overall Impact: This met 100% of the overall project goals. A clear understanding of the behavior of sporeformers and endospore, which cause major quality problems in milk processing and product manufacture, was established. The regression models developed will help the dairy industry plan processes by predicting population changes in the vegetative cells and endospores of sporeformers such as Bacillus coagulans, Geobacillus stearothermophilus, Bacillus stearothermophilus, and Bacillus licheniformis. We demonstrated the effectiveness of novel processes such as Hydrodynamic Cavitation in combination with pasteurization, as effective means to reduce counts of thermoduric sporeformers. The quality of dairy products is also adversely affected by the formation of bacterial biofilms, which are difficult to clean and cause cross contamination of processed products. We successfully studied the biofilm formation on both membranes and stainless steel, and generated evidence on the effectiveness of contact surface modifications for stainless steel surfaces, and enzyme cleaners for membranes to effectively clean biofilms. All these approaches provide a comprehensive approach to improving the quality of milk and cheese. In the area of dairy product safety, we have made significant progress in understanding the risk from Listeria, an important foodborne pathogen of recent concern. The cross contamination levels of the organism were observed to be predictor of injured cell recovery after the pasteurization. This will be helpful in Listeria control and therefore enhance food safety.

Publications

  • Type: Journal Articles Status: Published Year Published: 2017 Citation: Liu, D. Z., Jindal, S., Amamcharla, J., Anand, S. K., Metzger, L. (2017). Evaluation of a sol-gel based stainless steel surface modification to reduce fouling and biofilm formation during pasteurization of milk. Journal of Dairy Science, 100(4), 2577-2581.
  • Type: Journal Articles Status: Awaiting Publication Year Published: 2017 Citation: Marka, S., Anand, S. K. (2017). Feed substrates influence biofilm formation on reverse osmosis (RO) membranes and their cleaning efficiency. Journal of Dairy Science (accepted, under publication).
  • Type: Journal Articles Status: Published Year Published: 2017 Citation: Gupta, S., Anand, S. K. (2017). Induction of pitting corrosion on stainless steel (grades 304 and 316) used in dairy industry by biofilms of common sporeformers. International Journal of Dairy Technology. http://dx.doi.org/10.1111/1471-0307.12444
  • Type: Other Status: Published Year Published: 2017 Citation: Adamski, A., Bawa, D., Fernandez, N. G., Anand, S. K. (2017). Reported incidences of dairy pathogens. A review. MDFRC (Midwest Dairy Foods Research Center).
  • Type: Conference Papers and Presentations Status: Published Year Published: 2017 Citation: Jindal, S., Anand, S. K., ADSA Annual Conference, "Comparison of the adhesion characteristics of common dairy spore formers and their spores," Pittsburg, PA, United States. (Jun 25-28, 2017).
  • Type: Conference Papers and Presentations Status: Published Year Published: 2017 Citation: Khan, D., Anand, S. K., Gamma Sigma Delta, Honors Society of Agriculture Graduate Student Poster Contest, "Effect of rate of cooling and ripening temperatures on non-starter lactic acid bacteria in cheese.," SDSU, Brookings, SD, United States. Apr 03, 2017.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2017 Citation: Fernandez, N. G., Anand, S. K., ADSA Annual Conference, "Evaluating enzyme formulations for biofilm removal from dairy separation membranes," Pittsburg, PA, United States. (Jun 25-28, 2017).
  • Type: Conference Papers and Presentations Status: Published Year Published: 2017 Citation: Almalki, T., Anand, S. K., Gamma Sigma Delta, Honors Society of Agriculture Graduate Student Poster Contest, "Evaluation of the effect of cavitation on biofilm forming ability of sporeformers.," SDSU, Brookings, SD, United States. Apr 03, 2017.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2017 Citation: Fernandez, N. G., Anand, S. K., Hassan, A., ADSA Annual Conference, "Exopolysaccharides produced by dairy starter cultures impact biofilm formation on separation membranes," Pittsburg, PA, United States. (Jun 25-28, 2017).
  • Type: Conference Papers and Presentations Status: Published Year Published: 2017 Citation: Neha, N., Suliman, R., Anand, S. K., Djira, G., Kraus, B., Sutariya, S., ADSA Annual Conference, "Level of Listeria cross contamination in ice cream mix can serve as a predictor of its overall risk from injured cells. Graduate Poster Competition," Pittsburg, PA, United States. (Jun 25-28, 2017).
  • Type: Conference Papers and Presentations Status: Published Year Published: 2017 Citation: Neha, N., Anand, S. K., Djira, G., Sanford Annual Conference - It is all about science, "Listeria cross contamination levels and the associated risk in ice cream mix.," Sanford Research, Sioux Falls, SD, United States. (Jun 10, 2017).
  • Type: Conference Papers and Presentations Status: Published Year Published: 2017 Citation: Awasti, N., Suliman, R., Anand, S. K., Djira, G., ADSA Annual Conference, "Population dynamics of a common dairy sporeformer, Bacillus licheniformis, in spiked raw milk samples stored at low temperatures.," Pittsburg, PA, United States. (Jun 25-28, 2017).
  • Type: Conference Papers and Presentations Status: Published Year Published: 2017 Citation: Anand, S. K., NCCIA Annual Meeting, "Membrane biofilms formation and control," NCCIA, Rochester, MN, United States. (October 11, 2017).
  • Type: Conference Papers and Presentations Status: Published Year Published: 2017 Citation: Anand, S. K., MDFRC Annual Meeting, "Population dynamics of common dairy sporeformers," MDFRC, St. Paul, MN, United States. (July 31, 2017).
  • Type: Conference Papers and Presentations Status: Published Year Published: 2017 Citation: Anand, S. K., MDFRC Research Planning Meeting, "Resilient biofilms and their removal from dairy separation membranes," MDFRC, Ames, IA, United States. (May 9, 2017).
  • Type: Theses/Dissertations Status: Accepted Year Published: 2017 Citation: Dalia Khan, Master's Thesis Committee Chair, "Studying microflora of semi-hard cheese, and sporulation within contact surface biofilms," Completed (September 2015 - July 2017)
  • Type: Theses/Dissertations Status: Accepted Year Published: 2017 Citation: Taghreed Almalki, Master's Thesis Committee Chair, "Effect of ultrasonication on biofilm forming ability of common dairy sporeformers," Completed (January 2015 - May 2017)
  • Type: Theses/Dissertations Status: Accepted Year Published: 2017 Citation: Shivali Jindal, Master's Thesis Committee Chair, "Biofilm formation by common dairy sporeformers on native and modified stainless steel surfaces," Completed (January 2015 - January 2017)
  • Type: Other Status: Other Year Published: 2017 Citation: Nuria Garcia-Fernandez, Postdoctoral Research Supervision Report, "Evaluating enzyme formulations for biofilm removal from dairy separation membranes," Completed (2017)


Progress 10/01/15 to 09/30/16

Outputs
Target Audience:Food and dairy industry. Dairy producers. Researchers. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?The project has provided several opportunities at both graduate and undergraduate levels of teaching and research. One PhD and one MS students completed their degree programs while working on these projects in 2016. One Post Doc, three PhD and three MS students are currently working under my advising on these research projects. In addition, two undergraduate research projects were completed under this research program during 2016. Nuria Garcia-Fernandez, Ph.D. 2016. Role of exopolymeric substances in biofilm formation on dairy separation membranes. Completed. Dikshi Bawa, M.S. 2016. Inactivation of thermoduric sporeformers and spores in milk by continuous ultrasonication and hydrodynamic cavitation in combination with thermal treatments. Completed. Nuria Garcia Fernandez, Post Doc, 2016-17. Evaluating enzyme formulations for biofilm removal from dairy separation membranes. NDC funded project in collaboration with Eco Lab, Rochester, MN. Steve Beckman, Ph.D., 2013-18, Dairy Science, manufacturing. Application of ultrasonication and steam injection in controlling spore formers in milk concentrates. In progress. Neha Nain, Ph.D., 2015-18, Dairy Science, manufacturing. Listeria control during ice cream manufacture. In progress. Nancy Awasti, Ph.D., 2015-18, Dairy Science, manufacturing. Spore Germination and sporulation process. In progress. Taghreed Almalki, M.S., 2015-17, Dairy Science, manufacturing. Influence of cavitation on biofilm formation ability of injured sporeformers and spores. In progress. Shivali Jindal, M.S., 2015-17, Dairy Science, manufacturing. Biofilm development and control on modified SS surfaces. In progress. Dalia Khan, M.S., 2015-17, Dairy Science, manufacturing. Spore changes during cheese ripening and within biofilm matrices. In progress. Logan Johnson, 2016, Undergraduate research project completed. Microbial changes during various stages of ice cream manufacture and storage. Jae Young Sim, 2016, Undergraduate research project completed. Effect of cavitation on microbial quality of milk. How have the results been disseminated to communities of interest?Research community: Four peer reviewed publications, One invited Review, Eight scientific posters presentations in symposium of American Dairy Science Association, One PhD dissertation and one MS thesis completed. Stake holders: Midwest Dairy Foods Research Center Annual meeting, direct interaction with dairy producers, and dairy industry. What do you plan to do during the next reporting period to accomplish the goals?We plan to initiate work on the process scale-up for cavitation as a non-thermal application for inactivating the sporeformers and spores in milk. The studies on understanding the germination and sporulation mechanisms of thermoduric spores in the dairy environment to achieve the overall goals will be continued. The project work on the control of Listeria in ice cream, based on Response Surface Methodology, will be continued. In addition, under the USDA-HEC grant at the Honors College, a Food Safety colloquium will be offered, and further opportunities will be created for the undergraduate research in the area of food safety.

Impacts
What was accomplished under these goals? Objective 1 was supported by two National Dairy Council (NDC), and one Midwest Dairy Foods Research Center (MDFRC) grants. Under the first study, further trials were completed to demonstrate the effectiveness of cavitation as a means to inactivate thermoduric sporeformers in skim milk and concentrates. The findings led to submission of a new proposal to NDC for scale-up of cavitation for reducing the sporeformers. The second study was a continuation of our work on understanding the process of sporulation during various stages of milk powder manufacture. Studies were completed on the sporulation behavior of the most common dairy sporeformer Bacillus licheniformis during holding of raw milk. Under the third study, a fluorescent spore detection method will be evaluated. Objective 2 was supported by three National Dairy Council (NDC), and two Midwest Dairy Foods Research Center (MDFRC) grants. In the first study, experiments were completed to evaluate biofilm formation on modified plate heat exchanger (PHE) plates. Several types of coated coupons and PHE plates were tested for biofilm formation at both lab and pilot scale. The second study under this objective provided information on the biofilm forming ability of high heat resistant thermoduric sporeformers on modified surfaces. Under the third project, studies were initiated to develop an enzyme based cleaning solution for more effective removal of membrane biofilms. Under the fourth study, experiments were conducted to validate the inactivation of spiked Listeria innocua (a surrogate) to the ice cream mix pasteurization process, as influenced by total solid content. A Food Safety Colloquium is also being planned for offering during fall 2017 semester for Honors College under our USDA-HEC grant, Meeting the Grand Challenge', in order to promote recruitment in food safety related careers at the University. This will be in collaboration with two other food safety Professors and several invited specialists.

Publications

  • Type: Journal Articles Status: Published Year Published: 2016 Citation: Hegerfeld-Baker, J., L. Droke, P., Pallapu and S. Anand. 2016. Factors influencing choice of food safety related career path; an online focus group study. North American Colleges and Teachers of Agriculture Journal (NACTA) 60 (1): 1-8.
  • Type: Journal Articles Status: Published Year Published: 2016 Citation: Jindal S, Anand S, Huang K, Goddard J, Metzger L, Amamcharla J. 2016. Evaluation of modified stainless steel surfaces targeted to reduce biofilm formation by common milk sporeformers. Journal of dairy Science 99 (12): 9502-9513.
  • Type: Journal Articles Status: Accepted Year Published: 2016 Citation: Dylan Zhe Liu, Shivali Jindal, Jayendra Amamcharla, Sanjeev Anand, Lloyd Metzger. 2016. Evaluation of a sol-gel based stainless steel surface modification to reduce fouling and biofilm formation during pasteurization of milk. Journal of Dairy Science (accepted).
  • Type: Journal Articles Status: Under Review Year Published: 2016 Citation: Gupta S, Anand S. 2016. Induction of pitting corrosion on Stainless Steel (Grades 304 and 316) used in dairy industry by biofilms of common sporeformers. International J. Dairy Technology (submitted).
  • Type: Other Status: Published Year Published: 2016 Citation: Admsky Ashley, D Bawa, N Garcia-Fernandez, S Anand. 2016. Reported incidences of dairy pathogens. A review prepared for Midwest Dairy Foods Research Center.
  • Type: Theses/Dissertations Status: Published Year Published: 2016 Citation: Nuria Garcia-Fernandez, Ph.D. 2016. Role of exopolymeric substances in biofilm formation on dairy separation membranes. Completed.
  • Type: Theses/Dissertations Status: Published Year Published: 2016 Citation: Dikshi Bawa, M.S. 2016. Inactivation of thermoduric sporeformers and spores in milk by continuous ultrasonication and hydrodynamic cavitation in combination with thermal treatments. Completed
  • Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: Bawa D, S Anand, H Dhaiya, L Metzger. 2016. Study the effect of hydrodynamic cavitation on inactivation of thermoduric bacilli in skim milk. IFT Annual Meeting, Chicago IL July 16-19, 2016 Poster #021
  • Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: Jindal S, S Anand, Amamcharla J, and Metzger L. 2016. Comparison of Biofilm formation on Stainless Steel and Modified surface milk plate heat exchangers. ADSA 2016, Dairy Foods Division: Dairy Microbiology. Abstract # 35
  • Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: Jindal S, S Anand, Amamcharla J, and Metzger L. 2016. Evaluation of modified stainless steel surfaces targeted to reduce biofilm formation by common dairy related sporeformers. ADSA 2016, Dairy Foods Graduate Student Oral Competition. Abstract#702.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: Almalki T and S Anand. 2016. Evaluation of the effect of cavitation on biofilm forming ability of sporeformers. ADSA 2016, Dairy Foods Division: Dairy Microbiology. Abstract # 25
  • Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: Khan D and S Anand. 2016. Effect of rate of cooling and ripening temperatures on non-starter lactic acid bacteria in cheese. ADSA 2016, Dairy Foods Division: Dairy Microbiology. Abstract # 32
  • Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: Garcia-Fernandez N, A Hassan and S Anand. 2016. Evaluation of microbial enzymes for degradation of exopolymeric substances (EPS) within biofilm matrices for more effective cleaning. ADSA 2016, Dairy Foods Division: Dairy Microbiology. Abstract # 34
  • Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: Garcia-Fernandez N, A Hassan and S Anand. 2016. Slime production by Bacillus strains affects biofilm formation on dairy separation membranes. Gamma Sigma Delta Poster Competition, SDFSU April 04.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: Bawa D, and S Anand. 2016. Inactivation of thermoduric bacterial endospores in milk by combined effect of cavitation and thermal treatment. Gamma Sigma Delta Poster Competition, SDFSU April 04.


Progress 10/01/14 to 09/30/15

Outputs
Target Audience:Studies were directed towards the application of cavitation to inactivate thermoduric sporeformers, for producing nonfat dried milk with lower counts. In addition, coated plates in milk pasteurizer were evaluated as a means of reducing biofilm formation during milk processing. The results were shared with the US dairy industry through US Dairy Export Council, National Dairy Council, and the Midwest Dairy Foods Research Center meetings. The experiments involved active participation of dairy plants for immediate transfer of knowledge. A workshop was also conducted at the premises of a leading long life milk processor to share the information regarding control of sporeformers. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?The project has provided several opportunities at both graduate and undergraduate levels of teaching and research. Two MS students completed their degree programs while working on these projects in 2015. Four MS and three PhD students are currently working under my advising on research projects related to thermoduric sporeformers, and biofilm control. In addition, three undergraduate research projects were completed under this research program during 2015. How have the results been disseminated to communities of interest?Peer reviewed publications, invited lectures in conferences and national symposia, scientific posters presentations in symposium of American Dairy Science Association and Midwest Dairy Foods Research Center Annual meeting, direct interaction with dairy producers, and direct interaction with dairy industry. What do you plan to do during the next reporting period to accomplish the goals?We plan to continue work on the process of cavitation as a non-thermal application for inactivating the sporeformers and spores in milk. The studies on the control of biofilms by modifying the PHE contact surfaces, and understanding the germination and sporulation mechanisms of thermoduric spores in the dairy environment to achieve the overall goals as stipulated. A new project on the control of Listeria in ice cream, based on Response Surface Methodology will be initiated. In addition, under the USDA-HEC grant at the Honors College, further opportunities will be created for the undergraduate research opportunities in the area of food safety.

Impacts
What was accomplished under these goals? Objective 1 was supported by two National Dairy Council (NDC), and one Midwest Dairy Foods Research Center (MDFRC) grants. Under the first study, further work was taken up by collaborating with two dairy plants. Evidence was generated on the effectiveness of cavitation as a means to inactivate thermoduric sporeformers in skim milk and concentrates. The second study was a continuation of our work on understanding the process of sporulation during various stages of milk powder manufacture. Preliminary studies were conducted to study the generation times and conversion of vegetative cells to spores in selected milk sporeformers. The third study was a proof of concept experiment for the potential role of Direct Fed Microbials in contaminating raw milk at dairy farms. Objective 2 was supported by one National Dairy Council (NDC), and two Midwest Dairy Foods Research Center (MDFRC) grants. In the first study, experiments were continued to evaluate biofilm formation on modified plate heat exchanger (PHE) plates. Several types of coated coupons and PHE plates were tested for biofilm formation at both lab and pilot scale. The second study under this objective provided evidence on the bio-corrosion of stainless steel milk contact surfaces by biofilms of thermoduric sporeformers. Under the third project, studies were continued to evaluate the role of extrapolysaccharides in supporting the biofilm formation by EPS positive starter cultures. A Food Safety Colloquium was conducted for Honors College under our USDA-HEC grant, Meeting the Grand Challenge', in order to promote recruitment in food safety related careers at the University.

Publications

  • Type: Journal Articles Status: Published Year Published: 2015 Citation: KP Buehner, S Anand. G. D. Djira. 2015. Prevalence of Thermoduric Bacteria and Spores in Non-fat Dry Milk Powders of Midwest Origin. Journal of Dairy Science 98: 2861-2866
  • Type: Other Status: Published Year Published: 2015 Citation: S Anand, S Gupta and D Bawa. 2015. Cleaning and sanitation challenges in dairy industry. A review prepared for Midwest Dairy Foods Research Center. P1-49
  • Type: Conference Papers and Presentations Status: Published Year Published: 2015 Citation: Bawa D, S Anand, H Dahiya, H Patel. 2015. Inactivation of thermoduric sporeformers in milk by combined effect of cavitation and pasteurization. Oral Abstract #405. J. Anim. Sci. Vol. 93, Suppl. s3/J. Dairy Sci. Vol. 98, Suppl. 2. Page 525
  • Type: Conference Papers and Presentations Status: Published Year Published: 2015 Citation: Bawa D, S. Anand, H Dahiya, H Patel. 2015. Inactivation of thermoduric bacterial endospores with a combined effect of cavitation and thermal treatment. Abstract # T197. J. Anim. Sci. Vol. 93, Suppl. s3/J. Dairy Sci. Vol. 98, Suppl. 2. Page 382
  • Type: Conference Papers and Presentations Status: Published Year Published: 2015 Citation: Bawa D, S Anand, S Beckman.2015 Inactivation of thermoduric sporeformers in milk by continuous ultrasonication. Abstract # W144. J. Anim. Sci. Vol. 93, Suppl. s3/J. Dairy Sci. Vol. 98, Suppl. 2. Page 670
  • Type: Conference Papers and Presentations Status: Published Year Published: 2015 Citation: Beckmann S, L Alexander, S. Anand, LE Metzger 2015. Effect of ultrasound treatment on reconstituted deproteinized whey prior to lactose crystallization. Abstract #W145. J. Anim. Sci. Vol. 93, Suppl. s3/J. Dairy Sci. Vol. 98, Suppl. 2. Page 670
  • Type: Conference Papers and Presentations Status: Published Year Published: 2015 Citation: Garcia-Fernandez N, A Hassan, S Anand. 2015. Slime production by Bacillus strains affects biofilm formation on dairy separation membranes. Abstract # T141. J. Anim. Sci. Vol. 93, Suppl. s3/J. Dairy Sci. Vol. 98, Suppl. 2. Page 362
  • Type: Theses/Dissertations Status: Published Year Published: 2015 Citation: Khilendra Bhanduriya, Dairy Science manufacturing major for M.S., thesis program on Microbial aspects related to thermoduric bacteria during cheese manufacturing and ripening 2012-2015).
  • Type: Theses/Dissertations Status: Published Year Published: 2015 Citation: Somil Gupta, Dairy Science manufacturing major for M.S., thesis program on 'Studies related to microbially induced corrosion of stainless steel 304 and 316'(2013-15).
  • Type: Conference Papers and Presentations Status: Published Year Published: 2015 Citation: S Anand, 2015. Sporeformers and their control in milk. Annual meeting organized by the Associated Illinois Milk, Food and Environmental Sanitarians, at Bloomington, IL. May 6th.
  • Type: Other Status: Published Year Published: 2015 Citation: S Anand, 2015. What we learnt about dairy biofilms in the last 5 years? Annual Meeting Midwest Dairy Foods Research Center, MN, July 21.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2015 Citation: S Anand, 2015. Biofilms: development and control strategies. Abbott Nutrition, Columbus, OH. Aug 18.
  • Type: Other Status: Published Year Published: 2015 Citation: S Anand, 2015, Changing Trends in Foodborne Diseases and their Implications in Protecting Consumer Health, as part of the Meeting the Grand Challenges Honors Colloquium on Food safety, Oct 19.
  • Type: Journal Articles Status: Published Year Published: 2015 Citation: Hegerfeld-Baker, J., S. Anand, L. Droke and K. Chang. 2015. Factors influencing choosing food and agriculture related stem majors. North American Colleges and Teachers of Agriculture Journal 59(1): 34-40


Progress 10/01/13 to 09/30/14

Outputs
Target Audience: Food and dairy industry. Dairy producers. Researchers. The studies generated critical information related to incidence and control of thermoduric microorganisms and their biofilms during cheese manufacture and whey processing. The results were shared with the US dairy industry through a National Spore Seminar, DRI, and the 2014 Midwest Dairy Research Forum, NCCIA. Collaboration with dairy plants and dairy farms helped to resolve the microbial quality and spoilage issues due to thermoduric sporeformers and their biofilms. Information on the incidence of thermoduric sporeformers and spores in raw milk was also shared with the dairy producers around the 1-29 corridor to help improve the microbial quality of raw milk. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided? The project has provided several opportunities at both graduate and undergraduate levels of teaching. Three MS students completed their degree programs while working on these projects. Five MS and three Ph.D students are currently working under my advising on research project related to thermoduric sporeformers, and biofilm control. In addition, one undergraduate research projects was completed under this research program during fall 2014. How have the results been disseminated to communities of interest? Peer reviewed publications, invited lectures in conferences and national symposia, scientific posters presentations in symposium of American Dairy Science Association, direct interaction with dairy producers, and direct interaction with dairy industry. What do you plan to do during the next reporting period to accomplish the goals? We plan to continue work on the non-thermal applications for inactivating the sporeformers and spores in milk. The studies on the ability of sporeformers to cause corrosion on stainless steel surfaces will be completed. New studies have been initiated to control of biofilms by modifying the PHE contact surfaces, and understanding the germination and sporulation mechanisms of thermoduric spores in the dairy environment to achieve the overall goals as stipulated. In addition, we have been successful in getting a USDA-HEC grant under Honors College. Food safety colloquium and undergraduate research opportunities will be created under this grant.

Impacts
What was accomplished under these goals? Objective 1 was supported by two Dairy Research Institute (DRI), and three Midwest Dairy Foods Research Center (MDFRC) grants. The first study was a continuation of our work on the progression of thermoduric thermophiles in the manufacture of cheese and whey processing. The samples were collected from the continuous cheese making industrial runs. The studies helped to isolate and identify the causative organisms for cheese spoilage. The information was shared with respective cheese plants for necessary follow up. The work on membrane biofilms cleaning also resulted in identifying enzyme step modifications for more effective cleaning of membranes. These results formed the basis for a collaboration with industry to scale up the cleaning modifications under industrial setting. Under the second study further work was taken up related to the incidence of thermoduric sporeformers in raw milk and milk powders. Ten dairies around the I-29 corridor in South Dakota were sampled and analyzed for the counts of sporeformers and spores. The MALDI-TOF and molecular typing based identification revealed Bacillus licheniformis to be the predominant species in milk and milk powders of Midwest origin. The third study included application of a non-thermal technique, cavitation, in inactivating bacterial spores and sporeformers in milk. The effectiveness of cavitation in combination with heat treatment was demonstrated to inactivate thermoduric sporeformers. Objective 2 was supported by one Dairy Research Institute (DRI), and two Midwest Dairy Foods Research Center (MDFRC) grants. In the first study experiments were continued to further study the formation of resistant biofilms on reverse osmosis and ultrafiltration membranes. The lab developed biofilms were tested against several modification of the existing cleaning and sanitation protocol to remove the biofilms more effectively. The enzyme application and sanitation steps were successfully modified to achieve a greater inactivation of resistant microflora embedded under the biofilm matrices. The second study under this study focuses on the formation of biofilms on stainless steel surfaces by thermoduric sporeformers, and their ability to induce metal corrosion. Studies were continued to demonstrate the formation of thermoduric biofilms on SS coupons and the corresponding metal corrosion under lab conditions, as evident by scanning electron microscopy and atomic force microscopy. Studies were also initiated on a new project to evaluate the biofilm formation on modified plate heat exchanger (PHE) plates.

Publications

  • Type: Journal Articles Status: Published Year Published: 2014 Citation: SN Khanal, S Anand, K Muthukumarappan, M Huegli. 2014. Inactivation of thermoduric aerobic sporeformers in milk by ultrasonication. Food Control. 37: 232-239
  • Type: Journal Articles Status: Published Year Published: 2014 Citation: SN Khanal, S Anand, K Muthukumarappan. 2014. Evaluation of high intensity ultrasonication for the inactivation of endospores of some Bacillus species in nonfat milk. Journal of Dairy Science 97: 5952-5963
  • Type: Journal Articles Status: Published Year Published: 2014 Citation: KP Buehner, S Anand, GD Djira, A Garcia. 2014. Prevalence of Thermoduric Bacteria and Spores on Ten Midwest Dairy Farms. Journal of Dairy Science 97: 8009-8016
  • Type: Journal Articles Status: Published Year Published: 2014 Citation: Anand S, D Singh, M Avadhanula, Sowmya Marka. 2014. Development and control of bacterial biofilms on dairy processing membranes. Comprehensive reviews in food science and food safety. 13: 18-33
  • Type: Conference Papers and Presentations Status: Published Year Published: 2014 Citation: Buehner KP, S Anand, A Garcia. 2014. Incidence of thermoduric bacteria and spores on selected Midwest dairy farms. ADSA Annual conference, Kansas City, MO, July 20-24, 2014, abstract 1007, Poster T065, J. Anim. Sci Vol. 92, E-Suppl. 2/J. Dairy Sci. Vol. 97, E-Suppl. 1. Page 503
  • Type: Conference Papers and Presentations Status: Published Year Published: 2014 Citation: Gupta S, S Anand. 2014. Induction of Pitting on Stainless Steel 304 and 316 by Bacillus sporothermodurans. ADSA Annual conference, Kansas City, MO, July 20-24, 2014, abstract 1011, Poster T069. J. Anim. Sci Vol. 92, E-Suppl. 2/J. Dairy Sci. Vol. 97, E-Suppl. 1. Page 504
  • Type: Conference Papers and Presentations Status: Published Year Published: 2014 Citation: Beckman SL, S Anand, LE Metzger. 2014. Evaluation of a laboratory-scale batch crystallizer for lactose isolation from deproteinized whey. ADSA Annual conference, Kansas City, MO, July 20-24, 2014, abstract 1017, Poster T075. J. Anim. Sci Vol. 92, E-Suppl. 2/J. Dairy Sci. Vol. 97, E-Suppl. 1. Page 507
  • Type: Theses/Dissertations Status: Published Year Published: 2014 Citation: Sowmya Marka, Dairy Science manufacturing major for M.S., thesis program on CIP modifications for effective cleaning of biofilms (2012-2014).
  • Type: Theses/Dissertations Status: Published Year Published: 2014 Citation: Kim Buehner, Dairy Science manufacturing major for M.S., thesis program on The incidence of spores and thermoduric bacteria in Midwest dairies and commercial NFDM samples and their reduction via ultrasonication. (2012-2014).
  • Type: Theses/Dissertations Status: Published Year Published: 2014 Citation: Akimoto Ichinomiya, Dairy Science manufacturing major for M.S., thesis program on Bacteriocin producing lactic acid bacteria (2011-14).
  • Type: Other Status: Published Year Published: 2014 Citation: S Anand, K. Bhanduriya. 2014. Microbial quality of milk powders during manufacture and storage. A review prepared for USDEC. P1-7
  • Type: Other Status: Published Year Published: 2014 Citation: On invitation from Midwest Dairy Foods Research Center, six training modules (Camtasia based with narration) developed for the dairy producers and dairy industry for understanding dairy related thermoduric bacteria. These modules will be used by the US Dairy Industry to train dairy plant employees regarding the significance and control of thermodurics in dairy plants. The modules are copyrighted by Sanjeev Anand and were funded by MDFRC.


Progress 10/01/12 to 09/30/13

Outputs
Target Audience: Food and dairy industry. Dairy producers. Researchers. The studies completed under this project generated useful information related to incidence of thermoduric microorganisms during milk production and processing. The results obtained were also shared with the US dairy industry through a national spore symposium organized by the Dairy Research Institute, Chicago. Specific projects taken up in collaboration with dairy plants helped to resolve the microbial quality and spoilage issues due to thermoduric sporeformers during the manufacture of cheese and whey products. Information on the incidence of thermoduric sporeformers and spores in raw milk was shared with the dairy producers around the 1-29 corridor to help improve the microbial quality of raw milk. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided? The project has provided several opportunities at both graduate and undergraduate levels of teaching. Two MS students completed their degree programs while working on these projects. Four MS and one Ph.D student is currently working under my advising on research project related to thermodurics and biofilm control. In addition, nine undergraduate research projects were completed under this research program during 2013. How have the results been disseminated to communities of interest? Peer reviewed publications, invited lectures in conferences and national symposia, scientific posters presentations in symposium of American Dairy Science Association, direct interaction with dairy producers, and direct interaction with dairy industry. What do you plan to do during the next reporting period to accomplish the goals? We plan to continue work on the non-thermal applications for inactivating the sporeformers and spores in milk. The studies on the ability of sporeformers to cause corrosion on stainless steel surfaces will also be continued. We have submitted grants in the area of control of biofilms by modifying the milk contact surfaces, and understanding the germination and sporulation mechanisms of thermoduric spores in the dairy environment. If these grants are successful then we will be able to conduct this additional area to achieve the goals as stipulated.

Impacts
What was accomplished under these goals? Objective 1 was supported by two Dairy Research Institute (DRI) and one Midwest Dairy Foods Research Center (MDFRC) grants. The first study was focused on the progression of thermoduric thermophiles in the manufacture of cheese and whey processing. The samples were collected from the continuous cheese making industrial runs. Evidence was generated on the formation of thermoduric biofilms on plate heat exchangers and cross contamination of the final product by thermodurics. Mid-day wash was observed to be an effective means to reduce the carryover of thermoduric microflora beyond pasteurization. Similarly, thermodurics were also shown to persist during whey processing and caused spikes in the final product. Better cleaning protocols helped in reducing the spore counts in processed whey samples. Studies were also conducted to standardize spore testing protocols for whey protein concentrates and whey protein isolates. The second study was focused at the incidence of thermoduric sporeformers in raw milk and milk powders. Raw milk samples were collected from 10 dairies around the I-29 corridor in South Dakota and the samples were analyzed for the counts of sporeformers and spores. The isolates are currently being identified using MALDI-TOF and molecular typing. The third study included application of a non-thermal technique in inactivating bacterial spores in milk. Preliminary trials are focused at standardizing the process to achieve the desired levels of inactivation. Objective 2 was supported by two Midwest Dairy Foods Research Center (MDFRC) grants. In the first studies experiments were conducted to study the formation of resistant biofilms on reverse osmosis and ultrafiltration membranes. A Bacillus sp. obtained from an old membrane biofilm was used to develop biofilms under lab conditions. The lab developed biofilms were tested against several modification of the existing cleaning and sanitation protocol to remove the biofilms more effectively. The enzyme application and sanitation steps were successfully modified to achieve a greater inactivation of resistant microflora embedded under the biofilm matrices. The second study under this study focuses on the formation of biofilms on stainless steel surfaces by thermoduric sporeformers, and their ability to induce metal corrosion. Preliminary studies have been completed to isolate sulfate reducing proteolytic strains of thermoduric sporeformers from milk powder samples. Initial studies showed the formation of biofilms on SS coupons under lab conditions. Some of these organisms also resulted in initiating corrosion on these coupons, as evident by scanning electron microscopy and atomic force microscopy. These are just the preliminary findings and further studies are in progress.

Publications

  • Type: Journal Articles Status: Published Year Published: 2013 Citation: SN Khanal, S Anand, K Muthukumarappan, M Huegli. 2014. Inactivation of thermoduric aerobic sporeformers in milk by ultrasonication. Food Control 37:232-39. Anand S and Diwakar S. 2013. Resistance of the constitutive microflora of biofilms formed on whey reverse osmosis membranes to individual cleaning steps of a typical clean-in-place (CIP) protocol. Journal of Dairy Science. 96: 62136222.
  • Type: Conference Papers and Presentations Status: Other Year Published: 2013 Citation: S Marka, S Anand. 2013. Feed substrates influence biofilm formation on reverse osmosis (RO) membranes, and their cleaning efficacy. ADSA Annual conference, Indianapolis, Indiana, July 8-12, 2013, abstract 57166. T224. J. Anim. Sci. Vol. 91, E-Suppl. 2/J. Dairy Sci. Vol. 96, E-Suppl. 1. P 79. K Bhanduriya, S Anand, Lloyd Metzger. 2013. Causative organisms for slit defects in cheddar cheese samples-A case study. ADSA Annual conference, Indianapolis, Indiana, July 8-12, 2013, abstract 57155. W228. J. Anim. Sci. Vol. 91, E-Suppl. 2/J. Dairy Sci. Vol. 96, E-Suppl. 1. P 332 K Bhanduriya, S Anand, Lloyd Metzger. 2013. Tracking the progression of thermoduric bacteria during the manufacture of cheddar cheese-A case study. ADSA Annual conference, Indianapolis, Indiana, July 8-12, 2013, abstract 57136. W227. J. Anim. Sci. Vol. 91, E-Suppl. 2/J. Dairy Sci. Vol. 96, E-Suppl. 1. P 332 N Garcia Fernandez, A Hassan, S Anand. 2013. Role of exopolysaccharide-producing starters in biofilm formation on dairy separation membranes. ADSA Annual conference, Indianapolis, Indiana, July 8-12, 2013. T110. J. Anim. Sci. Vol. 91, E-Suppl. 2/J. Dairy Sci. Vol. 96, E-Suppl. 1. P 40. Graduate competition. Giovana, S Anand. 2013. Enumeration and Characterization of Thermoduric Spore Forming Bacteria in Nonfat Milk Powders. Undergraduate Research competition SDSU 2013. Undergraduate Research Scholarship and Creative Activity Day (URSCAD). Anand S, 2013. Micro 101 for cheese industry; Microbiological aspects of quality and safety of cheese. North Central Cheese Industry Association (NCCIA) annual conference, Rochester, MN. Anand S, 2013. Emerging thermoduric/ thermophilic bacteria and their inactivation by ultrasonication. Invited lecture, U.S. Dairy Spore seminar, organized by the Dairy Research Institute and US Dairy Export Council, SF, California. Anand S, 2013. Biofouling and cleaning of whey reverse osmosis membranes, Invited lecture, 15th Annual Dairy Ingredients Symposium, organized by the Dairy Products Technology Center, California Polytechnic State University, SF, CA. Anand S, 2013. Thermoduric thermophiles: progression during cheese making-Agropur Study. Agropur dairy plant, Iowa.
  • Type: Other Status: Published Year Published: 2013 Citation: Anand S and Som N Khanal. 2013. Significance of aerobic sporeformers in milk and dairy products. A review. US Dairy Export council and Dairy Research Institute. p 1-61.
  • Type: Journal Articles Status: Accepted Year Published: 2013 Citation: S Anand, D Singh, M Avadhanula, and S Marka. 2013. Development and control of bacterial biofilms on dairy processing membranes. Comprehensive Reviews in Food Science and Food Safety (accepted).