Bacterial Growth on Hamburger Meat

Philip Paquet Microbiology 305W Section J March 10, 2011 Introduction:

Hamburger meat contains many different types of bacteria many of which can be harmful to humans. The bacteria contained in the meat can, in many cases, cause sickness or, in extreme cases, death. In order to stop or inhibit growth of bacteria on meat, precautions are taken during processing in factories to decrease the likelihood of humans getting sick. Escherichia Coli is the most common type of bacteria found in hamburger meat and a particular species known as Escherichia Coli O157:H7 can even cause death. (Kassenborg, HD, ET AL). To test the amount of colonies found in an unknown hamburger meat source the meat was homogenized and was diluted by a ratio of 1:10 every transfer into spread plates and pour plates. The plates were left to grow for a week and the colonies of each of plates were counted. Using a consolidation of trials, a result of the amount of colonies grown for each dilution was reached. The concentration of colonies in each of the dilutions is correlated to the contamination of the meat. As the samples are diluted, the amount of colonies should decrease. Depending on the source of it, the results of the test should display the amount of bacteria that was present in the unknown sample of meat. The dilutions should show a decrease in growth once the most ideal and non-stressful conditions for bacterial growth is reached.

Materials and Methods: The hamburger meat was prepared by blending two grams of each of the five different unknown hamburger meats in 18 mL of 0.75% NaCl. 4.5 mL of 0.75% NaCl was transferred into tubes 10-2 to 10-6 then 0.5 mL of the prepared hamburger meat solution was transferred to the tube labeled 10-2, representing a 0.1 dilution from the original solution. 0.5 of the new solution in the 10-2 tube was then transferred to the 10-3 tube. The rest of the tubes were filled using the same method to ensure a 1:10 dilution from the tube preceding it. 0.1 mL of each of these solutions was then transferred to a spread plate labeled 10-3 and, using a hockey stick, was spread around evenly. This was repeated for tubes 10-3

to 10-2, the hockey stick sterilized with ethanol and flame between each spreading. A control was used by using a sterile spread plate to ensure accuracy. For the pour plates, 1.0 mL of the 10-3 diluted solution was moved to the pour plate labeled 10-3. This was then done for plates 10-4 to 10-6 using the appropriate tubes. For each of the pour plates, enough molten NA (nutrient agar) was poured on top to cover the bottom of the plate and mixed using a figure eight. The control used for the pour plate was 1.0 mL of sterile NA. The plates were incubated at room temperature for one week before they were counted. Titer, a method of determining the amount of bacteria in a culture, was calculated by taking the number of colonies and dividing it by the dilution into cfu/mL (culture forming units/mL). (Shand, R.F.) Results: Many of the plates were unable to be counted since they were NSS (Not Statistically Significant). The spread plates were NSS from 10-5 to 10-6 and the pour plates were NSS from 10-4 to 106

. Sample B had an average of 4.5x104 cfu/mL while sample C had an average of 2.06x106. Samples A,

D, and E had averages similar to each other within the range 1.32x105 to 1.5x105. (Table 1) The pour plates had a range of 4.85x104 to 1.16x105. (Table 2) Table 1 Table 2 Discussion: The objective of this experiment was to determine the amount of bacteria in five different hamburger meat samples from different unknown locations. It was hypothesized that the contamination of the samples would be determined by the source of the meat along with the handling of the meat and thus vary respectively. The amount of bacteria in the meat, measured in cfu/mL, did not differ widely except in samples B and C. Samples A, D, and E were similar with ranges between 1.32x105 to 1.5x105 which indicates that although they may not be from the same source, they are most likely from a similar environment or

taken from somewhere and similarly handled. This supports the hypothesis that although the meat may be similar, it does not change contamination and handling along with the source (the environment the animal that the meat came from lived in along with other factors) which will result in differing results from the other samples. The objective of the experiment was to show how hamburger meats differed due to the source and the handling of the meat. In order to better understand this, the sources should be more limited to three different sources and the source should be known by using a known source for the experiment along with more samples, it would be much easier to compare the results and see how the different sources contained varying amounts of bacteria. For future studies, two sources could be used and compared based on the environment the animal lived in allowing for further study of how the animals diet, climate, and native foliage affected the meats concentration of bacteria.

Citations
HD, Kassenborg, Hedberg CW, Hoekstra M, Evans MC, Chin AE, Marcus R, Vugia DJ, Smith K, Ahuja SD, Slutsker L, and Griffin PM. "Farm Visits and Undercooked Hamburgers as Major Risk Factors for Sporadic Escherichia Coli O157:H7 Infection: Data from a Case-control Study in 5 FoodNet Sites." Emerging Infections Program FoodNet Working Group. Web. 24 Mar. 2011. <http://www.ncbi.nlm.nih.gov/pubmed/15095199>. Shand, R. F. (2o11). Fundamental techniques and experiments in microbiology. Flagstaff AZ: Northern Arizona University. Vogt, R., & Dippold, L. (2005). Escherichia coli o157 H7: Outbreak Associated with the Consumption of Ground Beef June-July 2002. Public Health Records 174-178. Retrieved March 10, 2011, from www.jstor.org/stable/20056769