The zona pellucida (ZP) is a extracellular transparent coat surrounding the oocyte and preimplantation embryo in mammals. The ZP separates between the surrounding cumulus cells and oocyte. It plays a pivotal role in oocyte growth and fertilization and early embryo development. Zona pellucida formation and glycoproteins, roles of ZP on female fertility and infertility, ZP micromanipulation on embryo development and changes of ZP upon vitrification of oocytes is discussed. Zona pellucida is formed of glycoproteins during ovarian follicle growth and development. It protects the oocyte from physical or chemical damage, initiates fertilization, prevents polyspermy and supports early embryonic development. The ZP is responsible for recognition between oocytes and sperm. Mutations in glycoproteins cause female infertility characterized by empty follicle syndrome and oocyte degeneration. The ZP removal is detrimental for embryo implantation. Furthermore, there is a biophysical role of ZP during vitrification of oocytes. Hence, the current review article was designed to collect, discuss and highlight the characters of zona pellucida, roles in female fertility and infertility, ZP micromanipulation on embryo development and changes of ZP upon vitrification.

An Insight into the Roles of Zona Pellucida in Growth and Development of Mammalian Ocytes and Embryos: Changes of Age-related and Cryopreservation: A Review

Background: Affections of eye are commonly encountered in all the species of animals. The age-related changes of the retina primarily cause loss of visual acuity as well as reduction of the visual field. Hence, the current study was carried out to establish basic data related to ageing changes in the retina of buffaloes. 
 
Methods: The study was conducted on 63 eye balls of locally available buffaloes of different breeds. These buffaloes were categorized into 3 groups i.e., group I (1-5 yrs), group II (6-10 yrs) and group III (11 yrs and above). The eyeballs were isolated and fixed with Davidson&rsquo;s fluid. The paraffin sections were subjected for routine histological study. 
 
Result: The thickness of retinal pigment epithelium was increased from group I to III buffaloes from 6.06&plusmn;0.18 to 8.44&plusmn;0.32 and the quantity of melanin pigment was decreased with advancement of age. Rod and cone cells of the photoreceptor layer were tightly packed in young age, loosely arranged in old animals. The mean thickness (&micro;m) of photoreceptors and outer nuclear layers together was decreased from 64.22&plusmn;1.84 to 58.72&plusmn;2.1 with age advancement. The displacement of nuclei from outer nuclear layer into outer plexiform layer was significantly decreased in old animals. The outer limiting membrane was continuous and uninterrupted throughout the life. The thickness of outer plexiform layer was increased with advancement of age from 8.78&plusmn;0.58 to 10&plusmn;0.51 due to enhancement of synaptic fibers density. As age advances the number and density of horizontal, bipolar and amacrine cells were decreased and also the mean thickness (&micro;m) of this layer was decreased from 30.83&plusmn;1.48 to 22.56&plusmn;0.62 in the inner nuclear layer. The thickness of inner plexiform layer was increased from 42.44&plusmn;2.23 to 45.39&plusmn;0.81 with advancement of age due to increased cystoids spaces and thickening of retinal blood vessels. In ganglionic cell layer, the number of &alpha;-ganglion cells were more than the &beta;-ganglion cells, their common number were decreased approximately from 16-20 cells/sq.mm to 7-10 cells/sq.mm, whereas the average size was increased from 4.72&plusmn;0.49 to 15.83&plusmn;0.83 with advancement of age. In aged buffaloes nerve fiber layer showed corpora amylacea and thickened blood vessels. The inner limiting membrane became thick and uninterrupted in old buffaloes. The total thickness (&micro;m) of retina in group I, II and III buffaloes were 235.5&plusmn;7.25, 184.33&plusmn;3.64 and 201.05&plusmn;5.15 respectively.

Histological Studies on Ageing Changes in the Retina of Buffaloes (Bubalus bubalis)

<div>Background: Affections of eye commonly encountered in all the species of animals. If these not treated in time, the vision may be hampered, this may impair the physical ability, utility and productivity of animals leading to economic loss to the animal owners. Corneal dystrophies like basement membrane dystrophy, stromal dystrophy, endothelial dystrophy, corneal erosions and corneal ulcerations etc., seen in the aged buffaloes. The current study was carried out to establish basic data and to provide authentic information to the clinicians on ageing changes in eyes of buffaloes. 
 
Methods: The study on age related changes in the cornea was conducted on 63 eye balls of locally available buffaloes of different breeds in and around Proddatur. These buffaloes were categorized into 3 groups based on their age i.e., group I (1-5 yrs), group II (6-10 yrs) and group III (11 yrs and above) buffaloes. The eyeballs were isolated and fixed with Davidson&rsquo;s fluid. The paraffin sections of cornea were subjected for routine histological study. 
 
Result: The mean total thickness (&micro;m) of cornea was increased from group I to III animals and it was 747.22&plusmn;13.57, 853.33&plusmn;11.05 and 897.22&plusmn;8.06. Hence, The mean thickness (&micro;m) of all five layers of cornea increased with advancement of age. Superficial corneal epithelial layer was made up of non keratinized stratified squamous epithelium with 6 to 7 layers at early age and 12-14 layers of cells in adult animals. The mean thickness (&micro;m) of corneal epithelium in group I to III animals was 117.04&plusmn;3.37, 122.59&plusmn;2.23 and 133.71&plusmn;3.55 respectively. Bowman&rsquo;s membrane of cornea was measured 12.97&plusmn;0.86, 17.22&plusmn;0.55 and 18.71&plusmn;0.68 in group I, II and III animals respectively. The mean thickness (&micro;m) of corneal stroma in group I to III animals was 600.75&plusmn;10.33, 672.22&plusmn;10.46 and 690.00&plusmn;5.32 respectively. This was mainly due to progressive increase in number of collagenous lamellae. The keratocytes became thin in old animals. Thickness of posterior band zone (PBZ) of Descemet&rsquo;s membrane was increased with advancement of age. The mean thickness (&micro;m) of Descemet&rsquo;s membrane and endothelium together was in group I, II and III animals was 16.48&plusmn;0.91, 41.29&plusmn;2.25 and 54.82&plusmn;1.44 respectively. The corneal endothelial cells were enlarged and increased in size in old buffaloes. The age related pigmentation was also noticed in epithelium and stroma of cornea in animals of group III. Density of sub basal nerve plexuses between the stroma and the corneal epithelium was not altered with advancement of age. &nbsp;</div>

<div>Histological Studies on Ageing Changes in the Cornea of Indian Buffaloes (Bubalus bubalis)</div>

<div>Background: Transition period is a critical period&nbsp; in which most dairy animals undergo a period of negative energy balance (NEB), hypocalcemia, insulin resistance, hypoglycemia and susceptibility to infectious diseases. The critical changes in biochemical indicators are well reported in dairy cows whereas; in dairy buffaloes limited data related to alterations in hemato-biochemical changes have been reported so far. The current study aimed to evaluate the hemato-biochemical changes in buffaloes during transition period. 
 
Methods: A total number of 210 buffaloes aged between 3 to 14 years with parity ranged from 1st to 4th, divided in three groups (n=70; each) during transition period were used for the study. Three groups viz Group I (-30 days), Group II (0 days) and Group III (+30 days) were subjected for evaluation of hemato-biochemical parameters.&nbsp; 
 
Result: Our study showed significant increase (p&lt;0.05) in WBC (103/&micro;l), lymphocytes and granulocytes count at the time of calving. Significant decrease (p&lt;0.05) in the level of RBC, Hb and HCT were seen in Group-I and II. In biochemical analysis, parameters like albumin, total protein, triglyceride and A:G ratio showed significant (p&lt;0.05) decrease from Group-I to II. Calcium and Ca:P ratio decreased significantly (p&lt;0.05) along with suppressed levels of sodium and potassium levels in Group-III. Thus, the present study showed significance of hemato-biochemical analysis during transition period for possibility of occurrence of production diseases.</div>

Haemato-biochemical Profiling in Buffaloes in Relation to Metabolic Changes during Transition Period

The interaction between body weight, testicular growth and sperm production, is the key factor influencing the onset of puberty and sexual maturity. The present experiment was carried out to study the changes in testicular biometry and seminal parameters during pre and post pubertal period in Black Bengal bucks. Black Bengal males (n=8) of four months age were used in the study. Body weight and scrotal biometry parameters increased significantly as age advances. First ejaculates were collected from kids aged 4.5 months which contained mostly immature sperm cells, but semen ejaculates containing adequate numbers of matured motile sperm cells could be collected from six months of age onward.&nbsp; Semen ejaculate characters were optimal at 11-12 months of age with almost stable body weight and scrotal biometry. Based on the above facts, it could be concluded that the Bengal bucks attained puberty at 5-6 months and sexual maturity at 11-12 months of age.&nbsp;

Changes in Testicular Biometry and Ejaculate Characters during Pubertal Age in Black Bengal Kids

<div style="text-align: justify;">Changes occurring in seed during aging are very significant for determining seed quality, i.e. seed longevity. An experiment was conducted in Department of Seed Science and Technology, TNAU, Coimbatore to standardize suitable seed dry dressing treatments, using near nano size botanicals that can alleviate the deleterious effect of accelerated ageing and to improve the enzyme activities of soybean aged seeds. Five days aged seeds were treated with 1 h ball milled fenugreek seed powder, leaf powders of ashwagandha, tea and noni @ 2 g kg-1 with 60 min. shaking and evaluated for enzyme activities and field emergence (naturally and artificially aged seeds). Among the treatments seeds treated with fenugreek seed powder has the highest dehydrogenase activity (OD 10 min-1), protein (%), oil content (%), catalase activity, peroxidase activity (OD 10 min-1) (&micro;g H2O2 mg-1min-1) with minimum values of electrical conductivity (dSm-1), free fatty acid (%), lipid peroxidation (OD value), lipoxygenase activity (g mol s-1 mg-1). Field emergence (%) was also higher in fenugreek seed powder treated seeds (2 g kg-1 with 60 min).&nbsp;</div>

Changes of enzymes activities in botanical treated aged seed of&nbsp;soybean (Glycine max (L.) Merrill) cv. CO 3 seeds

<div>Background: Hansli chicken is reared in Mayurbhanj and Keonjhar districts of Odisha. They play a vital role in the economic upliftment of poor, landless and marginalised people in the rural areas besides providing them with nutritious egg and meat for consumption. 
 
Methods: Hansli chicks and Vencobb broiler chicks were divided into three age groups viz. group I (up to 1 month), group II (1-3 months) and group III (3-6 months) with six birds in each age group. On 4th week, 12th week and 24th week, six birds from each breed were used for the study of histomorphometrical features of the thymus, spleen and bursa of Fabricius. 
 
Result: The average thickness of the capsule, cortex and medulla of thymus, average width of thymic lobule and average maximum diameter of Hassall&rsquo;s corpuscles differed significantly (p&le;0.05) between the birds at specific ages. Similarly, the average thickness of the splenic capsule, average width of white pulp, average external transverse and internal longitudinal diameters of trabecular artery, average external longitudinal, internal longitudinal, external transverse and internal transverse diameters of central artery, average longitudinal diameter of capillary, average longitudinal and transverse diameters of splenic nodule, average thickness of PALS and PELS differed significantly (p&le;0.05) between the birds at specific ages. The average number of bursal follicles in larger and smaller plicae, average height and width of larger and smaller plicae, average length and width of bursal follicle, average height of columnar cell of pseudostratified FAE and IFE, average nuclear height of FAE and IFE columnar cells differed significantly (p&le;0.05) between the birds at specific ages.</div>

<div>Age Related Histomorphometrical Studies on the Major Lymphoid Organs of Chicken</div>

<div>Background: Banana is a commercial fruit of many tropical and sub-tropical nations. India is the largest producer of banana in the world. The fruit is a valuable source of carbohydrates, phenolics compounds, minerals and dietary fibre. Due to bumper production, seasonality and market gluts, huge post-harvest losses are recurring phenomena in many banana growing states in India. Hence, the present study was undertaken to evaluate the suitability of some commercial banana cultivars in the development of wine and to evaluate the qualitative changes during ageing. 
 
Methods: Wine was prepared from banana varieties (Poovan, Grand Naine, Yangambi (KM-5), Karpooravalli and Palyankodan) in 1:1 and 1:2 dilutions of pulp and water. The TSS content of the substrate was raised to 20&deg;Brix, followed by addition of yeast @ 1.25 g/l. The must was treated with Potassium Metabisulphite (KMS) @ 0.05 g/L to inhibit the growth of other microorganisms. Fermentation was carried out for 15 days, followed by clarification and subjected to ageing for 2 months. 
 
Result: During ageing of wine, the pH and alcohol content showed an increasing trend whereas titratable acidity, total soluble solids, ascorbic acid and phenols showed a decreasing trend. The flavour profile analysis of wine revealed that ethanol, ethyl hydrogen succinate and glycerin were the major flavour compounds in banana wine. After the completion of ageing, alcohol content of banana wine ranged from 8.40 to 9.64%, phenols were in the range of 40.50 to 56.25 mg 100 g-1 and total soluble solids varied between 3.6 and 4.5&deg; Brix. Wine prepared from the Poovan variety had the highest overall acceptability score throughout the ageing period.</div>

Qualitative Changes and Flavour Profile of Banana (Musa spp) Wine during Ageing

<div style="text-align: justify;">The gross studies were conducted on the thymus of 18 postnatal Surti goats. Three distinct parts of thymus were recognized viz., unpaired thoracic part, caudally situated paired body (cervical part) and right and left cranial parts from day old kid to 9 months. The cranial parts were not observed from the age of 12 months onwards. The caudal cervical part of thymus was gradually replaced by adipose tissue between 14 to 48 months, however, traces of thymus embedded within the adipose tissue was observed in cranial mediastinum even at the age of 48 months. The right limb of cervical thymus was slightly longer than the left limb in postnatal Surti goats. The maximum mean weight (26.850 &plusmn; 2.885 g) and volume (22.333 &plusmn; 2.275 cc) of thymus was recorded between 4 to 9 months. Relative weight of thymus was 0.39% in neonatal kids, which was much higher as compared to 0.015 % in adult Surti goats. Negative correlation was observed between age and weight of thymus during 12 to 48 months.</div>

Age related gross morphological studies on the thymus in postnatal Surti goat (Capra hircus)

Background: The productivity of livestock production has been greatly impacted by parasitic diseases. The infection prevalence of parasites in native and imported slaughtered animals has not received much attention, despite the fact that Saudi Arabia has data on animal parasites. This study aimed to determine the infection prevalence of parasites in slaughtered animals in relation to host origin and host age.&nbsp;

Methods: This investigation was conducted at Al Makhwah area in southwestern Saudi Arabia for six months from September 2021 to February 2022. The examined animals were 1208 in total, including goats (504), sheep (440), and cattle (264). All the animals examined were slaughtered at official abattoirs. The collected parasites were preserved, processed, and identified.&nbsp; 
 
Result: Out of 1208 examined, 72 were infected (5.96%). The parasite community consisted of six species, Dicrocoelium dendriticum, Fasciola hepatica, F. giagantica, Monezia expansa, Echinococcus granulosus (Hydatiid cyst), and Haemonchus contortus. Hydatidosis was a more prevalent parasitic disease.&nbsp;The infection prevalence of hydatidosis in the examined animals was high in cattle (4.85%) while were 3.76 and 4.77% in goats and sheep respectively. Significant differences in prevalence per host age in goats, sheep, and cattle were found. The prevalence of parasites in imported animals was higher (16.67, 17.5, and11.36 % in cattle, sheep, and goats respectively) as compared to that in native ones (5.42, 6.5, and 3.69% in cattle, sheep, and goats respectively). Based on our findings, Hydatidosis was identified as the most prevalent parasitic disease. Imported animals had a higher parasitic infection compared to local animals. It is essential to take necessary measures to prevent the transmission of Hydatidosis and improper organ condemnation from abattoirs. We recommend conducting further research to identify the reasons why imported animals are more likely to become infected than local animals.

An Insight into the Roles of Zona Pellucida in Growth and Development of Mammalian Ocytes and Embryos: Changes of Age-related and Cryopreservation: A Review

Histological Studies on Ageing Changes in the Retina of Buffaloes (Bubalus bubalis)

Histological Studies on Ageing Changes in the Cornea of Indian Buffaloes (Bubalus bubalis)

Haemato-biochemical Profiling in Buffaloes in Relation to Metabolic Changes during Transition Period

Changes in Testicular Biometry and Ejaculate Characters during Pubertal Age in Black Bengal Kids

Changes of enzymes activities in botanical treated aged seed of soybean (Glycine max (L.) Merrill) cv. CO 3 seeds

Age Related Histomorphometrical Studies on the Major Lymphoid Organs of Chicken

Qualitative Changes and Flavour Profile of Banana (Musa spp) Wine during Ageing

Age related gross morphological studies on the thymus in postnatal Surti goat (Capra hircus)

Parasite Prevalence in Slaughtered Animals in Relation to Host Origin and Host Age