Sujata Pandita

Age- and sex-related variability in physiological and immune responses to endotoxin challenge in Murrah buffaloes (Bubalus bubalis)

The host responses to disease agents differ between age and sex as is evidenced from epidemiological studies which have been ascribed to the effects of steroid hormones. The present study was carried out in 20 Murrah buffaloes of either sex at pre- and post-pubertal age having discrete sex steroid background to explore the variation, if any, in physiological and immune responses upon intravenous endotoxin challenge (E. coli 055:B5 @ 0.6µg/kg body weight). The rectal temperature, heart rate and respiration rate differed significantly from base values at only 1 h post challenge and reached peak at 4 h. The magnitude of responses determined as area under concentration × time curve (AUC) for these parameters was significantly low in post-pubertal animals of either sex. Plasma tumour necrosis factor alpha (TNFα, a pro-inflammatory cytokine) registered peak at 1 h post challenge which returned to base level value by 8 h post challenge. Responses determined as AUC of TNFα were significantly higher in post-pubertal males. Plasma NOx (an estimate of nitric oxide production) and XO (mediator of superoxide production) significantly differed from base value at 4 h and peaked at 8 h post challenge. Integrated AUC response of NOx was significantly higher in pre-pubertal males and that of XO in pre-pubertal males as well as females. Plasma cortisol (an anti-inflammatory hormone) was significantly different from base value at 2 h and peaked between 4 h and 8 h post challenge; its integrated AUC response was significantly greater in post-pubertal females. Endotoxin challenge resulted into neutrophilia (a stress marker), and the initial hyperglycemia was followed by hypoglycemia in all experimental animals irrespective of age and sex. Pre-pubertal buffaloes had no dominance of gonadal sex steroids in plasma. Post-pubertal males and females had plasma testosterone and oestrogen, respectively in significant quantity. The present findings in Murrah buffaloes recognise the age and sex as a source of variability in the magnitude of immune responses to endotoxin challenge like lipopolysaccharide (LPS).

Age-related changes and circadian variations in peripheral levels of thyroid hormones in Murrah buffaloes

Abstract The aim of this study was to investigate the variations in plasma triiodothyronine (T3) and thyroxine (T4) with the advancement of age and to determine their circadian patterns in prepubertal and pubertal Murrah buffaloes. The variations in plasma T3 and T4 with the advancement of age were observed from day 1 to 24 months of age. Significant higher levels of T3 and T4 were observed after birth and a gradual decrease in their concentrations occurred until 15 days of age. The mean plasma T3 and T4 ranged between 1.26–3.79 and 60.7–166 ng/ml, respectively, during 1–30 days of age. During 1–24 months of age, the variations in plasma T3 did not differ (p > 0.05) with the advancement of age, whereas significant (p < 0.0001) changes were observed in plasma T4. The circadian patterns of T3 and T4 were evaluated in prepubertal Murrah buffaloes (n = 8) aged between 14 and 16 months. The mean plasma T3 and T4 ranged between 1.04–1.85 and 43.0–76.1 ng/ml, respectively. Significant (p > 0.0001) changes in the secretory pattern of T3 were observed, whereas the secretory pattern of T4 did not differ significantly (p > 0.05). In addition, the circadian patterns of T3 and T4 in pubertal buffaloes (n = 4) aged between 28 and 30 months were observed and compared to that of prepubertal group (n = 4). The prepubertal group showed significant (p < 0.001) higher plasma T3 concentrations over 24 h than the pubertal group.

Effect of pre-partum supplementation of vitamin E to Murrah buffaloes on immune functions and viability of calves.

This study was undertaken in buffalo neonates born to vitamin E (dl-alpha-tocopherol acetate)-supplemented and non-supplemented Murrah buffaloes. Calves from vitamin E-supplemented buffaloes (n = 10; vitamin E -supplemented calves [VeC]) and non-supplemented buffaloes (n = 10; control calves [CC]) constituted the treatment and control groups respectively. Two colostrum samples were taken at the first post-partum milking and again after 12 h from dams for IgG estimation. Sampling of blood was performed on days 0 (before colostrum feeding), 1, 3, 7, 14, 21, 28, 42, 56, 70, 84, 98, 112 and 126 post-birth and analysed for apparent efficiency of absorption (%) of IgG and various immune parameters. Colostral IgG level was significantly higher (p < 0.05) in vitamin E-supplemented buffaloes. The calves in both groups were born hypogammaglobulinemic with IgG level <5 g/l. However, first colostrum feeding resulted in significantly elevated IgG levels (>10 g/l) in calves of both groups at 24 h, which remained high afterwards. Apparent efficiency of absorption (%) of IgG at 24 h was significantly higher (p < 0.05) in VeC than in CC. Plasma Nitric Oxide (NO) levels were significantly elevated in the calves of either group at birth, which declined significantly (p < 0.01) afterwards. Vitamin E feeding to dams had no added effect on NO levels in experimental calves. Total leucocyte counts did not differ significantly between the two groups. However, lymphocyte and neutrophil counts changed significantly between groups (p < 0.01) and days (p < 0.01), with lymphocytes increasing and neutrophils declining with age. This study revealed that the calves were immunologically immature at birth. Ante-partum supplementation of vitamin E did not influence plasma NO or IgG but had a significant effect on colostral IgG (p < 0.05). It also improved the apparent efficiency of absorption (%) of IgG at 24 h in VeC as compared to CC.

In Vitro Immune Competence of Buffaloes (Bubalus bubalis) of Different Production Potential: Effect of Heat Stress and Cortisol

Twelve healthy lactating Murrah buffaloes of similar parity (3rd) between 90 and 120 days of lactation, selected from the herd of National Dairy Research Institute (Karnal, India) and maintained at managemental practices as followed at the Institute they were included in this experiment. The animals were divided into two groups based on their production level in previous lactation. The average milk production level of group 1 and II was 9.3 and 6 lit/day, respectively. Blood was collected from these buffaloes on three occasions 10 days apart. The lymphocytes were separated and cultured in RPMI 1640 medium with PHA-P for 24 h at 37°C in a humidified CO2 incubator (95% air and 5%  CO2). The lymphocyte responsiveness was also evaluated in response to the in vivo heat stress and in vitro cortisol. Mitogen-induced stimulation index was not affected by production level (P < .01). Stimulation index was significantly reduced (P < .01) in both the groups when cortisol was added at 2.0 ng level in the culture. However, in heat-stressed buffaloes stimulation index did not vary despite increasing levels of cortisol, thus indicating that lymphocyte may become cortisol resistant during periods of acute heat stress. The results showed that lymphocyte proliferation response can be effectively used to study buffalo cell-mediated immunity in vitro.

Xanthine oxidase activity during transition period and its association with occurrence of postpartum infections in Murrah buffalo (Bubalus bubalis)

The aim of this study was to quantify xanthine oxidase (XO) levels during the transition period in Murrah buffalo (Bubalus bubalis) and determine its association with certain postpartum infections. For this, six healthy buffaloes were selected from the National Dairy Research Institute (NDRI) herd and managed under standard managemental practices as followed at the institute. Blood samples were drawn weekly from each buffalo from day -21 to +21 relative to parturition by jugular vein puncture. Additional blood samples were collected from buffaloes suffering from metritis (n=5), endometritis (n=6) and mastitis (n=8) on alternate days. XO activity followed a defined pattern with values gradually declining from day -21 up to the day of calving followed by an increase to day +21, but the difference was statistically not significant between pre- and post-partum stages. The activity significantly declined on the day of calving when compared to the pre-partum mean value (p<0.05). The activity was significantly enhanced in buffaloes with bacterial infections, endometritis, and mastitis compared to healthy controls (p<0.05). However, the levels were not significantly altered among buffalo with metritis. The results indicate that increased XO activity during the postpartum period was associated with some bacterial infections in buffaloes, which could be due to increased phagocytic activity as a part of the innate defense system. Keywords : Xanthine oxidase, transition period African Journal of Biotechnology Vol. 12(32), pp. 5101-5104

Analysis of In Vitro Effects of Sex Steroids on Lymphocyte Responsiveness in Murrah Buffaloes (Bubalus bubalis)

Present study was carried out on forty four apparently healthy Murrah buffaloes of different age groups of both sexes to investigate the effects of sex steroids on cell mediated immunity in vitro. Estrogen inhibited proliferation in mitogen-stimulated lymphocytes from prepubertal but not post pubertal buffaloes of either sex. Estrogen at 100 pg/mL concentration stimulating the proliferation significantly (P < 0.05). in all groups and had higher stimulatory effect in lymphocytes from day 10 than day 0 of estrous cycle. Progesterone inhibited lymphocyte proliferation, and inhibition was directly related to the dose, in all groups of either sex. Testosterone did not inhibit proliferation at any dose level and did not show any consistent and lucid effects on lymphocyte proliferation. Present study revealed that buffalo lymphocytes produce appreciable amounts of NO in culture system after treatment with estradiol. Significantly high levels of NO in culture supernatant were found in prepubertal buffalo calves and least in post pubertal buffaloes, which had an inverse relation with lymphocyte proliferation in presence of estradiol. NO in culture supernatant was high at the lowest dose of progesterone which was proportional to the lymphocyte proliferation when treated with progesterone. No significant difference in NO culture supernatant was observed between different concentrations of testosterone treatment.

Interferon-tau stimulated gene expression: A proxy to predict embryonic mortality in dairy cows

The embryonic mortality in cows is a growing concern for an ever-expanding dairy industry. The current study was an attempt to shorten the open period of dairy cows having suffered embryonic loss by diagnosing them at an earlier stage. The blood samples were collected from the Karan Fries (KF) cows on days 0 (day of AI/estrus), 4, 8, 12, 14, 16, 18, 21, 24, 28, 35 and 42 post insemination. The experimental animals were then categorized into pregnant (P), conception failure/early embryonic mortality (EEM) and late embryonic mortality cows (LEM), based on progesterone assay, ultrasonography and per-rectal palpation. There were 6 animals in each group. The plasma progesterone was higher in pregnant than EEM and LEM cows. Plasma Interferon-tau concentration was significantly (p < 0.05) lower in LEM than pregnant cows where it could be detected from day 14-21 but was non-detectable in EEM cows. The mRNA expression of ISG15, OAS1, MX1 and MX2 in blood neutrophils was significantly (p < 0.05) higher from day 8-42 as against day 0 in pregnant cows. The highest expression was observed around day 18-21 in pregnant cows. The ISG15, OAS1, MX1 and MX2 mRNA expression was significantly (p < 0.05) higher from day 4-42 as compared to day 0 in LEM cows, whereas in EEM cows the expression stayed close to that of day 0 (1.00 ± 0.00). The mRNA expression of ISG15, OAS1, MX1 and MX2 started to decline from day 24 onwards. The degree of expression of Interferon-tau stimulated genes was higher in pregnant and LEM cows than EEM cows. The study reveals that the Interferon tau stimulated gene expression in neutrophils can act as peripheral biomarkers for detecting the embryonic mortality in dairy cows.

CNCP Fractions and in Vitro Rumen Fermentation of Straw Based Total Mixed Rations Supplemented with Leucaena leucocephala Leaves

An in vitro study was conducted to evaluate the effect of the different ratios of Leucaena (Leucaena leucocephala) leaves (L), wheat straw (W) and concentrate mixture (C) on total gas production, methanogenic potential and other rumen fermentation parameters using in vitro gas production technique. Four total mixed ration (TMR) were prepared using dried ground samples of W, C and L: WC (90W: 10C), WCL10 (80W: 10C: 10L), WCL25 (65W: 10C: 25L) and WCL50 (40W: 10C: 50L). For estimation of fermentation parameters, TMR samples (200 mg) were incubated with 30 mL buffered rumen liquor at 39±0.5oC for 24 h. As the level of Leucaena leaves increased in the substrate, total gas production increased (P<0.05) while CH4 production (mL/100 mg DDM) decreased (P<0.05) by 15, 24 and 36% in WCL10, WCL25 and WCL50, respectively. In vitro DM and OM digestibility was also higher (P<0.05) as the levels of Leucaena increased in the ration. Thus, this study indicated that with the inclusion of Leucaena to straw based ration, there was improved utilization of nutrients and reduced methane production.

Growth performance, and enteric and manure greenhouse gas emissions from Murrah calves fed diets with different forage to concentrate ratios

The present study investigated the effects of different dietary forage to concentrate ratios on animal performance, and enteric and manure greenhouse gas emissions in growing calves. Fifteen male Murrah calves (153.5 ± 18.17 kg; 6 to 12 months) were randomly assigned to 3 dietary treatments and fed corn fodder, wheat straw and concentrate in 3 different proportions: 20:60:20 (C20); 20:40:40 (C40) and 10:30:60 (C60), for a period of 120 days. Increasing dietary concentrate proportion had no significant (P > 0.05) effect on dry matter intake (DMI) but increased crude protein (CP) and total digestible nutrient intake (P < 0.05). Average daily gain and feed conversion efficiency were significantly higher (P < 0.05) for C60 compared with C20 and for C40, these did not differ with C20 and C60 (P > 0.05). The apparent digestibility of dry matter, organic matter and CP were higher (P < 0.05), but acid detergent fiber digestibility was lower (P < 0.05) for C60 compared with C20 whereas, ether extract and neutral detergent fiber digestibilities were not affected (P > 0.05). Daily methane (CH4) emission (g/d), CH4 energy loss (MJ/d) and CH4 yield (CH4 g/kg organic matter intake [OMI], CH4 g/kg digestible OMI, and CH4 % of metabolizable energy intake) were significantly higher for C20 compared with C60 (P < 0.05). Methane yield as g/kg DMI although lower for C60 compared with C20 but the difference was not significant (P > 0.05). Manure CH4 (g/kg DMI) and nitrous oxide (N2O mg/kg nitrogen) emissions were not affected (P > 0.05), but N2O emission on mg/kg DM basis was significantly higher (P < 0.05) from the manure of calves fed C60 than that for C20. Thus, increasing dietary concentrate proportion improved animal performance, and reduced enteric CH4 emission (g/day) without any significant effect on manure N2O (mg/kg nitrogen) and CH4 emissions.

Inflammatory Mediators and Hormonal Changes in Subclinical and Clinically Affected Mastitis Cows

Dairy animals encounter mastitis as one of the most important disease causing hindrance in the development of dairy sector. Selective breeding of dairy cattle has led to a dramatic increase in milk yield over recent decades giving India an honour to become the highest milk producer country in the world but simultaneous increase in high incidence of mastitis and huge economic losses have been reported (Heald et al., 2000; Seegers et al., 2003; Oltenacu and Algers, 2005). Researchers agree that the economic impact of subclinical forms of mastitis is larger than clinical mastitis (Singh et al., 2016). The overall prevalence of sub-clinical mastitis has been reported to be 59.43% with quarter level prevalence of 34.78% (Bhat et al., 2016). The annual economic losses due to mastitis have been calculated to be Rs.7165.51 crores both cows and buffaloes almost with rupees 3649.56 and 3515.95 crores, respectively (PDAMAS, 2011). Subclinicacl mastitis alone causes economic losses of rupees 4151.16 crores (Bogni et al., 2011). This could be minimized by using certain makers in milk and plasma of mastitis animals. It has been found that nitric oxide (NO), interleukins (IL) and tumor necrosis factor (TNF-α) could play a vital role in the pathophysiology of this (Kushibiki et al., 2003; Hansen et al., 2004). International Journal of Current Microbiology and Applied Sciences ISSN: 2319-7706 Volume 6 Number 11 (2017) pp. 620-627 Journal homepage: http://www.ijcmas.com