Anil Kumar Puniya

Studies on the effect of particle size on solid-state fermentation of sugarcane bagasse into animal feed using white-rot fungi

Sugarcane bagasse, a complex substrate, when differentiated into four fractions of particle sizes (<1 mm, 1–3 mm, 3–5 mm, 5–10 mm) with a view to enhancing its nutritive value as animal feed rather than production of fungal protein for human consumption, showed a varying pattern of degradation by white-rot fungi and varying in vitro rumen digestions. The fractions of bagasse were assessed for their bioconversion efficiencies in terms of disappearance of lignin and change in digestibility using Pleurotus sp. P7, Agrocybe aegarita A1, Pleurotus eryngii, Pleurotus sp. P1 and Kuehneromyces mutabilis. The P. eryngii improved the digestibility of all the bagasse fractions. The mechanical separation of a substrate can be of significance when it is utilized as a material to be fermented by white-rot fungi to produce enriched animal feed.

Interaction of gut microflora with tannins in feeds

Tannins (hydrolyzable and condensed) are water-soluble polyphenolic compounds that exert antinutritional effects on ruminants by forming complexes with dietary proteins. They limit nitrogen supply to animals, besides inhibiting the growth and activity of ruminal microflora. However, some gastrointestinal microbes are able to break tannin–protein complexes while preferentially degrading hydrolyzable tannins (HTs). Streptococcus gallolyticus, Lonepinella koalarum and Selenomonas ruminantium are the dominant bacterial species that have the ability to degrade HTs. These tanninolytic microorganisms possess tannin-degrading ability and have developed certain mechanisms to tolerate tannins in feeds. Hence, selection of efficient tanninolytic microbes and transinoculation among animals for long-term benefits become areas of intensive interest. Here, we review the effects of tannins on ruminants, the existence and significance of tannin-degrading microorganisms in diverse groups of animals and the mechanisms that tannin-degrading microorganisms have developed to counter the toxic effects of tannin.

Molecular beacon : a multitask probe

The specificity of hybridization of complementary sequences in DNA is the basic strategy for identifying target genes. For this, stem loop oligonucleotide probes have been developed in order to enhance the specificity and selectivity to the target DNA. Among stem loop oligonucleotides, molecular beacons are the recent probes used for biomolecular recognition reactions. Molecular beacon-based assays are fast, simple, inexpensive, and enable real-time monitoring of nucleic acid reactions both, in vivo and in vitro. This review has been designed to provide a better understanding of the different aspects of molecular beacons, e.g. structure, designing and applications in real-time monitoring of nucleic acid amplification, detection of pathogens, nucleic acid–protein interaction, genetic analysis and array technology.

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.

Gut Microbiota Modulation and Its Relationship with Obesity Using Prebiotic Fibers and Probiotics: A Review

In the present world scenario, obesity has almost attained the level of a pandemic and is progressing at a rapid rate. This disease is the mother of all other metabolic disorders, which apart from placing an added financial burden on the concerned patient also has a negative impact on his/her well-being and health in the society. Among the various plausible factors for the development of obesity, the role of gut microbiota is very crucial. In general, the gut of an individual is inhabited by trillions of microbes that play a significant role in host energy homeostasis by their symbiotic interactions. Dysbiosis in gut microbiota causes disequilibrium in energy homeostasis that ultimately leads to obesity. Numerous mechanisms have been reported by which gut microbiota induces obesity in experimental models. However, which microbial community is directly linked to obesity is still unknown due to the complex nature of gut microbiota. Prebiotics and probiotics are the safer and effective dietary substances available, which can therapeutically alter the gut microbiota of the host. In this review, an effort was made to discuss the current mechanisms through which gut microbiota interacts with host energy metabolism in the context of obesity. Further, the therapeutic approaches (prebiotics/probiotics) that helped in positively altering the gut microbiota were discussed by taking experimental evidence from animal and human studies. In the closing statement, the challenges and future tasks within the field were discussed.

D1/D2 Domain of Large-Subunit Ribosomal DNA for Differentiation of Orpinomyces spp.

ABSTRACT This study presents the suitability of D1/D2 domain of large-subunit (LSU) ribosomal DNA (rDNA) for differentiation of Orpinomyces joyonii and Orpinomyces intercalaris based on PCR-restriction fragment length polymorphism (RFLP). A variation of G/T in O. intercalaris created an additional restriction site for AluI, which was used as an RFLP marker. The results demonstrate adequate heterogeneity in the LSU rDNA for species-level differentiation.

In vitro degradation of wheat straw by anaerobic fungi from small ruminants

Abstract Anaerobic ruminal fungi may play an active role in fibre degradation as evidenced by the production of different fibrolytic enzymes in culture filtrate. In the present study, 16 anaerobic fungal strains were isolated from ruminal and faecal samples of sheep and goats. Based on their morphological characteristics they were identified as species of Anaeromyces, Orpinomyces, Piromyces and Neocallimastix. Isolated Neocallimastix sp. from goat rumen showed a maximum activity of CMCase (47.9 mIU ml−1) and filter paper cellulase (48.3 mIU ml−1), while Anaeromyces sp. from sheep rumen showed a maximum xylanolytic activity (48.3 mIU ml−1). The cellobiase activity for all the isolates ranged from 178.0 – 182.7 mIU ml−1. Based on the enzymatic activities, isolated Anaeromyces sp. from sheep rumen and Neocallimastix sp. from goat rumen were selected for their potential of in vitro fibre degradation. The highest in vitro digestibility of NDF (23.2%) and DM (34.4%) was shown for Neocallimastix sp. from goat rumen, as compared to the digestibility of NDF and DM in the control group of 17.5 and 25.0%, respectively.

Isolation and characterization of methanogens from rumen of Murrah buffalo

Methanogens were isolated from the rumen of Murrah buffaloes (Bubalus bubalis). These isolates (BRM-1, -2 and -3) were found to utilize CO2 + H2 mixture, formate and acetate as substrate, but failed to grow on ethanol and methanol. Their physiological analysis showed that they could tolerate NaCl and bile salts up to 1.0% but 2.0% bile salt inhibited their growth. Based on 16S rRNA/mcrA gene sequence analysis, the isolates showed their phylogenetic relation with genus Methanobrevibacter and Methanomicrobium. BRM-1 and -3 showed 100% similarity with Methanobrevibacter smithii, while BRM-2 showed 100% similarity with Methanomicrobium mobile. The mcrA protein-based phylogeny also showed similar results to the mcrA gene, suggesting no apparent difference in the phylogeny between DNA and amino acid sequences of these isolates.

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.

Role of live microbial feed supplements with reference to anaerobic fungi in ruminant productivity: A review

Abstract To keep the concept of a safe food supply to the consumers, animal feed industries world over are showing an increasing interest in the direct-fed microbials (DFM) for improved animal performance in terms of growth or productivity. This becomes all the more essential in a situation, where a number of the residues of antibiotics and/or other growth stimulants reach in milk and meat with a number of associated potential risks for the consumers. Hence, in the absence of growth stimulants, a positive manipulation of the rumen microbial ecosystem to enhance the feedstuff utilization for improved production efficiency by ruminants has become of much interest to the researchers and entrepreneurs. A few genera of live microbes (i.e., bacteria, fungi and yeasts in different types of formulations from paste to powder) are infrequently used as DFM for the domestic ruminants. These DFM products are live microbial feed supplements containing naturally occurring microbes in the rumen. Among different DFM possibilities, anaerobic rumen fungi (ARF) based additives have been found to improve ruminant productivity consistently during feeding trials. Administration of ARF during the few trials conducted, led to the increased weight gain, milk production, and total tract digestibility of feed components in ruminants. Anaerobic fungi in the rumen display very strong cell-wall degrading cellulolytic and xylanolytic activities through rhizoid development, resulting in the physical disruption of feed structure paving the way for bacterial action. Significant improvements in the fiber digestibility were found to coincide with increases in ARF in the rumen indicating their role. Most of the researches based on DFM have indicated a positive response in nutrient digestion and methane reducing potential during in vivo and/or in vitro supplementation of ARF as DFM. Therefore, DFM especially ARF will gain popularity but it is necessary that all the strains are thoroughly studied for their beneficial properties to have a confirmed ‘generally regarded as safe’ status for ruminants.