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Special Features: HGT investigation

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It is necessary to know what the scientific researcher’s perspective of horizontal gene transfer is. Thus, we visited Peking University, Peking University Health Science Center, The Third Hospital of Peking University and National Institute of Biological Sciences (Beijing) in this summer, sent out hundreds of questionnaires, which were produced based on international biosafety manuals and domestic regulations. From results of the questionnaires, we got a rough idea of people’s awareness, and came to the conclusion that most of researchers knew little about HGT, resulting in their carelessness in experiment operation and worries.

The part one aimed at gathering people’s background information. Majority of lab workers received our questionnaires were graduate students, and the length of time they stayed in lab varied from several months to several years.

1.Do you know the biosafety level (BSL) of your lab?

2.Have you received any biosafety education when entered this lab? If yes, how?

3.Here are some descriptions of the genetic manipulation in the following, have you done it that way?

A.Standard precautions are always followed, and barrier protections applied (gloves, gowns, eye protection).

B.Make risk assessments when manipulate genetically modified microorganisms.

C.The objects or materials have been effectively decontaminated or disinfected by an approved procedure before discharge.

D.Air grills will not be blocked with notes, pipettes or other materials.

E.Open buckets or rotors after aerosols have settled or in a biological safety cabinet when using centrifuges. Use sealable buckets or sealed and tube breakage rotors.

4.When someone is doing inappropriate manipulations, would you stop him/her?

5.Do you know any national law concerning about biosafety?

Questions of part two investigate some general issues of biosafety. The first question was designed to investigate the lab members’ awareness of the biosafety level system and whether they paid attention to their own lab’s safety level.

The biosafety level is significant because it provides a detailed requirement for labs at different level and correspondently the experiment they can do when requirements were met. If the member did not know the system or did not know what level the lab they work in was, they would probably do experiment which were forbidden in the lab at this level. Thus, there will be potential safety risks. The second question revealed the lab’s emphasis on biosafety which is closely related to people’s awareness. Safety education for rookies is necessary to keep proper operation and prevent safety of both individuals and the laboratory. The graduate programs in China need to be improved a lot in this respect.

The third question is about genetic manipulation, and the five given items should all be conducted when doing experiment. This question investigated specifically how some manipulations important to biosafety were conducted in the labs. The results of this question indicate that many students were still lacking knowledge or good habits of experiment operation. Guidelines are necessary if new round of training and rectification starts. People would know which manipulation should be paid special attentions to.

The fourth question indicates whether people are aware of the serious results which will be caused by inappropriate manipulations. Many people knew the right way to conduct the experiment safely, but sometimes they would not do as they were taught or they might think that it made little difference to obey the standard for them. Luckily, almost all students would like to stop others’ improper actions and explain the reason. It is really important to improve people’s awareness of biosafety.

Similar with the first question in this part, the 5th question is designed to investigate people’s concerning degree for biosafety. It is also indicative for the nation’s publicity of their biosafety related law and the government’s attitude. Unfortunately, hardly any people knew any domestic law and regulation, while some international ones such as WHO biosafety manual and guidelines by NIH were better known. Thus both scientific institution and local government should intensify the publicity of the laws and regulations.

1.Which are the consequences of horizontal gene transfer?

A.Variation and evolution could be produced by the process of genetic transmission from parents to offspring

B.Some pathogenic bacterium will carry genes encoding antibiotic resistance

C.Evolutionary rate of indigenous flora would speed up.

D.The possibility of bacteria spontaneous lysis will be remarkably increasing after the gene horizontal transfer

2.Here are some stories happened in a laboratory, which ones do you think would possibly cause horizontal gene transfer?

A.Jim planned to do miniprep. When he stopped the shaker, he found that some of tubes fell down and liquid broth was releasing. He continued the experiment simply after spraying the shaker with 75% ethanol.

B.When the experiment was finished, Jim still had liquid broth left in tubes. Then he poured the liquid into the drain.

C.When doing the transformation, Mario was stuck because culture dishes are insufficient. However, he realized that with different gene reporters like gfp and yfp, samples could be differentiated by colors. Then Mario finished the coating job by mixing up some of samples.

D.Merlin wanted to analyze the toxin produced by bacteria. However, he noticed some contaminated microbes growing in culture dishes, and was curious about the characteristic of these bacterials. Merlin didn’t think they would carry any toxic gene and hardly took any protective measures in the following.

3.What do you think are appropriate ways to eliminate effects of HGT?

A.The physical containment of labs is divided into several classes.

B.Eating or storing food in labs is forbidden.

C.Waste of lab and the work surfaces are decontaminated daily.

D.Bacterial or biological waste would be autoclaved before disposal.

E.Take more biosafety considerations when inserted DNA sequences are not well characterized, such as preparing for genomic DNA libraries.

4.Have you ever taken HGT into consideration when design your experiments?

In part three, we investigate how much people know about the horizontal gene transfer. The first question could tell whether people know what horizontal gene transfer means. Only the second and the third statements are correct, and there were one-third people who cannot choose correctly. In the second question, we would like to know whether people are familiar with the consequence of horizontal gene transfer. Despite the slim chances , all of four choices might cause horizontal gene transfer, but few researchers could give the right answer.

Question three is about how to eliminate the effect of HGT. With this question we would like to remind people some easy ways to eliminate HGT and thus to better protect ourselves. All descriptions can eliminate the effect of horizontal gene transfer. Still, the accuracy rate calls for improvement. And the fourth question indicated that, few people were concerned with HGT when they made experimental design. Many of them had never thought about HGT or did not know what measures should be taken.

Many researchers mentioned that, in the last opening question, they didn’t really know what horizontal gene transfer exactly means, and wanted to know whether it was horrible and would affect themselves. Some students wanted to know the assessment measures, and doubted that whether this issue had been correctly clarified by biosafety agency. Many believed that more supervisions and improvements of law were necessary for scientific development.

To sum up, though horizontal gene transfer has been identified and characterized since the last century, many researchers are far from having a thorough knowledge of impact of HGT and factors that promote or limit HGT events. In-depth understanding will be helpful in implementing precautions against wrong-doings and the development of rational strategies to prevent the side-effect of HGT. Also, it would help researchers to better analyze and utilize national policies and regulations.

References

[1] Laboratory Biosafety Manuel. Third Edition. World Health Organization 2004

[2] Biosafety in Microbiological and Biomedical Laboratories. Fifth Edition. U.S. Department of Health and Human Services, Public Health Service, Centers for Disease Control and Prevention and National Institutes of Health. 2007