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Super-flies and parasites

5 Posts tagged with the fieldwork tag
2

Hello blood fluke enthusiasts,

 

Once again I am posting about my favourite parasite, the blood fluke called Schistosoma. I want to tell you about an exciting project that is going on on the beautiful archipelago of Zanzibar.

 

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Zanzibar is a semi-autonomous archipelago of Tanzania. The two main islands are called Unguja (or Zanzibar island) and Pemba. We are working on a very exciting project to stop schistosomiasis transmission on these islands.

 

This is a bit of a long post but please if you can bear it read on! If successful this project could revolutionize our approach to schistosomiasis (blood fluke disease) control.

 

Schistosomiasis control

 

As I explained in my first blood fluke post, infection with the blood fluke Schistosoma causes a disease called Schistosomiasis (aka Bilharzia).

This disease affects over 200 million people worldwide, the majority living in sub-Saharan Africa. It is strongly linked to poverty and does heart-breaking damage to children and adults in the poorest and most vulnerable communities.

 

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The clinical symptoms of schistosomiasis aka bilharzia, the blood fluke disease: (L)  bloody urine from children excreting the parasite eggs through urination and (R) a malnourished child with a hugely enlarged liver due to damage caused by the parasite eggs stuck in the tissue.

 

Depending on the species of the infecting schistosome worms the disease can cause:

•          Diarrhoea, bloody stool, blood in urine, painful urination.

•          Anaemia, stunted growth, enlarged liver and spleen.

•          Damage to the liver leading to liver fibrosis.

•          Damage to the genitals, kidneys and bladder potentially leading to bladder cancer.

•          Increased risk to sexually transmitted diseases like HIV.

 

Currently there is no vaccine. Schistosomes are masters of disguise when it comes to the immune system which means vaccines that rely on your immune system are difficult to develop. Researchers are trying though! Thankfully there is an effective oral drug called Praziquantel that kills the adult worms in humans. BUT it is the only effective drug against all species of this parasite, which raises concerns regarding drug resistance, and it does not stop people from becoming re-infected.

 

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A boy being treated for schistosomiasis. The treatment is an oral dose of Praziquantel. Although the side-effects are minimal the pill is quite bitter and can cause stomach upsets so making sure a child has some yummy juice and a bit of food with treatment is important.

 

Up until now efforts to control schistosomiasis in sub-Saharan Africa have focused on regular treatment of school children to reduce infections and prevent the severity of the disease. The theory being that if you treat regularly you can prevent the child from developing those nasty outcomes listed above. The drug is donated and there are excellent NGOs providing support to programmes wishing to deliver the drugs to schools. Hurrah!

 

However this regular treatment approach has NOT interrupted schistosomiasis transmission in a sub-Saharan African country. This means it requires a (very) long term commitment from the programmes and ministries of health. A lot of these countries have weak and struggling health systems burdened with many challenges (lack of water & electricity, clean needles & surgical equipment, painkillers, antiseptic cream etc) as well as a whole range of poverty-loving diseases to deal with. How long can a struggling health system keep up 'regular' treatments in difficult to reach areas? Once these are missed, or the programme is interrupted, the disease comes back.

 

What about stopping transmission?

 

Elimination = stopping local transmission

 

This is is exactly what is being attempted in Zanzibar through a multi-institute and major collaborative project led by:

 

There are three additional key players:

  • My friend and colleague Dr Steffi Knopp from the Museum and the Swiss Tropical and Public Health Institute. Steffi is tirelessly overseeing the details and daily running of this project as well as analysing the results and publishing whatever new insight we get into schistosomiasis elimination from this ambitious project.
  • SCORE (Schistosomiasis Consortium for Operational Research and Evaluation) funds this project with money from the Bill and Melinda Gates Foundation.
  • Schistosomiasis Control Initiative, a wonderful NGO based at Imperial College providing countries with all the logistical and implementation support needed for national treatment programmes (they do accept donations and fundraising so if interested just get in touch.

 

Together (and with a few other people whom I have not mentioned and I do hope will forgive me), they form (drum roll please...):

 

ZEST – the Zanzibar Elimination of Schistosomiasis Transmission

 

(And now superhero music, or better yet Vangelis’ Chariots of Fire)

 

This project aims to answer the question:

What tools do we have to stop transmission and what is the most effective way of achieving this?

 

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Transmission between humans and snails occurs in the local water bodies. In order to reach the water the parasite eggs come out with stool or urine. Because there are rarely toilets and no sewage system or human waste treatment facilities this human waste reaches the water that people frequent and the snails live in. The parasite is then able to continue its life cycle by first infecting a snail and then infecting a human.

 

So where on the life cycle can we intervene to stop transmission?

  1. We can kill the adult worms inside people by treating them with Praziquantel – Mass Drug Administration to communities at risk of infection.
  2. We can remove the intermediate host snail from the human water contact areas – Snail Control in local water contact sites.
  3. We can stop the eggs from reaching the water and warn people from going into known transmission sites – Behavioural Change Intervention.

 

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A typical transmission site for schistosomiasis. Families come to the water to wash, clean, fish, etc.

 

These are the three interventions we have available to us. What is the most effective way to eliminate schistosomiasis in an area?

 

In order to test this ZEST has randomly organised all the distinct community areas of Zanzibar and Pemba into our three intervention groups:

 

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A car full of donated Praziquantel treatment for schistosomiasis, about to head out to the communities.

 

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Collecting urine samples from children to test for the presence of schistosoma eggs. This is how we diagnose schistososmiasis.

 

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Spraying local water contact sites with a chemical that kills the aquatic snail host of schistosomes.


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This is a familiar face to you I’m sure, Dr Fiona Allan our resident schistosome snail expert surveying sites in Zanzibar. She has a sixth sense on where the snails will be and where transmission occurs. We are now calling her 'snail whisperer'.

 

1. Mass Drug Administration – Treatment of communities twice a year with Praziquantel. Now the truth is it would be unethical not to treat people we know to be suffering from the disease purely in the name of science. We may be scientists but we’re not evil scientists! So EVERYONE on BOTH ISLANDS IS GETTING TREATMENT. But in group 1 they are ONLY receiving treatment. No snail control, no behavioural intervention. This is to test the effectiveness of the current approach (treating people regularly).

 

2. Snail Control - Snail Control by spraying transmission sites with a safe and gentle dose of Niclosamide. The communities are receiving treatment as normal however their villages have been surveyed for human-snail water contact and schistosomaisis transmission sites. These sites then get sprayed with the molluscicide (chemical that kills snails) Niclosamide. Niclosamide is also used as parasite treatment for livestock and is safe for mammals and birds. It does kill all snails though so we only want to use it in the areas that have transmission, nowhere else. We also know that it quickly breaksdown in the environment. This is good because it means it does not linger around however it’s also bad because a good rain storm and off it goes down the river without killing any schistosome infected snails.

 

3. Behavioural Change Intervention – Mobilizing communities by teaching them about schistosomiasis transmission and supporting them to find their own solutions. Education teams go out to the communities, teach the village leaders, the religious leaders, the teachers about schistosomaisis and the blood fluke life cycle. They then help the communities to develop ways of raising awareness of schistosomiasis, educating parents and children and encouraging positive behaviour change that will prevent disease transmission. This has taken form of:

    • Special Kichocho (Swahili word for schistosomiasis) events, where safe games are played, little educational sketches are watched and fun is had.
    • Training teachers to teach children at schools about the schistosoma life cycle.
    • Building latrines and urinals for children and adults to use instead of urinating outside.
    • Making signs warning people of the presence of kichocho in the water and the risk of infection.
    • Other solutions like safe clothes washing areas etc.

 

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Big red signs warning the community about the presence of Kichocho (schistosomes) and konokono (snails - intermediate hosts of schistosomes) in the local water.

 

The behavioural intervention team on Pemba and Michael, an MSc student from the University of Tulane, with the help of the wonderful behaviour scientist Dr Bobbie Person, have created an amazing educational video in Kiswahili to show in villages and schools. Do take a look - it is fantastic!

 

 

A video made by the schistosomiasis behaviour intervention team on Pemba with the help of Michael Celone to teach communities about the life cycle of Kichocho (schistosomiasis). The video is in Kiswahili with English subtitles.


 

 

A second video teaching communities about behaviours that increase transmission and risk of infection as well as what they can do to prevent Kichocho (schistosomiasis). The video is in Kiswahili with English subtitles.

 

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The Zanzibar Elimination of Schistosomiasis Transmission study design. MDA – Mass drug administration of safe anti-schistosomal drug Praziquantel delivered to the villagers twice a year. Snail Control – removing snails in human water contact sites by spraying with safe molluscicide Niclosamide. Behaviour Intervention – Community-lead behavioural change intervention to stop behaviour that leads to transmission/infection of schistosomes.

 

Wish us luck and watch this space!

1

Hello honorary parasitologists,

 

I know there has been a bit of radio silence on my part and I apologise, summer was calling me and there was a lot of stuff to get through before I could escape on annual leave.

 

I'm back now and picking up where we left off. Perhaps you are wondering what happened to all those samples we collected in Tanzania in May (see previous posts). Well wonder no longer, I am about to reveal all.

 

A quick recap of our collecting in Tanzania:

  1. We collected miracidia, the parasite larval stage from infected children, and stored them onto special paper (called Whatman® FTA cards) that stores their genetic material.
  2. We also collected the intermediate host snail from potential transmission sites on the banks of Lake Victoria.
  3. Finally we collected cercariae, the larval stage from infected snails, and stored their genetic material on the Whatman® FTA paper.

 

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Visiting schools to identify infected children, collect the schistosome larval stage (miracidia) and treat the children.

 

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Snail collecting on the banks of Lake Victoria.

 

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Collecting cercariae, the Schistosoma larval stage, from infected snails.

 

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Whatman® FTA cards store the genetic material of schistosome larvae collected from infected children and/or snails.


 

Storing our collected schistosome larvae on Whatman® FTA cards is ideal for us because:

  1. We avoid storing them in flammable liquids like ethanol.
  2. We avoid having to bring back live, infected snails.

 

...two things that aiports and customs really don't like!

 

The Whatman® FTA cards lyse (break) open the parasite cells and lock the genetic material onto the card, keeping it stable and safe at room temperature until we need to use it. So this means we can bring back our samples safely in our suitcases. Hurrah!

 

The snails, on the other hand, have to be stored in glass tubes with ethanol, so these we have to leave behind in the safe hands of the National Institute for Medical Research, Mwanza. Our collaborators take good care of them until we can arrange a courier service to bring them to the Museum.

 

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Whatman® FTA cards with collected schistosome genetic material from Tanzania, catalogued and stored safely by SCAN in the Molecular Collections Facility at the Museum.

 

Once back in the UK I hand over all collected samples and forms to the wonderful SCAN,Schistosomiasis Collection at the Natural History Museum, team. SCAN takes care of the thousands of schistosome samples collected from all over the world and stored at the Museum. This colleciton is very precious and in high demand for lab-based scienists researching the genome of the parasite and host snail in search of new ways to understand and control the disease. 

 

SCAN cares for collected samples and manages all the associated data, such as:

  • GPS coordinates - so we know where the sample has come from.
  • Collection method - what technique was used to collect the sample?
  • Date of collection - how old is the sample?
  • Storage medium -  is the sample stored on Whatman FTA card or ethanol or any other storage tool?
  • Data on the parasite host - did the sample come from an infected human, cattle, snail? If a snail what species? If a human what age? Gender? 

And lots more.

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The fieldwork forms I fill in when collecting samples in Tanzania. I hand these forms over to the SCAN team along with all my collected samples. They then have the frustrating task of trying to decipher my handwriting.

 

You have met two members of the SCAN team in my previous posts; Fiona Allan, who acted as our fieldwork photographer whilst helping me in Tanzania and Muriel Rabone, who came to my rescue after Fiona had to head back to the UK. There is just one more person for you to meet; the ever-patient and resourceful Aidan Emery, who manages SCAN

 

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Fiona and I going through my fieldwork forms and samples - "what have you written here? It's illegible!" Oops!

 

As you can see there is A LOT of data that goes with each and every schistosome/snail collected and researchers need to have all this information when analyising a parasite or snail sample. SCAN ensures that all this information is properly entered into a database and linked with the samples stored in the Molecular Collection Facility (more on this in a bit).

 

The SCAN team has even created a wonderful online catalogue of all the collected samples they care for, along with all the data linked to each sample. This greatly assists researchers from all over the world, allowing them to have a look and see what samples are available to them. 

 

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Fiona and Aidan storing the Tanzanian schistosome samples collected onto Whatman® FTA cards. Fiona is showing off the little blue booties we have to wear in the Molecular Collections Facility to avoid bringing in contaminants or anything that could harm the hundreds of thousands of precious samples stored there. 

 

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The SCAN team takes a photo and measures the size of every snail that arrives from our African collaborators. In order to extract the DNA from the snail for molecular work the shell must be crushed and removed. It is good to have a picture of what the shell looked like before doing so.

 

The Molecular Collections Facility is a crucial facility in the Museum, as it has all the equipment necessary to keep molecular and genetic samples (such as our schistosome samples) stored safely, stabily and for a long, long time. What equipment am I talking about? I mean: -80 freezers, nitrogen tanks, air-tight cabinets, equipment to release/elute genetic material from stored samples, centrifuges, pipetting robots, you name it. It is run by the very helpful Jackie Mackenzie-Dodds.

 

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Jackie runs the Molecular Collections Facility where all our schistosome samples are stored. All the freezers and nitrogen tanks have alarms linked to them to make sure they continue to function correctly. If one fails an alarm goes off on Jackie's mobile phone so no matter where she is she is immediately notified and able to respond.

 

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Jackie is showing me the liquid nitrogen tanks in the Molecular Collections Facility. Whilst nitrogen in gas form is harmless, liquid nitrogen is very, very cold and any contact with it can cause severe frostbite, even freeze your arm off. Also as it boils it uses up a lot of oxygen in the air, which can lead to asphyxiation. So oxygen monitors are always used. Its incredible freezing ability means it is very effective at storing rare, degraded and old tissue samples.

 

So there you have it, all our samples are archived carefully until we are able to perform the molecular work we need to do for species identification and to determine how the genetic diversity of the parasites is being affected by treatment control programs. My next couple of blood fluke posts will be about the techniques we use to do this. So read up on Polymerase Chain Reactions (PCR)... it does feature!

 

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Fiona and I have managed to decipher my handwriting! Hurrah! The samples are saved!

1

Jambo from Mwanza, Tanzania

 

It's my last week here and as much as I have enjoyed the fieldwork I can't wait to get back home. But there are a few more days of work to do first!

 

Back to blood-fluke fieldwork

 

This time I want to tell you about our snail collecting work. Snail in Swahili is called Konokono. The snails we are interested in are aquatic, pulmonate little dudes belonging to the Biomphalaria genus.

 

They are the intermediate host of Schistosome mansoni, the blood fluke species responsible for intestinal schistosomiasis and it's detrimental health consequences in humans (see previous post - the Blood Fluke story).

 

We collect these snails in order to study the blood fluke parasites they carry.

 

The collecting process involves:

 

  • Scooping for snails on banks of Lake Victoria. We use protective waders to prevent blood fluke infection from the water.
  • Carrying the snails back to the lab, where we use microscopes to identify schistosome parasites.
  • Documenting the infected snails, which will be taken back to the Museum for DNA analysis.

 

 

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Aquatic snail of the Biomphalaria genus, host to the human blood fluke Schistosoma mansoni, the causative agent of schistosomiasis.

 

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Muriel and the team scooping for snails on the banks of Lake Victoria.


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Mr Revocatus and Mr James with the snail scoops and protective clothing (hip waders) to prevent blood fluke infection from the water. Credit Fion Allan.

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Village kids from local fishing village. Credit Fiona Allan.


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Mr Revocatus carrying the dredge to our next snail site. Yes this is a beach on Lake Victoria. Not the sea!


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Mr James with the dredge getting ready to collect those lake bottom snails. Credit Fiona Allan.


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Dredged up snails from the lake bottom. Now we have to find the small Biomphalaria species we are after. Credit Fiona Allan.

 

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Sometimes we have to work around the local fauna.

 

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More local fauna. Credit Fiona Allan.

 

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Activities by the snail collecting sites. This lady is drying small fish in the sun.

 

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Back in the lab, we sort through all our collected snails, put them in water and check for schistosome parasites.

 

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Biomphalaria snails in individual wells with water. We check each well for the presence of the parasite larval stage, cercariae.

 

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Blood fluke larvae (cercariae) under the microscope - those little white things in the water. They're looking for humans to infect!

 

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Identifying infected snails and giving them an ID number. We then preserve the snail in ethanol and bring them back to the Museum for genetic barcoding (species identification).

 

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After a hard days work, Muriel and James getting ready to tuck into some food.

 

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Me about to eat some Wali na Samaki (rice and fish).

0

Jambo from Tanzania,

 

I realise I'm a bit late with this post so lets get straight to it. Just a warning though, this will be a rather disgusting post so get ready to be grossed out.

 

You'll remember from my previous post about our school visits that we collect stool samples from infected children. This is because we collect the miracidia larval stage that hatches out of the parasite eggs. And these eggs come out with stool.

 

The blood fluke life cycle - a recap

 

Schistosoma. The worm pair releases schistosome eggs into the blood system. The eggs pierce through the wall of the intestinal/urinary tract and exit the host when he/she defecates or urinates. They reach fresh water and hatch out into a larval stage called miracidia.

 

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Life cycle of Schistosome, blood fluke parasite and the specimens we collect during our fieldwork (circled in yellow).

 

So in order to collect miracidia we need stool from infected children. Diagnosis of infection is achieved using the Kato Katz method: a specimen of stool viewed on a microscope slide. If schistosome (blood fluke) eggs are observed in the stool specimen then the person is infected with at least one pair of schistosomes. For more information on diagnosis have a look at this video.

 

Collecting eggs from stool

 

Once we know which kids are infected we go to the schools and get stool samples (see previous post). We take these back to the lab and then a long process of stool filtering begins. We filter the stool for schistosome eggs, these we place in water and light. This induces them to hatch out into miracidia. We collect the miracidia onto special cards that store their DNA. We transport these back to the UK.

 

We use a pair of filters called Pitchford funnels (devised by Pitchford & Visser). The inner smaller funnel has bigger pores that allow the schistosome eggs to pass through but stops larger pieces of stool. The outer funnel is made of a finer mesh with pores that stop schistosome eggs from going through, this allows us to pour lots of water through the funnel thereby washing the eggs of stool material that may stop them from hatching.

 

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Pitchford Funnels (devised by Pitchford & Visser). Credit Fiona Allan.

 

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Stool Samples. Credit Fiona Allan.

 

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Sieve to break up stool. Credit Fiona Allan.

 

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Using the sieve to break up the stool sample.

 

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Pouring stooly water through Pitchford Funnel. Credit Fiona Allan.

 

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Myself and Mr John processing stool samples. Credit Fiona Allan.

 

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Adding formalin to left over stool samples to kill of anything inside. These are disposed of safely later. Credit Fiona Allan.

 

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>Revocatus adding formalin to stool. Credt Fiona Allan.

 

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My Nagai releasing the eggs and some water into a petri dish.

 

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Petri dishes of eggs and water. Waiting to hatch.

 

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Fiona starts checking for miracidia swimming in the petri dish.

 

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Fiona and James in the lab in National Institute for Medical Research in Mwanza.

 

Sometimes out in rural areas where we use local hospitals to process our samples things can go wrong, such as a power cut. No electricity means no light through the microscope. Thankfully we rise to the challenge and strap our head torches round our microscopes as an alternate source of light. Not quite as clear but it still works.

 

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Even a power cut will not stop us, we use our head torches as a light source and continue working.

 

So that's it for now. Tune in for the next post - snail collecting on the banks of Lake Victoria.

0

Jambo (hi) from Tanzania!

 

We are now into our second week of the trip and the blood fluke parasite collection is going well. A few logistical hiccups but nothing we can’t handle (so far).

 

Last week we went to a few schools to collect schistosomes from infected children. Just to recap why and what we are collecting from schools, here is the life cycle with the stages we are collecting on this trip:

 

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The life cycle of blood flukes, Schistosoma, involving a vertebrate (e.g. human) host and an aquatic snail host. Transmission is through contact of infested freshwater. The yellow circles are the stages and specimens we collect when doing fieldwork.

 

So we have the delightful job of collecting the larval stage, called miracidia, that hatch out from eggs. How do we do this? We go into a school, collect stool samples from infected children and filter out the eggs. We then put them in some water in sunlight and wait for them to hatch. I will explain this in more detail in a subsequent post on lab work. For now let's stick to the first stage: visiting schools.

 

We visit state primary schools in the Mwanza region of Lake Victoria. To get to these schools we sometimes have to drive for hours through dirt tracks. All sorts of obstacles occur but the most common one is this: cattle!

 

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On our way to a school, a herd of cattle, goats and sheep block our path.

 

When we arrive we visit the head teacher and get a proper greeting from the school. The teacher then calls out our selected students - the ones we know are infected from a previous survey, more on this later.

 

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Children were practicing singing, dancing and music on the day we arrived. Credit: Fiona Allan.

 

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Up close and personal, the kids stare at us. Eventually we do get them to smile. Credit: Fiona Allan.

 

We’re a small team: two scientists from the Museum (Fiona and myself) and 3 research technicians from the National Institute for Medical Research in Mwanza - Mr John, Mr Nagai and Mr James. As well as our trusted driver – Mr Lenard.

 

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The team, Mr John, Mr Nagai and me. Getting our gloves on and our kit ready. Credit: Fiona Allan.

 

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My colleague Fiona Allan, a brilliant schistosome expert and our trip’s photographer.

 

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Me holding a football I am about to present to the headteacher as a present. Credit: Fiona Allan.

 

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Mr James is teaching the children how to give us a stool samples and most importantly to wash their hands afterwards! Good hygiene practice!

 

We give the kids a container to put a stool sample, and some toilet paper. They run off to the latrines and come back with a full container. How they are able to poop on demand always amazes me. We label the containers with unique identification numbers for each child. And then go back in the lab to process the samples. All the children in the school receive treatment a couple of weeks later. We always treat any infected child!

 

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Mr Nagai and Mr John handing out toilet paper to the kids. Credit: Fiona Allan.

 

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The children all grab for a container for their stool sample.

 

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School latrines. Credit: Fiona Allan.

 

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Mr James supervises the hand washing.


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We were very happy to see this in some of the schools: a warning about schistosomiasis, called Kichocho in Kiswahili, and an explanation about the life cycle. Credit: Fiona Allan.


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Some shots from the school. A little girl with a necklace of bottle tops, this actually serves as a abacus in the schools. Credit: Fiona Allan.

 

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Local abacus, device for learning arithmetic. Credit: Fiona Allan.

 

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Kids playing in front of a typical Mwanza rock.

 

This time I came with some gifts for the schools. Back in the UK I decided to get a football for each school. The footballs they use are often just rags and plastic wrapped into a tight ball and tied together, or completely deflated punctured balls. So I went shopping at Altimus. The staff and manager were curious about why I wanted 16 footballs. When I explained they very kindly gave me a generous discount. So this is a thank you to Altimus!

 

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Kids playing football with their old cloth ball.

 

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The new football next to the old football. You can see why the teachers and kids are delighted with the gift. Thank you Altimus.


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Girls playing basketball with the new ball from Altimus.


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Time to say Asante (thank you) and Kwaheri (good bye).

 

That's it for today. Next post - what do we do with poo and how to go parasite fishing with a microscope.

 

Asante sana (thank you very much in Swahili).