Virology Lab Technique Training

Virology Lab Technique Training

Basic Virological Techniques

18th November-23rd November

This past week I participated in a basic virology lab techniques course hosted by the Virology Research Laboratory at Manipal. Virology is a branch of science that deals with the study of viruses. Each day we learned about different techniques used in virology through lecture and hands on time in the lab. This week long course was one of my favorite experiences that I have had while in India. Since learning about scientific research on communicable diseases in South Africa, I have been inspired to investigate the field of virus research. I signed up for this course not sure what to expect but was blown away by the course material, which provided me with insight into a another whole area of academia. I had a blast working in the lab and look forward to doing more lab work in the future.
Although this blog post is a bit dense I wanted to provide you all with an idea of what we covered during the conference. I would be glad to talk with anyone that is interested about the specific techniques that we learned or the results that we obtained. I am still learning and hope that having the opportunity to talk about this experience on a scientific level will help me to understand it more myself! I have been told that teaching others is a great way to test your knowledge and better understand a concept.

Our schedule was as follows:

Monday

Introduction and Laboratory approach in Diagnostic Virology
Syndromic Approach in Diagnostic Virology and Sample Collection
Specimen Collection, Packaging, processing

Tuesday

Cell Culture
Virus Isolation

Wednesday

Haemagglutination Assay
Haemagglutination Inhibition Assay

Thursday

Quantification of Virus- TCID50
Neutralization Assay

Friday

Conventional PCR
Real time/ multiplex PCR

Saturday

Immunofluorescence Assay

In addition, there were a number of other techniques that I learned throughout the week including: how to preform a two-fold dilution, isolate RNA, navigate a level two lab, make liquids of a desired concentration and do a Plaque Assay. 

This image includes some of the instruments that we used on a daily basis in the lab. The bottles or flasks are used for cell culture, the trays with larger wells are used for techniques that involve staining such as a plaque assay which is used to  preform virus inoculation, the smaller tray in the back in a 96 well microtiter plate like the one we used for our hemagglutination assays, and the small tube on the right is a sample swab. 

Monday

On Monday we learned about the basic approaches to laboratory diagnostic virology, how to do syndromic diagnosis of diseases, and the processes for collection, packaging and transportation of samples. Diagnostic virology is used to increase awareness of viral diseases and is a major factor in: technological advances and modern management of infections. It is focused on the research of many public health issues such as AIDS, immunosuppressive therapy, and anti-virals.  There are a number of important factors that affect lab diagnosis of viral diseases including; specificity of the sample, transportation methods, selection of the appropriate assays, knowledge of disease kinetics, details of virus isolation, and clinical details. The basic approaches in laboratory diagnosis include microscopy (light and electron microscopy), antigen detection via. immunofluorescence assay or other assays, antibody detection through a neutralization assay or a haemagglutination inhibition assay and nucleic acid detection through PCR techniques.

Syndromic Diagnostic Approach in Virology:

Syndromic diagnosis is used by health care providers in order to quickly identify and treat a consistent group of symptoms that are easily recognizable. Treatment for such syndromic diseases is targeted towards the main organisms responsible for causing the syndrome. The syndromic treatment approach is preferred because it focuses on multiple symptoms at once, emphasizes direct doctor- patient interaction, and treatment is immediately initiated at the primary healthcare level. When we visited the public health facilities this semester we viewed the various guideline and protocol cards used by doctors to do syndromic diagnosis.

Major Clinical Syndromes-
1. Fever
2. Influenze like illness
3. Dengue like illness
4. fever and arthralgia (joint pain)
5. fever and rash (measles like)
6. Fever and parotitis (infection of the salivary glands)
7. Acute encephalitis syndrome
8. Adult respiratory distress syndrome
9. Acute jaundice syndrome
10. Acute diarrhea syndrome
11. Acute hemorrhagic fever syndrome

Specimen Collection, Packaging and Transportation

The specimen collection method is determined based on the syndrome that a patient exhibits.
When choosing a collection method there are a number of things to consider including; duration of viral shedding, type of virus, systemic involvement, the kinetics of the disease, and the age or immune system of the patient. Specimen collection techniques include respiratory samples, dermal samples, blood, CSF, urine, stool, saliva, conjunctival swab, and bone marrow. For the purpose of this course we focused on throat and nose swabbing. We were able to  practicing taking samples and packaging them for transport.

Tuesday

Tuesday we learned how to make a cell culture and perform isolation of a virus.

This is a video on cell culture.

Viruses are obligate intracellular parasites and require living cells to replicate. An essential step in propagating them in the laboratory is provision of cells in some way. The most widely used method is cell culture. Cell cultures are fairly easy to maintain and can have a wide host range. Cells in culture may be grown as a monolayer, a single layer of cells growing on a surface or in suspension in which cells multiply while suspended in a liquid media. Suspended cultures are used primarily for growing large volumes of viruses whereas monolayers are used widely in diagnostic virology for viral isolation. Though three types namely primary, diploid and continuous cell cultures are available, continuous cell lines are maintained commonly for the ease of maintenance and are most commonly used for experiments. Each cell line is chosen for a particular assay based on its specific qualities and the goal of the assay.
Cells grow best in the laboratory if the culture environment resembles the conditions experienced by the cells in vivo. Many types of synthetic media have been developed and modified to meet the needs of certain types of cells. Cell lines can be propagated or maintained dependent on the type of medium they are kept in. In virology, cell lines are chosen for their ability to act as hosts to specific viruses. For example, the Vero cell line that we used for many of our experiments was from the African green monkey kidney. We chose this cell because of its success in vitro.
Virus isolation can be done via animal inoculation, embryonated egg inoculation or cell cultures (the most common, safe, and ethically sound method and the method that we used).  Cell cultures are a convenient in-vitro method of virus isolation. There are three types of cell lines used in cell cultures; primary cell lines, diploid cell lines and continuous cell lines. In order to preform virus isolation one needs a tissue culture flask with confluent monolayer, a clinical sample with a suspected virus, maintenance media (solution with the necessary components for cell growth and division or life), and the necessary laboratory equipment (microscope, flasks, micro pipet, etc.).
In the lab we created a subculture of Vero cells. Subcultures are created when cells are taken from the maintained cell line and placed in growth medium in a new vial. After we had created a subculture we learned how to isolate a virus via cell culture inoculation. This means that we took a sample that was thought to contain a pathogen and added it to a cell line in a medium conducive to maintenace of cells. We then observed the cells over a number of days in order to determine whehter the virus had infected the cells. Cells that displayed the cytophathic effect; characteristic cellular changes such as ballooning and aggregation of cells, were suspected to have been infected by the virus and could be used for further studies.

Brain Break! (Don't worry we were using precautionary methods and inactive viruses) 

Wednesday 

On Wednesday we learned how to perform a hemagglutination assay and a hemagglutination inhibition assay.

Hemagglutination Assay (aka. virus titration)

The hemagglutination assay is a method used to quantify a virus in a sample or cell. Many viruses have a surface protein known as a hemagglutinin protein (HA) that binds to specific receptors such as those found on the plasma membrane of red blood cells. When red blood cells are mixed with the viruses containing HA protein (in the appropriate ratio) the virus bridges the red blood cells and changes their normal settling pattern creating a dense latex as opposed to a diffused monolayer. This processes is called hemagglutination. Hemagglutination assays are used after virus isolation and inoculation of a designated virus in order to determine the viral load.
Steps in a Hemagglutination Assay:

1. Prepare red blood cells  (RBS) and virus suspension
2. Add Phosphate buffer saline (PBS,  keeps the pH stable and prevents the cells from lysing) to all designated wells of a 96 well microtiter plate
3. Add the virus suspension to well 1
4. Preform a two-fold serial dilution, decreasing the concentration from left to right using the suspension in well 1 as initial source
5. Add fixed concentration and volume of RBS suspension to each well
6. Incubate the wells to promote binding of virus and RBS

Results:
The highest dilution of virus that causes complete hemagglutination is the endpoint of titration. One unit of HA is contained in the endpoint dilution of HA titration and is inversely equal to the total concentration of virus that was in the original sample. (aka. the endpoint is the highest dilution of virus that will still react with the red blood cells causing a latex to form, it is at this point that only one unit of HA protein is in the well therefore the concentration of the original sample is equal to the the number of times the dilution). A sample with an endpoint of 32 means that the original sample was diluted 32 times before there was only one HA unit and therefore originally the HA concentration was 32.

Hemagglutination Inhibition Assay:

If a person is infected by a virus bearing the HA protein, ant-HA antibodies may appear in his serum (sample collected may be blood, sputum, or other). These antibodies can block hemagglutination by binding to their target antigen and lead to the hemagglutination inhibition phenomenon. As a result no glutination will occur. Antibodies bind to the virus blocking binding sites on the viral surface and preventing the agglutination of the red blood cels. This assay is used to determine presence of antibodies in a patient serum by using an unknown serum and known virus or to determine the type or strain of virus by using a typing serum with known antibodies instead of a patient serum and an unknown virus isolated from a patient sample. In this case, the formation of the button or hallo is a negative result indicating that the serum did not have the target antibodies present. If the antigen antibody pair is a match, the red blood cells will not bind with the antigen and there will be no glutination.

This image is of the 96 well microtiter plate. If you look closely you can see the evidence of hemagglutination in the mirror; the red dots all the way to the right of the mirror image are "buttons" or congregates of red blood cells. The cells to the left are the more concentrated samples providing evidence that as the concentration of antibodies decreases the blood cells begin to bind with the antigens resulting in hemagglutination.  

Serial two-fold titration: a useful technique often used in diagnostic virology. Two fold titrations are used in order to manipulate the concentration of a sample (the antigen or antibody concentration).

Thursday 

Thursday we learned to preform a virus neutralization assay. We also learned how to quantify a virus using TCID50.
Neutralization of a virus is defined as loss of infectivity through reaction of virus with specific antibodies. Neutralization assay is primarily performed in order to quantify the concentration of antibody present in a sample serum. In vitro testing of virus neutralization is preformed by incubating the known virus concentration with the sera containing antibodies. Residual viral activity is detected by its ability to subsequently infect cell cultures or other host systems. Antibody concentration is equal to the reciprocal of highest dilution of serum that shown 50% or greater reduction in cytopathic effect (50% or less neutralization). The results of this study are viewed using specific staining techniques in which the the cells that have been infected by the virus are stained and the cells that have not been infected are washed away. This process is used to identify viruses, differentiate between closely related viruses, and determine specific antibody response following infection/ vaccination.
Tissue culture infective dose 50 (TCID50) is a viral quantification technique. It determines the titer of infectious virus present in a viral stock until neutralization occurs. The TCID50 technique determines the highest dilution of virus that infects 50% of cell cultures as evidenced by the cytopathic effect. The results of the TCID50 titration (infection ratio, cumulative # of infected and not infected cells, and percent infected) are entered into an equation that determines the quantity of viral load in the original sample.

This image is of a 96 well microtiter plate used in TCID50. The wells all the way to the right and left are cell controls. They are stained dark purple because they were not infected by the virus. Their purpose is to insure that the cells were viable and correctly fixed without any experimental variables. Wells 1, 2, and 3 do not have any cells because the cells that were present were infected by the virus. Infected cells are not fixed with the addition of ethanol and can be literally washed away by water whereas infected cells rows 5-10 are fixed and are therefore stained. Row 4 is evidence of the success of the TCID50 titer, some of the cells have been fixed and stained( were not infected) while others were washed away (infected by the virus). It is up to the experimenter to determine the % of infected cells in each well. Remember, the TCID50 assay determines quantity of a virus based on the viruses ability to infect > 50% of cells at different dilutions. 

This day was particularly challenging as I had to recall calculous from Mrs. Niller's math class in 11th grade (not my brightest moments). It was a lot of learned but I thoroughly enjoyed myself!

Friday

On Friday we did Conventional Polymerase Chain Reaction (PCR) and Real-time PCR. Polymerase chain reaction is a technique used to amplify a target gene sequence in vitro using the cells "built in" mechanism for replication of DNA. Conventional PCR is used for disease diagnosis, molecular epidemiology by genotype (hereditary DNA diseases), drug resistance studies, phylogeny analysis, DNA fingerprinting and studies of genetic variability.

A gel electrophoresis technique is used to view the results of conventional PCR. The amplified target sequence is loaded into an agarose gel and an electrical current is run through the gel propelling the negatively charged DNA away from the anion and towards the cation. Larger segments of DNA take longer to navigate through the agarose gel while small segments of DNA easily move through the well resulting in a separation of DNA segments by size. This technique is used to determine the success of PCR by identifying the target sample of known length. 

In comparison, real-time PCR is a specialized technique that allows PCR reactions to be visualized "in real time" during each cycle by using fluorescent dyes or probes. The advantages of real-time PCR include the ability to monitor the progress of the PCR reaction as it occurs, quantification of target sequence present based on the nucleic acid concentration, the specificity and sensitivity of the test, and the speed of procurement of results. The amount of target sequence during amplification is quantified by using fluorescent dyes which increase in a logarithmic pattern until the threshold concentration (point of maximum amplification) is reached. The concentration of virus is found by determining where the threshold and log phase intersect. This point, known as the Ct value, is inversely related to the amount of target sequence present.

Saturday 

On the final day of our course we learned how to preform an immunofluorescence assay. Immunofluorescence facilitates the rapid detection of viral antigen in clinical specimens and viral isolates. It is useful for the detection of respiratory viruses such as herpes sinplex virus, cytomegalovirus, and varicella zoster virus. This technique is based on the phenomenon of photo-excitation and subsequent emission of light molecules produced by fluorochromes or fluorophores (light absorbing and emitting molecules). Antibodies can be molecularly attached to fluorochrome molecules for experimental purposes. In immunofluorescence, antibodies are chemically conjugated to fluorescent dyes. Fluorochrome-conjugated antibodies against specific viruses can detect their protein antigens in clinical specimen and infected cells.  This concept is used in order to view antigen- fluorochrome-conjugated antibody complexes under UV lights. When molecules such as fluorochromes absorb light their electrons are excited to a higher energy state, as the electrons fall back to their original energy state they emit energy in the form of light. A high energy wavelength of light will be absorbed causing excitation of the electrons, the return of the electron to its original energy state emits excess energy in the form of a lower energy wavelength of light.  Immunofluorescence can be used to identify the presence and quantity of virus in a sample as the antigen(virus) will be bound to fluorochrome-conjugated antibodies and can be viewed using microscopy.

The end!

If you are interested in learning more about the lab that I was working in you can visit the departments webpage: http://www.manipal.edu/institutions/universitydepartments/departmentof%20virusresearch/pages/drgarunkumar.aspx

Inspirational Public Health Efforts by Peace Corps Volunteers

Malaria Eradication through Education: The Peace Corps Efforts in Uganda, Africa 

One of the reasons I love learning about public health is because I feel like I can be useful in a world where so much 'bad' occurs on a daily basis. Ever since junior year of high school when I completed a thesis project on the oppression of women in third world countries, I have been dedicated to figuring out ways in which I could 'help'. After taking a number of anthropology courses and talking with advisors, parents, and friends throughout college, I realized that this desire could be best fulfilled through a focus in public health. My public health education has been challenging and exciting and I have learned a lot about the reality of people's health and the disease burden around the world. This news update about work by volunteers in Uganda is inspirational. Despite the global initiative to eradicate Malaria, it is 90% endemic in Uganda and 100% of the population is considered at risk. The efforts by the Peace Corps are one of the many examples of ways to promote public health in the area through education.


The following links are for the article on the Peace Corps efforts in Uganda. They are great examples of creative ways to disseminate important public health information! (Hopefully the originals were written in a local language.)

Current News Update from the US Gov:
http://iipdigital.usembassy.gov/st/english/article/2013/11/20131107286094.html?CP.rss=true#axzz2jpIaOcb4

To see an online version of the book go to:
http://issuu.com/raegan_albright_spencer/docs/soa_and_the_moka_boky_english_versi#

On Malaria:

Malaria is caused by a parasite and transmitted vie the bites of infected females mosquitoes. Once in the human host, the parasite replicates in the liver and infects red blood cells decreasing their efficacy as oxygen carriers. Due to its dependence on the mosquito the disease is most often a problem in places where humans live in close contact with the environment and there is stagnant water. The best way to fight diseases such as Malaria are through education and adequate surveillance. Although much of the developing world has eradicated the disease through political and individual commitment, it is still a major burden for parts of the global south including parts of sub-Saharan Africa.

ISN'T IT FASCINATING!

Course work and research: The academic side of study abroad

As the last month of my time abroad begins it is bitter sweet. Over the past few months I have developed a life here and feel as if everything is just falling into place yet it is almost time to return home and I am looking forward to seeing family and friends.

The past couple of weeks, I have been extremely busy with classes (papers, presentations, midterm examinations, reading). In addition, I finally got approval to do independent research, and I signed up for a course in Basic Virology Lab Techniques. I feel as if each of these opportunities will provide me with the type of academic experience I was hoping to have and will be beneficial to my future goals. I am very excited about the work I am involved with in and out of the classroom!

My independent research is on the treatment seeking decisions of people in this region. We are focusing on the common alternative/ complementary therapies widely practiced in India including: Ayurveda, Yoga, Unani, Siddah, and Homeopathy (AYUSH). We will be conducting a qualitative study in the communities surrounding Manipal by interviewing individuals of known treatment regime in order to gain a better understanding of why they chose said system.

I have also started to do research for my final paper in my Contemporary Indian Culture Class for which I will be focusing on Sexuality in India in particular homosexuality among men. I have found some very interesting articles that discuss the theories on varying acceptance of homosexuality in different countries. I chose to do my final research paper on this topic because it is one area of society that is taboo and unapproachable yet is extremely apparent in everyday life. Unlike in the United States, it is common for men to share physical exchanges such as hand holding or close contact yet scandalous for a man and a woman to do the same. I am interested in learning about the historical context in which these social norms evolved and how they have changed with globalization and changes in religion. There is so much that is involved in this topic and I am looking forward to slowly peeling back the many layers to explore the problems and theories at the root of the issue.

If anyone is looking for a good book to read I highly suggest "Em and the Big Hoom". Em and the Big Hoom takes place in the urban city of Bombay (Mumbi) and addresses many of the taboo issues in a very unique way. If you have never read an Indian novel it might be beneficial to start with Karukku, a memoir by a Dalit woman, in order to get a better understanding of rural life. Both memoirs are easy reads and provide great insight into the lives of people on the ground. Of course, it is important to read with the understanding that it is just one persons experience and point of view and can not be accepted as the "norm". I would love to have a literary discussion with anyone who reads either book when I return.

I hope that everyone is keeping well and enjoying the crisp days of fall!
~Tata 

"Unity in diversity"

Unity in Diversity 

If there is one thing I learned about this trip it is that there is no "real india", only many faces.

Ten Days of Travel 
Official Schedule

"If you want to make God laugh, tell him your plans"- Woody Allen

Friday, October 4: Mangalore to Pondicherry 
A/C Bus to Mangalore 2:00 and then overnight train to Pondicherry from Mangalore Train Station. 

Saturday, October 5: Pondicherry

Stay in Pondicherry at Gratitude Guest House

Tour of French Quarters "White Town" 

Visit Heritage Sites 

Sunday, October 6: Pondicherry

Visit Sunday Market

Visit Aeroville Ashram

Heritage walk of Pondicherry's Tamil Quarter

Monday, October 7: Mahabalipuram 

(Take bus about 2 hours to Mahabalipuram)

Visit Krishna's Butter Ball, Arjuna's Pennance, Mahishasuramarthini Cave, Five Rahas, and Shore Temple (all near each other)

Tuesday, October 8: Travel Day from Mahabalipuram to Bangalore w/ stop in Vellore

(Take bus 3 hours to Vellore)

Visit the Golden Temple 

(continue 5 hours to Bangalore by bus, arrive 8:00 pm)

Dinner @ Hard Rock Cafe 

Wednesday, October 9: Bangalore

Tour of AOL India Huffington Post Office (Coolage!)

Visit Lalbagh Botanical gardens

Shopping in City Market

(Sleeper Class Night train to Coimbatore leaves bangalore 11:45 pm (7 hours) then bus to Ooty (3 hours)) 

Thursday, October 10: Ooty

Arrive and check into Kings Cliff HotelVisit Kota Village 

Friday, October 11:

Walking tour of Tea Plantation 

Visit Second Highest Peak in Southern India 

Tour Tea Factory (Nilgiri Tea)

Saturday, October 12: 

(Travel Day from Ooty to Mysore- Public A/C Bus 4 hours to Mysore)

Check into Sandesh the Prince Hotel

Visit the Mysore Palace at night and Devaraja Market 

Sunday, October 13: 

Visit Chamundi Temple

Tour Mysore Palace 

(Night train to Udupi, arrive at 10:18 AM Monday)

Monday, October 14: 

Arrive Manipal 11:00am 

Sleep...

Each day we were allotted free time to do whatever we wanted. My friends and I spent a lot of time exploring our surroundings, shopping, and enjoying good food! 

I learned that it best to travel with no expectations because then I am never disappointed. 

Pondicherry (Puducherry)

            

 

Pondicherry is a small city colonized by the French which retains much of its French influence due to the work of the historical society in the area. There are two sections to Pondicherry divided by what is now a sewage channel; White Town and the Tamil District. White town (the actual name since colonization), is well organized, quaint, and historic. It has well planned streets and many French flares such as architecture and cuisine. There is a historical society that works with the home owners in the area to plan renovations in order to maintain the historic feel of the city. The Tamil District is a bit more like some of the other cities that we have visited in India. The streets are filled with vendors, rickshaws, and cows all accompanied by the noise and smells of a city. This part of the city has not been maintained in the same way as White Town but it does have its own charm.

Heritage Site Buildings in the French Quarters of Pondicherry

During the evening the street next to the board walk along the coast is shut down and there are festivities including dance acts and talent shows. The board walk is similar to one that you would find in the US with vendors selling food, balloon animals, and trinkets. This was the first festivity of the sort we had seen since being in India and it was fun to see the crowds of people out and about enjoying the warm night and cool ocean breeze.
Our guest house in Pondicherry was my favorite location that we stayed in. The 
guest house was two stories with a veranda on the roof and an open court 
yard in the middle. Each of the rooms was styled differently and decorated with antique furniture and beautiful linens from FabIndia, one of our favorite stores. Each room smelt like lemon grass and was kept air-conditioned. The hotel was a wonderful place to return to after a hot day of walking around the city. Each morning we had a continental breakfast with curd (yogurt), granola, fruit, fresh croissants, omelets, and a few traditional indian dishes. We ate and slept like kings!   

On the board walk in Pondicherry

Chillin at Gratitude 

Mahabalipuram

Our visit to Mahabalipuram was quite a learning experience. Although it was cool to see the rock carvings we did not have much luck with the tour guides and wound up getting scammed. In addition our hotel was not a very pleasant experience. Despite the less than ideal conditions my friend and I managed to make the best of it and I have fond memories of staying up late playing cards and chatting. 

Krishna's Butter Ball

The Cat Story

Arjuna's Penance

  

Bangalore

As we drove into the big city it was as if we were in New York. It is a wonder how cities all over the world are so similar. The one morning in Bangalore I decided to go for a run. I left the hotel at about 7:30 and was told to just ask directions if I got lost then preceded to do exactly that. Running through a city is a novelty that I do not wish to become accostumb to. As a country girl I was unprepared for the amount of weaving and stopping necessary to navigate the busy streets and found it rather challenging to keep myself oriented. Although I didn't know where I was most of the time I did enjoy seeing a bit more of the city and exploring roads off of the beaten path.

Ooty

The Second Highest Point in Southern India

Ooty was may favorite town that we visited. In order to get there we took an overnight sleeper class train from Bangalore to a city about 3 hours away from Ooty then took a bus into the hills where the Western and Eastern Ghats meet. The drive into the mountains was one of the most eerie drives I have ever taken as each hairpin turn on the narrow road seemed almost impossible to make with our large bus and oncoming traffic. 
The town of Ooty is settled in the hilltops of the Nilgiri District. most of the economy is dependent on the vast tea plantations that are spread across the high elevation peaks. While staying in Ooty we had to opportunity to spend one morning walking through a plantation and the surrounding area. This morning was my favorite of the trip as we got to trek through the hillside at leisure enjoying the crisp mountain air. I have discovered that in India it is rare to find a place where the air feels clean and there aren't crowds of people and I greatly appreciated this literal breath of fresh air. 

Tea Plantation                                     

On our walk through town

                                                   
That afternoon we toured a tea factory where they processed the tea leaves grown in the region. The factory was extremely interesting. I was amazed by the simple process used to make tea and the quick turnover rate from plantation to package. 

The Second Highest Point in Southern India

Tea Production Process: 

Harvesting

Withering

Rolling

Fermentation

Cut, Tear, and Curl

Fermentation

Drying 

Green tea: The manufacturing of green tea is similar to manufacturing of black tea except the leaves are dried in pans over high heat of steamed in vats instead of being withered. These two alternate processing methods destroy the enzymes which cause fermentation.

White Tea: Is not machine processed or treated. It is simply fermented, hand rolled and sun-dried. Only the most delicate and newly formed buds are used to make white tea.

The Kota Tribe

We also spent a day in a Kota Tribal village. We ended up in this particular village through a retired Manipal professor who has been working with the village for a number of years. While in the village we played with the kids, were taught how to spin pottery on the traditional pottery wheel, saw the local temple, and listened to a traditional music performance and danced with the musicians. It was a great day and was, as always, wonderful to be on the ground interacting with people. Of course, as an anthropologist I cant help but question the consequence such visits have for members of the village. I only hope that they enjoy our company as much as we enjoy theirs.  

We stayed at the Kings Cliff Hotel in Ooty. The hotel has maintained its British inspired decor and atmosphere and was absolutely charming. We had every amenity including fire places in our rooms that were lit each evening. In addition, the restaurant at the hotel has been rated the best restaurant in Ooty (aren't we lucky)! Being in the chilly weather made me nostalgic for New England autumns and made me think about all of my friends and family back home! I hope everyone is enjoying the colorful leaves, apples, and golden 5:00 sun. :) Although it only lasted a couple of days, it was a wonderful respite from the heat and humidity of the lower elevations in Karnataka and Tamil Nadu. 

Mysore

Silk Shop!

Mysore is city that is smaller than bangalore however it felt more crowded because we spent more time in the main parts of town; walking around the palace, through the market, and around the center of the city. In addition there was an exceptionally large number of people due to the holiday, Dashara, taking place the day after we left. This was the first time during my stay in India where I had anxiety about the large crowds however, I managed to navigate the masses with my friends and made it out safely! 

The Mysore Palace

In the Market Place

       

Bangles on Bangles on Bangles!

I really enjoyed the first night we were in Mysore as we had time to walk around the palace lit up at night and explore the market, bargaining and testing our hands at haggling.

The Many Faces of India

One of the biggest "AHA" moments I have had thus far occurred during our travel week... 
After seeing so many parts of India it became apparent to me that contrary to common belief, India is made up of many states that each have a unique culture, people, and economy. On our trip we experienced the French Colony of Pondicherry with its planned city, french food, central park, and restored buildings. Mahabalipurham, a classic ocean side beach town complete with fishing boats and easy going "townies". Bangalore, a typical city and Ooty, a beautiful hill camp. Each place or 'face of India' was a different perspective of Indian life. Unlike the stereotyped portrayal of India, I learned that you don't have to be in a small tribal village to experience the 'real india'. With this revelation I realized that my experience at Manipal is just as 'real' as any other. I truly appreciated the opportunity to see the many faces of India on this travel week and enjoyed each of the places that we visited for the educational and social aspects they offered. 

Reflection

Reflection 

After almost two months of being in India I have done a lot of reflecting on my experience thus far. Traveling abroad is a very unique experience; you are immersed in a new place with a very different culture and you have to navigate new situations and overcome obstacles in a setting where you may not be able to communicate through verbal language. In addition, you are thrown in with a new group of people and have to figure out everyones personalities and navigate group dynamics. Many times throughout this journey I have felt alone and uncertain as to my purpose for being here, however; I have realized that it is exactly this scenario that makes studying abroad such a unique and beneficial experience. Only when one is completely out of their comfort zone is it possible to truly see the world from a different perspective. I have decided that this opportunity is worth pursuing and will be a learning experience even if it is not what I had expected. During the past couple of months I have found solace in friends; new and old, family, running, school work, and yoga. I greatly appreciate all of the support I have received and hope that you all know you are in my thoughts. As we are learning in yoga, the real challenge is to overcome any obstacles in life with grace and humility.  

~Namaste

Birthdays and Beaches

Birthdays and Beaches

WOW! I could not have asked for a more special way to spend my 20 birthday than to be in India. As one of the many September babies in our program I celebrated my birthday numerous times in different ways... 1st I was able to celebrate at Namma Bhoomi with the school children, 2nd I was able to celebrate with my (almost) birthday twin and lovely friend Coral, and 3rd I was able to celebrate on the day of my birthday.

Our trip to Namma Bhoomi was not only a time of celebration but also a time of making relationships. The first trip that we made to Namma Bhoomi was in August, during which we learned about the purpose and framework of the school (see previous post about NB). This time our purpose was to spend time with the children and work as a team to provide a unique and memorable experience for them.

During the morning (10:00am-12:45am) we were separated into 4 groups; measuring, coloring, games, and gardening. I chose to help with the gardening project. The school strives to be self sustainable and the garden is one step aimed at reaching that goal. For our part, we worked with a group of about 25 children to clear a plot of land, level out the surface, and line the edge of the garden bed with large stones. This work was both physically and emotionally rewarding. We not only were able to break land and see the garden bed come together but we were also able to bond with the children over our blisters and herpetophobia  (fear of reptiles or creepy, crawly things). There is no better feeling than getting a little dirt on your kurta in the process of giving back to an organization that has done so much for such wonderful children.

We broke bread with the students in their dinning commons and the meal was fabulous! After lunch we spoke with one of the program coordinators about the work done in the classroom. Namma Bhoomi focuses on democracy and giving children autonomy. Their education model is similar in some ways to the Montessori model in that; the children have the freedom to choose what they work on each day, the classrooms are mixed ages, concepts are learned through working with materials and teaching tools, and teachers are there for guidance not lecture style learning. We were invited to support the school by develop teaching materials such as flash cards, educational games, counting tools, and language props. I am thinking about developing tools for teaching health education or biology!

In the afternoon, we were divided into small groups and  explored our artistic sides. One group made masks, another did henna and the third danced. The children had an absolute blast and we did as well! At the end of the day we all met outside and the children sang us happy birthday in english and Hindi. Then we shared home-baked cookies and said our goodbyes. 

Beaches 

On Sunday, September 14th, a few friends and I traveled to Malpe beach, located in Udupi just around 20 minutes from Manipal. It was a beautiful and sunny day and I got my first bit of color as we walked up and down the beach. The water was lovely and it was a great way to celebrate. 

On my birthday I went on an adventure with my roommate, some mutual friends, and a friend that I met while traveling. We went in search of a large waterfall around an hour and 45 minutes away from Manipal. Once again we didn't ever make it to the falls but after a long trek we found a farmer who showed us to the river somewhere in the middle of the forest. After a challenging bus ride home via. public transportation, we went out to dinner at our favorite local restaurant and got oreo milkshakes at a local fast food joint. It was a fabulous way to spend my birthday; low key but it is always lovely to spend time with friends.  

the River

Local rice farm

Walking through a local rice farm following the farmer to the river. 

More than just a preschool: the Anagwadi Center

Anagwadi 

This past week we visited an Anagwadi in Udupi. Anagwadis are part of the primary level of health care and are community based. Their main goals are to: enhance the health and nutrition of children under 6 years, reduce infant mortality, morbidity and malnutrition, decrease school drop out rates, and lay a foundation for physical, mental, and social health. In order to meet these goals the Anagwadi provides a number of services to the community including; supplementary nutrition, health check ups, immunizations, basic preschool education, health and nutrition education for mothers, and referrals for specialists. Most of the work that is done at the Anagwadi and via outreach programs is done by the Anagwadi teacher, a honored and respected woman in the community.

The Anagwadi Center

The Anagwadi facility was clearly a place of learning and enrichment. It appeared well kept and safe and the two staff members; the teacher and the helper controlled the children in a friendly way. I was impressed by the respect the children displayed for the women’s authority and felt that it was clear that the teachers were looking after the best interest if the children. The facility was also academically stimulating. For example, the walls were covered in numerous visual teaching tools such as; images portraying important lifestyle choices associated with good health, the alphabet in Hindi and English, and the numbers 1-15 in the form of easy to remember pictures.

Some of the children at the center. After learning about the mission and organization of the center we were able spend time playing with the children. They showed us a number of songs and dances in Hindi then we taught them the makarina! 

This poster is a visual tool used to teach the children about important aspects of life in the village such as; agriculture, fruits and vegetables, work, and clothing. The children are taught both Hindi and English at the Anagwadi and most learn the local language while at home. 

The children learn their numbers 1-15 by drawing creative and easy to remember pictures. For example, the 5 is drawn as an apple and the 4 is drawn as a popsicle. 

In addition to education, the Anagwadi also provides health care for the children. While at the Anagwadi children are weighed, provided supplementary nutrition, and given the appropriate vaccines. In addition, a doctor visits on a monthly basis and referrals can be given to the community at any time. This is the weighing mechanism for the children. They were all excited to be put in the swing and told that they were a healthy weight!

This is a chart of all of the mothers, pregnant women, and women
of childbearing age in the community. It is the teachers
responsibility to keep tract of the members of the community
so that she can effectively provide healthcare for
the target group. 

I was also impressed by the community outreach coordinated by and through the facility. The food supplements to be distributed to the community were well-organized and kept in dry covered containers. In addition, the services offered to mothers’ pre and post delivery were quite impressive. The surveillance and monitoring of the mothers creates a relationship through which the Anagwadi teachers can relay important health messages to the community. Providing mothers with support during and after pregnancy provides the teachers with the opportunity to relay relevant information about health related to pregnancy and lifestyle, which benefits the community as a whole. Community education seems to be a major goal of the Anagwadi and is carried out through biweekly meetings and information sessions. I would be interested in learning about the attendance rate at these meetings.

These packets are examples of food supplements that would be given to the mothers on a monthly basis. There are certain packages that are given to women that are breastfeeding, women that are pregnant, and children that are malnourished. 

Although this Anagwadi was extremely well outfitted and effective, when we talked about the Anagwadis in other districts there was not the same success. Like many of the notional programs implemented on the community level, success varies greatly based on location and other social factors. There is a lot of effort through the instillation of top down management to try and homogenize success, however there are limits on the effectiveness of “outsider” intervention. I would be interested in testing the efficacy of a teaching movement in which respectable Anagwadi teachers traveled to other districts where there has been less success to teach the Anagwadi workers. The goal of their teaching would be to spread their passion for helping to achieve health in the community. In order for the movement to be successful, they would have to instill the seed of selflessness and pride for their work. If successful the movement would change the mindset of those who are exploiting the system and decrease maleficent behavior. If there was a way to decrease the corruption of the system by individuals through increasing education there might be more success stories in more districts across the country.