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Laboratory

The chart above shows the curve Crop growth of bacteria at 37 º C. Hourly samples were taken from the crop (black dots), and diluted in water so that all of them containing a final concentration of 10,000 organisms per milliliter. The samples were divided into three tubes of 1 ml each and treated as follows:

Tube 1: Nothing added. Control tube.
Tube 2: addition of chloramphenicol, an inhibitor of translation
Barrel 3: addition of penicillin

After leaving acting antibiotic for twenty minutes at 37 ° C, aliquots of 100 microliters of each tube and inoculated in Petri dishes with fresh medium and without antibiotic. After 24 hours at 37 ° C were counted and colony forming units on plates. The table below shows the results.

Note: The explanation is not due to the emergence of mutants resistant to antibiotics or any other type of genetic phenomenon

Explains

results

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Nature vs Biodiesel

A research team has isolated a microorganism of a pond whose average temperature is 25 º C. When studying the location of the microorganism in the water column of the lake with respect to day-night cycles, researchers found the following result (solid line on graph).

In the laboratory the research group working with the organism isolated and determined the following data on their physiology and metabolism. It is a microaerophilic bacterium strict heterotrophic metabolism, whose optimum temperature is 25 º C. This research group attempts to reproduce in the laboratory the results observed in nature simulating cycles day and night in a column of water inoculated with this organism, which is placed in a chamber incubated aerobically at 25 ° C temperature and in which by simulated solar light day and night cycles but you get a completely different result (dashed line on graph): Microorganisms are always at the same depth.


a. - Develop a hypothesis to explain the result.
b. - Design an experiment in which you can check this hypothesis

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inoculated with a population of 1,000 bacteria in a culture medium. The conditions are so ideal and perfect that microorganisms do not go through a conditioning phase or latency and begin to grow and divide from the outset.

At different times they are taken samples for analysis, measuring the optical density of culture (OJ, broken line), and the number of cells present in the crop (N, solid line). The values \u200b\u200bwere fed into a computer that is in a semilog plot.

Unfortunately, unconfigure a computer virus data table and produced two sets of graphs are different.

Which of the two graphs depicted above is correct and why?

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exponential growth, future promises

This problem appeared in the June review of the subject of Industrial Microbiology.



With the current energy crisis are looking for new sources of hydrocarbons. It is increasingly clear that biofuels produced growing yeast from grain or other human food sources are more cons than pros. One drawback is that they diminish the food supply, with subsequent higher prices for them.

One possible solution is the use of microorganisms to produce biodiesel, mostly a lot of interest in photosynthetic bacteria. Thanks to the bacteria produce lipid metabolism that are then extracted and processed to produce biodiesel.

But bacteria have been out a competitor. There are companies that are investigating eukaryotic unicellular algae. As photosynthetic bacteria, algae also produce lipids that are then processed into biodiesel.

growth parameters of both types of organisms are:

you think

• What are the advantages of using photosynthetic microorganisms to produce biofuels instead of yeast-based methods and food products?


• Which of the two crops is more advantageous or less inconvenience to the industrial production of biodiesel and why?

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wastewater treatment. Mass balance. Food Microbiology Bacteria

It is establishing a process wastewater treatment continuously. In the first stage of the total amount of suspended matter entering the bioreactor is 1,000 tons per day. This wastewater is treated with an inoculum of 10,000 kilos of activated sludge. After this first stage, suspended matter is reduced to 10% of the initial concentration. There is evidence that the batteries present in activated sludge completely degraded into CO2 and H2O to 75% of the material introduced.

also microorganisms in activated sludge have a growth rate of 8,333 kilos per hour and cell death rate of 30 kilos per hour. The cumulative mass of microorganisms within the bioreactor is of 2,470 kilos per hour. Write

mass balances accumulated in the bioreactor for suspended matter and bacteria in activated sludge coming out of it.

Cell Respiration Lab Ap Bio

A person who is going to hike an omelette prepared using egg farm. The tortilla is prepared the night before. Unfortunately, these eggs are contaminated in the shell with Salmonella enterica . By break to beat them there is an inoculation of these microorganisms in food. That person likes the tortilla curd is not everything, so that cooking does not end with the pathogenic bacteria found in the innermost part of the tortilla. When finished it, that person leaves the tortilla to cool to room temperature. At 9 am the next day, put the tortilla in a pan that you enter in your backpack and hiking.

Data: Number of

S. enterica in the shell: 1,000 cells per square millimeter. Eggs
used: 6 eggs
Inoculation: shows 25% of bacteria in 1 cm egg box for

Maximum cooking temperature reached inside the tortilla: 60 º C
actual cooking time inside the tortilla: 2 minutes
Decimal Reduction Time Salmonella under these conditions: 0.5 minutes

temperature overnight (12 hours time): 15 º C
specific growth rate at 15 ° C. μ = 0.23 h-1

average temperature during the trip: 28 º C

specific growth rate at 28 ° C. μ = 0.87 h-1

minimum inoculum should be taken to S. enteric infection occurs: 1 million bacteria.

Question: estimate the time that the intake of a spike of the tortilla will cause salmonellosis with 100% certainty.

Note for non-English speaking: 1 skewer tortilla is a quarter (approx.) of it

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genetically engineered a group has constructed a mutant Escherichia coli in the biosensor system that triggers the movement of the flagellum.

If inoculated wild type, call-1 strain in semisolid agar the bacteria can "Swim" and spread around:

wild strain:

(*) click on image to view larger

In the mutant strain, called strain-2, has the deleted gene chez encoding the protein Chez. Without this protein the flagellum movement control is lost.

First question: How do you behave the mutant? Does semisolid agar diffusion will be higher or lower than in the wild strain?

A the research team then takes the strain lacking Chez and I introduced a plasmid containing the gene for this protein, but under the control of a ribointerruptor (riboswitch) . This new strain is called strain-3.

The ribointerruptor works as follows:

• In the absence of a molecule, let's call it molecule A, the gene messenger mRNA chez can not be translated by ribosomes.


• In the presence of the molecule A, the mRNA of the gene chez can be translated by ribosomes.

Researchers Strain-3 inoculated in a semisolid agar plate where it has added a drop containing the molecule A in one side of the plate.

Second question: How will behave Strain-3 at the plate? Do or get closer to the molecule A?


This research group also introduced the gene for Green Fluorescent Protein or GFP ( F G reen luorescent P rotein ) in the strain-3, thus the strain-4. This bacterium emits green fluorescence by the protein

Third question: What practical use could be strain-4?

(*) Figure produced from the book " Brock. Biology of microorganisms " Pearson Publishing

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Biodeterioration of cultural heritage

A large number of caves containing prehistoric paintings ( Altamira Cave of the Bats, ...) have been closed to public visits to avoid damaging them. Despite the darkness and lack of nutrients from the environment, microorganisms can grow. The combination of carbon dioxide produced by the visitors and the microbes living on the surface of the rock lead to the degradation of paintings. In a first phase sought to avoid such degradation by using chemical biocides but these proved to be much more aggressive against the paintings against microorganisms. It was observed that the first stage of biodeterioration of a painting usually begins with the appearance of a blue-green stain. On this blue-green stain becomes available a biofilm composed of a variety of bacteria and fungi that will attack the paint.

Recently, the European Community has initiated a project that aims to prevent degradation and allow resumption of visits. There several research groups and each has developed a specific strategy to solve the problem. Here we describe the two most promising in terms of its preliminary findings and has failed:

• Strategy A: Lighting the caves with a blue-green light. It was observed that avoids the appearance of spots of the same color and therefore no subsequent biofilm develops.

• Strategy B: Use of synthetic siderophores. Siderophores are molecules for iron chelators. This strategy is used both in the initial stages of biofilm formation, as in the more advanced. It is observed reduced biofilm growth and even their disappearance.

• Strategy C: Use of specific antibiotics. The use of fungicides, bactericides or only attacked a certain group of microorganisms (antibiotics against Gram + or against Gram -) decreases the population of these microorganisms in the biofilm. However, the treatment does not appear to be effective or even initially, it is not observed any decrease in the growth of biofilm during treatment. To make matters worse, in the long term appear microorganisms resistant to these compounds, so that the treatment stops be fully realizable.

Questions:

1. What kind of organisms are affected by the strategy A?


2. Considering the answer to question 1, explain why they are the first organisms in the biofilm and how can these organisms live and reproduce in an environment like a dark cavern


3. Why is treatment with antibiotics ineffective even initially?


4. Why synthetic siderophores are effective in inhibiting the growth of the biofilm?


5. you show microorganisms resistant to siderophores?

Red Dot Scope Magnifier

biofertilization

A company that produces bio-fertilizers is testing the potential of two species of cyanobacteria as nitrogen fixers. One species in the genus Oscillatoria and other gender Anabaena. Below shows a graph comparing the efficiency of nitrogen fixation in both species over the day-night cycles.




explains why the behavior of both species is so different.

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Cyanobacteria and fecal contamination on beaches

In 1999, Huntington Beach in California had to be closed due to fecal contamination problem. Immediately there was a series of surveys to determine the source of pollution. This home was in the Santa Ana River water around the river waters, embracing an urban area, a power plant and a reclaimed marsh as a bird sanctuary (Talbert Marsh). Below is a map indicating contaminated water pipes to the river and the sampling stations (numbered circles).

In urban areas of the United States, the storm water network is separated from the network of black water (sewage). Additionally, the sewage is treated before being discharged into the Santa Ana River

The results of microbial contamination of the samples were as follows (numbers in CFU per 100 milliliters).

addition, a sampling of fecal microorganisms over several days at station 4 and it was found that:

Microbiologists postulated a hypothesis about the origin but this hypothesis did not convince the city council. It was decided to conduct a more comprehensive series of samples and after they were unable to determine exactly where the pollution occurred. It was found that the initial hypothesis was that by microbiologists correct.

Where contamination originated?

What data supports this conclusion?

Why is an oscillating pattern in station number 4?

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In the pictures above you can see a special transgenic plant. Arabidopsis belongs to the genus and is designed to act as a biosensor. It is produced by a Danish company for a very special purpose.

Normally the plant is green. But if the soil is the NO molecule 2, then the plant changes color. The NO molecule 2 is absorbed by the plant, and once inside, this molecule activates the synthesis of a red pigment.

The Davos Forum held in January 2007 called this pioneering and promising technology. The tests of this plant in Croatia and Serbia were quite successful and are planning to use in Africa. However, there is also criticism. One is that are transgenic plants and be released into the wild. The other is that according to the HALO Trust, is not a good biosensor, for if there is NO release 2, the plant is not dark.

Questions

1. What bacteria is used to obtain the transgenic plant? What
2. of genes have been introduced for this red pigment?
3. Why is using this plant?