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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

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.

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 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, 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 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 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 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 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 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.

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.

• 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”.

• 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).

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)

• 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)

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 • 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.

• 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

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.

• 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