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Tamarind seed husk, an agricultural waste, was evaluated bothin vitro&in vivoas a methane-ameliorating agent. Basal diets forin vitrostudy were formulated using finger millet straw and concentrate in 70:30 proportions, and tamarind seed husk was included as part of the concentrate with equal (w/w) replacement of wheat bran to achieve 2.5 (T1), 5.0 (T2) and 10.0% (T3) levels. Based on thein vitroresults, two levels (2.5%, T1and 5.0%, T2) of tamarind seed husk were selected for thein vivostudy in male cattle to investigate the effect on enteric methane emission. Fifteen crossbred cattle divided into three groups were assigned to the following diets: control (T0, no husk), test group 1 (T1, 2.5% husk) and test group 2 (T2, 5.0% husk). Thein vivostudy showed a decrease of about 17% in enteric methane emission with a higher level (5%) of tamarind seed husk supplementation. Thein vivostudy demonstrated a non-adverse impact of supplementation on dry matter intake and digestibility, while rumen methanogens notably decreased at 5% inclusion level. It may be concluded that tamarind seed husk supplementation achieved significant methane reduction via direct inhibition of rumen methanogens without affecting fibre degradability. [ABSTRACT FROM PUBLISHER]

Achieving stable, efficient and non-toxic pulmonary gene delivery is most challenging requirement for successful gene therapy to lung. Composite nanoparticles (NPs) of the poly(lactic-co-glycolic acid) (PLGA) and cationic polymer polyethyleneimine (PEI) is an efficient alternative to viral and liposomal vectors for the pulmonary delivery of pDNA. NPs with different weight ratios (0–12.5%w/w) of PLGA/PEI were prepared and characterized for size, morphology, surface charge, pDNA loading and in vitro release. The in vitro cell uptake and transfection studies in the CFBE41o–cell line revealed that NPs with 10% w/w PEI were more efficient but they exhibited significant cytotoxicity in MTT assays, challenging the safety of this formulation. Surface modifications of these composite NPs through PEGylation reduced toxicity and enhanced cellular uptake and pDNA expression. PEGylation improved diffusion of NPs through the mucus barrier and prevented uptake by pulmonary macrophages. Finally, PEGylated composite NPs were converted to DPI by lyophilization and combined with lactose carrier particles, which resulted in improved aerosolization properties and lung deposition, without affecting pDNA bioactivity. This study demonstrates that a multidisciplinary approach may enable the local delivery of pDNA to lung tissue for effective treatment of deadly lung diseases. [ABSTRACT FROM AUTHOR]

Gut microflora co-evolved with the human evolution, performing health-promoting functions and protection from the enteric pathogens. The long association of the microflora sustained through ages, which in the light of new research, needs specific nutrients that are not required by the humans. The present article revisits the ancient foods and crops, their availability during different prehistoric times and their contribution in maintaining the gut flora and accrual of the beneficial effects. Although the concept of prebiotics is new, the ancient diet components contained the molecules or precursors of the prebiotics. [ABSTRACT FROM AUTHOR]

Aim: The present study was undertaken to evaluate the effect of variable levels of silkworm pupae oil and roughage: concentrate ratio on in vitro methane production, fermentation characteristics, and rumen protozoa population. Materials and Methods: In vitro gas production study (24 h) was performed with graded levels of silkworm pupae oil, namely, 0.5, 1, 2, 4, and 5% of the basal diet and four variable dietary regimes consisting roughage and concentrate in different proportions (70:30, 60:40, 50:50, and 40:60). At the end of incubation, gas samples were analyzed for methane, while fermented rumen liquor was used for protozoa enumeration. A separate set of incubations was carried out for the determination of in vitro dry matter digestibility. Results: Results from the in vitro studies revealed no adverse impact of the silkworm pupae oil supplementation up to 2% level on total gas production. However, supplementation beyond 2% has shown a reduction in total gas production. Incubation with variable levels (0.5-5%) of silkworm pupae oil with different dietary regimes indicated negligible (3-5%) to a substantial reduction (25-30%) on methane production. A graded decrement in methane production was recorded with increasing levels of silkworm pupae oil. Similarly, the protozoal populations were decreased from 10 to 51.5% with graded levels of silkworm pupae oil in different dietary regimes as studies did not reveal any significant (p>0.05) variation between 2 and 4% of oil supplementation. Conclusion: The silkworm pupae oil supplementation at 2% level decreases methane production by 12-15% without any adverse impact on feed fermentation. Oil supplementation may have a more pronounced effect on methane reduction if added to high roughage diet at in vitro conditions. However, in vivo, studies in ruminants are warranted to confirm the methane reduction with silkworm pupae oil supplementation. [ABSTRACT FROM AUTHOR]

A study was conducted in eighteen sheep divided into three groups of six animals each and fed on a diet comprising finger millet (Eleusine coracana) straw and concentrate in 50:50. Animals in the control group (T0) received diet without tanniferous tree leaves, while in T1 and T2 groups, animals received diets with Syzygium cumini (source of HT) and Machilus bombycina (source of CT), respectively. Enteric methane emission from the animals was measured. Hydrolyzable and condensed tannin intake in the T1 and T2 groups was 14.08 and 4.29 g/kg dry matter, respectively. Results from the study did not reveal any adverse impact of hydrolyzable and condensed tannin-containing leaves on the dry matter (g/100 kg BW) or digestible dry matter intake. The study confirmed a significant reduction in enteric methane emission (g/d) due to the supplementation. The methanogen numbers in test groups T1 and T2 were lower than those in the control; however, the rumen protozoal population (× 105/mL) was adversely affected by Syzygium cumini. Supplementation of selected leaves would help in conserving additional energy for the animal's use and also to curtail the contribution of livestock to GHG emissions. Long-term studies are warranted with selected sources to confirm the persistency of methane reduction. [ABSTRACT FROM AUTHOR]

Objective: Ensiling of tannin-rich fruit byproducts (FB) involves quantitative and qualitative changes in the tannins, which would consequently change the rumen fermentation characteristics. This study aimed to evaluate whether ensiled FBs are effective in mitigating methane emission from ruminants by conducting in vitro assessments. Methods: Fruit byproducts (grape pomace, wild grape pomace, and persimmon skin) were collected and subjected to four-week ensiling by Lactobacillus buchneri inoculant. A defined feed component with or without FB samples (both fresh and ensiled material) were subjected to in vitro anaerobic culturing using rumen fluid sampled from beef cattle, and the fermentation parameters and microbial populations were monitored. Results: Reduced methane production and a proportional change in total volatile fatty acids (especially enhanced propionate proportion) was noted in bottles containing the FBs compared with that in the control (without FB). In addition, we found lower gene copy number of archaeal 16S rRNA and considerably higher levels of one of the major fibrolytic bacteria (Fibrobacter succinogenes) in the bottles containing FBs than in the control, particularly, when it was included in a forage-based feed. However, in the following cultivation experiment, we observed that FBs failed to exhibit a significant difference in methane production with or without polyethylene glycol, implying that tannins in the FBs may not be responsible for the mitigation of methane generation. Conclusion: The results of the in vitro cultivation experiments indicated that not only the composition but also ensiling of FBs affected rumen fermentation patterns and the degree of methane generation. This is primarily because of the compositional changes in the fibrous fraction during ensiling as well as the presence of readily fermented substrates, whereas tannins in these FBs seemed to have little effect on the ruminal fermentation kinetics. [ABSTRACT FROM AUTHOR]

Aim: The aim of the study was to explore the anti-methanogenic potential of phyto-sources from Northeast region of the country and assess the effect on rumen fermentation characteristics and protozoa for their likely inclusion in animal diet to reduce methane emission. Materials and Methods: Twenty phyto-sources were collected from Northeast state, Assam, during March to April 2014. Phyto-sources were analyzed for their tannin content followed by screening for methane mitigation potential using in vitro system. The effect of tannin on methane production and other fermentation parameters was confirmed by attenuating the effect of tannin with polyethylene glycol (PEG)-6000 addition. About 200 mg dried phyto-source samples were incubated for 24 h in vitro, and volume of gas produced was recorded. The gas sample was analyzed on gas chromatograph for the proportion of methane in the sample. The effect of phyto-sources on rumen fermentation characteristics and protozoal population was determined using standard methodologies. Results: Results from studies demonstrated that Litchi chinensis, Melastoma malabathricum, Lagerstroemia speciosa, Terminalia chebula, and Syzygium cumini produced comparatively less methane, while Christella parasitica, Leucas linifolia, Citrus grandis, and Aquilaria malaccensis produced relatively more methane during in vitro incubation. An increase (p<0.05) in gas and methane production from the phyto-sources was observed when incubated with PEG-6000. Entodinimorphs were prominent ciliates irrespective of the phyto-sources, while holotrichs represented only small fraction of protozoa. An increase (p<0.05) in total protozoa, entodinimorphs, and holotrichs was noted when PEG-6000 added to the basal substrate. Our study confirmed variable impact of phyto-sources on total volatile fatty acid production and ammonia-N. Conclusion: It may be concluded that L. chinensis, M. malabathricum, L. speciosa, S. cumini, and T. chebula are having potent methane suppressing properties as observed in vitro in 24 h. These leaves could be supplemented in the animal diet for reducing methane emission; however, in vivo trials are warranted to confirm the methane inhibitory action and optimize the level of supplementation. [ABSTRACT FROM AUTHOR]

Purpose: Cumulus cells (CC) play important roles in oocyte development and cumulus expressed genes can be used as markers for oocyte quality. This study aimed to investigate temporal changes in the expression of cumulus marker genes during oocyte maturation as possible biomarkers of embryo developmental competence in ovine. Methods: Gene expression was assessed in the CC of the BCB+ (developmentally competent) and BCB- (developmentally poor) oocytes at 0, 12, and 24 h of in vitro maturation (IVM). Further, the association between the temporal cumulus gene expression and in vitro oocyte and embryo development was assessed. Results: The maturation and blastocyst formation rates were found significantly greater for the BCB+ than the BCB- oocytes. At the 0 h of IVM, a significant upregulation in the expression of PTGS2, STAR, SDC2, LHR, FGF2, BCL2, IL7RA, HSPA1A, and IFNT was observed in the CC of the poor (BCB-) as compared to the competent (BCB+) oocytes. In contrast, it was observed that as maturation progressed, the cumulus expression of most of the favorable genes was reduced and was found significantly downregulated at the completion of IVM in the poor as compared to the competent oocytes. Conclusions: The study revealed noticeable differences in the cumulus gene expression profile at different stages of IVM between ovine oocytes of differential developmental ability. The results indicated that the loss of cumulus gene expression along the maturation period in the poor oocytes was related to their intrinsic poor quality in the ovarian follicle. [ABSTRACT FROM AUTHOR]

This study aimed to assess the effect of the insulin-like grow factor 1 (IGF-1) treatment during in vitro maturation on the gene expression and developmental ability of ovine oocytes. Ovine cumulus-oocyte complexes (COC) were matured in vitro without (control) or with the supplementation of IGF-1 (100 ng/ml) and then subjected to in vitro fertilization and culture. The rate of oocyte maturation and embryo development was recorded and expression of the selected genes (involved in the PI3K/Akt and apoptosis signaling) was assessed in the matured oocytes. The IGF-1 treatment significantly (p <.05) improved the oocyte maturation rate (%) as compared to the control (81.5 ± 2.40 vs. 73.6 ± 0.94). Similarly, as compared to the control, the IGF-1 treatment significantly (p <.05) improved the rate (%) of cleavage (54.7 ± 1.58 vs. 67.2 ± 3.65) and the formation of 4–8 cell embryos (30.7 ± 2.89 vs. 44.1 ± 4.01) and morula (20.7 ± 2.08 vs. 32.8 ± 2.78). The IGF-1 treatment significantly (p <.05) upregulated the expression of IGF1R, PI3KR1, AKT1 and BCL2 and downregulated the expression of GSK3β, FOXO3 and CASP9 in the matured oocytes. In conclusion, the IGF-1 treatment significantly improved the developmental competence of ovine oocytes through the regulation of the PI3K/Akt and apoptosis signaling. [ABSTRACT FROM AUTHOR]

Foot-and-mouth disease virus (FMDV) causes a severe infection in ruminant animals. Here we present an in-depth transcriptional analysis of soft-palate tissue from cattle experimentally infected with FMDV. The differentially expressed genes from two Indian cattle (Bos indicus) breeds (Malnad Gidda and Hallikar) and Holstein Friesian (HF) crossbred calves, highlighted the activation of metabolic processes, mitochondrial functions and significant enrichment of innate antiviral immune response pathways in the indigenous calves. The results of RT-qPCR based validation of 12 genes was in alignment with the transcriptome data. The indigenous calves showing lesser virus load, elicited early neutralizing antibodies and IFN-γ immune responses. This study revealed that induction of potent innate antiviral response and cell mediated immunity in indigenous cattle, especially Malnad Gidda, significantly restricted FMDV replication during acute infection. These data highlighting the molecular processes associated with host-pathogen interactions, could aid in the conception of novel strategies to prevent and control FMDV infection in cattle. • RNA-Seq transcriptome was compared in three breeds of cattle infected with FMDV. • The DEGs highlighted strong innate immunity in the indigenous Bos indicus calves. • Initiation of innate and cell mediated immunity contribute to a reduced viral load. [ABSTRACT FROM AUTHOR]

Abstract: The effect of alkali viz., sodium hydroxide and potassium hydroxide, on the recovery of xylan from sugarcane bagasse under two different conditions namely overnight incubation at room temperature and autoclaving was investigated. Conditions for the enzymatic cleavage of the extracted xylan leading to the best yields of xylooligosaccharides was investigated using a fixed substrate concentration (2%, w/v) and varied inputs of other critical factors like pH, temperature, enzyme dose and reaction time. The sugarcane bagasse constituted of 98.99±0.01% organic matter, 35.67±0.15% cellulose, 23.22±0.05% hemicellulose, 6.04±0.06% lignin and 2.3±0.01% protein. The highest xylan recovery of 85% was attained using 12% sodium hydroxide combined with steam application. Analyses of the enzymatic hydrolyzate showed the presence of XOS of varying degree of polymerization falling in the range of 2–3. The maximum conversion to XOS was observed at temperature of 40°C, pH of 4 with an enzyme dose of 2.65U and a reaction time of 8h. The response surface analysis predicted ideal condition viz., temperature 40.99°C, pH 4.92, enzyme dose 3.13U and reaction time 19.19h for maximizing xylobiose yield, while for maximum xylotriose yields the ideal variables were 40.37°C, pH 4.13, enzyme dose 5.89U and an incubation time of 18.14h. The findings established the suitability of sugarcane bagasse as a raw material for extraction of xylan vis a vis its conversion into XOS. [Copyright &y& Elsevier]

Deciphering sperm transcriptome is the key to understanding the molecular mechanisms governing peri-fertilization, embryonic development, and pregnancy establishment. This study aimed to profile sperm transcriptome to identify signature transcripts regulating male fertility. Semen samples were collected from 47 bulls with varied fertility rates. The sperm total RNA was isolated (n = 8) and subjected to transcriptome sequencing. Based on the expression pattern obtained from RNA profiling, the bulls were grouped (p = 0.03) into high-fertile and sub-fertile, and signature transcripts controlling sperm functions and fertility were identified. The results were validated using the OMIM database, qPCR, and sperm function tests. The sperm contains 1100 to 1700 intact transcripts, of which BCL2L11 and CAPZA3 were abundant and associated (p < 0.05) with spermatogenesis and post-embryonic organ morphogenesis. The upregulated genes in the acrosome integrity and functional membrane integrity groups had a close association with the fertility rate. The biological functions of these upregulated genes (p < 0.05) in the high-fertile bulls were associated with spermatogenesis (AFF4 and BRIP1), sperm motility (AK6 and ATP6V1G3), capacitation and zona binding (AGFG1), embryo development (TCF7 and AKIRIN2), and placental development (KRT19). The transcripts involved in pathways regulating embryonic development such as translation (EEF1B2 and MTIF3, p = 8.87E−05) and nonsense-mediated decay (RPL23 and RPL7A, p = 5.01E−27) were upregulated in high-fertile bulls. The identified transcripts may significantly impact oocyte function, embryogenesis, trophectoderm development, and pregnancy establishment. In addition, the study also reveals that the genes governing sperm functional membrane integrity and acrosome integrity have a prospective effect on male fertility. [ABSTRACT FROM AUTHOR]

Summary: The study aimed to investigate the utilisation of prebiotic D‐tagatose by probiotic lactobacilli (L. rhamnosus GG, L. casei, L. acidophilus and L. fermentum) and enteric pathogens (E. coli and S. typhimurium) to determine synbiotic potential of D‐tagatose and lactobacilli combination. The antimicrobial efficacy of the synbiotic preparations (D‐tagatose with lactobacillus) was assessed against selected pathogens in co‐culture assays. Evidently, D‐tagatose supported growth of selected lactobacilli, especially the L. rhamnosus GG and L. casei, but not the enteric pathogens. The tested synbiotic preparations completely inhibited growth of both the pathogens, even in the presence of D‐glucose in co‐cultures. The well‐diffusion assay demonstrated the presence of antimicrobial activity as recorded by drop of culture pH. The study substantiated that D‐tagatose could be an effective prebiotic component for formulation of potential synbiotic combinations with L. rhamnosus GG or L. casei that can be used as an alternative to antibiotics against enteric pathogens. [ABSTRACT FROM AUTHOR]

The rumen wharves conglomerate of symbiotic fibrolytic microflora with the potential to produce repertoire of carbohydrate-active enzymes to elicit the degradation of recalcitrant plant lignocellulosic complex into simpler forms that can be utilized by the host. A metatranscriptomics approach was adopted to explore the metabolically active microbiota and the transcripts involved in the deconstruction of lignocellulose in Indian crossbred cattle rumen. The experimental animals were fed with mixed diet for a period of twenty-one days and the total RNA was isolated from the rumen digesta using a modified total RNA extraction protocol. After mRNA enrichment, the cDNA libraries were subjected to sequencing under Illumina HiSeq platform and generated about 2.89 GB of raw sequences. The contig assembly was accomplished using two different tools and the redundant sequences were removed before further processing. A total of 133,930 orfs were predicted from the assembled contigs using MetaGeneMark tool and the taxonomic affiliation of orfs were achieved using MEGAN 6.0 Community Edition. A total of 17 bacterial, 5 eukaryotic and 1 archaeal phylum were identified from rumen metatranscriptomic dataset. Phylum Bacteroidetes were acknowledged as the most abundant and metabolically active rumen bacterial populations in cattle rumen with 36,414 orfs corresponding to it, followed by Firmicutes with 6,349 orfs. Further, MEGAN annotation of metatranscriptomic data at species level revealed that Prevotella brevis and Prevotella ruminicola as the most metabolically active bacterial populations representing 4,022 and 2,725 orfs each. The functional annotation of metatranscriptomic data using COG database revealed that a large number of transcripts (~16%) were corresponding to translation, ribosomal structure and biogenesis and carbohydrate transport and metabolism (15%). dbCAN annotation of rumen metatranscriptomic data identified and classified 5,267 transcripts belonging to 168 CAZyme families (GH-52%, GT-26%, CBM-14%, CE-6%, PL-2% and AA-0%). The microbial community analysis of the CAZyme encoding transcripts using M5NR database revealed that a significant proportion of CAZymes were contributed by genera Prevotella (37%) and Bacteroides (21%). The cattle rumen microbiome metatranscriptome analysis presented in this study facilitated the detection of large number of transcripts encoding diverse CAZymes that actively take part in the hydrolysis of plant lignocellulose complex that may be useful for improving livestock and biofuel generation. [ABSTRACT FROM AUTHOR]

The current study aimed to assess the effect of different seasons (summer, monsoon and winter) on the quality, preservability and fertilizing ability of spermatozoa in Bannur rams. The level of heat stress was significantly greater during the summer than monsoon or winter seasons. Spermatozoa concentration (×106/ml) in semen was found significantly (P < 0.05) lesser during the summer (2446 ± 80) as compared to monsoon (2759 ± 56) or winter (2769 ± 56). The progressive motility (%) of spermatozoa at the 72 h post-preservation was significantly (P < 0.01) lesser during the summer (47.5 ± 2.5) than monsoon (62.5 ± 2.5) and winter (57.5 ± 2.5). Significantly (P < 0.01 or P < 0.05) lesser liveability and acrosomal integrity, but greater total morphological abnormalities of spermatozoa were recorded at all the preservation time points during the summer as compared to monsoon or winter. The cleavage rate (%) was found significantly (P < 0.01) greater during the monsoon (56.6 ± 2.24) as compared to summer (41.7 ± 1.26) and winter (41.4 ± 0.99). The results indicate a definite seasonal pattern in the quality, preservability and fertilizing ability of spermatozoa in Bannur rams, which were found superior during the monsoon. [ABSTRACT FROM AUTHOR]

The ability to identify the sex of embryo and control of sex ratio has a great commercial importance to livestock industry. Prediction of embryonic sex could be useful in the management decisions of sex selection in breeding programs. Several methods have been attempted to determine the sex but the polymerase chain reaction (PCR)‐based sexing method is generally favoured, as it is cost effective, simple and reliable. The aim of the present study was to identify sex of sheep embryos produced in vitro through amplification of glyceraldehyde 3‐phosphate dehydrogenase (GAPDH), sex‐determining region Y (SRY) and amelogenin genes present in genomic DNA (gDNA) of embryos through PCR. To avoid false interpretation of the result by no amplification of SRY in female embryos, a duplex PCR was approached to amplify combinedly SRY and GAPDH genes. Sex‐specific blood was used in PCR as positive control. In vitro sheep embryos were produced as per standardized protocol of laboratory. Sexing of sex‐specific blood and in vitro produced embryos were approached though PCR to amplify the respective genes using gDNA present in the sample without its traditional isolation. The accuracy of sex prediction for embryos was 100% by this procedure. [ABSTRACT FROM AUTHOR]

The effect of interleukin-6 (IL-6) supplementation during the different phases of in vitro embryo culturing (IVC) on embryo development and embryonic gene expression was studied in ovine. IL-6 was added to IVC medium during the late phases (72–192 h; 5, 10, and 25 ng/ml IL-6) or entire period (0–192 h; 10 ng/ml IL-6) of IVC to determine its effect on embryo development. Further, the effect of IL-6 (10 ng/ml) supplementation at the 72 h of IVC on gene expressions associated with JAK/STAT signalling and pluripotency in 8–16 cell embryos (1 h post-supplementation) and compact morulae (48 h post-supplementation), and apoptosis and primitive endoderm (PrE) development in compact morulae was investigated. The supplementation of 10 ng/ml IL-6 during the late phases of IVC significantly (P < 0.05) increased blastocyst formation (35.2 ± 1.52%) compared to the control (21.1 ± 1.11%), and 5 ng/ml (25.9 ± 2.98%) or 25 ng/ml (16.5 ± 0.73%) IL-6 groups. Conversely, IL-6 (10 ng/ml) treatment throughout the IVC period significantly (P < 0.05) decreased the rate of cleavage (55.4 ± 1.57%) and blastocyst formation (14.5 ± 1.28%) compared to the control group (65.8 ± 1.35% and 21.5 ± 0.97%, respectively). In 8–16 cell embryos and compact morulae, the IL-6 treatment significantly (P < 0.05) affected the expression of genes associated with JAK/STAT signalling and pluripotency. Further, the treatment significantly (P < 0.05) downregulated BAX and CASP3, and upregulated GATA6 expression in compact morulae. In conclusion, IL-6 supplementation affected the in vitro development of ovine embryos in a dose- and time-dependent manner. The beneficial effect of IL-6 on the development of late-stage embryos was mediated through the changes in gene expressions associated with JAK/STAT signalling, pluripotency, apoptosis and PrE development. • Effect of IL-6 on development and gene expression of ovine embryos was assessed. • IL-6 had a dose- and stage-dependent influence on embryo development in vitro. • IL-6 promoted development of the late-stage embryos. • Beneficial effect of IL-6 was mediated through JAK/STAT and pluripotency signalling. [ABSTRACT FROM AUTHOR]

Graphical abstract Highlights • Exogenous IL-7 affected in vitro maturation of ovine oocytes. • Oocyte maturation was improved with low concentration (1 ng/ml) of exogenous IL-7. • Higher concentrations of exogenous IL-7 were detrimental for oocyte maturation. • Higher exogenous IL-7 likely caused ROS-mediated oxidative stress. • Higher exogenous IL-7 also stimulated apoptotic signalling. Abstract Interleukin-7 (IL-7) mediated signals are linked to development, proliferation, survival and differentiation of cells. Recent evidences indicate its role in oocyte maturation process as well. Nevertheless, the underlying mechanisms of IL-7 involvement in oocyte maturation are not well characterized. In addition, currently no information is available on the effect of exogenous IL-7 on oocyte maturation in ovine or any other species. In this study, the effect of IL-7 supplementation during in vitro maturation (IVM) on the maturation rate, production of reactive oxygen species (ROS) and gene expression of ovine cumulus-oocyte complexes (COC) was assessed. IL-7 (0.5, 1, 2, 5 and 10 ng/ml) was supplemented in IVM medium at the beginning (0 h) and maturation rate of COC was assessed at the completion of IVM (24 h). The maturation rate (%) was found significantly (P = 0.000) greater with the 1 ng/ml of IL-7 supplementation (69.5) than control (60.0). In contrast, the maturation rate was reduced significantly (P = 0.000) with the 2 (47.1), 5 (39.2) and 10 ng/ml (39.1) of IL-7 as compared to the control. The level of intracellular ROS in the matured COC was found considerably higher with the 5 ng/ml of IL-7 followed by 1 ng/ml of IL-7 and control. It was evident that in the presence of superoxide dismutase-inhibitor, 1 ng/ml of IL-7 did not stimulate oocyte maturation. In contrast, oocyte maturation was improved with 5 ng/ml of IL-7 supplementation in the presence of NADPH-oxidase-inhibitor. IL-7 supplementation influenced gene expression in COC in a dose and time dependant manner. The expression of genes related to ROS production and apoptosis were upregulated and the genes associated with antioxidant mechanisms were downregulated noticeably with the supplementation of 5 ng/ml of IL-7. In conclusion, IL-7 at low concentration was beneficial for oocyte maturation, which was likely mediated through the favourable level of intracellular ROS and antioxidant mechanisms. In contrast, the detrimental effects of greater IL-7 concentrations on oocyte maturation were possibly arbitrated through the ROS-mediated oxidative stress, compromised antioxidant mechanism and stimulated apoptotic signalling. [ABSTRACT FROM AUTHOR]

With artificial insemination (AI) and other precision dependent assisted reproductive technologies (ART) being followed in large scale in human and animal reproduction, assessing semen quality and fertilizability is under continuous scrutiny. Various tests have been developed to predict semen quality, but so far no single, highly reliable test is available. In this regard, transcriptomic profiling of spermatozoa assumes significance as it carries the information about spermatogenesis, sperm function, and paternal roles in post-fertilization events. Human spermatozoal transcriptome profiling has been carried out on a large number of individuals to predict the semen quality. A study in human indicated that the outcome of some idiopathic couples seeking reproductive care could be helped using transcriptomic profiling of spermatozoa. Such studies have a direct impact on the bovine dairy industry, wherein AI is practiced. Limited studies in bovine spermatozoal transcriptome profiling have revealed that the spermatozoa contain various classes of RNA, like in human. Approximately 13,000 bovine genes yield a series of spermatozoal transcripts, of which most are fragmented in nature. Their abundance is indicative of the timing of events associated with spermatogenesis, e.g., PRM1, IGF1, BMP2; sperm function, TSSK6, CRISP, HSFY2; fertility, UBE2D3, Integrin-β, LDC-1; and embryonic development, miR34c-5p, BCL2L11, BRCA1. The most abundant translated bovine transcripts are BSP3 and SPATA18, and are involved in regulation of germ cell development and the maintenance of chromatin integrity during spermatogenesis respectively. The presence of transcripts associated with placental development, e.g., placental associated glycoproteins (PAGs) have suggested their possible influence beyond early embryonic development. Changes in transcript levels like RPL31 and PRKCE that increase, and PRM1 that decreases, during cryopreservation need to be defined in order to optimize cryopreservation and fertility yield. Spermatozoal transcriptome profiling with validation studies are warranted in large numbers of animals to elucidate their significance for selecting fertile bulls for the breeding program. Abbreviations: AI: artificial insemination; BSE: breeding soundness evaluation; cfs-mRNA: cell-free seminal mRNA; piRNA: PIWI-interacting RNA; tRNA: transfer RNA; fg: femtogram; TPM: transcripts per million reads; RPKM: reads per kilobase million; rRNA: ribosomal RNA; mt-RNA: mitochondrial RNA; lncRNA: long non-coding RNA; sncRNA: small noncoding RNA; snoRNA: small nucleolar RNA; snRNA: small nuclear RNA; miRNA: microRNA; snaR: small NF90-associated RNAs; SINES: short interspersed nuclear elements; LINES: long interspersed nuclear elements; MER: medium reiterated sequence; F1 offspring: filial 1 offspring; PAGs: placental associated glycoproteins; TCP: Transcription factor T complex protein; BSP3: bovine seminal plasma protein 3; SCNT: somatic cell nuclear transfer; qPCR: quantitative (real-time) polymerase chain reaction; SSH: suppression subtractive hybridization; SNP: single nucleotide polymorphism; 2-DE: 2 dimensional gel electrophoresis; LC-MS/MS: liquid chromatography-tandem mass spectrometry [ABSTRACT FROM AUTHOR]

Nutraceuticals are gaining importance owing to their potential applications in numerous sectors including food and feed industries. Among the emerging nutraceuticals, d‐tagatose occupies a significant niche because of its low calorific value, antidiabetic property and growth promoting effects on beneficial gut bacteria. As d‐tagatose is present in minute quantities in naturally occurring food substances, it is produced mainly by chemical or biological means. Recently, attempts were made for bio‐production of d‐tagatose using l‐arabinose isomerase enzyme to overcome the challenges of chemical process of production. Applications of d‐tagatose for maintaining health and wellbeing are increasing due to growing consumer awareness and apprehension against modern therapeutic agents. This review outlines the current status on d‐tagatose, particularly its production, properties, biological role, applications, and the future perspectives. [ABSTRACT FROM AUTHOR]

The enzyme endo-inulinase hydrolyzes inulin to short chain fructooligosaccharides (FOS) that are potential prebiotics with many health promoting benefits. Although the raw materials for inulin production are inexpensive and readily available, commercial production of FOS from inulin is limited due to inadequate availability of the enzyme source. This study aimed to identify the fungi capable of producing endo-inulinase based on the in silico analysis of proteins retrieved from non-redundant protein sequence database. The endo-inulinase of Aspergillus ficuum was used as reference sequence. The amino acid sequences with >90% sequence coverage, belonging to different fungi were retrieved from the database and used for constructing three-dimensional (3D) protein models using SWISS-MODEL and Bagheerath H. The 3D models of comparable quality as that of the reference endo-inulinase were selected based on QMEAN Z score. The selected models were evaluated and validated for different structural and functional qualities using Pro-Q, ProSA, PSN-QA, VERIFY-3D, PROCHECK, PROTSAV metaserver, STRAP, molecular docking, and molecular dynamic simulation analyses. A total of 230 proteins belonging to 53 fungal species exhibited sequence coverage >90%. Sixty one protein sequences with >60% sequence identity were modeled as endo-inulinase with higher QMEAN Z Score. The evaluations and validations of these 61 selected models for different structural and functional qualities revealed that 60 models belonging to 22 fungal species exhibited native like structure and unique motifs and residues as that of the reference endo-inulinase. Further, these models also exhibited similar kind of interaction between the active site around the conserved glutamate residue and substrate as that of the reference endo-inulinase. In conclusion, based on the current study, 22 fungal species could be identified as endo-inulinase producer. Nevertheless, further biological assessment of their capability for producing endo-inulinase is imminent if they are to be used for commercial endo-inulinase production for application in FOS industry. [ABSTRACT FROM AUTHOR]

Sperm RNA can be used to understand the past spermatogenic process, future successful fertilization, and embryo development. To study the sperm RNA composition and function, isolation of good quality RNA with sufficient quantity is essential. The objective of this study was to assess the influence of sperm input concentrations and RNA isolation methods on RNA yield and quality in bull sperm. The fresh semen samples from bulls ( n = 6) were snap-frozen in liquid nitrogen and stored at −80 °C. The sperm RNA was isolated using membrane-based methods combined with TRIzol (RNeasy + TRIzol and PureLink + TRIzol) and conventional methods (TRIzol, Double TRIzol, and RNAzol RT). Based on fluorometric quantification, combined methods resulted in significantly ( P < 0.05) higher total RNA yields (800–900 ng/30–40 × 10 6 ) as compared with other methods and yielded 20 to 30 fg of RNA/spermatozoon. The quality of RNA isolated by membrane-based methods was superior to that isolated by conventional methods. The sperm RNA was observed to be intact as well as fragmented (50–2000 bp). The study revealed that the membrane-based methods with a cocktail of lysis solution and an optimal input concentration of 30 to 40 million sperm were optimal for maximum recovery of RNA from bull spermatozoa. [ABSTRACT FROM AUTHOR]

The highly pathogenic avian influenza (HPAI) H5N1 virus, currently circulating in Asia, causes severe disease in domestic poultry as well as wild birds like crow. However, the molecular pathogenesis of HPAIV infection in crows and other wild birds is not well known. Thus, as a step to explore it, a comprehensive global gene expression analysis was performed on crow lungs, infected with HPAI H5N1 crow isolate (A/Crow/India/11TI11/2011) using high throughput next generation sequencing (NGS) ( GS FLX Titanium XLR70 ). The reference genome of crow is not available, so RNA seq analysis was performed on the basis of a de novo assembled transcriptome. The RNA seq result shows, 4052 genes were expressed uniquely in noninfected, 6277 genes were expressed uniquely in HPAIV infected sample and of the 6814 genes expressed in both samples, 2279 genes were significantly differentially expressed. Our transcriptome profile data allows for the ability to understand the molecular mechanism behind the recent lethal HPAIV outbreak in crows which was, until recently, thought to cause lethal infections only in gallinaceous birds such as chickens, but not in wild birds. The pattern of differentially expressed genes suggest that this isolate of H5N1 virus evades the host innate immune response by attenuating interferon (IFN)-inducible signalling possibly by down regulating the signalling from type I IFN (IFNAR1 and IFNAR2) and type II IFN receptors, upregulation of the signalling inhibitors suppressor of cytokine signalling 1 (SOCS1) and SOCS3 and altering the expression of toll-like receptors (TLRs). This may be the reason for disease and mortality in crows. [ABSTRACT FROM AUTHOR]

Previous studies on mitochondrial DNA analysis of sheep from different regions of the world have revealed the presence of two major- A and B, and three minor- C, D and E maternal lineages. Lineage A is more frequent in Asia and lineage B is more abundant in regions other than Asia. We have analyzed mitochondrial DNA sequences of 330 sheep from 12 different breeds of India. Neighbor-joining analysis revealed lineage A, B and C in Indian sheep. Surprisingly, multidimensional scaling plot based on FST values of control region of mtDNA sequences showed significant breed differentiation in contrast to poor geographical structuring reported earlier in this species. The breed differentiation in Indian sheep was essentially due to variable contribution of two major lineages to different breeds, and sub- structuring of lineage A, possibly the latter resulting from genetic drift. Nucleotide diversity of this lineage was higher in Indian sheep (0.014 ± 0.007) as compared to that of sheep from other regions of the world (0.009 ± 0.005 to 0.01 ± 0.005). Reduced median network analysis of control region and cytochrome b gene sequences of Indian sheep when analyzed along with available published sequences of sheep from other regions of the world showed that several haplotypes of lineage A were exclusive to Indian sheep. Given the high nucleotide diversity in Indian sheep and the poor sharing of lineage A haplotypes between Indian and non-Indian sheep, we propose that lineage A sheep has also been domesticated in the east of Near East, possibly in Indian sub-continent. Finally, our data provide support that lineage B and additional lineage A haplotypes of sheep might have been introduced to Indian sub-continent from Near East, probably by ancient sea trade route. [ABSTRACT FROM AUTHOR]

Mammalian spermatozoa deliver various classes of RNAs to the oocyte during fertilization, and many of them may regulate fertility. The objective of the present study was to determine the composition and abundance of spermatozoal transcripts in fresh bull semen. The entire transcriptome of the spermatozoa from bulls (n = 3) was sequenced using two different platforms (Ion Proton and Illumina) to identify the maximum number of genes present in the spermatozoa. The bovine spermatozoa contained transcripts for 13,833 genes (transcripts per million, TPM > 10). Both intact and fragmented transcripts were found. These spermatozoal transcripts were associated with various stages of spermatogenesis, spermatozoal function, fertilization, and embryo development. The presence of intact transcripts of pregnancy-associated glycoproteins (PAGs) in the spermatozoa suggest a possible influence of sperm transcripts beyond early embryonic development. The specific regions (exon, intron, and exon-intron) of the particular spermatozoal transcripts might help regulate fertilization. This study demonstrates that the use of two different RNA-seq platforms provides a comprehensive profile of bovine spermatozoal RNA. Spermatozoal RNA profiling may be useful as a non-invasive method to delineate possible causes of male infertility and to predict fertility in a manner that is more effective than the conventional methods. [ABSTRACT FROM AUTHOR]