Lala Lajpat Rai University of Veterinary & Animal Sciences, Hisar

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Open Access
Effect of supplementation of zinc and nano zinc on Post thaw variables and oxidative status of bovine semen
(2023-06) Sudarshan Kumar; Sindhu, Sonia
The present study was undertaken to investigate the effect of supplementation of zinc and nano zinc on post thaw variables and oxidative status of bovine semen The concentration of ZnCl2 and nano ZnO was optimized for use in semen extender. 22 semen ejaculates from three indigenous cattle bulls were supplemented with the optimized concentration of ZnCl2 (40mg/ml) and ZnONP (10 and 20 µg/ml) and were cryopreserved as per the standard freezing protocol. Semen was evaluated for morphological, motility and antioxidant parameters. Microbial load was also evaluated. It was observed that the nanoparticles supplementation significantly increased pre and post freezing motility parameters of semen in groups 1 and 2 in comparison to control group and group 3. Post thaw seminal parameters viz., intact acrosome, plasma membrane integrity and percent live sperms were found to be significantly better in semen samples supplemented with nanoparticles with 20 µg/ml as compared group 1, 3 and control. Lipid peroxidation levels reduced, significantly in all supplemented groups as compared to control and ROS levels significantly reduced in samples supplemented with 20 µg/ml NPs. Level of glutathione peroxidase and reductase was significantly reduced in group 2 supplemented with 20 µg/ml NPs as compared to control. Microbial load of frozen thawed semen sample was evaluated to check the growth of common contaminants of semen samples by spread plate method. The NP supplemented semen samples had very less bacterial colonies as compared to control and group 3. Thus it can be concluded that nano ZnO as a supplement in the bull semen extender can be used to enhance the quality of semen as well as to replace the use of antibiotics in semen extender.
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Effect of plant based feed additives on rumen fermentation and blood biochemical profile in buffalo calves
(Lala Lajpat Rai University of Veterinary & Animal Sciences, Hisar, 2023-05) Sarthak; Madan, Jyotsana
The study was carried out using 10% aqueous and ethanolic extract of Emblica officinalis fruit powder ,Acacia nilotica leaves, Psidium guajava leaves to assess in vitro rumen fermentation parameters. The in vitro trial was conducted by taking different concentrations @ 0.2 ml, 0.5ml, 1ml and 2 ml of plant extracts with substrate media containing rumen fluid. After 6 and 24 hours of incubation pH, total gas production, ammonia- nitrogen, TVFA and total nitrogen was measured. The results revealed that the total gas production decreased in plant extract supplemented samples with respect to control. Total nitrogen production increased in aqueous E. officinalis and A. nilotica plant extract supplemented groups and increase in TVFA was observed in all plant extract supplemented samples. Ammonia- nitrogen production does not vary significantly, indicating no change in NPN degradation due to supplementation of different plant extracts at different concentration except A. nilotica (EE). In vivo rumen fermentation experiment included 3 months feeding trial, where 2 fistulated male buffalo calves were selected. In growth and biochemical studies, 12 male buffalo calves were divided into two groups of six animals in each group. The animals in treatment group were fed as per the control along with 30g of E. officinalis powder selected on the basis of in-vitro results. Growth parameters and haemato-biochemical parameters were analyzed in plasma samples at 0, 45 and 90 days. E. officinalis feeding indicated a promising effect on ruminal fermentation by enhancing VFA production whereas no significant effect was observed on growth parameters. Feeding of E. officinalis powder affects the nutrient metabolism, haematological parameters indicated by a significant increase in TEC and Hb concentration. Regarding biochemical parameters total protein and albumin concentration increased significantly (p < 0.05) in treatment group at 45day and 90 day. Cholesterol and LDL concentration increased significantly (p < 0.05) in both treatment and control group but they were in normal range. Considering all together, feeding of E. officinalis powder is likely to affect the rumen fermentation and haemato biochemical parameters in buffalo calves without any adverse effect on nutrient metabolism, growth and performance of buffalo calves.
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m6A modification of SARS-CoV-2 RNA: Exploring the epitranscriptomic regulation of proinflammatory cytokine production
(Lala Lajpat Rai University of Veterinary & Animal Sciences, Hisar, 2024-04) Lokender Singh; Shalini Sharma
A major risk factor of the severity/death of COVID-19 patients is associated with an increased level of inflammatory mediators including cytokines and chemokines also referred to as hypercytokinemia or cytokine storm, molecular mechanism of which is not well understood. Epitranscriptomic modifications are post-transcriptional modifications that occur on RNA molecules, which can have profound impacts on their structure, stability, and function and have emerged as the key players in regulating virus replication. In this study, it was demonstrated that SARS-CoV-2 infection to A549 cells results in methylation (m6A modification) of viral RNA and that, m6A modification in SARS CoV-2 genome is a dynamic event (switch on off phenomenon). SARS-CoV-2 infection to A549 cells also induces production of proinflamamtory cytokines (IL-6 and TNF-α). The m6A modification in SARS-CoV-2 genome and production of production of proinflamamtory cytokines genes appears to correlate with each other, indicating the role of m6A modification in induction of cytokines in the target cells. Further, the transfection of methylated- but not unmethylated RNA into A549 cells resulted in induction of IL-6, which suggests that IL-6 induction is m6A-depeendent. The levels of IL-1β and TNF α were comparable in cells that received either methylated but not unmethylated RNA, which suggested that induction of IL-1β and TNF-α is m6A-indepenedent. DZNep (a small molecule chemical inhibitor of methylation) suppressed SARS-CoV-2 yield by inhibiting synthesis of viral RNA. It also suppressed the levels of m6A-modidfes SARS-CoV-2 RNA, concomitant with decreasing the levels of IL-6. In conclusion, this study provides novel insights on epitranscriptomic control of proinflammatory cytokines (cytokine storm) production in SARS-CoV-2 infected cells. The cytokine storm may be therapeutically managed by inhibiting cellular enzymes responsible for methylation (m6A) of SARS-CoV-2 RNA in the target cells.
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Quantification of Urinary and Blood pregnanediol glucuronide and p-Cresol in buffaloes
(Lala Lajpat Rai University of Veterinary & Animal Sciences, Hisar, 2024-04) Choudhary, Renu; Ganguly, . Anita
PdG is one of the major immunoreactive compounds excreted in reproductively active female mammals across species including buffaloes. p-Cresol, also known as 4-methyl phenol, is a phenolic molecule that is created in the intestines of animals, particularly cattle, as a result of the microbial degradation of aromatic amino acids. An endocrinological study was performed to quantify pregnanediol-3α-glucuronide (PdG) and p-cresol in serum and urine simultaneously. The samples from forty adult breeding buffaloes(cyclic), 10 pre-pubertal(age<1.5yr), and 10 pubertal (age 1.5yr- 2 yr.) healthy Murrah buffaloes selected from the herds maintained at ICAR CIRB, Hisar, were collected on days 0 (day of oestrus), 10, 20, 30, 45, and 65. Immunoassay was performed on pooled serum and urine samples of representative days. The HPLC was used as a validation method for PDG quantification. A quantitative comparison of the immunoassay and chromatographic profiles revealed that progesterone and PdG in serum and urine were significantly correlated. At day 30, pregnancy was confirmed by gyno-clinical parameters, with the values in pregnant animals approximately twice those in non-pregnant animals. Serum progesterone concentration and urinary PdG concentration in pregnant animals increased sharply until day 45 and were maintained until day 65 when sampling was terminated. Based on the results, reproductive events, including oestrus cyclicity and pregnancy can be monitored by determining serum progesterone and urinary PdG in Murrah buffaloes. p-cresol in both serum and urine was generally higher in non-pregnant heifers compared to pregnant buffaloes in all three groups. p-Cresol conc. varied dramatically during oestrus in all groups on day 0 (day of oestrus), the mean value of p-cresol in serum and urine of nonpregnant buffaloes was higher than that of pregnant heifers. Therefore, PdG and p-cresol in buffalo urine could be used as potential markers for developing a non-invasive method for the reproductive assessment of dairy animals.
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Comparative studies on urinary metabotypes and scrotal thermal signatures in normal and cloned Murrah buffalo bulls
(Lala lajpat rai university Hisar, 2022-10) Krishan Kumar; Malik, Rakesh Kumar; Balhara, Ashok Kumar
The present study conducted on fourteen Murrah buffalo bulls - ten growing (five each, cloned and normal) and four adult (two each, cloned and normal), evaluated urinary metabolites, skin surface temperatures of different body parts (with particular focus on scrotum) and hemato-biochemical parameters in scrotal thermal signatures in normal and cloned buffalo bulls across four seasons (summer, rainy, autumn and winter) over one year. There was no insignificant difference in body surface temperature of normal and cloned animals except winter season. The scrotal skin surface temperature was significantly high in cloned animals as compare to normal growing (p<0.05) animals. Among different body parts recorded for surface temperatures, the eye temperature had highest correlation with rectal temperature and was also found closed to it at any point of time, suggesting its use as an alternate, non – touch method for recording internal body temperatures. The growth rate and blood-biochemical parameters recorded insignificant differences between normal and cloned animals. 1H-NMR spectroscopy suggested that there are qualitative and quantitative differences in the identified 136-140 urine metabolites, between cloned and normal animals in both groups (i.e. adult and growing). The differences were marked in summer and winter seasons in adult cloned animals. Of the 25 most abundant metabolites identified nine viz. L-arabinitol, xylose, glucose-1-phosphate, uridine diphosphate glucose (UDP-glucose), L-isoleucine, L-leucine, methylmalonic acid, keto-leucine and L valine were found in higher concentration in urine of cloned animals. Contrarily, four metabolites of these abundant metabolites viz. myo-inositol, glucuronic acid, taurine and glycine metabolite showed high relative abundance in the normal animals. The functional bioinformatics suggest significant differences in urinary metabolites between cloned and normal animals w.r.t. thermoregulation, energy metabolism, hormone synthesis and degenerative changes. Study also points towards higher scrotal temperature in growing cloned animals. The thermal screening conducted in the study also suggest that ocular temperature can serve as an alternate, non-touch method for estimating internal body temperature in buffaloes
Open Access
Transcriptome analysis of buffalo bull spermatozoa for identification of fertility associated gene(s)
(LUVAS, 2017) Vijayalakshmy.K; Virmani, Meenakshi
Keeping in view the importance of sperm RNA and its correlation with fertility, the proposed study was carried out to determine the gene expression between high fertile and sub-fertile buffalo bull spermatozoa and identify the genes potentially associated with bull fertility through transcriptome analysis. These fertilityassociated genes were further validated with qRT-PCR and their possible role in fertilization through in-vitro fertilization. Ten bulls were classified on the basis of conception rates (CR), where bulls having CR <40% and 50% were considered as sub- and high-fertile bulls, respectively. Total motility, rapid motility, average path velocity, curvilinear velocity, amplitude of lateral head, beat cross frequency, straightness, linearity and intactness of plasma membrane of sperm for high-fertile bulls were significantly higher than sub-fertile bulls. Semen samples from these bulls were utilized for sperm enrichment using three different density gradient methods viz., Percoll, BoviPure and Iodixanol. Purified sperm pellets were subjected to RNA isolation using conventional and kit methods with and without TRIZOL. Maximum yield of RNA was achieved when the semen sample was purified using BoviPure as density gradient followed by isolation of RNA by RNAqueous + heated TRIZOL method. Further, AKAP4, CRISP2, Fertilin- â and PLCZ1 transcripts showed significantly higher expression in high- fertile than sub-fertile groups. Transcriptome analysis was done by Next generation sequencing and a total of 33715 and 21788 transcripts were expressed in sub- and high-fertile groups, respectively. When these transcripts were annotated with the available sequence of buffalo, 1248 and 802 transcripts were uniquely expressed in the sub- and high-fertile groups, respectively and 2527 transcripts were identified as common in both the groups. When in silico translated these transcripts, 258 proteins were observed to be commonly present in both the groups. Apart from these, 6 and 18 proteins were specifically present in the high and sub-fertile group, respectively. The expression of AKAP4, CRISP2, Fertilin- â and PLCZ1 was evaluated in IVF derived embryos. All the four transcripts differentially expressed only in sperm and embryo up to cleaved and blastocyst stage, but these genes didn‟t express in immature and mature oocytes. Thus, the present study provides the first evidence of differential expression of AKAP4, CRISP2, Fertilin â and PLCZ1 genes in high and sub-fertile bulls and their transmission to embryo indicating the role of sperm RNA in fertility and embryo development.
Open Access
Relationship of residual feed intake with rumen fermentation and blood biochemical profile in growing buffalo calves
(LUVAS, 2017) Vikramjeet Singh; Madan, Jyotsana
The present study attempts to assess the relationship of residual feed intake (RFI) with rumen fermentation and blood biochemical profile in growing buffalo calves. Twelve healthy buffalo calves of seven to nine month of age were selected and fed with green fodder and concentrate mixture as per ICAR 2013 feeding standards. Weighed amount of feed and fodder was offered and residue was also weighed next day, early in morning. DM of feed and fodder offered and residue left was estimated on daily basis to assess DM intake. After completion of feeding trial, RFI value for individual animals was calculated. Experimental animals were weighed before starting the actual experiment and thereafter at fortnightly intervals. Using the data of fortnightly body weights ADG was calculated. Body growth parameters viz. body length, height, heart girth and abdominal girth of experimental animals were recorded in centimeters at the beginning, in the middle and at the end of experiment. A digestion trial of 6 days was conducted at the end of the experiment to know the nutritive value of feed and fodder. Representative samples of faeces, concentrate mixture, green and left over feed were taken daily and dried in hot air oven for determining dry matter content. The six days dried samples of feaces, feed and fodder were pooled, milled and analysed for proximate analysis according to AOAC (2005). A representative and homogenous sample of rumen liquor was collected through a stomach tube from the rumen of animals of high and low RFI groups and used for total nitrogen, ammonia nitrogen, total volatile fatty acids and individual volatile fatty acids estimation. Blood samples were collected thrice, once at the beginning, in the middle and at the end of trial, from all the animals for estimation of blood biochemical parameters in blood plasma. The results obtained regarding RFI values for high and low RFI groups were 0.235±0.04and -0.235±0.06, respectively. Mean body weight, mean metabolic body weight, average daily body weight gain and mean value of body growth parameters of the high RFI and low RFI groups were non-significant. The DM consumption was lower in low RFI group compared to high RFI group. Dry matter intake per 100 kg body weight (kg/100kg BW) and DMI per kg metabolic body weight was significantly lower in low RFI group (P≤0.01). Total nitrogen and ammonia nitrogen were non-significant in high and low RFI groups, while the TVFAs, acetic acid and butyric acid were negatively correlated with RFI (P≤0.01). Blood glucose, total protein and IGF-1 were negatively but non-significant correlated with RFI. Plasma albumin concentration was significantly positively correlated (r = 0.56) with RFI (P<0.01). BUN concentration was non-significantly positively correlated (r = 0.35) with RFI. RFI was positively correlated with AST (r =0.44) while negatively correlated (r = 0.35) with ALT.
Open Access
Effect of critical micronutrient supplementation during peri-parturient period on production and reproduction status of buffaloes
(LUVAS, 2017) Jyani, Vikas; Gupta, Meenakshi
Peri-parturient period is the critical phase in the life of dairy animals affecting future productive and reproductive performance. Present study was undertaken to evaluate the effect of critical micronutrient supplementation during peri-parturient period on production and reproduction status of buffaloes. Study was performed on twenty-two healthy Murrah buffaloes after dividing into two equal groups i.e. control and treatment respectively. Buffaloes in advanced stage of gestation were selected i. e. from 56 days prior to expected date of parturition. Buffaloes of both the groups were maintained on farm’s standard feeding practices as per their requirement. Additional micronutrient supplementation was carried out once in a week in buffaloes of treatment group having Zn, Cu, Co, Cr, Se, vitamin E, vitamin A, and niacin. Blood samples were collected pre-partum at day -56th, -28th, at day of parturition and thereafter at day 28th and 56th after parturition and subjected to plasma separation and estimation of blood biochemical profile and micronutrient status. Milk samples were collected weekly during initial 10 weeks of lactation. Ultrasonography was performed thrice during the study, i.e. on day 25-35, 55-65 and 85-95. Supplementation of critical micronutrient improved production performance of Murrah buffaloes in terms of total milk (18%) and fat corrected milk (P<0.05) production in addition to improvement in fat and protein percentage (P<0.05) of milk. Critical micronutrient supplementation did not affect blood biochemical and micronutrient profile significantly, due to uncompromised level of nutrients and health in buffaloes of control group, except for an improvement (P<0.05) in plasma inorganic phosphorus level in treatment group at day of parturition. Reproductive performance remained unaffected due to supplementation of weekly critical micronutrients in the ration of peri-parturient Murrah buffaloes.
Open Access
Effects of feed additives rich in essential oils on rumen fermentation, Methanogenesis and nutrient utilization in buffalo
(LUVAS, 2017) Krishan Kumar; Rose, Manoj Kumar
Present studies were conducted at the Indian Council of Agricultural Research- Central Institute for Research on Buffaloes (ICAR-CIRB), Hisar. Four feed additives (Eucalyptus leaves, Poplar leaves, Clove buds and Dalchini bark) rich in essential oils were selected to examine their effects on rumen fermentation, methanogenesis and nutrient utilization in buffalo. The extracts were prepared (15 g/ 100 ml) either in petroleum ether or n-hexane by Soxhlet’s apparatus. In in-vitro studies, the production of total gas, methane production, dry matter digestibility, ammonia nitrogen and production of volatile fatty acids were studied. The study revealed a significant (p<0.05) reduction in total gas, methane production, in-vitro dry matter digestibility, ammonia nitrogen and production of volatile fatty acids at high dose (2.0 ml/30ml buffered rumen fluid) as compared to control group. But at low (0.5 ml) and intermediate (1.0 ml) dose level the effects of extracts were comparable to control in all these parameters, with reduction (p<0.05) in methane production. The blends (extracts of poplar leaves, eucalyptus leaves and clove buds) of feed additives examined in this phase also showed a modulation in rumen fermentation with reduction in methane production. An in-vivo trial was conducted to study the effects of selected blend of feed additives on growth and nutrient utilization. Animals were distributed in three groups i.e. CON (basal ration), EOL-1(basal ration + 25 gm each of dry grounded poplar and eucalyptus leaves mixed with concentrate mixture), EOL-2 (Diet same as in control + 75 gm each of dry grounded poplar and eucalyptus leaves mixed with concentrate mixture). There was no significant (p>0.05) difference observed in growth rate and nutrient utilization among the in all treatments. In vivo methane productions were found to be inhibited in both treatment groups, as compared to control group after 3 months of dosing. The heamato-biochemical parameters remained similar (p>0.05) except significant (p<0.05) reduction in blood urea concentration as compared to control group. In conclusion, the essential oil rich feed additives could be used to reduce enteric methane emission and EOL-2 was found to significantly reduced methane production without any adverse effects on digestibility, growth and physiological status of buffaloes.

Lala Lajpat Rai University of Veterinary & Animal Sciences, Hisar

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