The objective of the present experiment was to investigate the effect of cysteamine and b-mercaptoethanol supplementation on in -vitro maturation, cleavage of oocytes and development of embryo in buffalo (Bubalus bubalis). Oocytes were aspirated from abattoir ovarian follicles of 3-10 mm diameter followed by maturation in the media in vitro containing cysteamine/b-mercaptoethanol (treatment) and without antioxidant (control). Matured oocytes were co-incubated with sperm (approx.1&times;106/ml) of Murrah bull in mSOF medium using heparin (10 &micro;g/ml). After 22 h of oocyte-sperm incubation, fertilized oocytes were stripped of cumulus cells and cultured in mSOF medium for 8 days to study embryo development. The oocyte maturation rate improved significantly (P&lt;0.05) following addition of &nbsp;50 or 100 &micro;M of cysteamine and 10, 50 and 100 &micro;M of &nbsp;b- mercaptoethanol (ME), respectively as compared to control. The cleavage rate was found to be significantly (P&lt;0.05) higher at 50 and 100 &micro;M of cysteamine and at all concentrations of b-mercaptoethanol as compared to control and development of embryos to morula stage was significantly (P&lt;0.05) improved with 50 &micro;M cysteamine/ b-mercaptoethanol. 
&nbsp;

Cysteamine and b-mercaptoethanol supplementation improves the in vitro&nbsp;embryo development in buffaloes (Bubalus bubalis)

<div>Background: Bovine anaplasmosis caused by Anaplasma marginale is highly endemic in tropical and sub-tropical regions of the world where it accounts for huge economic losses to the livestock sector. Approximately 30% of water buffalo (Bubalus bubalis) herds in India are located in Uttar Pradesh (U.P). However in India, most of the research studies on Anaplasma are focused on cattle whereas studies on this pathogen in water buffaloes have been neglected. Therefore, present investigation was undertaken for determining the prevalence of A. marginale infection in water buffaloes in U.P, North India.&nbsp; 
 
Methods: In the current investigation during 2017-2019, the prevalence of A. marginale infection in 228 water buffaloes in three districts of North-West U.P was studied by microscopy and by nested-PCR amplification of major surface protein 5 (msp5) gene.&nbsp; 
 
Result: Microscopic examination of blood smears of 228 water buffaloes revealed 7.89% animals positive for A. marginale. Conventional PCR of full length major surface protein 5 (msp5) gene detected 29.39% and nested-PCR amplification of this gene detected 64.0% of these animals as positive for A. marginale infection. A high incidence of the infection detected in this study area suggests that water buffaloes may serve as reservoir hosts of A. marginale and play a vital role in the spread of infection to other susceptible animals. However further studies are required on larger buffalo populations for determining the epidemiological status of anaplasmosis in these animals in India.</div>

Prevalence of Anaplasma marginale in Water Buffaloes (Bubalus bubalis) in the State of Uttar Pradesh, North India

<div style="text-align:justify">The objective of the study was to characterize the term Doka and to capture morphological changes in teat to find out the relationship between Doka and onset of estrus in Murrah buffaloes (Bubalus bubalis). The Murrah buffaloes maintained at of Livestock Research Centre (LRC), ICAR-NDRI, Karnal were classified into two groups based on the lactation number for the study of Doka characteristics. In group I, twenty animals up to 3rd lactation were incorporated, whereas thirty animals from 4th to 5th lactation were included in group II. The period when buffaloes showed teat engorgement was called as Doka period, is the local term used by the farmers in North India that can be taken as a visual sign for prediction of estrus. The length and diameter of teats, before and after milking were measured, for both the groups during morning and evening milking and subsequently analyzed. The present study confirmed that the teat diameter (mm) before milking during Doka was significantly (p&lt;0.05) higher as compared to pre Doka period in both the groups (group-I - 37.01 &plusmn;2.13 vs 28.42 &plusmn;1.64, group-II - 53.49 &plusmn;2.82 vs 41.096&plusmn;2.16). The teat diameter (mm) during Doka period after milking was significantly (p&lt;0.05) more in both the groups (group-I - 26.65 &plusmn;1.26 vs 24.44 &plusmn;1.16, group-II - 39.27 &plusmn;2.41 vs 35.31 &plusmn;2.17) as compared to pre Doka period. So, it can be concluded that the changes in teat diameter can safely be considered as a sign of Doka and the forthcoming heat in Murrah buffaloes. However, the difference in teat length between different stages of Doka was found to be non significant.</div>

<div style="text-align:justify">Changes in Teat Morphology (Doka Phenomenon) and Estrus Prediction in Riverine Buffaloes (Bubalus bubalis)</div>

<div>Background: Many years have been devoted to preserve fertility, but the effect of cryopreservation on gene functionality in primary, secondary and tertiary follicular stages; is still unclear. The present study was designed to assess the effect of vitrification on the histological structures and expression of follicular cells related genes.&nbsp; 
 
Methods: The buffalo ovarian cortical tissues were vitrified in two-steps. The sliced cortical tissues were incubated with VS1 (8.5% DMSO and 8.5% EG) for 10 min and sequentially incubated and stored in liquid nitrogen in VS2 (16.5% DMSO, 16.5% EG and 0.1M Sucrose). Morphological differences were assessed by hematoxylin and eosin staining, which indicated similar structures in both groups. Further, functionality of these follicles were evaluated by the relative gene expression of folliculogenesis-related genes; FOXO3, NLRP5, WNT4, SF1, VEGFA and HAS2. No significant difference was observed between the vitrified groups as compared to control. Moreover, toxicity in follicular cells during vitrification was rectified by MTT assay which also showed no significant difference.&nbsp; 
 
Conclusion: Present study can be considered as the key work that helps in filling the gaps regarding the growth of follicles after cryopreservation in buffalo species, as this imparts nonsignificant injury on follicular functionality and development.</div>

<div>Effect of Vitrification on Folliculogenesis-Related Genes in Ovarian Follicles of Bubalus bubalis</div>

The process of follicular development and features of morphological changes are essential to understand the factors involved during folliculogenesis and to improve fertility in buffalo. Hence the objective of the present study was to report ultrastructural features of atretic ovarian follicles in buffalo. In the early stage of atresia, the involvement of mitochondria was observed and whereas in the later stages of atresia, involvement of macrophages were observed. The granulosa cells of buffalo atretic tertiary follicles were observed to contain undulation of the nuclear membrane, crescent formation of the pyknotic nuclei and loopiness and undulations of the basal lamina. These features are similar to the reports in other species. The involvement of mitochondria in the early stage of atresia suggested the probable role of a reactive oxygen free radical induced damage.

Ultrastructural features of atretic ovarian follicles in buffalo (Bubalus bubalis)

<div style="text-align: justify;">Present investigation was carried out to determine the effect of calf weaning and natural suckling on the growth performance, immune status, health and behaviour of buffalo calves. Sixteen newly born calves (8 calves from weaned group and 8 calves from natural suckling group) were raised in two separate groups under loose housing system. The mean daily colostrum intake during first two days was higher in suckled calves as compared to weaned calves. The average daily body weight gain was significantly higher (0.506 &nbsp;vs. 0.438 kg/day) in suckled calves as compared to weaned calves. The mean of time spent on cross sucking, licking self and licking objects were significantly higher in weaned calves as compared to suckled calves. The level of oxidative stress of weaned calves were significantly higher as compared to suckled calves. It was concluded that the growth performance, immune status, health, behaviour and oxidative stress of natural suckled calves were better as compared to that of weaned calves.</div>

Influence of weaning on growth, health and behaviour of Buffalo (Bubalus bubalis) calves

<div style="text-align: justify;">The present study was conducted on hippocampus of six adult Surti buffalo. Hippocampus was a &nbsp;small organ located within the medial temporal lobe of the brain. The hippocampus made a curve from the deep face of the piriform lobe around the thalamus and formed the caudal part of the floor of the lateral ventricle. It was separated deeply by the hippocampal fissure from the dentate gyrus. The two hippocampi were connected at their highest parts by transvers fibers which constituted the hippocampal commissure. The hippocampus was as long &ldquo;C&rdquo; or small &ldquo;Y&rdquo;shaped structure. The ventro-medial hippocampal surface had a deep groove, the hippocampal sulcus, which divided this surface into a lateral and medial surfaces. The ventricular surface of the hippocampus was covered with a thin layer of white matter, the alveus, which arises from the crus of the fornix, and was therefore continuous with the fimbria. Moreover, the uncus was not observed. Several equations showed significant (P&lt; 0.5) and positive co-relationship between the weight of hippocampus and the weight of brain. The ratio of weight of the hippocampus and weight of the brain was about 1: 37.&nbsp;</div>

Gross anatomical and biometrical parameters of hippocampus in Surti buffalo (Bubalus bubalis)

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 style="text-align: justify;">An investigation was taken up to study the relationship of &nbsp;preovulatory follicle size at the time of first postpartum AI and CL biometry with conception using transrectal ultrasonography in 70 parous Graded Murrah buffaloes maintained under rural conditions. POF sizes were measured ultrasonographically and grouped in to small preovulatory follicle (SPF), medium preovulatory follicle (MPF) and large preovulatory follicle (LPF). Serum progesterone concentration at the time of AI and pregnancy status was negatively correlated indicating that when progesterone level declined to &lt; 0.3 ng/ml (basal level) at the time of AI, the chances of the animal becoming pregnant was increased. The size of the POF was positively correlated to the size of the CL on day 10 in pregnant buffaloes. The overall POF diameter at the time of estrum was 12.31 &plusmn; 0.29 mm (Range 9 to 16 mm) in Graded Murrah buffaloes and 50 % of the buffaloes had POF size &gt; 12 - &pound;14 mm. They conceived only if the POF diameter was more than 9 mm at the time of AI however there was no significant correlation between the POF size at the time of AI and conception. Thus, it was concluded that physiological maturity rather than the diameter of the follicle influenced the fertility in Graded Murrah buffaloes under field conditions.</div>

Effect of preovulatory follicle on fertility in Graded murrah buffaloes&nbsp;(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>

Cysteamine and b-mercaptoethanol supplementation improves the in vitro embryo development in buffaloes (Bubalus bubalis)

Prevalence of Anaplasma marginale in Water Buffaloes (Bubalus bubalis) in the State of Uttar Pradesh, North India

Changes in Teat Morphology (Doka Phenomenon) and Estrus Prediction in Riverine Buffaloes (Bubalus bubalis)

Effect of Vitrification on Folliculogenesis-Related Genes in Ovarian Follicles of Bubalus bubalis

Ultrastructural features of atretic ovarian follicles in buffalo (Bubalus bubalis)

Influence of weaning on growth, health and behaviour of Buffalo (Bubalus bubalis) calves

Gross anatomical and biometrical parameters of hippocampus in Surti buffalo (Bubalus bubalis)

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

Effect of preovulatory follicle on fertility in Graded murrah buffaloes (Bubalus bubalis)

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