eChapters

Introduction Sea food processing is one of the major contributors in food processing sector. The purpose of seafood processing is to prevent the wastage and prolong the shelf life of highly perishable commodity. It also increases the economy of the country through exports (Gopal et al., 2014). India produce 9.6 million MT of sea food during the every year and is one of the largest fish producers in the world. 23% of marine products are processed currently in India (Sathianandan, 2017). Pasteurized crab meat is one type of processed marine commodity exported from India. Globally, the pasteurized blue crab and swimming crabs are the major contributors to the economic wellbeing of the seafood industry. In India, pasteurized crab meat industries are significantly contributing to the export markets. Among the crab export, 70-80% is exported as pasteurized crab meat (Jebadurai17, 2014).

Introduction Aquaculture is one of the world’s fastest-growing food industries, providing a sustainable source of income for millions of people (Mohanty et al., 2018, Troell et al., 2014; FAO 2018). From the total world’s production around 33% of the all harvested fishes are not only utilized for human utilization for different reasons like size, species, quality, amount, non-accessibility of handling or transportation office. Today, waste generated from fish processing is a major source of pollution in the environment. Even when excellent grade fish is produced, only 50% of it is consumed by humans, with the remainder being discarded as waste. These wastes, which are dumped in an open environment without being treated, produces foul odour, which, as previously said, might be a reason of environmental pollution, water pollution and other municipal difficulties. Fish meal and oil made from low-grade fish and discarded fish parts not only solves many of these health and environmental issues, but also boosts the fishing and fish processing sectors’ overall revenue. With the rapid build-up in yearly extraction since 2000, global usage of natural resources has increased dramatically in recent years, reaching 92.1 billion tonnes and resulting in a 254% rise from 27 billion in the nineteenth century (Coppola et al., 2021). As a result, the research is stepping up its attempts to develop a circular bio-economy that aspires to increase the value of material flows while also achieving sustainable consumption and production. It is a new idea that is gaining traction for the goal of ensuring human quality of life through the effective and sustainable use of resources, energy, and infrastructure.

Introduction Noodles have received importance worldwide due to their particular taste, flavor, convenience, consumer acceptance, and low cost. The noodles generally come under the category of unhealthy and junk food because of high carbohydrate content and low vitamins, minerals, and fiber content (Park et al., 2011). Due to the increasing popularity of instant noodles, many people are becoming more prone to various health problems. The substitution of various ingredients to wheat flour noodles is increasing by incorporating different pulses, cereals, and pseudocereals to improve the nutritional properties and also make them health food. Among the pseudocereals, the use of amaranth and chia seeds in food products has been increasing because of their high nutritional value and good health benefits. Their proteins have well-balanced amino acids with a high content of essential amino acids and also good bioavailability (Lopez et al., 2019). These pseudocereals are also gaining importance due to their high content of phytochemicals and other macromolecules. The amaranth (Amaranthus sp) seeds are spherical and small, having approximately 1 mm diameter with a creamish to light reddish color. The composition of amaranth flour includes amylose (7.05%), proteins (15.27%), fat (7.33%), crude fiber (2.04%), and ash (2.93%) (Siwatch et al., 2019).

Introduction Pulses are a vital source of protein (20-25%) in comparison to cereal proteins (10-12%); which cater to the most vegetarian population (Venkidasamy et al., 2019) and other countries which are scarcity of animal protein availability. In the category of pulse crop, chickpea is considered the second utmost significant crop, which is grown in almost 35 countries globally. Chickpea mainly contains carbohydrates, protein, minerals, vitamins, dietary fiber, folate, β-carotene and health-promoting fatty acids (Badshah et al., 2003; Tosh & Yada, 2010) and consuming these grains will repeatedly help in lowering the risk of chronic diseases (Jukanti, Gaur, Gowda, & Chibbar, 2012). Ozone is often regarded as a potent oxidant capable of penetrating the grain (Tiwari et al., 2010), and it can control the bio-agents such as insects, molds, and mycotoxins (Wang et al., 2016). Due to the fact that ozone gas is unstable, it convert back into oxygen without leaving any residues in short time (McClurkin, & Maier, 2010), so, it can be used as a fumigant without leaving any residue. The Food and Drug Administration (FAO) has approved it for fumigating grains, fruits, and vegetables.

Introduction In today’s environment, the greatest challenge for the food industry is to develop health promoting foods; it is determined by the raw materials used and its impact on consumer health. Furthermore, people lifestyle has changed greatly, resulting in a change in food eating patterns. In today’s world, people are moving towards eating foods that are nutrient base and tasty (Bonciu, 2018). of all the nutrient based products available in market, expanded snacks have become a popular snack all over the world because of its crunchy texture and low density making it consumer’s acceptance (Grasso, 2020). The basic idea for developing any expanded snacks is utilizing a starch-based material for a better crispy and crunchy product, but to make it more nutritious incorporation of protein and fibre rich raw materials is recommended (Gu et al., 2017). Along with the raw materials, the other important parameters for developing any extruded product is the particle of the flour, its physicochemical and rheological properties of the extrudates are much concentrated (Bouasla & Wójtowicz, 2021) Thereby, extruded snack is considering as functional food with major health benefits.

Introduction In latest years, there has been an upsurge in customer situation over using artificial colorants in foods, especially on the subject of healthiness and safety issues (Amchova, Kotolova, & Ruda-Kucerova, 2015). Dietary colorants have been banned or regulated by the European Food Safety Authority (EFSA) and various other national authorities throughout the world because to their confirmed or suspected link to enhanced cancer progression or adverse reactions. For the same motives, various pigments are being investigated and are most effective provisionally authorized at this time. Consequently, the use of natural additives in food production is becoming increasingly popular among food companies (Schwartz, Joachim, & Giusti, 2007). Red C-PE is the most extensively used colour and has various application in food, cosmetics, and pharmaceutical industries; however, because they are rare in nature, they must be synthesized. Therefore, the food sector is expressing an increasing interest in the finding, application, and stabilizing of natural red colorants, which are currently in high demand. A growing number of commercial companies are turning to cyanobacteria for colour extraction, including Spirulina sp. for the extraction of Phycoerythrin, Haematococcus sp. for the extraction of astaxanthin pigments, and Dunaliella sp. for the extraction of beta-carotene pigments (Sharma et al., 2021).

Introduction Flavour has a significant impact on consumer pleasure and subsequent food intake. Chemical synthesis or extraction are used to create the majority of accessible fragrance molecules. Synthetic flavourings are frequently shunned by customers who believe that hese compounds are harmful or pose a health risk to them. (Teixeira et al., 2004). The association between flavour stability and food quality and acceptance has piqued researchers' interest, yet it's tough to control. Food manufacturing and storage procedures, packaging materials, and ingredients frequently alter overall flavour by lowering the intensity of aroma compounds or introducing off-flavor components (Lubbers et al.,1998). Because there are so many variables in flavours, they constitute extremely complicated systems. Some are more stable in water soluble carbohydrates, whereas others are more stable in lipid-based coatings. Physico-chemical characteristics, concentration, and interactions of volatile aroma molecules with food components are just a few of the aspects that determine the overall quality of the food. It's a good idea to encapsulate volatile components before using them in foods or beverages to prevent fragrance degradation or loss during processing and storage. Encapsulation is the process of coating or encasing one material or a mixture of components into another material or system. The coating substance is known as shell, wall material, carrier, or encapsulant, and the coated material is known as active or core material.

Introduction The onion (Allium cepa) is one of the world's oldest cultivated crops, with a history dating back over 5,000 years (Gorrepati et al., 2014). At present, India is one of the world's largest producers and exporters of onions. At the time, India ranked second in onion production with a yearly production of around 23262 TMT and an average yield of 18.10 MT/ha (Prajapati and Saini, 2019; Horticulture Statistics at a Glance, 2018; Tripathi and Lawande, 2016). Maharashtra, Madhya Pradesh, Karnataka, Gujarat, Rajasthan, Bihar, Andhra Pradesh, Haryana, West Bengal, Uttar Pradesh, Chhattisgarh, Odisha, Tamil Nadu, Jharkhand and Telangana are the major onion producing states in India. Maharashtra alone covers 55-60% of countries total production. Solapur, Satara, Dhule, Jalgoan, Osmanabad, Beed, Latur, Aurangabad are the major onion producing districts in Maharashtra (Tripathi et al., 2005). Unfortunately, India witnessed around 40-50% post-harvest losses of onions every year (Prajapati and Saini, 2019).

Introduction A functional food is a food that provides alimentative beyond their actual nutritional value & are claimed to have an extra function by the addition of new ingredients or more of already present ingredients into it. Functional food have also become a part of our everyday daily diet & are told to offer potential various health benefits other than the widely accepted nutritional effects among them. (Roberfroid MB 2009). Many bioactive active compounds are contained by them like roughage, OS, vitamins, inorganic elements, & bacterium probiotic, having scented whiff, relish & fineness, which sustain the balance & balance of the enteric system. Functional may also be said as artificer‘s foods, NC, restorative foods, curative food, & super food etc because of the roles & functions they perform. (Wlodzimierz Grajek et al., 2008). There are many utilitarian sequels of functional food & beverages with added bug & probiotics on the health of humans which are increasingly promoted & recommended by the nutritionists & other we-being professionals. (Aditya Chaudhary 2019). For a time being, the most salient & over & over again used functional foods amalgam are probiotics & probiotics, or they altogether called as symbiotic‘. he dwell Microbes or foods microbes, which are beneficial for human, are probiotics. (Armachius James &Yousheng Wang 2019).

Introduction Sea buckthorn berry belongs to Elaeagnaceae having 6 species (Hippophae rhamnoides, Hippophae neurocarpa, Hippophae salicifolia, Hippophae gonicarpa, Hippophae tibetiana and Hippophae litangenis) and 12 subspecies grown all over the world. India, is having its plantations in chilly and barren conditions of Ladakh, Himachal Pradesh, Uttarakhand, Sikkim, and Arunachal Pradesh. Berries and fruits are believed to have such bio-active compounds like antioxidants in them that it has constructive consequences on health of human beings. Moreover, they are been proven in lessening the cardiovascular diseases, age related issues,cancer etc. Berries of Sea buckthorn is an abundant provenance of these bio-active compounds. One of the major lipophilic compounds is carotenoid pigments. And the concentration of the same has difference depending upon origin of plant, harvesting period and weather conditions. Nutritionally, the fruits have a substantial amount of carbohydrate, protein, organic acids, amino acids, and vitamins. Moreover, it also contains bioactive compounds like carotenoids, dietary minerals, antioxidants, flavonoids, vitamin C, minerals, monosaccharide, organic acids, tannins, etc. Sea buckthorn plant parts are becoming one of the useful and promising nutraceuticals for the food industry and all over the world.

Introduction Rice cultivation is a means of livelihood for a large number of people from the state of Assam, India (Saikia, 2019). It is related to the cultural aspects in rural Assam where people indulge themselves in traditional as well as modern ways of farming. Like any other north-eastern states in India, rice majorly dominates the Assamese cuisine. It is the primary source of diet in Assam. There are more than 100 varieties that are genetically modified breeds in which indigenous varieties contribute to 20 germplasm (https://www.rkbassam.in/). The different rice varieties of Assam have different culinary applications in both savoury and sweet cuisines. The intermediate and high amylose rice, parboiled or nonparboiled are consumed as staple for major meals while the low amylose rice is mostly consumed as snacks, breakfast cereals and desserts. The major constituent of rice is starch consisting of about 90% of the mass. Thus, the major compositional changes occurring in rice is due to starch. Starch is a polymer of glucose linked by glycosidic bond. It consists of amylose (linked by alpha 1- 4 linear chains) and amylopectin (alpha 1-4 & alpha 1-6 branched chains). On the basis of composition, rice is classified as high amylose (>25%), intermediate amylose (20 -25%), low amylose (7-20%) and waxy or glutinous (1–2%) types (Juliano, 1979).

Introduction Consumer demands for convenience and mildly processed meals with long shelf lives have led to unique and current developments in food packaging solutions (Dobrucka and Cierpiszewski, 2014). Additionally, the development of unique and creative packaging solutions that do not compromise food safety or quality is being fueled by the current retail trend and changing lifestyles (Dainelli et al., 2008). The widespread use of packaged meals has raised consumer demand for prepared foods like microwaveable dinners, and the growing popularity of smaller food packages all help to fuel the food industry's explosive growth in creative packaging (Restuccia et al., 2010). The rise in food-borne microbial outbreaks, which calls for the usage of packaging that possesses antimicrobial qualities while retaining food quality, is another major driver of new food packaging (Appendini and Hotchtkiss, 2002). The initial advancements in packaging were electrically powered packing equipment, flexible packaging, metallic cans, aseptic packaging, aluminium foils, and flexographic printing. A considerable movement away from metal, paperboard, and glass packaging in favour of plastic and flexible packaging was also brought about by the discovery of various materials such as polyester, polypropylene, and ethylene vinyl alcohol polymers.

Introduction India is a country of festivals and celebrations. Indian sweets enjoy a legendary reputation across the world. India is home to a number of delicious sweets that play a vital role in every Indian celebration. While gulab jamuns, rasgullas and kaju katlis may be the most popular sweets in the country (Pal Sanchari, 2016). Functions and events in India are celebrated with intake of sweets. Also, it is customary to sweeten the mouth in social gathering like marriage, birthday, poojas, etc. (Gulati and Misra, 2014). In the past few years, guidelines are formulated based on analyses of all published scientific studies on the consumption of sugars. In view of the rising prevalence of overweight, obesity and related disorders globally, in 2002, the WHO recommended that sugar should be less than 10 percent of daily calories. The consensus dietary guidelines for Indians in 2011 recommend less than 10% of total calories from free sugars per day. In May 2014, the expert panel of WHO recently recommended decreasing sugar intake to 5% of total calorie intake to combat obesity. A high intake of added sugars is a primary risk factor for diabetes. It is these added sugars in sweets that makes them palatable, but also does the physical damage. (Willet Brian, 2011). People have developed taste towards food in which fat and sugar contents are high.

Introduction Solid–liquid extraction process, has been extensively used for extraction of bioactive compounds from plant food matrices by using appropriate solvent, which can be used as functional ingredients and additives in food processing, pharmaceutical sectors, and others industries. Usually this extraction/recovery process is based on unsteady diffusion of compounds targeted for extraction, through the matrices followed by the dissolution of the extractable compounds in the bulk of the used solvent (Milic, Rajkovic, Stamenkovic, & Veljkovic, 2013). The concentrations of bioactive compounds in food matrices as well as in the extracts are generally low in percent, so extra energies are required to improve the yield of the extraction process while minimizing the overall cost. From the technological approach, extraction process is mainly associated with the operating variables such as solvent used, extraction technique, operating temperature, concentration of solid in solvent, and extraction time, and by optimizing these parameters extraction yield could be enhanced (Milic et al., 2013; Orphanides, Goulas, & Gekas, 2014).

Introduction Diet and dietary component are proved to be the first line of intervention in prevention and treatment of non-communicable diseases by many studies (R. Vrolix and R.P. Mensink, 2010). Hence the interest has grown in development of functional food product in recent years which provides not only basic nutrients but also has health promoting and diseases preventing properties (Rozek et al., 2010). The Functional Food Center (FFC) defines “functional food” as natural or processed foods that contain known or unknown biologically-active compounds; the foods, as defined, effective, and non-toxic amounts, provide a clinically proven and documented health benefit for the prevention, management, or treatment of chronic disease (Martirosyan et al., 2015). Fiber is classified into two separate groups i.e. soluble which forms gel like substance when comes in contact with water and insoluble is a solid form that does not break down when exposed to water (web 4). Soluble fibers increase total transit time by delaying gastric emptying and also slow glucose absorption, which includes pectins, β-glucans, galactomannan gums, mucilages, and some hemicelluloses (J.H.Cummings and A.M.Stephen 2007). The American Society for Nutrition stated that cereal fibers are modestly associated with a reduced risk of type II diabetes, obesity, and cardiovascular disease (Cho Qi, Fahey Jr & Klurfield, 2013).

Anatomically the male reproduction system comprises of testes, scrotum, duct system, accessory sex glands, prepuce, and penis. Production of spermatozoa and male sex hormone are the main functions of male reproductive physiology. The Male reproductive system can be categorized as: • Primary sex organ – Testicles • Secondary sex organ – Epididymis, vas deference, urethra, and penis. • Accessory sex glands – Seminal vesicles, prostate, and bulbourethral glands/cowper’s gland. 1. Testes (Paired male gonads) (Singular: Testis or Testicle) • Testes are the primary sex organs. They are situated in the scrotum at the inguinal region with the help of tunics and spermatic cord. Opening of the scrotal pouch into the abdominal cavity is called as inguinal canal. The testis is covered with an extra abdominal tunic named as tunica vaginalis, tunica albuginea, and tunica vasculosa from outside to inward. The fluid filled in between the parietal and visceral layers of tunica vaginalis is responsible for the movement of the testis within the scrotum. The upper end of a testicle is occupied by the caput epididymis and the lower end is connected to the cauda epididymis.

• In male animals, endocrine regulation of reproduction is mainly controlled by pituitary gonadotrophins and gonadal androgens. It is not the same as in females as there is only negative feedback controls in case of male while it is positively as well as negatively feedback control in females. In other words, the synchrony of pituitary gonadotrophins with gonads in male is non cyclic while in female it is cyclic i.e. even after castration (absence of androgen) the pulse frequency of LH and FSH is retained. • Sex Hormones are sex specific e.g. estrogen and progesterone work primarily in females and testosterone in males but sex specificity is limited as far as sexual behaviour is concerned. As estrogen injection in castrated ram recovers the maleness while testosterone injection in ovariectomized female maintain the receptivity for male up to a certain extent.

• To identity the normal fertile, infertile (Temporary loss of fertility), subfertile (reduced fertility), or sterile (no fertility) male animals makes the requirement of breeding soundness evaluation necessary. Infertile and sterile animals prove to be harmful for the economic survival of the dairy industry and should be eliminated from the breeding management system. • The breeding soundness examination is very necessary for the breeding management system as the ultimate goal is the achievement of healthy pregnancies. It is also useful for successful artificial insemination (AI) programs that involve the collection and cryo-processing of quality semen.

Puberty • It is the age at which sexual organs are functionally developed and gonads start responding to GnRH to start steroidogenesis and spermatogenesis. Animal at puberty develops secondary sexual characters with proper libido. • At puberty, the bull is able to produce 50 million spermatozoa with 10% progressive motility in 1st seminal ejaculates. It should be 100 million spermatozoa with 10% progressive motility in 1st seminal ejaculate in case of stallion. • At puberty, gonadotrophic cells are functionally developed in the mature pituitary tends to gonadal maturation. The steroidogenesis starts properly which leads to the development of genital organs and secondary sexual characters.

Libido • It is the willingness & eagerness of male animal to mount and perform complete service. It is sexual desire or drive which is an innate character of male animals. • Libido and mating ability have no direct relation with fertility. Libido is not related directly to sperm production and semen quality. • The libido of an animal depends on androgenic steroid hormone. Vasectomy or cryptorchidism does not affect libido in male animals. • Animals have no or reduced libido that are castrated before puberty. If the animals, that know the copulation, are castrated after puberty, they display the copulation behaviour for longer period. • Reaction time – It is the time between the contact of a male animal with female animal to 1st false mount with erected penis. Reaction time is the good predictor of libido.

• 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.

Fertility is the ability of a male to impregnate the fertile female. The reduced or temporary loss of fertility is called infertility whereas permanent loss of fertility is called sterility. A male with low libido or poor sex drive may have excellent semen quality or a male with good libido may have poor seminal quality. So, libido has no direct relation with the male fertility. Male infertility can be categorized into Impotentia coeundi and Impotentia generandi. 1. Impotentia Coeundi • The condition of reduced or lack of sex drive or Inability to copulate (failure of normal service) is referred to as impotentia coeundi. • It may occur due to immaturity, poor experience to perform coitus, inability or unwillingness to mount, inability to perform intromission or failure of ejaculation. • The following conditions may reflect the immaturity or poor/bad experience of the male animal which leads to impotentia coeundi:

• These are the injuries occurred to male animal at the time of natural mating. These injuries do not occur only to genital organs but also to other organs or muscles. • Most coital injuries occur to male animal with good libido (high testosterone level) but having aggressiveness and poor experience of mating. Such animals tend to quick response in the presence of estrous female. Some of the coital injuries are: 1. Balanitis and Posthitis • Trauma to the penis and prepuce which later on leads to infection and inflammation. • This occurs especially in young inexperienced bulls with great sexual desire • Adhesion may lead to phimosis or paraphimosis condition • IBR-IPV infection from cow genitalia after mating results in pus formation and necrosis of the penis and prepuce. • Prepucial eversion leads to laceration and bleeding.

• These are the bad habits of the male animal due to which ultimately their maximum reproductive potential cannot be utilized. • There are so many vices seen in male animals like stall walking, weaving, cribbing, wind sucking, crib biting, viciousness etc. These behavioural disorders are mostly due to improper managemental practices like abusive handling, poor exercise, irregular semen collection and poor exposure to female etc. These may be also due to hormonal imbalance or nervous disorders. - Some important vices which are more related to reproductive behaviour are: 1. Masturbation (Onanism) • Bulls masturbate by rubbing their prepuce and penis against any object. • Boar masturbate and ejaculate by inserting the penis into the preputial diverticulum called Balling up • Boar may show rectal copulation and leads to ejaculate : Pederasty

Objectives • Seminal evaluation and processing • Evaluation of bull fertility on the basis of its seminal characters • To make the Artificial insemination programme successful History Related to Semen Collection • The use of artificial vagina was started by Milovanov for semen collection. Before that method of placing the cotton ball in vagina was used. • AV for Dog was discovered by Amantea in 1914. Milovanov discovered AV for Bull, ram and stallion. Mckenzie discovered AV for boar (1931). • The electroejaculatory method of semen collection was initiated in a bull by Laplaud & Cassou and in ram by Gunn. • Burrows and Quinn (1937) developed the method of abdominal massage and pressure to collect semen in poultry.

• Mature sperm cells are undifferentiated cells which are the testicular product as well as indicators of testicular spermatogenic function. A spermiogram reflects the testicular activity and fertility status of a male animal. • Disease conditions, age, stress factors, vaccination or any altered physiological conditions may affect the seminal quality. Objectives of Semen Evaluation • To provide essential information about the fertility status of the male animal, though the fertilizing capacity of the animal cannot be completely defined by these tests. • To diagnose various pathological conditions of testes and male genitalia. • To determine the eligibility for further processing of semen cryopreservation • To determine the potential of semen for AI purposes

Semen • Semen, a fluid released from the male reproductive system during ejaculation, is made up of a liquid phase called seminal plasma and a cellular component called spermatozoa. • The term “spermatocrit” refers to the percentage of spermatozoa(v/v) in semen, which varies from less than 2% in pigs to more than 30% in sheep. It is about 10% in bull. Semen= Spermatozoa + seminal plasma Spermatozoon • In 1677, the scientist Van Leeuwenhoek first discovered the spermatozoa and called them ‘‘animalacula’’. • Spermatozoa or sperm cells or male gametes, develop within the wall of the seminiferous tubules as a result of a complex process known as spermatogenesis. • The spermatozoon is a highly specialized cell that performs the sole function of fertilizing an oocyte.

Sperm abnormalities may be genetical or morphological. They are directly related to the male infertility. • The wet mount method and phase contrast microscopy facilitate sperm visualization for abnormalities. Semen stained with vital stain can also be used for evaluating sperm abnormalities. Sperm morphology or related abnormalities can be evaluated by using eosin-nigrosine stain, carbolfuschin, aniline blue-eosine etc. • Generally, for head and tail abnormalities rose Bengal and William’s stain can be used while for acrosome and head abnormalities Giemsa stain can be used. Categorization of Sperm Abnormalities 1. On the basis of site of abnormality on the spermatozoa • Head abnormalities e.g. microcephalic head, macrocephalic/giant head, narrow or short sperm head, pyriform head, detached glea capitis (acrosomal cap), detached head and double head etc

• Semen cryopreservation – It is the process of preserving the semen in a suitable medium at an ultra-low temperature which reduces spermatozoa metabolism with the least production of metabolic waste products of sperm cells to prolong their life span. • Every fall of 100 C temperature decreases the metabolic rate of spermatozoa to half. Principle of various components involved in cryopreservation 1. Low temperature • Reduce metabolism (negligible) and least metabolic waste product • Decrease solute solubility leads to increased viscosity and physical resistance • Reduce motility • Prolong life span 2. Protection from cold shock and cryoinjury • When sperm cells are subjected to a reduction in temperature during the freezing process, there are: • Sperm membrane damages and lead to leakage of intracellular components like enzymes, ATP, K+ , cholesterol, lipoproteins etc. • Irreversible abnormality in motility pattern (bending of sperm tail)

Semen is needed to be extended with a suitable medium and cryopreserved without affecting its survivability and fertility for the purpose of inseminating female artificially. Qualities of Ideal Extenders • Semen extender is a nutrient media which provide energy to spermatozoa, protection from osmotic shock, thermal shock and various metabolic wastes for prolonging the life span of sperm cell with maintaining the acceptable fertilizing capacity. A semen extender should have qualities to protect spermatozoa during dilution and cryopreservation. These qualities may be : 1. Preserving sperm cell integrity (membrane, cellular, mitochondrial and acrosomal integrity etc) 2. Maintenance of osmotic pressure (isotonic; 280-300 mOsm/l) similar to seminal plasma 3. Providing nutritive elements as energy source for sperm metabolism (aerobic & anaerobic metabolism)

• After equilibration, a pre-freeze evaluation of semen is done. Pre-freeze motility should be more than 70% at 370 C which makes the semen eligible for freezing. Semen Packaging • Semen is packaged in suitable container like pellets, glass ampoules, tupol (polyethene tubes) and semen straws. • In the Ampoule method, semen is packed in glass ampoules and stored in LN2 . Here labeling and handling is easier with less chances of contamination. But more storage space, more time taking to thaw and the use of glass catheter for insemination make their the use less convenient. • In Pellet methods, diluted semen is frozen by pouring it into the holes (0.1, 0.2 ml) made on dry ice (Solid CO2 at 790 c) and these pellets (frozen semen balls) are stored in LN2 . Here glycerolization is not needed as well as process is also quick but labeling is difficult and the chances to get contaminated are more. Handling pellets for insemination is also difficult (breakage or getting stuck to forceps during retrieval).

• Artificial Insemination (AI) is the technique of deposition of semen (Fresh, Diluted or frozen thawed) in the female genital tract by artificial means for the purpose of conception. • It is first generation biotechnology technique. It is the most economical and best technique which has been widely accepted for genetic improvement and production (calf crop and milk production) in farm animals. • All the aspects like selection of bull, semen collection, semen evaluation, processing of semen, freezing, cryopreservation, thawing and insemination are equally important for making AI successful.History Related To AI • In the fourteenth century an Arabian placed a cotton ball in the mare's vagina and collect it after mating (soaked in semen) with a high-grade stallion; then by inserting this cotton ball into the vagina of desired mare and got it conceived. • Anton Van Leeuwenhoek (Dutch scientist, 1678) and his assistant, Hamm, discovered sperm cell and called it as “Animalcule”.

There are some methods to control male reproductive behaviour for better breeding and managemental practices i.e. Vasectomy and Caudal epididymectomy, Penile surgeries, and castration etc. • Surgical and non surgical interventions in the male reproductive system are carried out for: • To make teaser bull • To make bull sterile • To prevent the transmission of venereal diseases • Population control or Breeding management • To rectify any ailment or deformity Teaser Bull • A male animal whose capability of spermatogenesis as well as libido is maintained but it is not able to impregnate a female. • For the preparation of teaser bull or Gomer bull, Male animal should be selected well before the start of breeding season. Bull should be young vigorous and fertile. It should be of moderate adult size with mild temperament. Heavy and large sized bull is tough to handle, needs more time for surgery, has chances of more hemorrhage, and may have more post surgical complications.

1.1 Introduction Pearl millet [Pennisetum glaucum (L.) R. Br.] is a warm-season, C4 , annual cereal crop mainly grown for food, feed and fodder purpose. It is diploid species having chromosome number 2n = 2x = 14, belongs to genus Pennisetum, tribe Paniceae of Gramineae family. It has several species e.g., americanum, typhoides, and glaucum. Pearl millet originated in northern Africa in the region from western Sudan to Senegal. It is the sixth most important cereal crop in the world and third most important crop in India after wheat and rice.1.2 Area and Production Pearl millet is cultivated in arid and semi-arid regions of the world. India is the largest producer of millet in the world. In India, pearl millet occupies 6.93 million ha with an average production of 8.61 million tones and productivity of 1243 kg/ hac. The major pearl millet growing states are Rajasthan, Maharashtra, Gujarat, Uttar Pradesh and Haryana which account for more than 90% of pearl millet acreage of the country and are commonly grown in kharif season. It is also cultivated during the summer season in parts of Gujarat, Rajasthan and Uttar Pradesh and during the rabi season at a small scale in Maharashtra and Gujarat.

1.1 Introduction Millets were one of the first crops to be domesticated by mankind in Asia and Africa which later spread across the globe as critical food sources to the evolving civilizations. The earliest evidence of millets has been found in the Indus civilization around 3000 BC. Millets are a collective group of small-seeded annual grasses that are grown as grain crops, primarily on marginal land in dry areas of temperate, sub-tropical and tropical regions (FAO). However, due to the popularity of the cereal crops like wheat, rice and maize; millets have lost dominion and have become an underutilized crop in last few decades. Millets have the potential to be the crop of the future, as our major crops have reached yield plateau, also changing climate has introduced several new biotic and abiotic stresses to the major crops, so to feed the ever-growing population we need to shift towards other potential crops. India is the largest producer of millet as it contributes about 41% of the total millet production of the world (APEDA, 2020).

1.1 Introduction Sorghum [Sorghum bicolor (L.) Moench], a cultivated diploid (2n = 20) tropical cereal C4 grass plant, is currently the fifth most important cereal crop grown in the world and the first important crop in Sudan where drought stress is a major productivity-limiting factor since most of the sorghum is grown under rain-fed conditions. It is a monocotyledon plant of tropical origin, belonging to the Poaceae family. Starch, followed by protein, fat, and fibre, makes up the majority of sorghum’s nutritional makeup, which is similar to maize in terms of its nutritional content. Compared to other cereals, including wheat and maize, the crop exhibits comparatively good water use efficiency. Sorghum is a valuable botanical model crop for other tropical grasses with complex genomes that use C4 photosynthesis due to its vast adaption to severe settings, resistance to stress conditions, diverse germplasm collections, and short (710 Mb) genome size. Sorghum, a historically important source of food and a staple crop, is becoming a crop with a variety of applications, including food, fuel, fodder, and a model for the functional genetics and genomics of tropical grasses.

1.1 Introduction Millets are the most ancient grains that have been cultivated since time immemorial and consumed as food, feed and pastures. Millets are considered nutraceuticals’ since they possess significantly higher nutritional qualities compared to other cereals. Millets are grown across the world for grains and human food due to their nutritional advantages and ability to withstand adversities. Millets are preferred over other grains due to their short life cycle, low input requirement and ensured production with in-built genetic tolerance for abiotic stresses. Major millets include Pearl millet (Pennisetum glaucum L.) and sorghum (Sorghum bicolor L.), while there are numerous minor millets classified as small millets. With an extent of over 0.87 lakh ha and a production of 0.66 kg ha-1, India is the world leader in small millet production (Maitra and Shankar, 2019). Small millets are primarily grown on rainfed uplands in arid and semi-arid climates with sub-optimal management. Following the green revolution due to extreme demands for food, the main focus in India was on the production of fine cereals mainly rice and wheat to overcome hunger and poverty, while millets became ignored grains. Furthermore, urbanization, increased affluence and dietary changes relegated millets to the status of poor man’s food. However, due to a re-evaluation of nutritious characteristics, millets have recently regained their lost prestige (Maitra, 2020).

1.1 Introduction Among the C4 grasses known as panicoids (subfamily: panicoideae), which also includes nutrient-rich cereals like millets and sorghum as well as commercially valuable crops like switchgrass, napier grass, and guinea grass, are bioenergy feedstocks like sugarcane and miscanthus. Among all, millets overpowered others in terms of biotic/abiotic stress resilience, resource sustainability as well as nutritional benefits. Minor millets, often known as nutritional cereals, are a type of small-seeded grains that are members of the Poaceae family. Up to 35 kinds of grass from 20 genera are well-known for being minor millets. Finger millet, foxtail, proso millet, barnyard millet, kodo millet, and little millet are the most significant small millets that are grown worldwide. India is the leading producer and consumer of millet crops and their products. The total area of small millets in India is around 1.63 m ha, with a production of 1.82 m tones during the year 2016- 17 (Annual progress report 2018-19, ICAR-AICRP on small millets).

1. 1 Introduction, Origin and Taxonomy Barnyard millet (Echinochloa species) is one of the oldest domesticated small millet crops which can be grown in both warm and temperate regions of the world and it is widely cultivated in Asia. It is distributed predominantly in the Indian subcontinent, China, Japan, and Korea and also to some extent in Africa and United States. In India, this crop is mainly grown in Maharashtra, Orissa, Tamil Nadu, Madhya Pradesh, Bihar, Punjab, Gujarat and the hills of Uttarakhand (Kumar et al., 2000). It ranks fourth in terms of production amongst the small millets. Indian Barnyard millet (Echinochloa frumentacea) and Japanese Barnyard millet (Echinochloa esculenta) are the two important species of the barnyard millet. Barnyard millet is majorly grown for human consumption, though it is also grown for livestock feed and fodder (Sood et al., 2015).

1.1 Introduction Millets are nutri-cereals that have been the oldest food since time immemorial and are decisively the first cereal grain to be used for domestic purposes. Millets generally include small seeded members of the Poaceae family globally used for food and fodder purposes. The word “millet” emanated from the French word “Mile” which means “thousand” signifying that a small quantity of millet may contain thousands of grains. Millets were erstwhile produced in marginal or degraded terrains with very low soil nutrients and semi-arid conditions with very little rainfall. After the green revolution, there was a shift away from millet in favour of wheat, rice, and maize as a result of more land becoming irrigated to alleviate extreme poverty and malnourishment in the country and to feed millions. Most of the millet crops possess the C4 photosynthetic pathway and are therefore able to withstand frequent changes due to environmental stresses. Consequently, millet production has declined and within a few decades post-green revolution, crops that were once eaten by every household were transformed into fodder crops. Millets are considered as ‘good for the planet crops’ as they take less to give more, due to such multi-faceted qualities millets are considered as miracle crops.

1.1 Introduction Millets are one of the world’s earliest food grains, arguably the first cereal grain utilized for household uses. It has been a cherished crop in India for ages and constitutes a staple food for over one-third of the world’s population. Recently ICAR has designated millets as “Shree Anna”. Several types of small-seeded cereals are grouped together as small millets. These crops offer a reliable and affordable harvest from the regions where the primary cereals fail to produce a sustainable yield. Small millets are insignificant in terms of food production, but they are vital as staple foods in their particular ecosystems. Small millets are more than just finger millet (Eleusine coracana) and include kodo millet (Paspalum scrobiculatum L.), little millet (Panicum sumatrense), foxtail millet (Setaria italica L.), proso millet (Panicum miliaceum L.) and barnyard millet (Echinochloa frumentaceae). The rest of the two small millets namely, fonio millet (Digitaria exilis) and teff (Eragrostis tef) are of local significance in Africa. Small millets have a high nutritional value, but also their demand has deteriorated due to a lack of standard processing techniques to compete with fine cereals. Small millets have a huge potential to satisfy our nutritional needs, therefore we must push ourselves and make efforts to enhance its utilization by incorporating them into our daily diet.

1.1 Introduction Millets are small-grained ancient cereal crops that were renowned as one of the oldest sources of nourishment in prehistoric times. They are believed to have been cultivated since 8000 BC. Apart from providing nutritious grains, these crops are well adapted to the marginal and dryland ecosystems due to their climatic resilience, biotic stress tolerance, and capacity to flourish in the wild. Among the millets, small millets make up a distinct subgroup and comprise of finger millet (Eleusine coracana), little millet (Panicum sumatrense), foxtail millet (Setaria italica), proso millet (Panicum miliaceum), kodo millet (Paspalum scrobiculatum), barnyard millet (Echinochloa frumentacea) and brown top millet (Brachiaria ramosa L.) in India along with a few more cereals like teff, fonio, job’s tears and guinea millet in some other parts of the world. As fine cereals gained popularity after the 1960s, the acreage and significance of these crops began to decrease. In recent years the drastic climate change has made us to focus on underutilized crops.

Introduction Millets are a group of annual grasses which are small seeded, hardy, drought tolerant, low maintenance and mainly found in semi-arid regions. The historic importance of millets can be derived from various types of traditional dishes which uses millets like roti and chapati flatbreads in India, uji (a type of porridge), injera (a fermented flatbread commonly eaten in Ethiopia) and beer in Africa, and kasha and fermented drinks in Eastern Europe. Millets are the 6th most important cereal grain in the world. India, China and Nigeria are the largest producers of millets, and the Research Institute for the Semi-Arid Tropics estimates that in Africa and Asia more than 90 million people depend on millets in their diets. There are hundreds of millet types, but the major types grown commercially are: proso millet, foxtail millet, pearl millet, Japanese barnyard millet and browntop millet. In this book chapter, we will mainly focus on two minor millets i.e., Teff and Fonio

Introduction The world recently celebrated the birth of its eighth billionth baby, a major milestone in the development of the human race and the doubling of our numbers in 47 years. This tremendous growth has been driven by ambitious advances in technology and health care, soaring achievements of global prosperity and a longer life expectancy that has come to a staggering 72 years. However, the glory of these achievements is marred by how we are failing the lion’s share of the global population in meeting their basic requirements. Almost one in seven people on our planet do not have enough to eat. Food and nutritional insecurity are detrimental to human development and are a major brake on overall socio-economic development which downplays any major accomplishments we have made since not having enough to eat is an international security issue. Adequate food production, easy access and the capacity to buy food, relevance concerning nutrition and stability of these elements are all necessary for sustainable food security (Helland and Sorbo, 2014).

1.1 Introduction In parallel with the expansion of organically cultivated land and low-input agricultural system, there is an increasing demand for crops that have promising potentials to be grown successfully in these systems. Buckwheat is generally sown in smaller areas, which may be found suitable for low-input agricultural systems. Buckwheat is a fast-growing crop with a variety of utilizations. Buckwheat, one of the pseudo and minor-cereals, has the ability to satisfy the rapidly rising population’s ever-increasing food need in the nation’s hill regions in the context of changing climatic conditions. It is also known as common buckwheat or Fagopyrum esculentum, it is not a cereal grain, but rather a seed that is related to rhubarb and sorrel. It has a unique nutty flavour that makes it an excellent ingredient in a variety of dishes. It is known in India by a variety of names, including ogal, phaphar, bresha, mittahe dyat, dro, and brotitae.

1.1 Introduction Amaranth is considered as one of the “superfoods” of the coming century due to its exceptional qualities in various aspects. However, the crop is not a novel discovery, and it has faced quite a number of obstacles in order to get to where it is today in terms of nutritional status and acceptance by modern day consumers as a food which is able to provide nutrition. Despite the setbacks and disuse, Amaranth has had three redeeming qualities that have earned its standing as a “superfood.”In recent years, the grain amaranths have garnered increased attention because of their nutritional qualities and, in particular, the nutritional value of the seed protein. Amaranth seed is higher in fiber (8%) than the grain of most cereals.

Introduction Cabbage (Brassica oleracea var. capitata) belongs to the family Cruciferae or Brassicaceae having chromosome number 2n=18. ‘Cole wart,’ a wild non-heading variety of Brassica oleracea var. sylvestris, is the ancestor of cabbage. The majority of the 100 species that make up the genus Brassica are indigenous to the Mediterranean region. It is also believed that Germany is where the true-headed cabbage originated. Currently, mostly cabbage is grown in the country like Caribbean, Indonesia, Malaysia, Central Asia, and Western and Eastern America.The third-largest producer of cabbage worldwide is India. The majority of the nation’s states where it is grown include UP, Orissa, Bihar, Assam, WB, MH, and Karnataka. Annually in India, cabbage was produced around in the area of 418 thousand hectares and production of 9.72 million metric tons with the productivity of 22.7MT/hectares (NHB, 2021-22). The other main producers in the globe, in addition to India, are China, Russia, Japan, Korea, the USA, Indonesia, and Ukraine.

Introduction Cauliflower Brassica oleracea var. botrytis) is a cold season vegetable cropbelongs to family cruciferous or Brassicaceae family with somatic chromosome number 2n = 18. Cauliflower is derived from the Latin word i.e., ‘caulis’ means stem and ‘floris’ means flower and wild progenitor is Brassica cretica. In India, cauliflower is grown in two different varieties: snowball type and tropical type. However, at first, other varieties of cauliflower, such as the European and Cornish types from England, were also brought to India. Later, the Cornish and European forms of cauliflower were intercrossed to develop the tropical type of cauliflower. The Indian type or tropical type of cauliflower can be grown in hot weather and high rainfall condition therefore, such type of cauliflower is mostly grown in Indian plains during the month of May to January followed by temperate type or snowball group based on the temperature based varietal classification.

Introduction In India, knol-khol (Brassica oleracea var. gongylodes) is belongs to family brassicaceae with chromosome number is 2n=18. It is grown in north eastern states of India, J &K, West Bengal, Assam, Punjab, and other regions of southern India also. It is distinguished by the formation of knobs(tuber) is a thickening of the stem tissue above the cotyledons. This knob is picked for human use as a raw or cooked vegetable, while young leaves are also utilized in some areas. Economic Importance & Uses Knol-khol, also known as kohlrabi or German turnip, is a biennial vegetable that belongs to the same family as cabbage and broccoli. It is a popular crop in many parts of the world, including Europe, Asia, and the Americas.

Introduction The ‘Brussels sprouts’ (Brassica oleracea L. var. gemmifera) coming under Brassicaceae family with chromosome number 2n=18 preferred by farmer for its edible green bud or sprouts (small heads that resemble miniature cabbages). It is a robust, slow-growing, longseason vegetable. The “sprouts” are formed in the leaf axils, beginning at the base of the stem and progressing upward. Brussels sprouts are high in vitamin A and C, as well as folic acid and dietary fiber. It also has anticancer effects due to the presence of sulforaphane. Similarly, to other Brassicas, Brussels sprouts contain indole-3-carbinol, which inhibit cancer cell growth. The temperate regions in the hills have the greatest potential for commercial production of brussels sprouts, followed by coastal regions with prolonged cool falls.