eChapters

Definition

Any fracture or discontinuity in bony structure like scapula and pelvic girdle present above the shoulder joint and hip joint respectively are called as highest level fracture.

Aim

To know the method of repair of highest level fracture.

Procedure

As there is no satisfactory method of external immobilisation till date, generally animal is kept under restricted movement. Sometimes application of charge (glue bandage) externally may give some satisfactory result.

Fracture of scapula, shoulder dislocation, fracture of pelvic girdle, hip dislocation have been grouped under highest level fracture.

For large animal, immobilization of bone & joint have not been developed satisfactorily/ till date. Hence after correction of dislocation or rearrangement of fractured the site Is kept immobilized with application of gum bandage.

Aim

To know the use of Conjoined ‘U’ Plate with Padded ring for fracture side immobilization.

Site to be immobilized

Upper 3^rd metacarpus, Carpo - meta carpo joint dislocation, Radio - Carpal Joint dislocation. Lower Radius & Ulna fracture are can also be immobilised with this device

Definition

Handing pin cast is used for management of longbone fracture. Pin which is used for immobilisation of fracture of proximal radial and tibial bones is called hanging pin cast.

Special equipment

Intramedullary pin, hand driven drill, plaster of paris bandage Procedures :,

Clsssification

Fracture of upper third humerus, shoulder dislocation and scapula has been grouped under highest level fracture. (Fig.1)

Purpose

To know the method of casting the highest level fracture by modified valeau sling application.

Requirement

Thick 8 layer cotton padding, adhesive gum, 6"-8" long bandage rolls.

Strong bamboo

4 feet long of 4 inch diameter

Gl.Wire

6 mm diameter of 34 kg G.l. wire

Aim

To know the methods of preparation of side bars for large animal Thomas splint fabrication

Procedure

Two strong dry matured bamboo splints are of more than actual length of Thomas splint are prepared, (figure 1)

Three nicks at a distance of 2 inch are grooved on the side of the side of the splint at one end of the splints for keeping the tying end of the ropes in position (figure -2)

The bamboo splints at the lateral ends are thinned to some extent. For knob kniiting of splint to the metal ring by making an arc (figures).

All the steps are depicted in figure 1 -4.

Before fixing the metal ring with the bamboo extention splint the thinned ends are emersed inside water for easy arching of the bamboo splint.

Purpose

Method of tying with bamboo splint with ring is very important because of the following points.

A string fitting with the bamboo splint is required as with loose fitting with it may come up at any stage before actual bone healing.

Aim

To knw the method of softening the ring of the perpendicular Thomas splint to avoid direct trauma of the metal wire with the skin.

Materials required

Jute /thin gunny bag,bandage roll,cotton threads

Aim: Bending of the lower half of the ring of Thomas splint to an angle 45- 60degree to comfort the limb and to provide free circulation.

Importance

After tieing the ring with bamboo splint and padding the ring with jute and cotton .bending the lower half of the ring upto more than 45 degree angle is highly essential because of placement of the fractured limb within the ring of Thomas splint comfortably, so as to the flow of blood circulation in fractured limb uninterruptedly.

Aim: To know the actual length of a Thomas splint & measurement for an animal.

Procedure

Animal is kept in lateral recumbancy with affected limb upward.

After padding & medical bending of Thomas splint, it is fitted upon the fractured limb.

At the time of measurement, the limb should be kept vertical to body for accurate measurement, otherwise the length of Thomas splint may be shorteror longer if the limb is kept forward or backward respectively.

The ring of the Thomas splint is kept close to the axilla (forelimb), groin (hind limb).

Limb is put downward & Thomas splint is pushed upward keeping both the extension bamboo of Thomas splint in crossed manner, measurement of length of Thomas splint is taken up to the bulb of the heel & mark is recorded.

The excess length is trimmed short (figure-1). Two nicks are made on the distal end for retaining the thread in position.

Both the distal ends are tied together with thread, soft padding is placed at the site where hoof is to be kept & retained in position with bandage (figure-3).

Aim: To know the method of securing thomas splint, in the forelimb.

Description

After immobilising the fracture site covering the distal as well as proximal end properly the thomas splint is fitted over the forelimb.

The foot is immobilised to the distal end of Thomas splint.

The immobilised forelimb kept closed and straight with the posterior extension of bamboo of the thomas splint. The limb is fixed with posterior bamboo splint with bandage roll at several place to avoid anterior, posterior movement of the limb.

At the time of fixing the limb with posterior extension bamboo of the Thomas splint, bandage are rapped along with the netting of thomas splint, to avoid medullary lateral movement of the immobilised limb.

Finally the immobilised forelimb is rapped along with the bandage roll including both the anterior and posterior extension of bamboo splint.

To avoid outside movement of the immobilised limb, the method of immobilisation of fracture of forelimb has been shown in fig-1.

Purpose:- To know the method of fitting the immobilised fractured hind limb in Thomas splint.

Description

The fractured site is immobilised properly covering the area joint below and joint above from the fractured site.

The immobilised limb is kept within the prepared Thomas splint.

The immobilised limb is kept straight and come close with the front extension bamboo of the Thomas splint.

The foot is fixed to the distal end of the Thomas splint with bandage roll.

In the first attempt the immobilised limb is secured with the front bamboo splint with bandage roll at several places to avoid posterior movement of the immobilised limb.

At the time of fixing the immobilised limb with anterior extension bamboo of Thomas splint, the limb is fixed with the netting of Thomas splint to avoid medial movement of immobilised limb.

Finally the immobilised limb is covered with bandage roll wrapping around both the anterior and posterior extension bamboo of the Thomas splint to avoid outer movement of the immobilised limb.

The method of immobilisation of the hind limb with Thomas splint has been shown in the figure.

Defination

Dislocation of carpometarcarpus and tarso metatauses along with upper fracture of MC and MT bone (Fig. 1 and Fig.2). It is considered as difficult fracture and may not be healed appropriately by the usual procedure of lower level fracture repair method. Due to constant movement of fracture ends its treatment should be similar to middle level fracture repair method.

Materials Required

Thick padded cloth

Adhesive gum (fevicol)

Netted bamboo splint

Thomas splint

Purpose

As the immobilisation of upper third part of radius ulna is very difficult ,the purpose of the immobilisation tech. Is to fix the upper third of radius-ulna with the posterior half of ring of Thomas splint with bandages are very important for achieving no movement of fracture fragment during healing.

Indication

Correction of upper third fracture of radius-ulna by Thomas splint application indicated in higher level fracture.

Definition

Fracture of Upper 3rd of tibio-fibula Presence as a difficult Problem for the Surgeon to immobilise it Perfectly. As the fracture site generally moves within the imobiliser. Its healing becomes difficult. The aimof immobilisation technique of upper 3rd tibio-fibula fracture is to fix the upper 3rd of tibio-fibula with the anterior half of ring of thomas splint with bandages. This is an important step for achieving no movement of fracture fragment during healing.

Indication

Correction of upper 3rd fracture of tibio-fibula (Fig.1) by thomas splint application.

Use of conjoined Thomas splint for bilateral tibia fibula fracture for large animal.

Aim

To know the immobilization technique for both tibia-fibula fracture in large animal practice.

Instruments

Conjoined Thomas splint (Fig 1).

Definition: Mandible forms the skeleton of lower jaw and comprises of a body and two rami. The two halves of this bone fuse incompletely at the mandibular symphysis, situated at the midline within the body.

Indication: Immobilization of fractured mandible, minimally or non displaced fractures with an intact opposite hemimandible, vertical ramus fracture.

Anasthesia: Local anaesthesia for large animals. A20 gauze 2.5 cm needle is used to deposit 5-1 Omi of lidocaine at the exit site and into the canal.

Objectives

Immobilization of fracture of mandible.

Requirements

‘U’ Plate 4 Pins Electric drill Orthopaedic hammer Pin cutter

Procedure

Under local infiltration of an anesthetic & parenteral sedatives the 4 pins are drilled 2 on either side of the fracture site from one side mandible to other side mandible. During drilling the pin, the holes, importance should be given for the passage of the pin through prefix of the drill hole in metal frame

The fracture end should come accurate apposition during pinning procedure & the symmetry of the fractured mandible should be as norma! as possible.

Sterilised antiseptic gauge bandages are covered, the pin between frame and the body. The pins are retained in position either with nut-bolt or by passing a plastic tube after which the hole of the plastic tube is filled with resin.

Aim: To know the method of immobilization of the limb after repair of the flexor tendon.

Procedure

The immobilization procedure of limb after flexor tendon repair is done in 2 stages.

Aim

To depict the detail method of tendon suturing of transected gastrocnemious tendon for union.

Procedure

After cleaning, debriding and flushing the transected site thoroughly local antiseptic and antibiotic is applied.

1. To expose the tendon ends clear visual as well as proper exposer the site was enlarged by two vertical incision placed at dorsal and ventral part of the transected site.(Fig.1).
2. The three different tendons and their locations is identified. The both end of tendon of a particular group are brought close together and joins end to end and retained with strong polyester suture putting to interrupted knots at 90 degree to each other.(Fig. 2).
3. Their peritendon cover are pulled over the suturing site from their either ends and are also retained in position with additional interrupted suture running at 90 degree to each other.(Flg. 3). In this way 4 interrupted suture are required for approaching one tendon.
4. In this way all the 3 tendons are joined with their respective other ends with suture. Total no of 12 interrupted suture are required for joining all the 3 structure of the gastrocnemious tendon. At the end the suture tendons are strengthened by placing to interrupted stainless steel suture. (Fig. 4).

Finally a flushing catheter was placed at higher level and the suture site is covered with a thick dry antiseptic absorbent cotton padding.

Hip joint is a ball and socket joint which involves cotyloid cavity of os-coxae and head of femur. Hip dislocation can be corrected by external immobilization of the area with POP cast and keeping the animal in a mobile sling for several weeks.

The dislocation involves cotyloid cavity and head of femur and the affected limb deviate outward. (Fig.1)

The fracture site is immobilized extending joint below and joint above level initially with plaster of Paris.(Fig.2)

Cloth bandage is applied over it for support involving joint above and joint below which should not be too tight or too loose.(Fig.3)

Second layer of cloth padding is provided over the first layer to provide more strength and further immobilization.(Fig.4)

Final immobilization is done by application of rubber belt or suspensory bandage. (Fig.5)

At the end the entire animal is kept in mobile sling for proper immobilization for some weeks and housed in limited space(Fig.6)

Aim: Regular flushing of compound and fracture using topical antibiotic or antiseptic required to bring the open wound and fracture either to a aseptic stage or to reduce the degree of infection and pus formation.

Procedure

After through cleaning the exposed bone as well as the wound of the compound fracture the wound is placed inside its part.

A thick walled rubber tube / plastic tube threaded with two end of a suture with a straight needle is carried to a higher site from the fracture within the wound.

The suture and needle is brought out, the sutures separated and again threaded and the flushing catheter is fixed with suture. Fig.1.(a) and Fig. 2. (a), (b).

The wound is protected with a thick absorbant sterile cotton padding, keeping the flushing catheter out through a small hole within the bandage. Fig. 1 (b) and Fig.2.(c)

The limb is immobilized as per the recommended procedure depending the level of fracture, outer end of the flushing catheter is kept outside, of the immobilization for carrying out regular flushing of the wound.

Objetive

To completely exercise a cancerous hoof in presence with cauliflower like growth with regular bleeding & myosis.

Material Required

Amputation saw, thick padded with bandage roll, a mixture of powder containing resin and naphthalene ball powder, bicycle tube protection sleeper (Fig-3), local anesthesia.

Indication

Irreparable injury.

Frost bite, burn.

Malignant neoplasia.

Incurable peripheral vascular disease.

Necrosis, gangrene, extensive injury of nerve of fore limb.

Indication: Irreparable damage of the meta tarsal joint.

Introduction

A downer cow is an animal In sternal recumbency and isn’t showing obvious clinical sign of hypocalcemia,hypomagnesmia or any limb or spinal Injury. Most causes are related to parturition, occurring in few days before or upto a week after calving.

Causes for Making an Animal Downer

Animals Inability to stand may be caused by illness or injury. Following may be the reasons

Purpose

To make a list on the requisites on materials required for preparation of rehabilitation mobile sling for large animals.

It consists of 2 parts-A) fabrication of fixed supporting frame B) fabrication of animal swing frame.

Fabrication of fixed supporting frame -

Materials required - Bamboo pole and rope

Fabrication Technique - Size a frame 3m for large animals, 2.5m for medium sized and 2 ml for small sized animals.

Fabrication of animal swing frame-

Materials required - Strong fixed rectangular iron frame

Fabrication technique - The required no. of bicycle tyres are fastened upon the frame. All individual tyres are tied with jute threads at 3-4 places.

Indication: Rehabilitation of downer animal

Recommended Duration for Slinging Large Animal

Aim: To know slinging duration for large animal.

Indication: Downer cow, sternal redumbency, hypocalcemia, hypomagnesemia, limb or spinal injury, cases,related to parturition.

Metals are toxic and non biodegradable pollutants released into the environment through effluents from agriculture waste, mining, metal processing industries, pharmaceutical industries as well as domestic and natural activities. Metallic pollutants tend to persist indefinitely and eventually get accumulated in the ecosystem. Metals especially arsenic, copper, cadmium, chromium, lead, mercury, nickel, selenium and silver etc. possess a risk to living systems. Conventional processes applied to remediation of metallic pollutants are often restricted due to technical and economical constraints. Particularly when the metal contamination is at very low concentration the conventional methods become inefficient. In this situation application of bioremediation techniques is a technology of choice for environmental protection and clean up.

Several substrates, like proteins (enzymes), are very sensitive to alteration in the concentration of the solution they are placed in. Hence is the importance of a buffer which controls the chemical properties of the medium. Literally a “buffer” has the ability to resist changes in the hydrogen ion concentration. Although, buffers do contain other molecules, that might, in addition to the hydrogen ion concentration, influence the ionic strength, the activity of enzymes, and other parameters of the experiment.

In any biological experiment, appropriate buffer must be selected based on their effect on the experiment. Ideal buffers control pH and ionic strength without interacting adversely with the experiment undertaken. For example, although phosphate is a common buffer, it may not be suited when phosphate is a substrate or product of the reaction being studied. Also, some proteins interact poorly with some buffers. For example, Tris is not ideal as it exhibit high pKa and the large change in pKa when temperature changes, although it is cheap and most proteins are stable in Tris buffers, moreover it seldom reacts with reaction substrates hence, Tris buffer is commonly used in biochemical experiments.

Transfer 1.5 mL overnight culture to a 1.5 mL microfuge tube, centrifuge for 30 sec, and decant supernatant.

Resuspend cells in 400 µL TE by vortexing, add 50 µL 10% SDS, 50 µL proteinase K (20 mg/mL in TE). Incubate for 1 hour at 37^oC.

Shear DNA by 3-5 passages through a 26 G needle.
Extract twice with 500 µL phenol: chloroform (1:1), and twice with 500 µL chloroform. 
Precipitate nucleic acids by adding 25 µL 5 M NaCl and 1 mL 95% EtOH, vortex, and centrifuge for 10 min, decant supernatant. Dry pellet.

Reagents

Nuclease-Free Water: 14µl

10X Restriction Buffer: 2µl

Acetylated BSA (1mg/ml): 2µl

DNA (~1µg): 1µl

Restriction Enzyme (10u): 1µl

Final Volume: 20µl

Reagents

Restriction enzymes of choice, such as BamH1 and EcoRI

Restriction enzyme reaction buffer,

70 % Ethanol

100 % Ethanol

3 M Sodium acetate (pH 5.2)

Distilled water

Plasmid DNA isolated from bacteria, such as pGFP(R) and pBC(R)

Start with a 1:2 molar ratio of vector: insert DNA when cloning a fragment into a plasmid vector. This ratio will vary with other types of vectors, for example, cDNA and genomic cloning vectors. The following example illustrates the conversion of molar ratios to mass ratios for a 3.0kb plasmid and a 0.5kb insert DNA fragment.

Principle

Plasmids requires hosts to replicate, hence it has to be inserted into bacteria. But efficiency of uptake of plasmid DNA by bacteria is low and will not do so without getting degraded. Therefore, to enhance the probability of uptake of the plasmid DNA by bacteria it is necessary to make the bacterial cells “competent” to absorb the plasmid DNA. One of the methods to increase efficiency of cell competence is given below. It should be noted that competent cells are more fragile than normal bacteria. Vortexing the cells, heating the cells above 42ºC or to 42°C for prolonged periods, or exposure of the cells to abusive treatments may kill them

Transformation

The uptake of DNA by competent cells is called transformation. Transformation involves:

Mix a small volume of DNA (usually 1-3 µl) with 100 µl of competent cells.

The mixture is incubated on ice for 30 minutes to allow the cells to take up the DNA.

The cells are then heat-shocked at 37 to 42°C for 1 minute, and cold shocked on ice for 2 minutes.

The heat shock/cold shock reverses the effect of the competent cell process, by healing the damage inflicted by the competent cell solution.

Add 0.5 ml of LB culture mediumz

Incubate the cells for 10 - 30 minutes at 37°C to aid the healing process.

This cell/medium mixture is then spread onto bacterial culture plates containing agar, LB, and a selection mechanism.

Principle

The principle of Electrophoresis involves the motion of a particle in a fluid medium, where the gravitational attraction (on a flowing liquid) is the sedimenting force, and the frictional force is the opposing force and is proportional to the velocity of the particle. These forces balance each other acting in opposite directions and hence balance each other. This will lead to the uniform motion of the particle in the mobile liquid medium (i.e. the particle moves at constant velocity). In case the external field is an electric field instead of a gravitational field, a macromolecule will respond to the external field in two ways. In case the molecule is charged, it will migrate in an electric field to the electrode of opposite charge. Under this principle the technique of electrophoresis functions.

All electrophoresis operations are governed by the principle: