We were not able to go to the hospital to look around the microbiology laboratory. This is something Patan hospital is strict about and so last week Sabina and Rajendra from the Oxford Clinical Research Unit came to us instead. They brought Dr Tom Darton too who works in the Oxford Vietnam research unit and was visiting Nepal to help set up a big study to understand the numbers of people carrying and effected by the bacteria within Patan in Nepal, but also in parts of Malawi and Bangladesh. Our visitor's experienced the full force of Jeewan Jal curiosity.
Each shared stories of the part they play in the story of global health research from the scale of the petri dish to that of a study spanning continents. Conversations were so in depth we didn't complete all of what was planned and so this week Sabina returned with photos of the lab to explain to us some of the specific materials and techniques they use to diagnose and research disease. This was a good chance for the group to show some of what they remembered.
Things that Anu remembered were:
That some people can be carriers of Typhoid but not be sick and so drew a carrier bag...and a gall bladder where the typhoid bacteria like to head to in the body and multiply.
That for an unknown reason Typhoid preferentially affects men so she drew a man becoming infected by drinking contaminated water.
In the UK in some drug trials they are injecting consenting research participants with Typhoid bacteria!
We know the age of typhoid because there is a molecular clock. There are only tiny differences in the genetic code of bacteria and by seeing that differences and analysing it we know that Typhoid is only 30,000 to 60,000 years old.
Devashree remembered the story of Typhoid Mary, an Irish cook in the U.S who was a carrier of Typhoid but had no symptoms herself. Despite this she was able to infect others and was held in quarantine for much of her life.
Jemi remembered that one of the ways of knowing what people are ill from is to do a blood culture. And that for this we have to collect blood of 5-10 ml. Sabina told us that this was 3ml for children.
She remembered that scientist find it difficult to grow typhoid bacteria from water even if it is there but that it can be cultured in the blood and then identifies.
She also drew one of the big drivers of the epidemic of Typhoid that occurred in 2002, the maoist insurgency. This drove people to Kathmandu from the hills and the population rapidly grew without the infrastructure to support it. This is when the research unit started doing a lot of research. There was a lot of concern because the antibiotics that were normally used (Ciprofloxacin) were not working any more. The bacteria had mutated into drug resistant strains!
Jemi also remembered that Dr Tom had told us that evidence suggests that Typhoid bacteria are iron loving. If you drink alcohol you have more iron in the blood and might be more susceptible to infection. This could explain why women and men have different risk of contaging the disease. Women are less likely to drink as much as men and also, menstruation might mean that they have less iron and so are slightly protected.
Jari drew some of the ways in which we might treat water from sources like the well and stone tap through boiling and filtering. Boiling kills all the bacteria. There aren't many studies done but it is thought that filtering kills the bacteria too and some types of filtering with iron particles might even trap viruses.
Drew the structure of the Salmonella typhi: its rod shape and its lipopolysaccharide (fat and sugar) wall. He remembered that there were Y shaped things...which Anu helped to name as antibodies.
Devashree pointed out that it was ciliated (with little molecular tails) to help it move around and that the thing that looks like an eye was actually a nucleus...the place where the DNA can be located all squished up. Sabina told the group that the DNA is actually in a ring shape in bacteria and if we cut it and stretched it out it would be about three times the length of the bacteria itself.
Bindu talked about typhoid being bile loving (which is why it heads for the gall bladder in the body) and is transferred through faecal oral routes and so mainly through water and food such as pani puri!
But we were all in for a surprise because Sabina told us that in fact a study done by Dr Karkey in the research unit had found that pani puri seems to be protective against contracting Typhoid! We don't know why but speculated that it could be something to do with the spices in the juice. I am still contemplating whether or not to risk it next time I pass a pani puri vendor on the street.
Bindu also remembered the protective measures that scientists use in the lab such as lab coats, gloves and biohazard boxes (which have their own airflow to stop airborn diseases like TB from entering the atmosphere of the lab).
We then went through some photos from the lab to hear about some of the other things we might find there.
This is a test to see which antibiotics bacteria are sensitive to or not. The white disks are antibiotics which seep into the agar media. Where there is no white on the plate the bacteria are unable to grow.
Different bacteria give off different gases. These can be used to identify the bacteria. If you see a little black then you can say this is Salmonella Typhi. If it has a lot of black it might be another organism. No black means paratyphi.
The Biohazard Box
Agar medium being cooked.
An autoclave is used to sterilise the things that will be reused in the lab. They work like big pressure cookers like the ones we use for cooking dal. This is the lab assistant who is cleans everything.
This is bacteria down the microscope from a stained slide. The staining process is called Gram Staining after a man that invented it called Christian Gram.
First a purple dye is used then iodine and alcohol and acetone are used to rinse off the dye. If the bacteria on the plate are gram positive then they will be purple at this point. If not then they will go clear again. Another dye has to be used to dye the bacteria if they are clear. If you look down the microscope and the bacteria are dark purple then you have a gram positive bacteria. If you see that they are pink then they are gram negative. Typhoid bacteria are gram negative.
This is a sputum sample! Used to look at lung infections (not Typhoid)
This is an incubator where the agar plates that are growing the bacteria from samples for identification are kept.
This is the broth used to do children's blood culture. They have a seal on the top and so samples have to be injected into the bottles with needles. These cost more than regular bottles and so are not used normally in Nepal. At the moment there is a paediatric study and so the bottles have been bought for this.
We finished off our discussion by thinking about our story. In it we are given a medicinal plant by the Goddess Bagmati (the river that runs to the North of Patan). Sabina suggested that maybe the plant could have antibiotic that hasn't been discovered before.
"What should we call it?" asked Anu. The group came up with suggestions: Peneoxyfloxycillin?...Blah Blah cillin?...Siancillin? (I vetoed that one)
Finally we decided on Wasacillin. Wasa is Newari for medicine...With a new fantasy antibiotic on our hands we realised that we have a lot of fantasy responsibility...suddenly our story needs to incorporate drug companies, lawyers and patent agreements...We are starting to realise that things are never simple!