Frog dissection

University Animal Research Day, 29.3.17

On 29th March, I attended a day at The University of Manchester’s school of biological sciences to learn about the use of animals in science and the role of veterinarians.  

Introductory lecture

After registration, I sat in a lecture theatre for an introductory talk about animals in biomedical research and how this is utilised. We did not sit down for long before being asked to stand up, and when an applicable personal statement was called out we had to sit down. ”Have you ever had general anaesthetic, have you ever used an inhaler, have you ever taken antibiotics, have you ever taken allergy tablets”. Unsurprisingly, everyone was sat back down again. This highlighted the significance that animal research has on our lives, because they have an integral role in drugs trialling.

But with the videos and photos (anti-testing propaganda?) repeatedly emerging on social media, why on earth would we choose to test on animals? The academics I met had grown up aspiring to have a role caring for animals, they are not emotionless beings who want to inflict pain on another creature. It is important to test drugs in the complexity of a living body, with multiple complicated systems. In vitro alternatives are overly simplified models and cannot show any adverse side effects to a whole organism. The law also states that before clinical trials, the drugs must be tested on at least 2 different species. This is a valid argument against the suggestion that animal testing may be ‘unnecessary’, because pharmaceutical companies need to know if there are any side effects, what dosages people should use and most importantly if they are safe. As our anatomy and physiology is similar to animals, animals are relatively accurate models. In addition to this, mice have a quick generation turnover and are small and easy to handle. At 6 weeks old they are sexually mature and gestation is only 3 weeks. I was pleasantly surprised to hear how controlled the testing is, with each project requiring 3 licenses from the Home Office with regular inspections under the Animal Scientific Procedures Act 1986. By law, if there is a realistic alternative than the animal research cannot commence. Researchers also have to follow the 3 Rs (replacement, reduction and refinement).

NC3R implement the 3Rs as framework to support science, innovation and animal welfare. It was established in 2004 by the government. I learned that guidelines depend on the neurophysiology development of the species so this is no ‘speciesism’ or favouritising certain animals. This is why fish are lower order than rodents which are lower order than primates, which have a higher level of protection.

The 3Rs are from a Russell and Butch book, and we were briefly taught about the meaning and purpose of each R.

Replacement: methods that avoid the use of animals should be explored, from established animal cell lines to mathematical and computer models. Immature vertebrate forms (embryonic/foetal) are not protected under ASPA regulations.

Reduction: this is to minimise the number of animals used in testing. If researchers share data and resources, smaller populations can be used to give reliable data. Imagining for longitudinal studies can be used to follow disease progression, so a test can have an earlier termination time. As a result of the reproducibility crisis with 85% of research being wasted, the ARRIVE guidelines were produced. This is a 20 item check list of what should be included when a research paper is published such as gender and weight of specimens. A lot of stakeholders endorse ARRIVE guidelines to report in researchera’ manuscripts in order to receive their funding. This reduces the animal research waste and promotes experimental design.

Refinement: this is to minimise the pain, suffering and/or distress of an animal. This includes the basic husbandry and environment to the scientific procedure itself. I later saw that the fish had artificial coral and the rodents had tubes, as enrichment. This keeps them mentally stimulated. EDA is one of NC3R’s projects to make the research process easier by producing flow diagrams of an experimental design. This helps scientists be more conscious of animal numbers, and the logistics of their plan.

In addition to promotion of the 3Rs, NC3R fund research projects. At Newcastle University, Grimace Scales were produced as a new way to assess pain in animals. These currently widely distributed posters are a series of photos of rats, mice and rabbits with indicators of facial expressions associated with pain. In order to alleviate pain, researchers need to asses pain. Therefore, this product benefits millions of laboratory animals worldwide.

We were then given an insight into the UK’s animal testing industry through a set of statistics. Rodents constituted 72% of the testing, in 2016 1.1 million mice were tested on. This sounds like a large statistic but it was put into perspective because 220 million animals are killed by domestic cats each year, 2.5 BILLION animals are killed for food each year. One opinion is that we would not restrict life saving drugs if a loved one was going to die, but we could choose not to consume animal products.

Before the open day, I thought animal testing was mainly invasive surgeries but I was informed that ‘mild’ procedures such as a change in diet and behaviour observation are the most common.

Introductory lecture
After registration, I sat in a lecture theatre for an introductory talk about animals in biomedical research and how this is utilised. We did not sit down for long before being asked to stand up, and when an applicable personal statement was called out we had to sit down. ”Have you ever had general anaesthetic, have you ever used an inhaler, have you ever taken antibiotics, have you ever taken allergy tablets”. Unsurprisingly, everyone was sat back down again. This highlighted the significance that animal research has on our lives, because they have an integral role in drugs trialling.
But with the videos and photos (anti-testing propaganda?) repeatedly emerging on social media, why on earth would we choose to test on animals? The academics I met had grown up aspiring to have a role caring for animals, they are not emotionless beings who want to inflict pain on another creature. It is important to test drugs in the complexity of a living body, with multiple complicated systems. In vitro alternatives are overly simplified models and cannot show any adverse side effects to a whole organism. The law also states that before clinical trials, the drugs must be tested on at least 2 different species. This is a valid argument against the suggestion that animal testing may be ‘unnecessary’, because pharmaceutical companies need to know if there are any side effects, what dosages people should use and most importantly if they are safe. As our anatomy and physiology is similar to animals, animals are relatively accurate models. In addition to this, mice have a quick generation turnover and are small and easy to handle. At 6 weeks old they are sexually mature and gestation is only 3 weeks. I was pleasantly surprised to hear how controlled the testing is, with each project requiring 3 licenses from the Home Office with regular inspections under the Animal Scientific Procedures Act 1986. By law, if there is a realistic alternative than the animal research cannot commence. Researchers also have to follow the 3 Rs (replacement, reduction and refinement).
NC3R implement the 3Rs as framework to support science, innovation and animal welfare. It was established in 2004 by the government. I learned that guidelines depend on the neurophysiology development of the species so this is no ‘speciesism’ or favouritising certain animals. This is why fish are lower order than rodents which are lower order than primates, which have a higher level of protection.
The 3Rs are from a Russell and Butch book, and we were briefly taught about the meaning and purpose of each R.
Replacement: methods that avoid the use of animals should be explored, from established animal cell lines to mathematical and computer models. Immature vertebrate forms (embryonic/foetal) are not protected under ASPA regulations.
Reduction: this is to minimise the number of animals used in testing. If researchers share data and resources, smaller populations can be used to give reliable data. Imagining for longitudinal studies can be used to follow disease progression, so a test can have an earlier termination time. As a result of the reproducibility crisis with 85% of research being wasted, the ARRIVE guidelines were produced. This is a 20 item check list of what should be included when a research paper is published such as gender and weight of specimens. A lot of stakeholders endorse ARRIVE guidelines to report in researchera’ manuscripts in order to receive their funding. This reduces the animal research waste and promotes experimental design.

Egg Practical

The first practical was intriguing as I have already opened up the incubated eggs that were unsuccessful in hatching, to identify the stages of foetal development. I learned that in a laboratory environment, for experimental purposes such as the egg practical, that it should be younger than half of the gestation of the animal. Chicken eggs have a large role in learning about the development of humans in the womb. They are a developmental model; our genomes share homology, cellular developmental processes can be examined and it is easy to obtain the specimen.

In this specific practical, we were opening eggs and timing the heart beat of a chick embryo to investigate how temperature affects heart rate. After plotting all of the groups’ results, there was a directly proportional relationship with the x axis of temperature and y axis of heart rate. This is due to the increased metabolic activity. If we had conducted an experiment with a wider range, exceeding the highest temperature, then the enzymes would have been denatured.

Animal Lab

After changing into an all-in-one and placing shoe covers on, our group was given a tour of the animal laboratory facilities at the university. Although other students who are not accustomed to the smell of animals thought the rodent area had a stench, it was incredibly clean and smelt far less than anticipated due to the tight sanitation controls. I saw the rodents and zebra fish before going to the farm area where there were 5 sheep in a pen, with sufficient space. I asked lots of questions about controversial topics such as the euthanasia methods, hearing that it is all heavily controlled my regulations and qualifications.

Ethical Debate

After the tour, we sat back down in a lecture theatre to be briefly presented with the history of the animal testing ethical issues and how these have changed over time.

The emotional connection to an animal used to be the determining factor for animal testing. Before a review of the law, octopi were considered as an ideal animal to test on due to the ease of catching them on coasts. It wasn’t until their intelligence and mental ability was discovered that they became a protected animal.

Fly Practical
I was intrigued to hear that fruit flies (Drosophila melanogaster) had a key role in the development of human medical knowledge. They are inexpensive and low maintenance. There is a large fly facility in the university. In order to identify mutations, they have genetic markers. We had to look down a microscope and identify a series of genetic markers on fruit flies.

We then did an experiment to see which flies would walk up a test tube faster (the younger or the older flies). Another set were epileptic, so by shaking the test tube, these flies were knocked out. It amazes me that these tiny creatures have made such a huge impact in medicine and understanding diseases and development.

Abattoir

Being a veterinarian is about facing life and death every day. A day in a slaughterhouse opened my eyes to the rigorous legislation and practices that go on behind the scenes.

Due to the increasing demonstrations, and tightened bio security, many vet school students are turned away when locating their compulsory abattoir placement. Therefore, as someone with animal welfare at the forefront of their life, I will share my experience.

Enforcing regulations to safeguard public health to ensure food is safe for human consumption is one of the main roles of an OVS (official veterinary surgeon). There are many rules and regulations under UK and EU laws. RSPCA welfare standards apply and the animals must be slaughtered humanely. Therefore a vet has to be on site during slaughter.

When I arrived at the abattoir, I put a boiler suit and hair net on before walking out of the office. When I was walking between buildings, it was required to put my coat on over the white boiler suit. This is to prevent food contamination. Thoroughly washing hands and wellies was very important.

The first load for slaughter were two Dexter calves as the pigs being unloaded in the side pen was not allowing the calves to remain calm. So the workers decided to take the cattle in first, this was to minimise distress, because animal welfare comes first.

Under the 96/23/EC directive, a selection of samples must be sent to the VMD every month for residue testing under the National Residues Control Plan. The vet had a final sample to take for March- a cow blood sample, so this was collected during the first slaughter of the morning. This was to test the testosterone levels. Monitoring levels in animals sent for human consumption is extremely important to police suppression times which is the period after an animal was last administered a veterinary medicine before it can be sent to an abbattoir/milk can be consumed. It is important to safeguard food with the responsible use of medicine. Use of antibiotics also results in antibiotic resistance and someone may be allergic to for example penicillin.

However, before the slaughter process, once the animals are loaded off the wagon, the correct paperwork must be presented. Animals should be correctly identified.
The vet then conducts an ante-mortem inspection to assess animal welfare, if an animal is safe for human consumption along with other things. The livestock must be clean enough to reduce the risk of meat contamination too. The animal can then be moved from lairage to slaughter.

A penetrating captive bolt was used into the skull and the calves dropped to the floor unconscious. They then rolled out of the bottom of the box and a slaughter man cut the animal’s throat which led to rapid blood loss. A hind leg was then shackled to hoist the calf up. The head was then cut off and placed on a hook to be inspected. The feet were removed and the calf was skinned. Another worker then used a large saw to cut through the brisket. The organs were removed and the meat inspector looked at the organs and carcass before stamping them. But before this, the meat inspector opened the lymph nodes in the head to look for signs of disease, for example bTB. If it had not advanced and was localised, then a region could be removed and the carcass would be safe for human consumption. All of these inspections are done as legislation outlines.

I then stood in the stunning pen whilst four pigs were stunned with a large pair of tongues which clamp either side of the brain, I saw that they were immediately unconscious. There was a 20 second time period that the pig had to hoisted up and bled out whilst they are unconscious. I saw how animal welfare was paramount, the pigs were very calm and the process lasted seconds. After they have bled out, the pigs are placed in the hot bath. Then rolled about to remove the hair, a torch is then used to burn any remaining hair off before having their testicals removed. This is to prevent any contamination with the meat. They are then butchered and the removed organs are assessed by the meat inspector. I saw how pneumonia affects the lungs of a pig which was extremely interesting as I have treated calves for pneumonia. A pig also had an abscess removed, so the leg will be butchered off.

When a wild boar came down the line, a sample of striated muscle was taken from its diaphragm for trichinella. The weanlings that came down the line also had samples taken. Trichinosis can affect humans and can lead to death if consumed in under cooked meat. If the animal is too small to provide an 11g sample, the tongue can be taken instead. Before the carcass can be butchered, the vet needs to hear back from the test for an all clear.

When more cows came down the line, a deceased cow was brought in with documentation of a farm vet of being shot on farm. Although animals should be alive when they arrive, there are exceptions, for example if an animal is unfit to travel. However, it must be butchered within 2 hours. And if it is older than 48 months, it is tested for Bovine Spongiform Encephalopathy.

Vet course, Avian flu team task, 12.3.17

VetQuest was an opportunity to visit one of the universities offering Veterinary Science. The first lecture was by an academic working in admissions, exploring the range of careers in veterinary medicine as well as the academic and work experience expectations.

From teaching, to pharmaceuticals to working for APHA… there are many roads you can go down following a veterinary degree.

My first timetabled workshop was preventing disease spreading, about the topical avian flu. Following the work I have done on the farm, this was a great opportunity to see the working relationship between a private vet and an APHA vet. As a group, we had to decide what we would do in hypothetical situations where vets are under immense pressure to make choices regarding controlling outbreaks. We decided what protective clothing a vet should wear: a face mask to prevent inhalation of pathogens, of dust and toxic ammonia from hen faeces, 2 boiler suits, boot covers and gloves. Two of our team members had 3 minutes to get changed into the appropriate clothing, preparing to go into the pretend shed 4. The given scenario was that a farmer raised an alarm over his shed 4, as he believes many of them are presenting clinical signs.

Avian influenza is extremely infectious, and kills birds very quickly, so it is important for a vet to react swiftly. Before they ‘went to the farm’, the vet asked whether vets should assess the sheds in the order 1234 or 4321, I raised the issue of going to 4 first as this is the suspected infected group so there is a risk of spreading this to to others so for bio security reasons, go to 4 last.

The two team members acting as the initial call our private vets then left the room and a phone rang. I picked up the phone and had the role of the APHA vet, speaking on behalf of the rest of the group. I asked the vet to provide information on the number of animals on the farm, the living conditions and structure, if all of the sheds were affected, how many animals have recently died, if there has been a movement of new animals in etc. I concluded that the hens needed to have blood taken before confirming this is avian flu and to ensure the hens remain quarantined in their shed to prevent spread in the case of only one shed being affected at this moment in time.

This is what it is like for farm vets, being outside of a closed practice means communication is key.

As soon as I put the phone down, a question with a timer came up on the interactive whiteboard… to cull all of the hens in all four sheds or to wait 24 hours until the blood tests are back. The group were split, but I strongly believed in waiting for 24 hours given the circumstances. Culling would not happen over night, so the 24 hours is not a preventable measure for the suffering for the animals.

The blood tests came back and showed positives for viruses but negative for the ELISA antibody test. This is because avian flu would kill the hen before their body produced the antibody immune response.

Cronkshaw Fold Farm, Cupboard clear out, 8.2.17

In preparation for the kids to be born, I cleared out, cleaned and organised the medical cabinet finally… it is a job I have been wanting to do for a while as I love to be organised.

First of all, I removed everything out of the and sorted through what needed to be thrown away.

I then labelled boxes and arranged all of the equipment and medication into sections before writing a stock list.

  

I then found the wormer ‘Superverm’ and estimated the weight of two of the goats. Using a syringe, I measured 14ml of the wormer. Holding the goats head upwards, I put the syringe into their mouth and tried to stop any of the medication running out… I can definitely get better at that!   
  

Cronkshaw Fold Farm, Biology + Goats = Awesome, 6.2.17

I am very fortunate to have a great biology department who provided me with the tube to collect some of the goat lice to take in to study under a microscope. 

Opportunities to combine my scientific skills and goats are great, so I am looking forward to taking the lice in tomorrow.

After finishing the evening jobs, the first task was to get the lice into the tube, which sounds easier than it is. I was shocked at how effective the spot on treatment was, and this is the only time I would be gutted that lice had gone. There were a few remaining dead lice, however, so using Dot’s tweezers (let’s hope she doesn’t get goat lice in her eyebrows), I carefully picked out some lice.



Mildred certainly did not appreciate being held on my knee. It’s times like this that I wish I had three hands… One hand holding the goat still, one hand using the tweezers with phone torch balanced so I could see where the lice were.

  
I also got my first fan mail, which was very cute.

*7.2.17 update*

I took the lice into sixth form and looked at them under the microscope. It was really interesting!

Cronkshaw Fold Farm, Goat health checks, 4.2.17

When I arrived at the farm I did the usual morning jobs of feeding the goats a mix of granola and sheep nuts, changing their water, strawing down their pens and filling up the hay nets.

After piling up the straw in the isolated goat pen, I learned to not let my projected human empathy overrule the animal’s needs; it is important to think like the animal. As I could see that the goat had lost condition, it was natural to feel sorry for the poorly goat and attempt to provide it with more comfort. However, goats require hard stand to keep their hooves in tip-top condition. Accumulation of faeces and moisture in the straw is a recipe for poor hoof condition, something that needs to be particularly avoided in a weak goat. Lesson learned: think like a goat.

When I walk into the pens to pour the feed into the troughs, the goats like to jump up to try and take the bucket out of my hands. As great as goats are, they do not have manners. Also, the billy goats are beginning to fight more, they will establish a pecking order especially as there is a pregnant doe in their pen.

Although the doe (Bella) is a big softie who likes human attention, she was separated from the other females for being a bully. She threatened the other does’ pregnancies as it is important that they are not butted or stressed.  She is still a lovely goat in my eyes, unlike Gretchen and Regina who would happily drag me across the barn in an attempt to lead them into hoof treatment baths!

Holly and I then fed the guineapig and took layers pellets up to the duck shed.

Due to the recent outbreaks of bird flu on a game farm in Preston, the ‘prevention zone’ has been extended until 28th February 2017. Extra biosecurity measures are necessary in order to prevent contact with wild birds (potential carriers of avian influenza). So this morning the hens got bread rolls and vegetables as a treat, spreading these around the hen house is stimulation for them. Surprisingly, they prefer the granola-filled feeder to the layers pellets.

Today’s main job was to begin to make individual profiles for the goats whilst doing thorough health checks. This was the perfect opportunity to give the awesome goats some awesome names and the not-so-nice goats a matching evil name.

Starting in the main pen, we caught the goats one by one- which is harder than it sounds, especially when half are pregnant and stubborn. Never underestimate goat power.

Here is an example of the general health check before I go into details of the interesting findings and a funny misdiagnosis.

Health check

As my camera roll of 2,000 animal pictures takes up my phone memory, Holly was responsible for taking all of the photos and they are brilliant! Each goat had side profiles, a head-shot and a teeth photo. This is so we can I.D the goats, it is important to know your livestock so we agreed that naming the animals would help.

Firstly, we wrote about their colouring and markings, whether they are disbudded and any distinguishing features (e.g. beard or wattles).

Teeth

I held the goat’s mouth open whilst Holly  took a picture. We used the number and type of teeth to age the goat, so health checking all 15 goats gave us considerable practice.

In the first year of a kid’s life, the 8 teeth that grow are small and sharp. Just like humans, these fall out to be replaced by ‘adult’ teeth. A yearling loses its two front teeth and the permanent teeth grow. At two years old, the second pair of permanent teeth will grow (adjacent to the two middle teeth). This pattern continues until the goat reaches four years old, when there are 8 permanent teeth.

Exceeding 4 years old is when ageing a goat gets more difficult, because an estimation is made on the wear on the teeth. The gum will recede and the teeth appear longer, hence the phrase “long in the tooth”. 
Eyes

Continuing with the facial features, we checked the goats eyes.

A few things to look out for:

•Blindness: a goat will seem disorientated and lack blink reflexes

•Irritation: this may be caused by an external irritant such as dust, but in the case of infectious irritants a goat should be isolated as the infections are extremely contagious

•Ulcers

•Discharge

•Redenned scelera (area around the cornea): pink eye


Nose and ears

A goat’s nose is an indicator of some upper respiratory tract infections, such as pneumonia and nasal tumours which can cause sinus infections.

A few of the goats had slightly crusty areas on their nose but none had significant discharge that would indicate a problem.

However, some more research is required to find out what the small brown patches on the inner ear skin are. I suspect it is lice droppings after discovering the lice. Vaseline will help to moisturise the dry skin and after the case of the scaly legged hens we have plenty of Vaseline in the medical cupboard!


Hooves

We examined the hooves for foot scald and foot rot by looking at the interdigital space.

I have never seen it so white before, in the past the skin has been pink and infected. Therefore, the zinc sulphate baths have been highly successful.

The hoof outer wall has started to grow under the sole but the overall shape of the hooves were good. It was a pleasant change to not stink of rotten hooves.
Coat

As a whole, the coat condition of the goats was okay- all bar one had no bald patches or thinning and no cuts or red skin.  

On close inspection, we could diagnose pediculosis (lice infection). Although Holly and I knew that there was an ectoparasite infection, we compared photos to examples online to confirm that these are in fact lice rather than mites. I will write a more detailed blog posts about lice and the treatment plan on a later date.


Bum

When the goats’ diet was changed to granola from sheep nuts, a few of the kids had dietary scour (diarrhoea triggered by this change). This can cause dehydration but infectious scour damages the gut villi. By holding the goat’s tail upwards, we looked for signs of scour.

Apart from a few scabby bumholes, the goats were clear!

We then looked around the pens at the faeces to see the colour and consistency. As soon as we caught a few of the goats, they pooed on command which was helpful.

Finally we assessed their behaviour.

These photos definitely highlight the different personalities of the goats. Now we have named the goats, I plan to write a ‘meet the goats’ blog post!

Chestnut is very friendly and enjoys human contact, compared to other timid young goats.

Carlos is crazy and crazily in love with Holly. However, he is overly confident and boystrous. I do love how he wags his tail and acts like a dog… That’s why I named him after my crazy border terrier.

This is Regina. Yes, like Regina George she likes her own way! She falls to the ground when anyone tries to move her. 

Findings

On close inspection, I could see little parasites crawling across the goats. Every goat in each pen had a lice infestation.  

 

I administered 5ml of spot on to treat the lice. I will then use powder as a preventative. 


Unfortunately, Scabius has taken a turn for the worse and has lost condition. Although her facial scabs have healed, her coat is thinning and her skin has deteriorated.   

Floss the sheep dog also bit her up the bum after a goat v dog altercation! So she has a bright blue backside…

Regina the evil goat’s side profile looked abnormal when we were taking the side shot so I felt the lump to discover that it moves. It felt like fluid.

This could be the start of bottle jaw which is a symptom of anemia. In goats, the most common cause of anemia is Barberpole worms. The FAMACHA is an indicator of the severity of anaemia based on the colour of the eye mucus membranes.

Therefore, the goats will be wormed!

When I assessed the pen for the second time, I saw that the cause of the fluid build up will have been from the goat hitting her jaw on the divider between the two goats.
As they have not been outdoors, there is no way that she could have been infected by Barberpole worms.
However, I still wormed some of the goats in poor condition.

 

 


Dissection, 26.1.17

As part of the biology specification, our class did a required practical of a lamb heart dissection.

We used scissors to cut through the heart and a glass rod to find routes through the blood vessels.

First of all, I looked down the top of the heart to see the semi-lunar valves in the aorta and pulmonary artery. I also located the pulmonary vein and vena cava.

Using scissors, I cut 3cm above the apex of the heart. The two holes are where the ventricles are. The hole with the large amount of surrounding muscle is the left ventricle. This is required to maintain the blood pressure as it is pumped around the whole body.


I then cut through the pulmonary vein to see the inside of the heart more clearly. 
During the lesson, I was allowed to dissect two chicks I had brought in. It was interesting to see the differences in the lungs between the two chicks as one had died after hatching and one could not break the membrane to hatch. I cut the legs off to make it easier to open up the chick body. 

Here is the tiny heart…


As we had just learned about animal gas exchange, I located the trachea. I knew by the cartilaginous rings around it.


Finally, I cut the eye out and located the brain!