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Investigation of threonine dehydratase feedback control using enzyme mutants

Posted by Lisa @ Medlink Students on December 7, 2017 at 1:25 PM Comments comments (0)

Investigation of threonine dehydratase feedback control using enzyme mutants

Introduction:

Negative feedback inhibition is a process where the end product in a biochemical pathway binds with the allosteric site of the first allosteric enzyme in the pathway,the the enzyme then undergoes a conformational change causing it to become inactive (Storey, 2005). This ensures that whilst the end product is available in the surroundings, it is unnecessary for the cell to continue with the process and so it is innactivated until the end product is not available and blocking the allosteric site. This is required to regulate cellular biochemical pathways in order to maintain a healthy cell. Previous studies have explored the importance of understanding the allosteric mechanisms in these biochemical pathways, they have shown that it is of scientific and industrial importance in the development of effective production strains (Lin Chen, 2012). Studies on regulating biosynthesis of isoleucine were one of the first examples of this mechanism. Isoleucine is the end product of a pathway of which threonine is the initiator which is catalysed by threonine dehydratase to form α-Ketobutyrate (I. T. Szamosi, 1994).Threonine dehydratase binds to a seperate site on threonine than isoleucine.TD activity is regulated by negative feedback and is inhibited by the end product isoleucine, (Möckel B, 1994) and activated by the metabolite valine which is from a competing branch . Valine is from a competative pathway where pyruvate reacts with hydroxyethyl-TPP to go through steps and form leucine and valine, valine can also activate threonine dehydratase by binding to its allosteric site (D Travis Gallagher, 1998), this can be shown in experiment by using enzyme mutants.

 

Zoo Keeping: Risks, PPE (Equipment), Caring for Animals Well-being

Posted by Lisa @ Medlink Students on November 24, 2017 at 4:30 PM Comments comments (0)

South American coati.

Equipment How to use it? What to avoid? Tips

Gloves/gauntlets

Gauntlets must be worn at all times to prevent bites and scratches. Gauntlets are for direct restraint of the coatis. Do not restrain animal if the gauntlets are damaged or are loose and could fall off. Gauntlets can be tricky to use because you lack sensation and this can make it difficult to determine how hard to grip the animal.

Crush cage

Crush cages would be used to administer medication such as injections. The crush cage is a versatile piece of equipment because it can be used as a carry cage but also has a mechanism that allows the handler to push the animal to the back of the cage to prevent them

Only use the crush cage mechanism when it is necessary, such as for administering medical treatment that requires the animal to be still. Crush cages are a stressful method to use and should only be used as a last resort for important medications.

Towel

Use the towel to wrap around the coati so that its legs are restrained and nothing but the head is exposed. The tail can hang freely but may get in the was so if you do wrap the tail as well then care must be taken to not damage it. Best done by someone who is experienced in this. Need 2 people to carry this out.

Royal Python.

Equipment How to use it? What to avoid? Tips

Snake Hook

Place the hook so that it is on its side and slide this under the middle part of the snake’s body. Gently lift the snake from the ground and for extra control you can use our other hand to hold on to the snake’s lower body.

If you are trying to handle the snake by the head you must place the side of the hook at the back of the snakes head where the neck joins the head and hold firmly but not too hard. Avoid by accidently crushing the snakes neck if you are trying to restrain its head. Hold the hook with your dominant hand.

Tube

To place the snake into the tube you need to have its head restrained so you can navigate it into the tube. Do not let the snake pass all the way through the tube by always holding onto its body when its head is inside the tube. Hold the tube with your dominant hand and allow the snake to slide through your other slightly so that its head is somewhat in the middle of the tube before gently gripping the snakes body.

Pillow case

A pillow case is a simple way to restrain a royal python as its one of the least stressful ways to go about restraining them. To use the pillow case you need to simply place the snake inside the case and twist the top so that the snake cannot escape. Make sure that all of the snake’s body is inside to case before you tighten or twist the end. Label the case so that it is clear to anyone that could see the pillow case that there is a live animal inside.


A. Explain the importance of keeping up to date and accurate records in the zoo – with examples. What records are kept, why, how, why are they critical? Explain clearly how record keeping relates to animal welfare.


Keeping up to date records for animals and staff in zoos is essential for the health and welfare of zoo animals and staff alike.

Examples of what can be recorded for zoo animals.

• Pregnancies.

This notifies keepers whether the animals are able to breed.


• If they have been fed or not.

This is important for the keepers to know as it prevents other keepers over feeding or under feeding animals if they are kept up to date.


• Veterinary treatment administered, if any?

Keeping an accurate recording of this prevents an overdoes as a keeper could give an animal medication twice because its record was not updated when it last had medicine.

• Suitable restraint equipment.

 

• Additional notes that may want to be made by staff.

There are so many miscellaneous details that may be mentioned by zookeepers but do not have a category for them to place it.

 

• Training staff.

Recording the training of staff lets head keepers know whether staff are competent enough to work with certain species or not. This prevents inexperienced keepers from working with species their not use to and potentially hurting themselves.

 

• Animal training.

All recordings of animal training must be up to date to ensure that the keepers know what the animal has learnt and not learnt.

 

• H&S

Health and safety measures used must be recorded to show what has been done to protect staff, public and the animals.

 

• Risk assessment

A risk assessment must be carried out for all the areas that the staff work to prevent hazards. This can also be done for the animals enclosures to prevent hazards for the animals in their enclosures.

 

• Next of kin detail what if staff are injured? Slips n trips.

Details of staff family or partners should be recorded in case they are injured at work to notify them what has happened and what action has been taken for example: sent to hospital.

B. Next you need to describe records of restraint procedures specifically and relate this to chemical sedation in a zoo. You could use examples and show that you know what is recorded, when and why.

Chemical sedation is a form of restraint where a drug is used to restrict the movement or sedate a zoo animal for ease of transportation and/or to carry out medical examinations.

What to record:

• Chemical used

What chemical being used is important to know because then it can be estimated how much to use and what the effects will be on the animal.

• Amount of chemical used

Recording the amount of the chemical used is important because it can then be looked back on in case an emergency occurs to prove that the correct amount was used.

• Time

Time of day is important because it would be easier to sedate an animal that is nocturnal during the day because they are less active at this time and helps time when they wake up with when they would naturally wake up in the wild which can lower stress experience when they do wake.

• Licensing

Dart guns require special licensing for someone to be qualified to use it. This is known as a firearms license and must be up to date otherwise a prison sentence of 6 months could be faced.

• Licensed Veterinarian.

It is essential that whoever is administering the chemical must be a licensed veterinarian as this licensing proves that the veterinarian is skilled enough to be able to identify the appropriate type of chemical used and how much for a specific animal.

 

C. Finally describe records for animal movement/transportation, animal transfer between collections and legal record requirements in the zoo.

 

In the UK zoo animals will be monitored by either the European Endangered species breeding programme (EEP) or the European Studbook (ESB).

 

The EEP gathers information from the zoos which have the species which come under its authority. The point of monitoring these species is to maintain genetic diversity and coordinate where each animal goes depending on what species the zoo is keeping.

An EEP coordinator will look at the results from their data that has been collected and will establish recommendations to zoo on where to transfer the animals. The zoos must cooperate with these recommendations as the EEP owns these animals.

 

When an animal is being transferred it is usually done by an external transport company, a trained member of staff must be present to supervise the transfer. They may assist with unloading and loading the animals and monitor its environment.

 

 

 

 

When transporting animals there is legislation involved that all zoo must abide by:

BALAI Directive

Balai registered zoo – good disease control.

Sweeps up all the other animals not under EEP and ESP and keeps them all on an up to date database that is similar to EEP and ESP.

 

 

 

Welfare of Animals Transport (England) Order

This legislation insists that certification is provided for the animal and certain recordings are made before transport is carried out to ensure that the animals five needs are being met before, during and after transport. The recordings should state:

• Time is kept to as low as possible.

• The animals are fit to travel

• The staff are trained and competent

• If the vehicle is suitable

• When Water, feed and rest were last given

• Is there sufficient space for the animal?

 

Because this legislation is an extention of the Animal Welfare Act England 2006 it enforces the five animal needs to be met in as best way possible during transport, which are:

1. The need for a suitable environment.

2. The need for food and water.

3. The need to be housed with or without other animals.

4. The need to be free from injury, disease or pain.

5. The need to express natural behaviour.

 

 

 

 

 

 

 

 

International Air Transport Association (IATA), Live Animals Regulations

This legislation has species specific guidelines which state how to conduct the transport for the animal.

This includes:

1. Design and construction materials used for the transport crate such as walls, floor, doors, ventilation, perches, labelling, handles, locks etc

2. Feeding and watering guide

3. General care and loading

e.g. is it best to keep the animal in the dark?

Convention of the International Trade in Endangered Species.

When transporting animals, the zoo must have a license for transporting the animal under this legislation. Animals that come under this legislation are categorised under Appendix I, II, and III.

If an individual animal being transported comes under Appendix I then the zoo must have an article 10 document for that one specimen.

Animal health licenses.

Rabies license import licenses.

Import notification.

Export certificate.

 

Conclusion.

Recording everything is essential for the benefit of everyone involved in zoo keeping as there is a very strong risk when sedating any animal that they could be harmed. When we make recordings we are then able to look back on everything that has taken place, if for example an animal is harmed or even killed during sedation we are then able to look back at records and could potentially find what was wrong with the procedure when it was carried out such as an overdose.

 

HOW TO UNDERTAKE ANIMAL HANDLING AND SAFE WORKING, ADVANTAGES AND DISADVANTAGES

Posted by Lisa @ Medlink Students on November 22, 2017 at 11:45 AM Comments comments (0)

ANIMAL: PIG

EQUIPMENT: Trailer, feed bucket and a board.

ADVANTAGES: Trailer: trailers are secure and allow the animal to feel safe. Trailers are well ventilated and have a ramp to make loading easier.

Feed bucket: feed buckets are good for enticing the animal and making the pig have a positive association with the handling.

Board: boards are good for keeping the handler safe and allows more control of the pig.

DISADVANTAGES: Trailer: can be stressful for the pig going into the trailer, overcrowding can occur, requires planning, expensive to buy and hard to store.

Feed bucket: the pig might not have any interest with the feed because it is stressed or the pig might not be used to being fed at an unusual time.

Board: Boards can be very intimidating for pigs and are heavy.


ANIMAL: DONKEY

EQUIPMENT: Head collar: gives the user good control of the donkey, has an adjustable size and can come in different colours for identifying animals.

Feed bucket: Gives the donkey a positive association with handling.

Board: protects the handler and easy to use.

ADVANTAGES: Head collar: the donkey might not be used to head collars and may become distressed when it is being put on. This will make it hard to put on the donkey. Head collars can slip off and break easily.

Feed bucket: the donkey may associate the bucket with negative experiences of being handled in the past and may become restless.

Board: donkeys may run into the boards and harm themselves and they can be heavy.

DISADVANTAGES:


ANIMAL: SNAKE

EQUIPMENT: Snake hook, Pillow case, tubes.

ADVANTAGES: Snake hook: very useful for

Pillow case: calms the snake as it is dark inside the pillow case.

Tubes: when a snake is in the tube it is completely restrained and easy to handle.

DISADVANTAGES: Snake hook: requires training to use.

Pillow case: snakes can bite through the fabric.

Tubes: it is hard getting the snake into the tube.


ANIMAL: GOAT

EQUIPMENT: Head collar, feed bucket and board.

ADVANTAGES:

Head collar: head collars can be hard to put on the goat if the goat is not accustomed to it and could stress the goat.

Feed bucket: the goat may associate the bucket with negative experiences of being handled and become restless.

Board: goats that are not accustomed to a board may panic and run into it harming themselves and maybe the handler.

DISADVANTAGES:

Head collar: head collars can be hard to put on the goat if the goat is not accustomed to it and could stress the goat.

Feed bucket: the goat may associate the bucket with negative experiences of being handled and become restless.

Board: goats that are not accustomed to a board may panic and run into it harming themselves and maybe the handler.


ANIMAL: GUNEA PIG

EQUIPMENT: Carry cage, soft mat and table.

ADVANTAGES: Carry cage: very secure.

Soft mat: very easy to use and prevents the risk of unwanted contamination on surfaces such as tables, pet carriers and operating surfaces.

Table: easy to move, stable surface, requires no training and is a suitable height for ease of handling.

DISADVANTAGES: Carry cage: carry cages can knock into things and scare the guinea pig.

Soft mat: can tear easily.

Table: there is a risk that the guinea pig could fall off the table and harm itself.


ANIMAL: AGGRESSIVE DOG

EQUIPMENT: Mikki muzzle, Catch pole and gloves.

ADVANTAGES: Mikki muzzle: prevents the dog biting and protects the handler.

Catch pole: gives good control to the handler and allows for there to be distance between the handler and dog.

Gloves: protects the user from potential bites and scratches.

DISADVANTAGES: Mikki muzzle: hard to put on a dog if it is aggressive and is very difficult for the dog to drink, eat and pant. Requires different sizes.

Catch pole: requires training to use and can harm the dog if it is used incorrectly.

Gloves: reduces sensation for the handler and may lead to the handler being too firm or gentle when handling a dog.


ANIMAL: Cats, Rabbits, small dogs and small mammals.

EQUIPMENT: CARRY CAGE

ADVANTAGES:

• Animal cannot escape once they are in the carry cage.

• Can easily see where the animal is.

• Well ventilated.

• Easy to clean.

Allows the animal to see their environment.

DISADVANTAGES:

• May require training for the animals to become accustomed to it.

• Not insulated.

• Can be heavy.

• Can be stressful for some animals as the cage may make the animal feel exposed.


ANIMAL: Horses, Goats and Pigs.

EQUIPMENT: TRAILER

ADVANTAGES:

• Suitable for a range of animals.

• Well ventilated.

• Can move more than one animal at a time.

• Have ramps which make loading easy.

DISADVANTAGES:

• Requires planning.

• Can become overcrowded.

• In a confined space which doesn’t allow natural behaviours.

• Expensive.

• Hard to store.


ANIMAL: Snakes, Lizards and Birds.

EQUIPMENT: PILLOW CASE

ADVANTAGES:

• Calms the animal.

• Easy to use.

• Cheap.

• Warm, breathable and soft.

• Easy to wash.

DISADVANTAGES:

• Animals can bite through the pillow case.

• Can become soiled easily.

• Hard to see where the animal is.

• Not very secure.


ANIMAL: Snakes, Lizards and Tortoises.

EQUIPMENT: POLYSTYRENE BOX

ADVANTAGES:

• Cheap.

• Insulating.

• Secure.

• Able to write on the box.

DISADVANTAGES:

• Requires breathing holes.

• Has to be taped.

• Lids can break easily.

• Doesn’t absorb any spills.

How to feed Animals and accommodate their diets

Posted by Lisa @ Medlink Students on November 20, 2017 at 10:05 AM Comments comments (0)

Species: All animals require different nutrients from different sources and at different times of the day or night.

Time of feeding: Some animals are nocturnal which means they can only be fed at night while some animals are diurnal and so means they must be fed during the day to meet natural rhythms.


What to feed: Animals are ordered into three main dietary categories. Carnivore (Eats meat), Omnivore (Eats both animal and plant matter), Herbivore (Eats plant matter) and other sub groups of those three categories. This is the biggest factor affecting an animals feeding plan.


How to feed: Depending on an animals diet will depend on how the feed should be given:

Carnivore: Carnivores like lions will usually hunt for their food in the wild but because it is illegal to feed live vertebrates it is possible to feed pre killed animals. If the animal eats insects like a Leopard gecko then the feed can be live insects. Carnivorous animals require high protein in their diet.

Herbivore: Herbivorous diets are easy to present to an animal because there are many ways the feed can be given. Herbivorous animals need high carbohydrate and fibre in their diet.

Omnivore: Omnivores like Bearded dragons need a moderate amount of protein and a moderate amount of carbohydrate and fibre in their diet.


Social behaviour: Some animals live in groups and so feed will need to be spread evenly to make sure no animal is being deprived or having an excess of food.


Enclosure: Depending on the animals natural habitat will affect how feed is presented because some animals are arboreal, terrestrial and/or aquatic.


Age: Age affects the diet because animals at different ages require different amounts of nutrients. Life stages in animals is categorised into:


1. Neo natal – If a mammal it will take all its nutrients from its mother’s milk. But if not then an increase of protein is needed to accommodate the rapid growth rate at this point in the animal’s life.

2. Juvenile – Same diet as a neo natal animal that is not nursing as they are still growing.

3. Adult – An adult will need a balanced diet of protein, fat and carbohydrate suited to its species to maintain body condition.

4. Senior – Senior animals are less active and so will need less energy and fat because they are not using as much calories as they used to but keep protein the same as if feeding an adult. Feed needs to be presented in a way that makes it easier for the senior animal to access as they may be suffering from old age related diseases that could inhibit the animal’s access to feed. Senior animals tend to develop age related problems such as “Hip Dysplasia” which means they may need to be supplemented with medications like Glucosamine to lubricate their joints.


Life stage:

• Pregnant – Pregnant animals need to be able to eat as much as they want to ensure the proper growth and development of their offspring prenates.

• Lactating – A lactating animal needs to be fed as much as they want to nutritionally satisfy themselves and their neonates.

• Sloughing – Animals that are sloughing will not need any food as they will not have interest.

• Recovering – A recovering animal requires more energy and protein.

• Health: Animals have different nutritional requirements depending on the status of their health. An animal in impeccable health will need a balanced diet suited to its species to maintain its body condition. An animal that is extremely ill may need details on how to present its feed in a feeding plan as its feed may need to be presented in a way so that it is easier to get to. This is so they do not spend as much energy trying to get to it. Another way health can affect a feeding plan is if an animal has a health condition such as allergies which could inhibit what it can actually eat. This would need to be specified in a feeding plan to prevent the animal eating food that could cause it harm. Some animals may need lower protein in their diet due to impaired Kidney or Liver function. This is most common in older animals and would need to be specified in the feeding plan to prevent harm to the animal.

• Neuter status –Animals that have been neutered such as dogs will generally needs 25% less calories than an animal that is intact and 24-33% less calories in neutered cats.

• Work Level: Working animals need a higher amount of energy, protein and fat in their feeding plan to accommodate the demands of their body to replenish lost energy. Working animals are more vulnerable to injury so they need more protein in their feeding plan to repair tissue.


Comparing Diets (Wet & Dry):

Golden retriever

Inactive/not working

Adult

Male

32KG

3½ years old


Behaviour of the animal: Active, displays natural behaviours for life stage, age and species.


Condition of the animal: Body score = 3. In good condition with no health problems.


Amount of food eaten: Was given 680g of Pedigree, Adult Dry Dog Food and all the food was eaten.


Cost of feeding: £5.73 for a 22KG bag.


Evaluation

The current feeding plan is a cheap way of providing a complete diet that gives a dog the minimum amounts of each nutrients that they need. However the ingredients in the feed are of low quality and are mainly of plant origin. This is indigestible for a dog and so will have little use nutritionally and a lot of the food contains starches that the dog can digest but it is mostly stored as fat.

The feeding plan has maintained an ideal body condition of the dog due to the amount given and his health is in good condition. His behaviour is normal in terms of age, species and life stage except he should be more active overall. This could be diet related due to the fact it contains a high amount of sugars that would make his energy levels spike and the drop drastically. I have witnessed some coprophagia which is a direct link to a diet that is of low quality and leads to dietary deficiencies.


Current diet. (Dry)

To satisfy the Dog’s nutritional needs you need to feed it 680g of the dry feed which costs £0.17 per a serving.


Alternative diet.

To satisfy a dogs nutritional needs with a wet diet you need to feed it 2.078KG (3 tins+1/3). This is because it is so high in water content (82%) that most of the tin is useless for the dog nutritionally. This would cost £2.18 per a serving.

A dry diet is more economical than the wet diet as you need to feed more of the wet diet to fulfil the nutritional needs of the dog which costs 4X more than the dry diet to feed.


Bibliography

www.drsfostersmith.com/pic/article.cfm?aid=459#answer_4

www.wisconsinhealth.org/diet-for-working-dogs-and-service-dogs.php

www.petinsurance.com/healthzone/pet-articles/pet-health/Pet-Nutrition.aspx

rotrain.hs.llnwd.net/e1/sitefiles/642/Documents/en_VNAChapter5_MAS.pdf

www.petdiets.com/Library/Pet-Nutrition/Lifestages

How to treat an egg bound hen/chicken (home remedy)

Posted by Lisa @ Medlink Students on November 15, 2017 at 1:15 PM Comments comments (0)

Treatment of an egg bound hen.

Chickens can become egg bound due to a number of reasons which include calcium deficiency, malnutrition, dehydration, obesity and because they are too weak from illness.

If a chicken is suspected to be egg bound then it is vital that immediate action is taken to relieve the hen and help her pass the egg before it becomes fatal. Death can occur in around 48 hours if the egg does not pass.

Preparation

1. Hand washing is essential! As illness could be the cause of the egg binding for the hen. If this is the case then the hen already has a compromised immune system and will be vulnerable to pathogens that could be passed on from you.

2. Long hair must be tied up!

Equipment required.

Gloves. (Latex)

Overalls.

Suitably sized bowl of warm water.

3. Check records to confirm that no one has tried help the hen or if it has been recognised and a veterinarian is on their way.

Approach

1. If the hen is in a large paddock then you need to move her into a smaller space such as a chicken coop to make it easier to catch. This avoids stressing the hen when trying to catch her.

2. You must consider if there are any hazards to yourself when you approach the hen and make sure it is safe to approach. Is there any debris in the way of your path you could trip on? Are there any spills you could slip on?

3. Make sure you remove any other occupants of the enclosure to a secure location to avoid accidents.

4. Approach the hen slowly and quietly. The hen will already be experiencing a lot of stress which makes a calm approach essential to avoid making it worse.

5. Try to avoid cornering the hen.

6. When you are in a reachable distance to the hen you need to crouch to make yourself look smaller to minimise stress to the hen.

7. When you are crouched you can then slowly reach for the hen making sure you go from the side.

Method

1. When you reach for the hen you need to make sure you go in from the side rather than the top to avoid startling the hen.

2. Pick the hen up with both hands either side of the wings so that they are flat against the hen’s body and she is unable to flap her wings. Make sure you have a firm but gentle grip so that the hen is restrained but comfortable.

3. You can now walk to the water container.

4. When you are near the water container then you can slowly lower the hen into it.

5. Make sure the water reaches the hen’s cloaca but does not exceed this. The water may need to be replaced to keep it warm, if this is the case then use the same method of handling as stated above when removing the hen.

6. The hen should be monitored while she tries to pass the egg. The egg should pass within 2 hours using this method. If she does not pass within 2 hours and still shows symptom of egg binding then continue treatment for another 2 hours.


7. If she still has not passed the egg then you must seek a veterinary surgeon immediately for further advice. If egg binding is confirmed then she can receive an injection of calcium gluconate which should make her pass the egg.

*Never try to remove the egg by hand or by crushing it!

Finishing

1. When you can confirm the hen has passed an egg you can then remove her from the water and keep her in a separate enclosure where she can be monitored for the next 24 hours.

2. Disinfect and dry all equipment used.

3. Update records mentioning what has happened, method of treatment and results of the given treatment. Please state the date and time.

4. Make sure the hen has access to fresh water and food.

Health and safety considerations

Chickens are known carriers for zoonotic disease such as Salmonella, E.coli and Clostridium. Gloves, overalls and thorough hand washing must be used to prevent the risk of the handler contracting the disease.

Do not eat, drink or put your hands near you face until your hands are thoroughly washed after handling.

Do not eat, drink or put your hands near you face until your hands are thoroughly washed after handling.


Ethical and Non-Ethical Sourcing of Animals

Posted by Lisa @ Medlink Students on November 15, 2017 at 2:30 AM Comments comments (0)

Ethical Sourcing:

 

Ethical sourcing means to procure an animal in an ethical way.

 

Ethical:

 

Captive bred (CB): Captive-bred means that the animal has been born in a captive environment. This is the most ethical source of exotic animals as captive bred animals are more acclimated to their captive environment and will experience less stress than a wild caught animal.

 

I feel that CB is the most ethical source of obtaining exotic animals as to get an animal to breed in captivity in the first place, the animal will have to be in the correct environment and have its needs met. Animals born in captivity experience less stress because they don’t go through being captured and going through transit like wild caught specimens do. Captive bred animals are healthier because they are less likely to experience health problems that they would in the wild such as parasites and diseases.

 

Long Term Captive (LTC): Animals that are LTC are animals that have been in captivity for more than 3 months and so are more established in captivity than a wild caught animal because they have been in captivity longer.

 

I think that this is an ethical source of obtaining an exotic animal because the animal has been in captivity long enough to acclimate which means the animal will experience less stress.

 

Non-Ethical:

 

Wild Caught (WC): Wild caught animals have been taken from their natural habitat and put into captivity. This is a non-ethical way of obtaining animals as they experience a lot of stress from being captured and going through transit. Wild caught animals have a high mortality rate where they can’t acclimate to captive life and become sick from the stress. Wild caught animals can have internal parasites that can spread rapidly to other animals in a collection.

 

 

 

The five animal needs:

 

1. Need for a suitable diet.

 

The animal needs to be fed a diet that is suitable for its species ie: a dog will need to be fed meat-based food because it is primarily a carnivore.

 

2. Need for a suitable environment.

 

The animal must be provided with a suitable environment that it would have in the wild ie: a Bearded dragon will need to be kept in an enclosure that is heated as it is an exothermic reptile which means it relies on its environment to maintain its body temperature.

 

3. Need to be housed with, or apart, from other animals.

 

This means that the animal must either be with or without other animals of its own species depending on its natural behavior ie: Guinea pigs are very gregarious and must be kept with other guinea pigs to behave naturally and be stress-free.

 

4. Need to be able to exhibit normal behavior patterns.

 

5. Need to be protected from pain, suffering, injury, and disease.

 

How does the captive environment meet the animal’s needs?

 

Leopard Geckos (Eublepharis macularius) are a large gecko native to the Middle East which are widely captive bred and thrive in captivity. Leopard geckos are insectivores which means they exclusively eat insects and are crepuscular meaning that they are active in the early evening.

 

1. Need for a suitable diet.

 

Live insects are bred in large numbers and can be purchased at most exotic pet stores to feed.

 

2. Need for a suitable environment.

 

Leopard geckos need a constant heat source as they can’t regulate their own body temperature and so rely on their captive environment for this. Heat can be provided using a heat mat and

 

3. Need to be housed with, or apart, from other animals.

 

Leopard geckos are solitary and so will need to be housed on their own otherwise they may fight with other geckos leading to injury.

 

4. Need to be able to exhibit normal behavior patterns.

 

Leopard geckos actively hunt their food in the early evening and so must be provided with live insects in the evening so they can behave naturally.

 

5. Need to be protected from pain, suffering, injury, and disease.

 

Leopard geckos need to be provided with vitamin and mineral supplementation that can be provided by frequently coating the live feed of vitamin and mineral powders like Nutrobal.

 

The effectiveness of the legislation for keeping exotic collections or animals?

 

Cites (Convention on the International Trade of Endangered species) the purpose of cites is to monitor the trade of wild plants and animals and make sure that their survival is not threatened by their trade. Cites is an international agreement amongst governments.

 

DWA (Dangerous Wild Animals act): This act is for ensuring that people who own an animal that is deemed dangerous do so in a manner that protects the public and ensures the animal’s welfare is of the highest standard.

 

SSSMZP (Secretary of state’s Standards of Modern Zoo Practice): Gives zoos in the UK guidance on how to run their zoo. This legislation requires that a zoo is inspected every four years by an inspector that refers to the SSSMZP which entails how to run the zoo.

 

 

Toxic Chemicals in Your Everyday Products

Posted by Lisa @ Medlink Students on November 14, 2017 at 4:55 PM Comments comments (0)

Toxic chemicals in your products?

It is important to know what you are using on your skin, this goes from shampoos to moisturizers, perfume and makeup. Here is a list of just some of the ingredients found in many cosmetics and skincare products to watch out for:-

• Fragrance/parfum (unless essential oils), many chemicals are used and disguised as fragrance on the ingredient list

• Parabens (methylparaben, ethylparaben, and propylparaben) – easily penetrates the skin and has been shown to have hormone-disrupting abilities

• DEA,MEA and TEA – can cause allergies and are toxic if absorbed over a long period of time

• Mineral oil and petroleum

• Oxybenzone (generally found in sunscreens as SPF) – has shown endocrine disrupting abilities

• Toluene (found mainly in nail polish removers) – harmful when inhaled and is been linked to the disruption of the central nervous system

Have a look at the ingredient list on your shampoo bottle, moisturizer, perfume and makeup products and see how many of these ingredients are listed.

If you are interested in non-toxic beauty products that don’t contain any harmful ingredients that have been linked to allergies, cancer and have hormone-disrupting abilities. Check Amera's blog where you will find reviews and swatches of organic, vegan and non-toxic makeup products and brands.

How to look after a Cheetah

Posted by Lisa @ Medlink Students on November 12, 2017 at 7:05 PM Comments comments (0)

Cheetah

The five animal needs:

1) Need for suitable accommodation; cheetahs are the fastest land mammal, so accommodations must ensure they have the space to be able to run and have their own territory, if a cheetah were not to have this space they would show stereotypical signs like pacing along the enclosure. They also live in open grassy areas where it is extremely hot during the day, so their accommodation will reflect this, with open area and a few trees to shade, a small pond/lake will ensure the cheetah has access to water as well.

2) Need for suitable diet; in the wild a cheetah will run to catch their prey, however they cannot run for long distances, they will eat other animals ranging from gazelle to small mammals, to ensure this is met they would be fed the same types of prey and in a way that would reflect their natural hunting style, this is often from race line, the food is attached to. Cheetahs will rarely go for stationary food, so if they are not fed in a way that allows them to run, they may even not eat the food at all, they will also become restless from lack of enrichment.

3) Need to be able to exhibit normal behaviour; as previously stated, a cheetah will naturally run for their meals, so they must have the same enrichment when being fed.

4) Need to be housed with or apart from other animals; cheetahs are naturally solitary animals, with an exception of some males will stay together, normally brothers, in a zoo environment, you would either have a single female, or a couple of males, if females were to be housed together they would fight over territory and food.

5) Need to be protected from pain, suffering, Injury and disease; naturally in the wild cheetahs are at risk of getting injured of suffering from disease, however in captivity this can be reduced greatly, by ensuring all measures are taken that the enclosure is safe and clean, and preventing at the highest standards any risk of disease from effecting them. (needs reference - https://www.rspca.org.uk/whatwedo/changingthelaw/whatwechanged/animalwelfareact )

Good and bad health in cheetahs: (reference- https://bigcatrescue.org/animal-health/ )

Working with your cheetah every day you will learn their natural behaviours, this will make it easier to tell if they are in ill health, from not acting the same as normal, to their faeces being different, some examples of good and poor health include;

Good health signs; Bad health signs

Showing natural behaviours Change in normal behaviour(active animal being lethargic)

Good coat condition

(glossy/shiny, intact) Poor coat condition(fur missing, dirty)

Eating well Change in eating habit

Maintain a good weight Loss of gain of weight unexpectedly

Healthy faeces and urine Unhealthy faeces or urine( blood, discoloration)

Clear ears, eyes, nose and mouth Discharge around ears, eyes, nose and mouth

Moving correctly Limp, unable to move

Feline Aids: FIV- Feline immunodefiency virus:

FIV is an infectious virus caused by a retrovirus in the lentivirus, it will attacks the cells of the immune system, making it difficult to fight of any secondary infections. FIV will be carried to the reginal lymph nodes, once it has done this it will then replicate itself in the T Lymphocytes (white blood cells) it will then spread throughout all other lymph nodes in the body, due to this lack of T Lymphocytes the cheetahs immune system will be unable to defend itself against other diseases.

Symptoms:

• Weight loss.

• Poor coat condition.

• Gastroenteritis.

• Gingivitis.

• Stomatitis.

• Diarrhoea.

• Infections in the skin, eyes, urinary tract

Transmitted: FIV is present in cat saliva, and is most often transmitted through bite wounds, meaning either through territorial fighting, or biting when mating, can transmit it, at can also be transmitted onto cubs through an infected mum.

Diagnosed: there are few ways to diagnose FIV in cheetahs and cats alike, however running the blood test ELISA(enzyme-linked immunosorbentassary) is one of the most common ways, however if a cheetah has been vaccinated against it the blood results can come up as false positives, the other ways to diagnose are by looking out for any symptoms common to FIV and through looking at the cheetahs genetic history.

Treatment: there is no cure to FIV once the cheetah has it, however it can be manged through regular vet check-ups, parasite control and certain anti-bacterial and anti-fungal drugs. They can live for many years.

Prevention: the best way to prevent FIV is through vaccinating against it, do not house infected cheetahs with others, and to not breed any infected.

FIV references- http://www.cat-world.com.au/feline-immunodeficiency-virus http://www.catsg.org/cheetah/05_library/5_3_publications/B/Brown_et_al_1993_Exposure_of_cheetahs_to_FIV_and_FIPV.pdf

Crypotococcosis:

Crypotococcosis is yeast like fungal infection, which is contracted through the nasal passages, from this it will then start to affect the brain, eyes and lungs. The fungal spores are found in bird droppings and rotted vegtables, it can also infect the stomach and intestines.

Symptoms: the symptoms vary on the organs affected, these symptoms range from;

• Fever.

• Nasal discharge.

• Seizures.

• Sluggishness.

• Altered sense of balance.

• Nodular tissue seen at nostrils.

• Disorientation.

• Swelling at bridge of nose.

• Lesions on head.

• Enlarged lymph nodes.

 

Diagnosis: nasal passages samples, along with blood and urine cultures, can also to biopsy’s of skin lesions.

 

Treatment: treatments include antifungal medication, and surgery may be necessary, although normally only if the nervous system has been affected.

 

Prevention: to prevent crypotococcosis ensure that any foods given are safe and that the enclosure is regularly disinfected and kept clean, try to reduce the risk of birds flying over.

 

Crypotococcosis references - http://www.petmd.com/cat/conditions/infectious-parasitic/c_ct_cryptococcosis http://www.jsava.co.za/index.php/jsava/article/viewFile/432/418

 

How to De-flee cats at home (remedy)

Posted by Lisa @ Medlink Students on November 12, 2017 at 10:10 AM Comments comments (0)

Administering flea spot – on treatment to a tame cat.

Cats need to be treated for fleas every 3 months to prevent them becoming overwhelmed and ill. Applying the treatment requires restraint to effectively apply the treatment or you run the risk of using the treatment incorrectly. I have made a step by step guide on how to effectively and correctly administer spot on flea treatment for a cat.

Preparation

1. Wash hands.

2. Long hair must be tied back.

3. Check records to confirm that no one else has already given the cat spot on treatment

4. Prepare the spot on treatment and leave it somewhere you can easily pick it up for when you need it.

Gloves. (Latex)

Overalls.


Pet carrier.


Treats suitable for cats.

Towel.

Table.


Approach.

1. Watch the cat’s body language to make sure it isn’t showing signs of stress.

2. Do not make sudden movements as you could frighten the cat.

3. Stay calm.

4. Move any objects that obstruct your path to eliminate tripping hazards.

5. Begin approach by slowly walking towards the cat.

6. Place the pet carrier near the cat with the door open.

7. Give the cat a treat and keep it calm with a reassuring voice so it feels safe.

8. Place some of the treats inside the pet carrier so the cat is enticed and walks into the pet carrier.

9. When the cat is inside the pet carrier, close the door and secure it.

10. Using correct handling technique, slowly pick the pet carrier up by the handle and hold your other hand underneath it for support. Make sure you do not bend your back when you pick it up. Practice good handling techniques such as crouching and maintaining a straight back to avoid injury.

11. Walk over to the desired destination in a calm manner and keep the pet carrier still to avoid stressing the cat.

12. Place the pet carrier on the table and check that the cat is calm. Do not open the pet carrier until you are certain the cat is calm and won’t run out.

Method.

1. Place the cat on the table so that it is facing away from you.

2. Hold the cat by its shoulders with both hands.

3. Stand near the table so that there is no space between you and the table’s edge. This prevents the risk that the cat could fall if it rears back

4. With one hand holding the cat by the shoulders use your other hand to pick up the treatment.

5. Using the same method of restraint pictured above, administer the treatment by parting the fur at the base of the neck.

6. Using the same method as the picture below, apply the contents of the treatment by pressing the tube gently onto the cats skin and emptying the contents of the spot only on the skin.

7. Make sure that the contents of the spot on only goes onto the skin of the base of the neck and nowhere else on the cat’s body. If any of the treatment goes into the cat’s eyes or mouth then call a vet immediately!

Finishing.

1. After the treatment has been given, make sure you do not touch the application site for at least 24 hours after use. If the treatment makes contact with your skin then wash your hands thoroughly with soap and water.

2. Dispose of the treatment in a hazardous chemicals bin.

3. Disinfect and dry the work surface.

4. Give the cat a treat after treatment and use the same method mentioned in preparation to get the cat into its pet carrier.

5. When the cat is in its pet carrier close and secure the door.

6. Hold the handle with one hand and support the base with your other hand.

7. In a calm manner walk back to where you first got the cat and slowly place the pet carrier on the ground.

8. Open the pet carrier door and allow the cat to leave at its own pace.

9. Update records stating that the cat has been treated for fleas.

10. Monitor the cat for 24 hours after treatment to confirm there are no side effects.

11. Do not wash the cat for 48 hours after treatment.


Health and safety considerations.

Cats are known carriers for Toxoplasmosis which leaves pregnant woman vulnerable to this parasite. It is strongly advised that if you suspect you may be pregnant that you do not carry out this procedure as Toxoplasmosis can cause miscarriages.


Cats are very alert animals and can be easily frightened. In rare occasions cats can bite or scratch when startled or uncomfortable which is why PPE is essential during the handling. There is a small risk the cat may become aggravated and cause injury to you when you are applying the treatment so you must be prepared for this to occur.


If you are bitten or scratched during the procedure, make sure you thoroughly wash the injury with water and antibacterial soap to prevent infection. If the injury is bleeding, use a bandage to wrap around the wound to stop the bleeding.


Do not eat, drink or put your hands near you face until your hands are thoroughly washed after handling.


Control of Gene Expression in Yeast: Heat Shock Promoters

Posted by Lisa @ Medlink Students on November 10, 2017 at 7:10 PM Comments comments (0)

Transcriptional Regulation Due to Environmental Stresses

Eukaryotic and prokaryotic cells react to environmental stimuli by activating the expression of certain genes, which encode protein, enzymes that will help the cell endure the environmental change. Examples of environmental stimuli include Heat shock factor, which expresses genes for survival at high temperatures, hypoxia inducible factor, which expresses genes for survival in low oxygen conditions, and sterol regulatory element binding protein, which maintains proper lipid levels within the cell.


Heat Shock in Eukaryotes and Their Response

This experiment aims to study the effect of gene expression in a cell responding to high temperature stress using heat shock promoters. Heat shock gene expression is activated as a response to increased temperature. Increasing the temperature activates the transcription of the cell’s multiple heat shock genes. The proteins synthesised help reduce the damage from thermal denaturing of important cellular proteins. A large number of proteins is formed rapidly in a response to heat shock which act as ‘chaperones’ aiding other proteins to retain their 3 dimensional form and function properly. The heat shock genes are activated by the binding of the protein called heat shock transcription factor to the heat shock response element. The heat shock transcription factor is not active in non-heated cells, however increasing temperature cause a structural change of the protein that allows it to bind to the heat shock response element in DNA. Then the protein is modified by phosphorylation, allowing it to express gene transcription. (Hardin, 2012)


Reporter Genes

A reporter gene is a gene that is attached to a regulatory sequence of another gene that’s being studied, in this case, in a cell culture. A gene is chosen as a reporter because its characteristics that are expressed on an organism are easy to identify and measure or because they are selectable markers. Selectable markers are genes introduced into a cell that grants it a specific characteristic for artificial selection. Reporter genes are usually used to specify whether a specific gene has been expressed or taken up by the cell. In this case, the promoter hsp26 gene from yeast has been attached, artificially, to a gene that’s coding for the enzyme Beta-galactosidase. Beta-galactosidase is an enzyme present in bacteria and its function is to breakdown Beta-galactoside sugars. For example, lactose is converted to galactose and glucose. Replacing the lactose with the substrate o-nytrophenyl-beta-galactoside (ONPG), instead results in the formation of galactose and o-nitrophenol (ONP) – a yellow discolouration which is due to high pH. Hence, the gene for Beta-galactosidase, named LacZ, has an easily assayable product. LacZ is the reporter gene, as the promoter is Beta- galactosidase, whose activity is being measured. 2


The strain of yeast (Saccharomyces cerevisiae) being used is W303. It contains a plasmid (pUKC414) which consists of the promoter; hsp26, and the Beta- galactosidase coding sequence. Approximately, one copy per cell was present in each plasmid.


In this particular experiment, placing the yeast under heat shock conditions (30-45 degrees Celsius) caused the hsp26 promoter to activate hsp26 and Beta-galactosidase. Beta-galactosidase converts lactose into galactose and glucose. Beta-galactosidase also hydrolyses the ONPG molecule into galactose and o-nitrophenol (ONP). The end solution was yellow, due to the presence of ONP, which was used to inspect enzyme activity using a colorimetric assay. 1


The hypothesis was that as temperature increases, the intensity of yellow colour increases as ONP presence increases as it tries to protect the cell, hence B-galactosidase activity is increased. The null hypothesis was that increase in temperature would have no effect on the B-galactosidase activity, no matter what incubation time was.


Method

The protocol provided in the in-lab hand-out was used, however one modification was intentionally made. The first incubation time was changed from 15 to 20 minutes to allow more time for B-galactosidase to convert the disaccharides, producing more ONP. This didn’t seem like enough time because only the 39 deg. Celsius tube had a very tinted yellow colour. All the tubes were returned to the water baths for an extra 5 minutes. Furthermore, too much SDS was accidentally added, 200 instead of 20ul, to one of the tubes, however this was assumed to have no significant impact on the results. This error was ignored. Some spillage occurred with tube 37 deg Celsius however there was nothing that could have been done to fix this as it was too late to start the process again.


Results

The specimen results were used because the individual results seemed very random and chaotic. Individual results are shown below but they were regarded as an anomaly and were assumed to be inaccurate.

The specimen results were examined and the Beta-galactosidase activity was calculated. Beta-galactosidase activity is measured by working out ONP (nmol) /cells assayed (ml)/time left in bath (mins). The absorbance was converted into nmol via the following equation;

0.0045 at 420nm = 1 nmol of ONP, which was then divided by volume and time left in water bath (15 rather than 25mins, because specimen results were used). Hence, B-galactosidase activity was calculated for all the results. An example of the first calculation is given below:


B-galactosidase activity (nmol/ml of cells/min) : ( ({0.247⁄0.0045}⁄1.5))⁄15

=2.440 nmol/ml of cells/min

The following table shows Absorbance for both incubation temperatures and the calculated activity of B-galactosidase, for both time durations.

Absorbance at 420nm B-galactoside Acitivty (nmol/ml cells/min)

Temp

(deg. C) 20 mins 45 mins 20 mins 45 mins

30 0.247 0.235 2.440 2.321

37 0.330 0.556 3.259 5.491

39 0.313 0.924 3.091 9.126

42 0.208 0.254 2.054 2.509

45 0.242 0.153 2.390 1.511



Discussion

Hsp26 gene from yeast was an artificially attached gene whose purpose was to code for the enzyme Beta-galactosidase which eventually lead to the production of ONP, the yellow colour being measured. This was introduced into the cell via Plasmids, instead of modifying a gene in the yeast chromosomes, because plasmids are easier to manipulate.

The results showed a sensible pattern. When the tubes were incubated for a longer period, 45 minutes, optimal enzyme activity was shown at 37-39°C. However, enzyme activity dropped significantly afterwards from 9.1 at 39°C to 2.5 nmol/ml of cells/min at 42°C, which could be due to the enzymes not being able to cope with the heat shock anymore as they begin to denature. At 30°C, not many enzymes were active as the temperature could be considered moderate or low.

The hypothesis was fairly true since the B-galactosidase activity did increase as temperature increased however to a certain point. After 42°C, activity dropped significantly for both incubation times. It was also found that the longer the incubation the faster the activity rate of B-galactosidase. The null hypothesis was regarded as false.

B-galactosidase activity was highest for the cells at 37°C as peak activity was reached, for the 20 min incubation. However, peak activity for incubation of 45 mins was at 39°C. This is above the yeast cells’ optimum growth temperature; 37°C, which makes sense for heat shock proteins to be very active providing the cell with thermotolerance, as the proteins have more time (45 compared to 20 mins) to be produced.

When the tubes were incubated for only 20 minutes, a fairly similar pattern occurred however at a much slower rate. Optimal temperatures were also at 37-39 with a B-galactosidase rate ~3 nmol/ml of cells/min. This could be due to the heat shock response being more receptive when allowed a longer incubation time period, so all the circular DNA plasmids having more time to penetrate the yeast cell membranes.

The heat shock response was also active at 30°C which is surprising since there’s no high temperature that the cell is competing with; however this is normal due to the increase in temperature during incubation. Usually there is some heat shock activity in the cell due to a the presence of a small number of heat shock proteins that exist in the cell to cope with other functions of repair, altering protein confirmation, modulating protein activity, modifying protein degradation and folding peptide chains correctly during protein translation. (Takayama, 2003)

This study could be improved by measuring enzyme activity at a more precise range of temperatures and different incubation times to find the optimum B-galatosidase activity as each strain of yeast performs differently at each individual temperature. However the cells cannot be kept in heat shock for too little time or too long, as the cells need a recuperation period to recover but also need time to replicate the plasmid DNA while incubated, and produce the proteins required for heat shock.

Furthermore, it’s predicted that the individual experiment failed to produce any sensible results due to several factors. Some of which could be due to inconsistencies in the age of the yeast cells being used, amount of circular plasmid DNA used, the duration and intensity of the heat shock and the time allowed for recovery period. These factors could be taken into account if the experiment was to be repeated to produce a more accurate result. Some complex factors that cannot be controlled in cells include the stability of mRNA and other factors that affect the expression of the promoter being studied; hsp26.

In conclusion, this experiment showed the heat shock factor was an important natural phenomenon that helps the cell cope with one of its many environmental stresses. It is essential for life, and its complex activity on a molecular level needs to be studied in more detail to determine accurate results in how far the cell’s natural state can be stretched while still functioning with no complications. Artificially introducing the promoter hsp26 to the plasmid showed that it was an essential factor that lead to the production of B-galactosidase, which lead to the production of ONP – the yellow product that was measured, indicating a physical indicator of activity of heat shock proteins.

Full report with tables and graphs can be downloaded at  www.dropbox.com/s/js2r4uxtecqq3em/Transcriptional%20regulation%20due%20to%20environmental%20stresses.docx?dl=0 



Bibliography

Hardin, J. (2012). World of the Cell (Vol. 8th Ed). Pearson Education, Inc.

Takayama, S. (2003). Heat shock proteins as regulators of apoptosis. Oncogene, 9041-7.

1Barman, kumar (n.d.). Heat shock at 37°C with plasmid ligated at 37°C yields more number of Escherichia coli transformants than plasmid ligated at 16°C: a possible role of ligated plasmid conformation during heat shock. Retrieved April 8, 2014, from /www.currentscience.ac.in/Volumes/104/06/0747.pdf

2Seidenfeld, J. (2010, June). The Heat-Shock Response. Retrieved April 8, 2014, from /www.stanford.edu/group/hopes/cgi-bin/wordpress/2010/06/the-heat-shock-response/

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