R. C. Upadhyay

New aspects and strategies for methane mitigation from ruminants

The growing demand for sustainable animal production is compelling researchers to explore the potential approaches to reduce emissions of greenhouse gases from livestock that are mainly produced by enteric fermentation. Some potential solutions, for instance, the use of chemical inhibitors to reduce methanogenesis, are not feasible in routine use due to their toxicity to ruminants, inhibition of efficient rumen function or other transitory effects. Strategies, such as use of plant secondary metabolites and dietary manipulations have emerged to reduce the methane emission, but these still require extensive research before these can be recommended and deployed in the livestock industry sector. Furthermore, immunization vaccines for methanogens and phages are also under investigation for mitigation of enteric methanogenesis. The increasing knowledge of methanogenic diversity in rumen, DNA sequencing technologies and bioinformatics have paved the way for chemogenomic strategies by targeting methane producers. Chemogenomics will help in finding target enzymes and proteins, which will further assist in the screening of natural as well chemical inhibitors. The construction of a methanogenic gene catalogue through these approaches is an attainable objective. This will lead to understand the microbiome function, its relation with the host and feeds, and therefore, will form the basis of practically viable and eco-friendly methane mitigation approaches, while improving the ruminant productivity.

Changes in methane emission, rumen fermentation in response to diet and microbial interactions

To evaluate relative contributions of different microbial groups in rumen, the mono-culture (i.e. bacteria, protozoa and fungi) and co-cultures (i.e. bacterial-protozoal, fungal-protozoal and bacterial-fungal) were tested in vitro using high and low roughage diets. Total gas and methane were higher in bacterial-fungal and bacterial-protozoal co-cultures, while lower in fungal-protozoal than controls (high and low roughage with complete rumen consortia; control 1 and 2, respectively). Digestibility and total volatile fatty acids were lower in bacterial-fungal co-culture with both high and low roughage diets. Methanogens decreased in bacterial-fungal co-culture with high roughage. With high roughage, counts were lower for bacteria with bacterial-protozoal, protozoa with fungal-protozoal, and fungi with the bacterial-fungal co-cultures. Total gas was higher in bacterial mono-culture with low roughage, but methane was not detected in any mono-culture. Digestibility and total volatile fatty acids were significantly lowered with protozoal mono-culture. Methanogens reduced significantly in mono-cultures with high roughage diet than control 1. Defaunation reduced methanogens without significantly affecting rumen fermentation.

Heat stress and animal productivity

CHAPTER 1 Thermoregulation.- 1. Introduction.-2. Thermoneutral zone.- 3. Heat stress indicators.- 4. Adaptation to high temperature.- 5. Thermoregulatory mechanisms.- 6. Response to heat stress.- 6.1Water vaporization.- 6.2 Physical responses.- 6.3 Sweating and panting.- 7.Heat production.- 8. Heat Increment.- 9. Assessment of Adaptability.- 10.Genetic adaptations/cellular changes during heat stress.- 11.Genetic improvement for adaptation.- 12. Genomic responses during acclimation.- CHAPTER 2 Heat stress and Hormones.- 1.Introduction.- 2.Thyroxine (T4) and triiodothyronine (T3).- 3.Cortisol.- 4. Insulin.- 5. Insulin like growth hormone 1 (IGF-I).- 6.Growth hormone.- 7.Aldosterone.- 8.Adiponectin and leptin.- 9.Reproductive hormones.- 10.Catecholamines.- 11.Prolactin.- CHAPTER 3 Heat Stress and Milk Production.- 1.Introduction.- 2.Effect of climatic variables on milk yield.- 3. Behavior of cows during heat stress.-4.Effect of heat stress on bovine somatotropin (bST) administered cows.- 5.Metabolic heat production.- 6.Role of Acclimation during heat stress.- 7.Metabolic Adaptations to Heat Stress.- 8.Mechanism by which heat stress reduces milk production.- 9.Mechanism of regulation of milk secretion and mammary function.- 10. Heat Stress Effects on Heifers.-11.Genetic factors regulating response to heat stress.- 12.Reducing Heat Stress in dairy animals.- 13.Effect of heat stress on milk production in Buffaloes.- CHAPTER 4 Heat stress and Reproduction.- 1. Introduction.- 2. The effect of heat stress on reproductive functions.- 3. The effect of heat stress on the hypothalamic hypophyseal ovarian axis.- 4.Female reproduction.- 4.1 Ovarian follicle.- 4.2 Steroid production.- 4.3 The oocyte.- 4.4 Embryonic development.- 4.5 Fetal development.- 4.6 Uterine environment.- 5.Male reproduction.- 6.Effect of heat stress on reproduction by altering energy balance.- 7. Genetic plasticity controlling the magnitude of heat stress effects.-8.Consequences of action of climate change on reproduction for species survival and distribution.- 9.Reproduction in buffaloes .- 10.Management and production systems that can improve fertility during summer months.- CHAPTER 5 Heat stress and immune function.- 1 Introduction.- 2 Innate and adaptive immunity.- 3 Heat stress and Cell-mediated immunity.- 4 Regulation of stress response.- 4.1 Role of glucocorticoids and catecholamines.- 4.2 Role of cytokines.- 4.3 Role of stress hormones.- 5 Acute and Chronic Stressors .- 6 Mechanism of action of heat stress on immunity .- 7 Nutrition and immunity.- 7.1 Vitamins.- 7.2 minerals.- 8 Effect of free radical production on immunity and role of antioxidants.- CHAPTER 6 Biological Rhythms.- 1. Introduction.- 2. Circadian timekeeping mechanisms.- 2.1 Biological clock.- 3. Daily rhythms.- 4. Annual rhythms.- 5. Functions of sleep.- 6. Pineal gland and melatonin.- 6.1 Synthesis and metabolism of melatonin.- 6.2 Melatonin receptors.- 6.3 Daily rhythm of melatonin .- 6.4 Annual rhythm of melatonin.- 7. Cortisol.- 7.1 Daily rhythm of cortisol.- 7.2 Annual rhythm of cortisol.- 8. Leptin.- 8.1 Daily rhythm of leptin.- 8.2 Annual rhythm of leptin.- 9. Lipid metabolism.- 9.1Lipid metabolism in ruminants.- 9.2 Daily rhythms of free fatty acids (FFA) and glycerol.- 9.3 Seasonal variation of lipid metabolism.- 10. Feeding entrained circadian rhythms.- 11. Circadian clocks as mediators of the homeorhetic response to lactation.- 12. Oxidative stress.- 13.Circadian rhythm of cardiovascular parameters.- CHAPTER 7 Shelter Management for Alleviation of Heat Stress in Cows and Buffaloes.- 1. Introduction.- 2. Heat transfer and concentration and emission of harmful gases in dairy buildings.- 3. Cooling methods for alleviation of heat stress.- 3.1 Provision of shade.- 3.2 Ceiling fans.- 3.3 Evaporative cooling.

Expression profiling of major heat shock protein genes during different seasons in cattle (Bos indicus) and buffalo (Bubalus bubalis) under tropical climatic condition.

Heat shock proteins consist of highly conserved stress proteins, expressed in response to stress and play crucial roles in environmental stress tolerance and adaptation. The present study was conducted to identify major types of genes under the HSP70 family and other HSPs and to evaluate their expression pattern in Sahiwal and Tharparkar breeds of zebu cattle (Bos indicus) and Murrah buffalo (Bubalus bubalis) with respect to different seasons. Quantitative real time polymerase chain reaction was performed to analyze the transcript variants of three HSP70 family genes (HSPA1A, HSPA1B, and HSPA8) and HSP10, HSP60, HSP90 and HSF1 in each breed. The major finding of this study was the higher abundance of all the studied HSP genes during summer and winter compared to spring season, but the magnitude of increase was higher during summer as compared to winter. HSPA1A and HSPA1B genes showed maximal induction (P<0.001) during summer and winter while HSP60 and HSP10 were found to be the second most abundantly expressed HSPs. The relative mRNA abundance of HSF1 significantly increased (P<0.001) in Murrah buffalo compared to Tharparkar and Sahiwal cattle during summer and winter. Expression pattern of heat shock protein genes indicated that amongst the breeds, the expression was higher in Murrah buffalo compared to Sahiwal and Tharparkar cattle, thereby indicating the more adaptive capacity of later during periods of stress. Hence, this study suggests that heat shock protein genes may be conveniently used as biomarkers for assessing stress response in cattle and buffalo and the expression is species and breed-specific. Furthermore, the variation in expression is associated with heat tolerance and adaptation to different climatic conditions.

Seasonal variations in physio-biochemical profiles of Indian goats in the paradigm of hot and cold climate

To explore the heat and cold adaptation in Indian goats by the physiological, haematological, blood biochemical parameters and their seasonal variations, this study was conducted on heat- and cold-adapted Indian goats maintained in their natural habitat. Study was carried out in three different phases coincide with the three seasons (winter, spring and summer). The levels of physiological responses, that is rectal temperature, respiration rate and pulse rate, were observed to be significantly (p < 0.01) lower in heat-adapted breeds and higher in cold-adapted breeds, whereas the levels of Hb, PCV and TEC were significantly (p < 0.01) higher in cold-adapted goats. Significantly (p < 0.01) higher levels of plasma thyroid hormones (thyroxine and triiodothyronine) and plasma stress enzyme (AST and ALT) were also observed in cold-adapted goats. Significant (p < 0.01) seasonal variations in physiological responses, haematological and blood biochemical parameters in both heat- and cold-adapted breeds were reported in this study. Physiological responses, plasma enzymes and plasma cortisol levels significantly (p < 0.01) increased during summer in all the goat breeds. The levels of haematological parameters (Hb, PCV, TEC and TLC) and plasma thyroid hormones (thyroxine and triiodothyronine) decreased during summer. The changes in physiological parameters during summer due to heat stress were higher in cold-adapted goats whereas the levels of changes in these parameters during winter due to cold were higher in heat-adapted goats. High neutrophil/lymphocyte ratio during summer in cold-adapted breeds is an indicator of higher level of stress. Decrease in plasma electrolytes (Na and K) during summer also observed in cold-adapted breeds during summer. The variations in physiological, haematological, blood biochemical parameters in heat- and cold-adapted goats may be due to their adaptation to different environmental and geographical conditions essential for their survival.

Peripheral blood leukocytes transcriptomic signature highlights the altered metabolic pathways by heat stress in zebu cattle.

High temperature during summer greatly affects animal production due to altered reproductive and metabolic functions. However, information regarding high throughput analysis of change in gene expression in diary animals are relatively nil. In present study, gene expression profiling by microarray was done in peripheral blood leukocytes of heat exposed (42 °C, 4h) cattle (n=3), Tharparkar (Bos indicus). A total 460 transcripts were differentially expressed with a fold change of ⩾ 2. Randomly selected real-time validation showed that 73.08% correlation with microarray data. Functional annotation and pathway study of the DEGs reveals that, up-regulated genes significantly (P<0.05) affect the protein processing and NOD like receptor pathways, while down regulated genes were significantly (P<0.05) found to associated with Glycolytic pathways. In conclusion, the present study showed that heat stress affects expression of significant number of genes in peripheral blood leukocytes and further analysis is required to understand their functional role in livestock.

Impact of Climate change on Milk production of Murrah buffaloes

Abstract Global warming is likely to impact productivity of buffaloes due to their sensitivity to temperature changes. Air temperature, humidity, wind velocity and solar radiation are the main climate variables that affect buffalo production in tropical climate. In the present study sensitivity of lactating Murrah buffaloes to sudden temperature (Tmax, Tmin) change and THI have been analyzed from milk production and climatic records (1994-2004) of Karnal. Algorithms were developed and validated on lactating buffaloes during 2005-2006 at the Institute. A sudden change (rise or fall) in Maximum/Minimum temperature during summer and winter was observed to affect milk production. The decline in minimum temperature (>3°C) during winter and increase (>4°C) during summer than normal were observed to negatively impact milk production upto 30% on the next or subsequent days after extreme event. The return to normal milk production depended on severity and time period of thermal stress/ event occurrence. The R² was very low for cool period observed during Feb- April/Sept-Nov and actual effect on milk production was minimum. This indicated that low THI had a relatively small effect on milk production performance. The lactation period of animals are shortened during extreme summer when THI were more than 80 and reproductive functions were also adversely affected. Thermal stressed buffaloes did not exhibit estrus or exhibited estrus for short period. The potential direct effects of possible climate change and global warming on summer season milk production of Murrah buffaloes were evaluated using widely known global circulation model UKMO to represent possible scenarios of future climate. Both milk production and reproductive functions of Murrah buffaloes are likely to be affected due to warming effects.

Effect of Vitamin E and Zinc Supplementation on Energy Metabolites, Lipid Peroxidation, and Milk Production in Peripartum Sahiwal Cows

The study was conducted to evaluate the effect of vitamin E and zinc supplementation on energy metabolites, lipid peroxidation, and milk production in peripartum Sahiwal cows. For this, thirty-two pregnant dry Sahiwal cows were selected at sixty days prepartum and divided into four groups viz control, T1, T2, and T3 of eight each. Group T1 were supplemented with zinc at 60 ppm/d/cow, group T2 were supplemented with vitamin E at 1,000 IU/d/cow and group T3 were supplemented with combination of vitamin E at 1,000 IU/d/cow and zinc at 60 ppm/d/cow during d 60 prepartum to d 90 postpartum. Blood samples were collected on d −60, −45, −30, −15, −7, −3, 0, 3, 7, 15, 30, 45, 60, 90, and 120 with respect to day of parturition and analysed for glucose, non esterified fatty acid, and thiobarbituric acid reactive substance. Body condition score was maintained significantly better (p<0.05) in T3 than in the control, T1 and T2 groups. Overall glucose level was higher (p<0.05) in T3 than control, T1, and T2 groups. Levels of nonesterified fatty acid, and thiobarbituric acid reactive substance were lower (p<0.05) in T3 than control, T1, and T2 groups. Milk yield was higher (p<0.05) in T3 than control, T1, and T2 groups. In conclusion, the present study indicated that the supplementation of vitamin E and zinc in peripartum Sahiwal cows enhanced milk production by reducing negative energy balance.

Diversity of cultivable bacteria associated with fruiting bodies of wild Himalayan Cantharellus spp.

The cultivable bacteria associated with fruiting bodies of different Cantharellus species collected from the forests of the north-western Himalayan region were studied. Repetitive extragenic palindromic (REP) and BOX-PCR fingerprinting analyses revealed that different strains were distributed within Cantharellus species. The number of bacteria varied from 0.5 to 1.1 × 103 CFU/g of fresh tissue for different fruiting bodies. Thirty different operational taxonomic units (OTUs) were found among the bacteria analyzed. 16S rRNA sequence analysis revealed that most of the bacteria associated with different Cantharellus species were Gammaproteobacteria belong to the genera Hafnia, Enterobacter, Ewingella, Rahnella, Stenotrophomonas, and Pseudomonas, and γ-proteobacterium, followed by Betaproteobacteria (Alcaligenes) and Firmicutes, (Bacillus). The most common bacteria associated with the majority of Cantharellus species were Hafnia and Stenotrophomonas species.

Evaluation of physiological and biochemical responses in different seasons in Surti buffaloes.

Aim: This study was conducted to evaluate the impact of hot dry, hot humid and comfortable season on physiological, hematological, biochemical, and oxidative stress parameters in Surti buffaloes. Materials and Methods: Ten lactating Surti buffaloes of similar physiological status were selected. Based on the temperature-humidity index (THI), their natural exposure to the environment was categorized as hot dry (THI1), hot humid (THI2) and moderate winter/comfort season (THI3). Blood/serum samples were collected and analyzed for physiological, hematological, biochemical, and oxidative stress parameters. The results were analyzed using standard statistical methods. Results: With increase in THI, significant rise in physiological parameters such as respiration rate (RR), hematological parameters such as red blood cell (RBC), hematocrit, hemoglobin (Hb) and mean cell Hb concentration (MCHC), biochemical parameters such as alanine aminotransferase (ALT), Na, K, creatinine, blood urea nitrogen, Mn, Cu and Zn, hormones such as cortisol and oxidative stress parameters such as glutathione peroxidase (GPx), superoxide dismutase (SOD), lipid peroxide (LPO) and total antioxidant status (TAS) and significant decline in glucose, cholesterol and triiodothyronine (T3) was observed. Conclusion: It was concluded that THI is a sensitive indicator of heat stress and is impacted by ambient temperature more than the relative humidity in buffaloes. Higher THI is associated with significantly increased RR, total RBC count, Hb, hematocrit, MCHC, ALT, urea, sodium, creatinine, triiodothyronine, SOD, GPx, LPO and TAS and with significant decrease in glucose, cholesterol and triiodothyronine (T3).