eChapters

Sexual behavior

The pattern of courtship, display, motor activities and postures are directed to bring the male and female gametes together to ensure fertilization and pregnancy. The components of copulatory patterns are sexual arousal, courtship (sexual display), erection, penile protrusion, mounting, intromission, ejaculation, dismounting and refractoriness. The duration of courtship and copulation varies with the species both events are shorter in cattle and sheep than in swine and horses. The males sniff and lick the genitalia of female are more common and an important way of chemical communication through olfaction. Except in swine, the males smell the female’s urine and then raise head, with lips curled, in the ritualized “Flehmen’s reaction”. In sheep, goat and cattle, tactile stimulation of the female is made by nuzzling and licking the perineal region, whereas with the horse, the stallion often bites the female’s neck and in swine, the boar noses her flanks.

Introduction
Good Laboratory Practice (GLP) has been a cornerstone of laboratory operations, ensuring quality, reliability, and reproducibility in research and development across various fields, including pharmaceuticals, biotechnology, and environmental science. As we move further into the 21st century, the landscape of GLP is evolving rapidly, influenced by technological advancements, regulatory changes, and the increasing complexity of scientific research. This article explores future trends and innovations in GLP, delving into key areas such as automation, data management, regulatory shifts, and sustainable practices.
1. The Role of Automation in GLP
1.1 Increased Efficiency and Accuracy
One of the most significant trends in GLP is the integration of automation. Laboratory automation includes the prompt use of technology to perform tasks in a much efficient way with minimal human interventions, thus significantly increasing efficiency and accuracy of laboratory practices. Automated systems can handle repetitive tasks such as liquid handling, sample preparation, and data analysis, reducing the potential for human error.
Benefits of Automation
• Reduction of Human Error: Automation minimizes the chances of mistakes that can occur when humans perform these repetitive tasks, thereby improving the reliability of the results.
• Speed up the Research Process: For example, robotic systems can be used in drug discovery. These systems can conduct high-throughput screening, which means they can quickly test thousands of different compounds for potential effectiveness as drugs.

(A) Anatomy of Lens

The lens is anoptically clear, avascular, biconvex structure of crystalline appearance with an anterior surfaceless curved than the posterior surface, placed between the iris and the vitreous. It is suspended by the suspensory ligament of the lens which is attached to the ciliary body and equator of the lens.

The lens consists of a central nucleus surrounded by the lens fibres and anterior epithelium, encapsulated in an elastic lens capsule. The anterior surface of the lens is in contact with the posterior surface of the iris whereas posterior surface is in contact with the vitreous. As the adult lens does not have a blood supply, its metabolic needs are met by the aqueous humour. The lens is composed of 65% water, 35% protein with scant amounts of electrolytes. The proteins can besubdivided into soluble proteins,

Introduction

While experimenting with chemicals, sometimes we need to dry the chemical/ solution (solid, liquid or gas) using a process like evaporation or distillation, mainly to remove water or different solvents. Drying is the process of evaporation or distillation to remove water/solvent from a solution, solid or even from gas. Generally, a drying agent (usually an anhydrous inorganic salt) is used for this process, which reacts with the water (present in the chemical/ solvent to be dried) to form a hydrate. For example, anhydrous MgSO4 is one of the drying agents. The drying agents are recognized by their capacity (the rate at which they absorb water) and their intensity (the amount of water left behind in the solvent at equilibrium). No chemical reaction or change in phase occurs during the drying process.

Type of Solvents

Apart from water (a universal solvent), there are three families of organic solvents used in laboratories.

1) Oxygenated solvents: Alcohols, Ketones, Glycol ethers, Glycol esters, etc.

2) Hydrocarbon solvents: Aliphatic and Aromatic hydrocarbons.

3) Halogenated solvents: Chlorinated hydrocarbons.

There is no rapid direct test to determine fertility of semen sample. During freezing and thawing of semen, 20-80% of the spermatozoa become immotile or dies. In addition to this, there may be damage to spermatozoa by disintegration of sperm cell membrane and acrosome leading to loss of intracellular enzymes. The only reliable method to evaluate the semen is to inseminate a cow and get a calf at the end of gestation period. Now-a-days new methods have been developed for in-vitro assessment of fertility of spermatozoa. Some of these methods are exhaustive, time consuming, need for a full-fledged analysis and difficult under field conditions.

The progress of AI organization depends upon the type of service over and above that initially expected by the members. A well-conducted field service is very important in expanding the growth of any breeding organization. It includes activities as publicity, cooperation in health control, production testing and culling and progeny appraisal, the holding of demonstrations and open house and participation in cattle shows. The salary of a technician with bonus incentive encourages to do work of a high order. The semen costs per inseminated cow can be reduced by maintaining highest quality standards in producing, processing and shipping, by increasing number of cow serviced and by cutting all unnecessary costs. When breeding efficiency is high, the required amount of semen is kept at a minimum. An increase in cows inseminated is a key factor in keeping an organization in the back.

Introduction

For agricultural applications as well as for research work, often we need to prepare solutions of different agrochemicals of varying strength, as per the requirements. Agrochemicals are used to help crop plants grow, produce, and protect them from biotic/abiotic stresses. Many of the agrochemicals (insecticides, herbicides, fungicides, rodenticides, molluscicides, nematicides, etc.) are used to protect the crop from insect pests and diseases. However, some of them (fertilizers and soil conditioners) are used to augment crop yield. While in the previous chapter (Chapter 8) we discussed about preparation of laboratory solutions, in this chapter we will briefly discuss about the preparation of different agrochemicals.

Agrochemical/agrichemical is a generalized term used for the chemicals applied in agriculture. In general, it also includes the feed-supplements used in f isheries and animal husbandry.

Agrochemicals

Fertilizers

Fertilizers are used to augment growth of crop plants and to mitigate nutrient deficiency in the soil. Fertilisers can be organic (containing natural substances, prepared through natural processes) and inorganic fertilisers (synthetic fertilizers, manufactured artificially using chemical processes using natural deposits, by chemical alterations).

Soil Conditioners

Soil conditioners include manure, livestock and leaf compost, peat that help making the soil in good condition. They are laid on the top soil and then mixed properly. They not only increase nutrient contents in the soil but also improve aeration and water holding capacity.

Introduction
Fertilization is sequence of events that begins with contact between a sperm and an oocyte, resulting in fusion of two gametes and formation of a zygote (unicellular embryo). Site of fertilization is distal end of ampulla of fallopian tube (ampulla-isthmus junction) in mammals and in birds, is infundibulum. Only 1% of sperms deposited in vagina enter cervix. Movement of sperm from cervix to uterine tube occurs by muscular contractions of uterus & uterine tube and by their own propulsion. Immediately after ovulation, ovum is caught by the fimbriae of the infundibular part of oviduct and is brought to the ampulla.Cilia along epithelium of oviduct moves OCc. In farm animals (except swine) corona radiata cells drop off in ampulla. In swine, they persist and help in attachment of ova to each other in forming egg-clumps. The optimum fertilizable life of egg in most mammals is 12-24 hours after ovulation, but in dog it may extend up to 4 days.
Site of ejaculation is
Cow, primates, sheep, goat: Anterior vagina
Horse, pig: Body of uterus
Capacitation
Spermatozoa are not able to fertilize the oocyte immediately after arrival in female genital tract. To acquire fertility, they must reside there for a certain period of time. The changes that occur during this period constitute capacitation. Ejaculated spermatozoa must be exposed to secretions of female reproductive tract for a variable period of time before they attain capacity to fertilize oocyte. The secretions of uterus & oviduct participate in capacitation. Important processes during capacitation are: 1. Surface of spermatozoa in region of epididymis acquires certain molecules (proteins & carbohydrates in nature) and in seminal plasma, these molecules are masked by surface proteins. In female reproductive tract, these molecules along with surface proteins are removed. Removal of cholesterol and GAGs sperm cell surface.

Bone is an integral part of musculo-skeletal system of body. In the living animal it is bluish pink, whereas, fresh dead bone is yellowish-white in colour. It is hard and resistant to pressure but also have certain amount of elasticity. Its tensile strength is equal to the cast iron, though bone three times lighter and ten times flexible to cast iron.

Number of bones: The number of bones of the skeleton varies with age and species. Number of bones in cattle-210; Horse-208; Dog-271; Fowl-166 and human-208 (207-212)

Composition of bone: Bone contains different bone cells, organic and inorganic matrix and water. Organic matrix is about one-third and Inorganic matrix is two-third weight by volume. The organic material consists of 90% collagen [type-I (maximum), type V and type XII] and 10% non-collagenous glycoprotein and bone specific proteoglycans (osteocalcin, osteonectin, sialoprotein, bone phosphoproteins and small proteoglycans) and bone growth factors [TGF-beta family; Insulin-like growth factor-1, 2; bone morphogenetic protein (BMP); platelet derived growth factor (PDGF); Interleukin -1 (IL-1); Interleukin-6 (IL-6); colony stimulating factor]. Organic matrix of bone resembles the matrix of dense fibrous tissues such as tendons, ligament and joint capsule. It is not certain whether all of these proteins are synthesized by bone cells or whether they are synthesized by cells outside of the bone and then incorporated into the bone matrix. However, their presence within bone and their potential to affect the activity of bone cells suggests they have important role in controlling bone cell functions. Inorganic matrix of bone or mineral component of bone serves as an ion reservoir and gives bone of its stiffness and strength. It reserves 99% of body calcium, 85% body phosphorus and 40-60% body sodium and magnesium. Inorganic matrix of bone is composed of 85% calcium phosphate, 12% calcium carbonate, 3% of magnesium phosphate, calcium fluoride and magnesium carbonate. It also contains traces of chlorine, sodium, potassium. These minerals remain as hydroxyapatite crystal form within bone. However, bone mineral crystal is not pure hydroxyapatite, instead they contain both carbonate and acid phosphate groups (HPO₄) and bone crystal does not contain OH groups. Bone matrix has such great durability and stability that it can remain essentially unchanged and retain much of its strength for centuries after death. Removal of organic matters renders the bone fragile, brittle and shattered, whereas, decalcification renders bone soft and pliable and resistant to fracture.

Fracture healing is a specialized type of a wound healing response in which the regeneration of bone leads to a restoration of skeletal integrity. Because healing of a fracture in a living organism is accomplished by cells, it can be modified by almost any endogenous or exogenous factors like nutrients, vitamins, hormones, growth factor etc. that has an influence on the metabolic function of bone cells. Fracture healing may be induced by malnutrition that involves certain nutrients, notably calcium (Ca), phosphorus (P) and vitamin D, but also iodine (I), copper (Cu), zinc (Zn), manganese (Mn), fluorine (F), silicon, vitamin A and perhaps vitamin C as well as different endocrines. The number of these nutrients and their interaction is so large that dietary intervention in fracture management is likely to be the most effective if directed at consumption of a good diet that is completely balanced for growth, maturity, old age or stress. Substitution of a good diet for a bad one is also likely to be more convenient and safer than most attempts at partial supplementation of a faulty diet in order to bring it into balance. Underfeeding a good diet decreases size-for-age of bones during growth and delayed fracture healing. Underfeeding may engender osteoporosis in older animals. Underfeeding of Ca, P and vitamin D, causes delayed fracture healing. Over feeding a balanced diet induces rapid growth of newly formed bone and sometimes-accelerated fracture healing but it also causes obesity in adults.

Three major component of bone are osteogenic cells, organic matrix and inorganic matrix (mineral). The osteogenic cells include osteoblasts, osteocytes and osteoclasts, while the matrix consists predominantly of collagen and proteoglycans and constitutes approximately one-third of bone mass (on dry basis). The inorganic/mineral component that makes up approximately two-third of bone and it performs two essential functions : it serves as an ion reservoir and it gives bone most of its stiffness and strength. Bone is a reservoir for 99% of body calcium, 85% of body phosphorus and 40-60% of body sodium and magnesium. The main mineral composition of a long bone is: 85% calcium phosphate as hydroxyapatite, 12% calcium carbonate and 3% magnesium phosphate, calcium fluoride and magnesium carbonate and trace amount of chloride, potassium and sodium. For collagen synthesis, hydroxylase enzymes requires ferrous ion and vitamin C. Impaired collagen synthesis also occurs during deficiencies of vitamin D and E and zinc. The organic intercellular bone matrix is rich in mucoplysaccharide. Their synthesis involves enzymes that require manganese, zinc and vitamin A.

Bone differs from other tissues not only in physio-chemical structure, but also in its extraordinary capacity for growth, continuous internal remodelling and regeneration throughout post fetal life. Bone formation is a complex process regulated by diet, vitamins, hormones and growth factors. Growth factors are polypeptides that increase cell replication and have important effects on differentiated cell function. Growth factors are initially considered as systemic agents, but current evidence indicates that they act primarily as local regulators of cell growth. The transforming growth factor-β family of proteins, which comprises polypeptide growth factors that have diverse effects on the growth, differentiation, and function of cells, is receiving considerable attention for potential clinical applications and is likely of physiological and orthopedically significance.

Transforming growth factors

It is one of the most important growth-promoting osteo-inductive substances, which have been identified at the site of fractures and other places. Transforming growth factors (TGFs) have been categorized into TGF-β (alpha) and TGF-β (beta). TGF-α has not been isolated from bone tissue and cannot be considered a local regulator of bone remodelling, although it is mitogenic for bone cells and stimulates bone resorption. TGF-α shared amino acid sequence, receptor-binding and functional activity with epidermal growth factors but had only weak transforming growth factors activity. Furthermore, TGF-β by itself is completely ineffective, but together with TGF-α, it potently induced growth of colonies of cell-line.

Introduction
Introduction: The digestive system, or gastrointestinal (GI) tract, plays a fundamental role in the overall health and well-being of animals. In veterinary anatomy, understanding the structure and function of the digestive system is essential for diagnosing and treating various disorders that affect food intake, digestion, and nutrient absorption. This complex system is responsible for the mechanical and chemical breakdown of food, the absorption of nutrients, and the elimination of waste, making it crucial for maintaining homeostasis in animal species. Structural Organization: The digestive system can be divided into two main categories: the gastrointestinal tract and the accessory digestive organs. The gastrointestinal tract is a continuous tube extending from the mouth to the anus and includes the oral cavity, esophagus, stomach, small intestine, large intestine, rectum, and anus. Each segment of the GI tract is specialized for specific functions, reflecting the dietary habits and evolutionary adaptations of different species. Mouth and Esophagus: The digestive process begins in the mouth, where food is mechanically broken down by teeth and mixed with saliva, which contains enzymes like amylase that initiate carbohydrate digestion. The esophagus serves as a conduit for food, transporting it to the stomach through peristaltic movements. Stomach: The stomach is a muscular organ that serves as a site for food storage and initial digestion. It secretes gastric juices, including hydrochloric acid and pepsin, to denature proteins and activate digestive enzymes. The morphology of the stomach can vary significantly across species. In ruminants—such as cattle, sheep, and goats—the stomach is uniquely adapted for processing fibrous plant material through a complex structure consisting of four compartments: the rumen, reticulum, omasum, and abomasum.
1. Rumen: The largest compartment, the rumen, acts as a fermentation chamber where microbial digestion occurs. It is lined with a stratified squamous epithelium and contains millions of microorganisms, including bacteria, protozoa, and fungi, that help break down cellulose and other complex carbohydrates. The rumen’s large surface area, facilitated by papillae, enhances absorption of volatile fatty acids (VFAs) produced during fermentation, which serve as a primary energy source for ruminants.

The knowledge and principles from interconnected disciplines of bioengineering, material science and life sciences are combined in tissue engineering with the aim of developing a construct, that can wholly or partly restore/maintain/augment the function of a damaged tissue or organ. Tissue engineering construct generally comprises cells on a suitable scaffold. The ultimate functionality of a tissue engineering construct is mainly dependent on the cell behavior on scaffolds. The proliferation and cell viability on scaffolds varies from one cell type to another and from one species to another. So it is important to investigate individual cell-scaffold combinations. The similarity in structure and composition to bone mineral, osteoconducive properties, ability to integrate with the bony tissue and absence of immune response makes hydroxyapatite [Ca₁₀ (PO₄)₆(OH)₂] the choice of calcium phosphate biomaterial with only disadvantage of slow rate of resorption. On contrary, silica–calcium phosphate composite in comparison to calcium phosphate is rich biomaterials having a faster resorption rate. Studies have shown that coating of hydroxyapatite with a calcium silicate containing layer encourages cell proliferation and osteogenic differentiation of human bone marrow-derived stromal cells. So in this chapter, a triphasic composite scaffold (calcium silicate, hydroxyapatite and tricalcium phosphate), namely HASi with elements in the following percentages: 66.36 % - calcium, 25.35 % - phosphorus and silicon - 8.29 % and porosity of 50 – 500 µm is reported to evaluate the rBMSC proliferation and differentiation properties in vitro.

The present chapter is therefore, included with the aim of assessing cytocompatibility of HASi and rBMSC in terms of cell attachment, cell morphology, cell proliferation and osteogenic differentiation.

Introduction
The integumentary system is a vital organ system in animals, encompassing the skin, hair, nails, and associated glands. This complex system is essential for protecting the body, regulating temperature, providing sensory information, and facilitating communication among individuals. The development of the integumentary system is a remarkable evolutionary adaptation that reflects the diverse environmental challenges faced by different species. The integumentary system has evolved to meet the specific needs of various species, playing a crucial role in survival. As organisms transitioned from aquatic to terrestrial environments, the integument had to adapt to prevent dehydration, protect against UV radiation, and manage temperature fluctuations. For example, the development of a thicker, keratinized epidermis in terrestrial animals minimizes water loss and provides a barrier against environmental stressors. In mammals, the integumentary system exhibits even greater complexity. The evolution of hair, fur, and other adaptations allows for enhanced thermoregulation, sensory perception, and social interaction. For instance, many mammals have evolved specialized hair types, such as the insulating undercoat found in Arctic species or the short, coarse hair of desertdwelling animals that aids in heat dissipation. Anatomical Structure The integumentary system is composed of several key components: the epidermis, dermis, hypodermis (subcutaneous layer), and associated appendages such as hair follicles, sebaceous glands, and sweat glands.
1. Epidermis: The outermost layer of the skin, the epidermis, is primarily composed of keratinocytes, which produce keratin, a protein that provides strength and waterproofing. Melanocytes, located within the epidermis, produce melanin, the pigment responsible for skin color. The thickness of the epidermis varies significantly among species, with adaptations reflecting the ecological niches of each animal.

Fracture healing is a specialized type of wound healing response in which the regeneration of bone leads to restoration of skeletal integrity. Though the process of fracture healing is usually considered to be biologically optimum; the healing of 5-10% is delayed or impaired. For management of these types of fractures, use of biological methods for the enhancement of fracture healing is essential. Furthermore, fracture-healing can be modified by almost any endogenous or exogenous factor that has an influence on the metabolic function of cells. These factors include transforming growth factor-beta, bone morphogenetic proteins, hormones, vitamins, prostaglandin and calcium containing feed supplement medicine.

Keeping in view the above, in this chapter, efficacy of herbal product-Magacal in promoting osteogenesis during fracture healing in experimental animals is discussed. Magacal optimizes absorption, bioavailability and utilization of dietary calcium, phosphorus and magnesium. It is claimed to have advanced chelating agent to form organic complex chelate with dietary Ca, P and Mg, besides containing natural Vitamin D metabolites in highly bioavailability organic form, naturally protected and stable biotin and modern delivery system for optimum translocation of Ca, P and Mg ions to the sites of requirements.

Clinically healthy, adult New Zealand White rabbits of either sex (10) are divided equally into two groups A and B each consisting of 5 animals. All animals are kept on standard diet and management throughout the experiment. In all animals, midshaft simple transverse fracture is created in both the ulnae under thiopental (2.5%) anaesthesia. In test group (Group A), Magacal is given @5ml/animal/ day, orally from day 5-20 post fracture, whereas, in group B, no drug is given which serve as control. Antibiotic dressing of the skin wound is done daily for one week with povidone iodine and himax ointment. All the animals are administered streptopenicillin @ 0.5 gm/animal/day for 5 days postoperatively. Skin sutures are removed on the10th postoperative day.

Clinically all the animals are observed for inflammation, pain and edema at the fracture site, surgical wound healing, appetite, general health, muscular atrophy of the limb, if any, at different intervals postoperatively.

Minimally invasive percutaneous osteosynthesis (MIPO), or minimally invasive fracture repair, was introduced into human orthopedic surgery during the 1990s because of several distinct advantages over traditional open reduction techniques. In MIPO, large incisions to facilitate open reduction are avoided and implants are introduced through small stab incisions in the skin. This led to advent to more biology friendly techniques. These biological techniques lay stress upon maximal preservation of blood supply around the fractured bone by minimal direct handling of the fracture environment. MIPO offer advantages which include preservation of the fracture hematoma; less surgical trauma to the surrounding soft tissues, reduced operative time therefore, decreased risk of infection and ultimately leading to indirect bone healing with abundant callus. Minimally invasive plate osteosynthesis (MIPO) is a novel technique for application of principles of biological fracture healing with a philosophy of dealing with soft tissues with utmost respect. Following advancements in implant technology and development of techniques of indirect fracture reduction this technique gained more popularity. The evolution of MIPO began with the use of bridge plating. Initially a conventional plate is applied for comminuted femoral fractures using long incisions but with preservation of the vastus lateralis muscle resulting in healing with abundant callus. Subsequently, the size of incisions got decreased, rather two small incisions are given at proximal and distal ends and plates are bridged sub muscularly over the fracture fragments. This technique causes minimal distress to soft tissues and bone, provides access to the bone through soft tissue windows, with minimal or no contact with the fracture by indirect reduction tools and leaving behind minimal foot prints.

One of the most common fractures seen in the dog is comminutuion of femoral shaft, often related to sever impact from an automobile accident or jump from a height. Comminuted fractures are characterized by one or more butterfly fragments and assorted small bone chips that are especially difficult to align accurately and stabilize completely. These complicated fractures require sophisticated repair technique that are capable to resisting compression, bending and rotational forces. Rebuilding of comminuted fractures so that all fragments are accurately aligned and gaps are eliminated is often impossible. Non-union and osteomylitis have their highest incidence in these fractures. Over the years, several techniques are tried to treat comminuted femoral fractures with different success rate and there is a lack of uniform fracture presentation. Hence, it will be difficult to compare different techniques in a particular type of fracture. The present chapter is, therefore, undertaken to evaluate radiographically and histopathologically the efficacy of three different internal fixation techniques in the management of comminuted diaphyseal femoral fracture in dogs.

Clinically healthy, adult mongrel dogs (15) of either sex are randomly divided into groups A, B and C of 5 animals each. In all the animals unilateral comminuted diaphyseal femoral fractures is created under general (thiopental - 5%) anaesthesia and is immobilized with intramedullary (IM) pinning with cerclage wiring (Gr. A), neutralization bone plating (Gr. B) and neutralization bone plating with cerclage wiring (Gr. C) respectively. All the animals are administered broad spectrum antibiotic (streptopenicillin @ I g intramuscularly (dog/day) and analgesic (diclophenac sodium @ l mg/ kg b. wt. daily) and surgical wounds are dressed daily with povidone iodine and antibiotic cream for 5 days postoperatively.

Lateral and oblique views of radiography are taken at the operated site just after the operation and subsequently on days 15, 30, 45 and 60 postoperatively. The radiographs are observed for status of fracture reduction and fixation, progress of healing at the fracture site and complications like fixation failure, osteomylitis etc., if any, at different intervals. Bone sections are collected from the fracture site including adjacent normal bone after sacrificing the animals on day 60. The sections are decalcified using Goodling and Stewart’s solution (80% distilled water, 15% formic acid and 5% formalin) and then processed routinely for H & E staining for histopathological studies

Healing of fracture is a consequence of cellular behaviour in a living organism and as such can be modified by almost all endogenous or exogenous factors that affect the metabolic function of cells. These factors include transforming growth factor-β, bone morphogenetic proteins, hormones, vitamins and prostaglandins. The anabolic hormones like thyroxine and insulin accelerate bone formation in experimental animal. Hence, this chapter is proposed to evaluate the effects of thyroxine and insulin therapy in the management of tibial metaphyseal fracture in dogs.

Clinically healthy, adult mongrel dogs (15) of either sex are randomly divided into three groups (A₁, A₂ and A₃) of 5 animals each. In all the animals, unilateral metaphyseal fracture is created in the proximal tibia and then immobilized with cross intramedullary pinning with interfragmentary wiring under thiopental (5%) anaesthesia. All the animals are administered broad-spectrum antibiotic (streptopenicillin @ 1 g intramuscularly /dog/day) and analgesic (5 ml I/M daily) and surgical wounds are dressed daily with Povidone iodine and Lorexane cream for 5 days postoperatively. In group A₂, thyroxine sodium tablets (0.013 mg/kg body wt, orally) and in A₃, insulin injection (2 IU/kg body wt, s/c) are given on day 7 and then on every alternate day till day 45 postoperatively, at one specific time. Group A₁, is remained as control, in this group no hormonal therapy is given.

Craniocaudal and mediolateral radiographs of the fracture site are taken immediately after surgery and subsequent on days 15, 30, 45 and 60 postoperatively. The radiographs are observed for status of fracture reduction and fixation immediately after surgery and subsequently for the maintenance of reduction, progress of fracture healing and complication, if any, at different intervals.

Sodium Clinical Significance Compared to potassium, sodium is distributed differently throughout the body. Due to the sodium pump mechanism, most sodium is extracellular. Compared to this, just 2% of potassium is extracellular; the remainder is intracellular. Regarding the plasma concentrations of sodium and potassium, renal function is the single most significant homeostatic mechanism.

• Sexual activity or mating behaviour is innate character of the males. Unlike the female animal, male animal has a nature of sexual receptivity permanently. • Male gonadal steroid androgen controls sexual behaviour. At regular interval of 2-4 hrs, there is secretion of testosterone which converts into estradiol due to aromatization. Due to this very frequent rise in estradiol level, male shows sexual behaviour permanently. In female animals, a rise in estradiol occurs only during estrous. • When in a group of animals, any female can be mated with any male, it is called promiscuous sexual behaviour. Sexual behaviour in some animals is according to their social structure and territorial/home range i.e. male and female share some of their home range for the purpose of mating e.g. Roe deer, Muntjak antelope etc. In some animals, both males and females share whole of their territory for the mating purpose called a harem e.g. wild rabbit. Some animals come into a temporary territory meant only for mating in breeding season e.g. seal.