Dairy production plays an integral part in supporting smallholder farmers’ livelihoods. The desire to increase the number of dairy cattle is not feasible due to the reduced output of feed resources occasioned by clim...Dairy production plays an integral part in supporting smallholder farmers’ livelihoods. The desire to increase the number of dairy cattle is not feasible due to the reduced output of feed resources occasioned by climate change. Consequently, the need to increase productivity per cow is inevitable. Conventional protein supplements are costly;hence, the need to explore affordable nutrientdense alternative feed resources. Liquid brewers’ yeast (LBY), a by-product of the brewing industry, is a rich protein supplement in dairy production. This study aimed to assess the dairy farming conditions and utilization levels of LBY as a feed supplement in Githunguri Sub-county, Kiambu. Semi-structured questionnaires were administered to 457 dairy farmers in a cross-sectional survey. The findings revealed that most farmers (94.2%) fed their cattle on established forage/fodder and crop residues with supplementation. Even though 53.1% of the respondents were aware of the use of LBY, only 30.6% utilized it to supplement dairy cows, most of whom (96.0%) used it fresh without preservation. Membership in farmers’ organizations increased awareness of LBY (r = 0.732). Principal component analysis indicated that the benefits of using LBY outweigh the challenges involved with a loading matrix of 0.891 - 0.954 and 0.681 - 0.807, respectively. The low adoption and use levels of LBY as a source of protein supplements were due to low awareness. There is a need for concerted efforts by stakeholders in the industry to increase farmers’ knowledge base on the utilization and effectiveness of LBY in dairy production.展开更多
Using probiotics as animal feed additives instead of antibiotics is gaining momentum to avert adverse negative effects on human health. Liquid brewers yeast (LBY) is an industrial by-product containing probiotic micro...Using probiotics as animal feed additives instead of antibiotics is gaining momentum to avert adverse negative effects on human health. Liquid brewers yeast (LBY) is an industrial by-product containing probiotic microorganisms and is also used as a protein supplement for dairy animals. Nevertheless, value chain actors lack of appropriate handling practices compromises the by-products quality and safety. This study aimed to determine the effect of variation in temperature on microbial diversity and probiotic effects during the storage time of LBY sampled from distributors and farmers from Githunguri sub-county of Kenya. The samples were stored at 20C, 25C and 30C, then tested on 0, 5, 10, 15 and 20 days. The studys parameters involved determining the pH levels, lactic acid bacteria (LAB), total coliform count (TCC), mould, and yeast in LBY. The rate (k) of the reaction kinetics model was used to extrapolate the expected probiotic shelf life. The LAB and yeast populations were reduced in a first-order reaction at all storage temperatures. The rate of reduction in the numbers of LAB reduced with an increase in temperature (k = 0.019 and 0.023) at 20C and 30C, respectively. Yeasts highest rate of growth reduction was 25C (k = 0.009) and least at 30C (k = 0.043). The minimum effective concentration for probiotics of 106 CFU/mL needed to observe the beneficial physiological impact on farm animals was achieved between 34.9 and 35.5 days at the tested storage temperatures. The study provides insight into the unexploited low-cost probiotic potential of LBY in dairy production. Conversely, handling practices and environmental microbial contamination along the value chain can compromise product quality and safety. There is a need to advocate its use in dairy for improved productivity and sensitize farmers to appropriate hygienic measures along the LBY value chain.展开更多
Ladakh’s cropped and non-cropped areas suffer greatly from weedy invasion under subsistence agricultural system where weeding is not a priority but total crop + weed biomass together contribute to livestock feeding s...Ladakh’s cropped and non-cropped areas suffer greatly from weedy invasion under subsistence agricultural system where weeding is not a priority but total crop + weed biomass together contribute to livestock feeding significantly. As agriculture along with livestock rearing is major activity of livelihood options, thereby contributing significantly to Ladakh economy and income generation for supporting resource poor farmers. Amongst various invasive weed species, the common reed (Phragmites australis) has become a serious ecological threat for agri-silvi-pastoral system biodiversity and accelerated its expansion in cropped lands including many wetland and salt-affected habitats of cold arid region due to its intermediate plant growth habit of C3 - C4 photosynthetic ecotypes. This character makes Phragmites a climate-resilient species, which is a major challenge for cold arid agriculture in the changing climatic patterns. Simultaneously, it is one of the best fodder substitute available under harsh climate. This paper deals with its unique characters and potential threats to the agrobiodiversity of Ladakh.展开更多
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 o...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.展开更多
Background: The Y chromosome in mammal is paternally inherited and harbors genes related to male fertility and spermatogenesis. The unique intra-chromosomal recombination pattern of Y chromosome and morphological dif...Background: The Y chromosome in mammal is paternally inherited and harbors genes related to male fertility and spermatogenesis. The unique intra-chromosomal recombination pattern of Y chromosome and morphological difference of this chromosome between Bos taurus and Bos indicus make it an ideal model for studying structural variation, especially in crossbred (Bos taurus x Bos indicus) bulls. Copy Number Variation (CNV) is a type of genomic structural variation that gives information complementary to SNP data. The purpose of this study was to find out copy number differences of four Y chromosomal spermatogenesis-related candidate genes in genomic DNA of crossbred and purebred Indicine bulls. Result: Four Y chromosomal candidate genes of spermatogenesis namely, sex determining gene on Ychromosome (SRY), DEAD box po/ypeptide 3-Y chromosome (DDX3 Y), Ubiquidn specific peptidase 9, Y-linked ( usPgY), testis-specific protein on Y chromosome (TSPY) were evaluated. Absolute copy numbers of Y chromosomal genes were determined by standard curve-based quantitative real time PCR. Copy numbers of SRYand TSPYgenes per unit amount of genomic DNA are higher in crossbred than Indicine bulls. However, no difference was observed in DDX3Yand usPgYgene copy numbers between two groups. Conclusion: The present study demonstrates that the structural organization of Y chromosomes differs between crossbred and Indicine bulls which are reproductively healthy as observed from analysis of semen attributes. The absolute copy numbers of SRY and TSPY genes in unit mass of genomic DNA of crossbred bulls are significantly higher than Indicine bulls. No alteration in absolute copies ofDDX3Yand usPgYgene was found between the genome of crossbred and Indicine bulls. This study suggests that the DDX3Yand USPgYare likely to be single copy genes in the genome of crossbred and Indicine bulls and variation in Y chromosome length between crossbred and Indicine bulls may be due to the copy number variation of SRY gene and TSPYarray.展开更多
Direct-fed microbials(DFM), generally regarded as safe status, are successfully used in improving rumen ecology, gastro-intestinal health, feed efficiency, milk production and growth rate in ruminants. On the other ...Direct-fed microbials(DFM), generally regarded as safe status, are successfully used in improving rumen ecology, gastro-intestinal health, feed efficiency, milk production and growth rate in ruminants. On the other hand, methanogenesis in rumen, which accounts for a significant loss of ruminant energy and increased greenhouse gas in environment, is of great concern, therefore, use of DFM for improving productivity without compromising the animal health and ecological sustainability is encouraged. The present study was conducted to investigate the methane reducing potential of bacteriocinogenic strain Pediococcus pentosaceus-34. Since, the culture showed no hemolysis on blood agar and DNase activity, hence, it was considered to be avirulent in nature, a prerequisite for any DFM. The culture also showed tolerance to pH 5.0 for 24 h with 0.5% organic acid mixture, whereas when given a shock for 2 h at different p H and organic acids concentrations, it showed growth at pH 3.0 and 4.0 with 0.1 and 1.0% organic acids, respectively, as having good animal probiotics attributes. The total gas production was significantly(P〈0.05) higher in live pedicoccal culture(LPC) and dead pedicoccal culture(DPC) both with wheat straw, when compared to the control. In sugarcane bagasse, gas production was significantly lower(P〈0.05) with LPC compared to the control and DPC both. Methane was reduced by the inclusion of LPC in sugarcane bagasse(0.07 mL CH4 mg–1 dry matter digestibility) with no effect on other rumen fermentation parameters. However, with wheat straw and LPC total gas, in vitro dry matter digestibility, total volatile fatty acids increased significantly but no reduction in methane production was observed in comparison to the control. Therefore, further research is warranted in this direction, if the bacteriocinogenic strains can be used as DFM for ruminants to improve the ruminant productivity.展开更多
Study was conducted to assess the possible transfer of heavy metals from livestock feeds to animal products. In experiment-l, samples (n = 503) collected from conventional farms in Central Greece were analysed for h...Study was conducted to assess the possible transfer of heavy metals from livestock feeds to animal products. In experiment-l, samples (n = 503) collected from conventional farms in Central Greece were analysed for heavy metal Copper (Cu), Zinc (Zn), Cadmium (Cd), Lead (Pb), Nickel (Ni) and Chromium (Cr) contents in muscle tissues, livers, kidneys, excremental and basic mixtures in livestock such as cows and sheep. Results showed that transfer of heavy metal contaminants from feed to animal products fluctuated below the permissible risk levels. In experiment 2, samples (n = 440) of feedstuffs used in different feeding regimens and seasons at different livestock farms [sheep (n = 140), dairy cows (n = 180), pigs (n = 120)] were collected. Results showed that relatively high concentration of Cu in the excrements of pigs (155 + 9.13) and Zn in the excrements of sheep (144.56 + 5.78) are explained by the use of Cu in the diet of developing piglets and sheep. Ovine showed higher concentration ofNi, Cr, and Pb in their excrements, while, the concentration of heavy metals in forages was under the allowed range to the EU legislation.展开更多
Components with strong adsorption capacity for cholates from buckwheat proteins were screened, separated and purified by several methods, and the effects of ultra-high-pressure(UHP) on the structure and function of bu...Components with strong adsorption capacity for cholates from buckwheat proteins were screened, separated and purified by several methods, and the effects of ultra-high-pressure(UHP) on the structure and function of buckwheat 13S globulin(BW13SG) were studied. Samples were treated by UHP at different p H(3.0 and 7.0) value(s) and at 100–500 MPa for 10–30 min. The results showed that the tertiary structure of BW13SG was partially denatured and aggregated. The decrease in the unordered structure indicated that UHP resulted in a looser secondary structure of BW13SG. UHP treatment also increased solubility, emulsion activity and stability, foaming capacity and stability. The samples treated at 500 MPa, p H 3.0 for 30 min had the most enhanced functionality. Moreover, under this condition, the sodium cholate and sodium deoxycholate adsorption capacities of BW13SG were both higher than 98% and the adsorption capacity of sodium taurocholate, which can be difficult to adsorb, was higher than 60%.展开更多
Objective:To understand the level of progesterone (P4) in different quality of buffalo cumulus oocyte complexes (COCs) and further to evaluate the effect of exogenous P4 supplementation on maturation and subsequent de...Objective:To understand the level of progesterone (P4) in different quality of buffalo cumulus oocyte complexes (COCs) and further to evaluate the effect of exogenous P4 supplementation on maturation and subsequent developmental ability of poor quality brilliant cresyl blue (BCB-)COCs.Methods: Progesterone secreted by different quality of buffalo oocytes was estimated by enzyme linked immunosorbent assay and the concentration differences were translated into P4 doses to be incorporated in the maturation medium of BCB-ve COCs followed by expression analysis of genes involved in the cumulus expansion, extracellular matrix disintegration and progesterone receptor signalling. In addition, the study also evaluated the effect of exogenous P4 on sperm-cumulus interaction. Results: More than 10-fold up-regulated expression of progesterone receptor in P4 supplemented oocytes signified that P4 might be acting predominantly through this receptor. Also, exogenous P4 supplementation had significant effect on transcatheter arterial chemoembolization protease regulated by P4-progesterone receptor pathway which in turn had an important role in extracellular matrix disintegration. On the contrary, cumulus expansion genesHAS2,TNFAIP6,AREG were not altered upon P4 supplementation. Also, it was observed that P4 addition did facilitate passage of significantly more number of spermatozoa through P4 treated cumulus cells. Further, incorporation of different doses of P4 did not improve significantly the cleavage and blastocyst rates of BCB-ve COCs.Conclusions: Different qualities of buffalo COCs secrete substantially diverse levels of P4, and its supplementation has a role in oocyte maturation via modulation of cumulus characteristics but perhaps not fertilization.展开更多
The well documented source for adult multipotent stem cells is spermatogonial stem cells (SSCs) of mammalian testis. It is foundation of spermatogenesis in the testis throughout adult life by balancing self-renewal an...The well documented source for adult multipotent stem cells is spermatogonial stem cells (SSCs) of mammalian testis. It is foundation of spermatogenesis in the testis throughout adult life by balancing self-renewal and differentiation. SSCs isolation from mammalian testis is difficult because of their scarcity and the lack of well characterized cell surface markers. Thus, the isolation of SSCs is of great interest for exploration of spermatogonial physiology and therapeutic approaches for fertility preservation. CD9 is a surface marker expressed in mouse and rat male germline stem cells. In this study, CD9 positive SSCs were successfully isolated from the goat testis using enzymatic digestion followed by three step purification: Differential plating, Percoll discontinuous density gradient followed by Magnetic activated cell sorting (MACS). Percoll discontinuous density gradient showed significant differences in the percentage of CD9+ SSCs across individual fraction. The fraction 36% and 40% gave the highest percentage of CD9+ SSCs i.e. 82% ± 1.2 and 9.2% ± 1.3 respectively. Magnetic activated cell sorting of CD9+ cells in the magnetic fraction of goat testes was in the range of 15% - 18% which is upto threefolds. CD9+ SSCs were further recovered with appreciable efficiency after immunomagnetic isolation by using various bead: cells ratio in which 4:1 ratio gave the highest yield of 69.06 × 105 with 18% of CD9+ SSCs. Magnetic activated cell sorting using anti-CD9 antibodies provides an efficient and fast approach as compared to conventional approaches such as differential plating and percoll discontinuous density gradient for enrichment strategy for spermatogonial stem cells from goat testes for undertaking research on basic and applied reproductive biology.展开更多
PH-20 (Spam-1) antigen appears to be a bi-functional sperm plasma membrane protein with hyaluronidase activity and its role is related to cumulus penetration as well as zona binding. (LDH-C4) is a key enzyme, distribu...PH-20 (Spam-1) antigen appears to be a bi-functional sperm plasma membrane protein with hyaluronidase activity and its role is related to cumulus penetration as well as zona binding. (LDH-C4) is a key enzyme, distributed specifically in testis and is highly immunogenic, function of which relates to energy metabolism and capacitation of the sperm. Therefore, in this study, we observed the effect of purified native PH-20 and LDH-C4 antigens on sperm function and morphology in dog. Native PH-20 (46, 32 kDa) and LDH-C4 (36, 30, 28) sub units, purified from dog sperm SDS- extracts were used to test as immunocontraceptive candidates in stray dog. Dogs 1, 2, 3, 4, 5/6 and 7/8 were immunized with 46, 32, 36, 30/28, 46/32, 36/30/28 kDa sub units respectively by i/m route and semen was analysed at weekly intervals. High titre of 3200 - 6400 was achieved in all dogs after three boosters. Immune response in treated dogs was confirmed by increase in Ig level in blood serum and reaction of partially purified serum Ig of immunized dogs with respective antigens in double immune diffusion and immunoblots. Immunization of dogs either with individual PH-20/ LDHC sub units or pooled PH-20/LDHC4 sub units showed a significant effect on sperm count, percent motility, viability, HOST and sperm morphology. All the sperm parameters declined to a significant level (p < 0.05) between 45 - 115 DPI and remained low than that of the recommended values for fertile dog semen till 175 DPI. The incubation of spermatozoa in HOS solution in the presence of anti-PH-20 and anti-LDHC Ig significantly reduced the percent HOS positive spermatozoa. A significant decline (p ≤ 0.05) in acrylamide penetration assay was also observed in the presence of anti 46 and 30/28 kDa Ig. Since percent motility, sperm count, HOST and percent normal spermatozoa in immunized dogs were significantly less than that of recommended fertile dog semen, it can be concluded that PH-20 (46/32 kDa) and LDHC4 (36/30/28 kDa) sub units had an immunocontraceptive potential in dog.展开更多
The production of cells capable of expressing gene(s) of interest is important for a variety of applications in biomedicine and biotechnology, including gene therapy and a novel method of stem cell therapy in the vari...The production of cells capable of expressing gene(s) of interest is important for a variety of applications in biomedicine and biotechnology, including gene therapy and a novel method of stem cell therapy in the various diseases. Achieving high levels of transgene expression for the longer period of time, without adversely affecting cell viability and differentiation capacity of the cells, is crucial. In the present study, we investigated the efficiency of plasmid vector for the production of transgenic cMSCs and examined any functional change of cells after transfection. To do so first we have collected bone marrows from the adult goats and cultured them for isolation of mesenchymal stem cells (cBM-MSCs). These cells were characterized using MSC specific markers including differentiation into osteocytes and adipocytes. Transfection with plasmid vector did not adversely affect cBM-MSCs morphology, viability or differentiation potential, and transgene expression levels were unaffected beyond passage 12th. The results indicated that we have been able to generate transgenic caprine MSC (tcBM-MSC) and transfection of cBM-MSCs using plasmid vector resulted in very high and stable transfection efficiency. This finding may have considerable significance in improving the efficacy of MSC-based therapies and their tracking in animal model.展开更多
Background: An oviduct-specific glycoprotein, OVGP1, is synthesized and secreted by non-ciliated epithelial cells of the mammalian oviduct which provides an essential milieu for reproductive functions. The present st...Background: An oviduct-specific glycoprotein, OVGP1, is synthesized and secreted by non-ciliated epithelial cells of the mammalian oviduct which provides an essential milieu for reproductive functions. The present study reports the effects of recombinant buffalo OVGP1 that lacks post-translational modifications, and native Buffalo OVGP1 isolated from oviductal tissue, on frozen-thawed sperm functions and in vitro embryo development.Results: The proportion of viable sperms was greater(P 〈 0.05) in the recombinant OVGP1-treated group compared to the native OVGP1-treated group at 2 h, 3 h, and 4 h of incubation. The proportion of motile sperms at3 h and 4 h of incubation; and membrane-intact sperms at 4 h was greater(P 〈 0.05) in the native OVGP1-treated group compared to the control and recombinant OVGP1-treated groups. The proportion of capacitated and acrosome-reacted sperms was greater(P 〈 0.05) in the native OVGP1-treated group compared to the recombinant OVGP1 group at 4 h. The rates of cleavage of embryos and their development to the blastocyst stage were greater(P 〈 0.05) in the presence of either native or recombinant OVGP1 in comparison to control at 10 μg/mL concentration as compared to 5 or 20 μg/mL.Conclusions: The study suggests that both native and recombinant OVGP1 impart a positive effect on various sperm features and in vitro embryo development. However, native OVGP1 was found to have a more pronounced effect in comparison to recombinant non-glycosylated OVGP1 on various sperm functions except viability. Hence,our current findings infer that glycosylation of OVGP1 might be essential in sustaining the sperm functions but not the in vitro embryo development.展开更多
Objective:To study the effect of L-ascorbic acid supplementation on the in vitro development of buffalo embryos and evaluate the relative mRNA abundance of some pro-apoptotic,anti-apoptotic,and embryonic development-r...Objective:To study the effect of L-ascorbic acid supplementation on the in vitro development of buffalo embryos and evaluate the relative mRNA abundance of some pro-apoptotic,anti-apoptotic,and embryonic development-related genes.Methods:In experiment 1,we evaluated the effect of the addition of 0(control),50,and 100μM L-ascorbic acid to the in vitro maturation medium on the developmental competence in terms of blastocyst rate and relative mRNA abundance of some pro-apoptotic(BAX,BID),anti-apoptotic(BCL-XL,MCL1),and embryonic development(GDF9,BMP15)related genes.Based on the results,we chose 50μM as the suitable dose of L-ascorbic acid for the subsequent experiments.We further evaluated the blastocyst rates following the addition of 50μM L-ascorbic acid to the in vitro culture medium(experiment 2),and in vitro maturation and in vitro culture media(experiment 3).In all three experiments,the maturation and culture media devoid of L-ascorbic acid served as the control group.Results:The blastocyst rate after adding 50μM L-ascorbic acid to the in vitro maturation medium was significantly higher than the control group(P<0.05),whereas 100μM L-ascorbic acid exhibited a negative effect on the blastocyst rate.The blastocyst rates for embryos cultured in 50μM L-ascorbic acid in the in vitro culture medium alone and both in vitro maturation and in vitro culture media were significantly higher than their corresponding control groups(P<0.05).The relative mRNA abundance of BAX significantly decreased in blastocysts produced after the addition of 50μM L-ascorbic acid as compared with the control group(P<0.05),whereas,for MCL1,it significantly decreased in blastocysts produced after the addition of 100μM L-ascorbic acid(P<0.05).Conclusions:The supplementation of 50μM L-ascorbic acid to in vitro maturation and in vitro culture media supports in vitro embryonic development in buffaloes by improving developmental competence and altering the expression of apoptosis-related genes.展开更多
Objective:To assess the stability of 10 candidate internal control genes(ICGs),namely GAPDH,ACTB,RPL23,RPS15A,ATPSF1,GLUT5,HMBS,ATP2B4,PPIA,and BRP to normalize the transcriptional data from testes samples of Zebu and...Objective:To assess the stability of 10 candidate internal control genes(ICGs),namely GAPDH,ACTB,RPL23,RPS15A,ATPSF1,GLUT5,HMBS,ATP2B4,PPIA,and BRP to normalize the transcriptional data from testes samples of Zebu and crossbred bulls.Methods:Total RNA was isolated from testicular tissue of Zebu and crossbred bulls(n=6 each)between 2-8 years of age.cDNA was synthesized,and the quantitative real-time polymerase chain reaction(PCR)was performed.The cycle threshold values were used for the analysis of the stability of ICGs.Four different statistical algorithms:geNorm,Normfinder,BestKeeper,and RefFinder,were used to assess the stability of these genes.Results:ATPSF1,HMBS,PPIA,and RPS15A were the most reliable and stable ICGs for Zebu testes,and ATPSF1,RPL23,and PPIA for crossbred testes.Conclusions:A panel of stable ICGs(ATPSF1,HMBS,PPIA,RPS15A for Zebu and ATPSF1,RPL23,and PPIA for crossbred)for normalization of gene expression data in testes samples can be helpful for researchers to conduct functional genomics studies at the testicular level in cattle bulls.展开更多
Quality semen production remains the main focus and objective of semen processing laboratories throughout the world. Bacterial and other microbial contaminants affect the semen quality and hence the fertility, and als...Quality semen production remains the main focus and objective of semen processing laboratories throughout the world. Bacterial and other microbial contaminants affect the semen quality and hence the fertility, and also lead to reproductive disorders as well as lower conception rates and increased embryonic mortality, abortion and other complications in females. Microbial contamination affects the semen adversely, by exerting direct spermicidal effect, formation of reactive oxygen species, toxin production, adherence with spermatozoa, deriving nutrients and oxygen from the medium and thus competing with spermatozoa for the factors of growth and normal functioning. Despite hygienic measures, several ubiquitous and opportunistic microbes find their ways into semen during collection, processing, and storage of semen, and survive even during freezing. Stringent sanitary precautions are therefore required at every step of collecting semen and its processing. Preputial cavity is considered as main source of semen contaminating microorganisms. Flushing the preputial cavity with normal saline or any suitable liquid combination with antimicrobial activity, prior to semen collection reduces the microbial load and thereby improves the semen quality.展开更多
Low sperm numbers in artificial insemination (AI)-doses are being used widely to make the best use of high genetic value bulls as well as sex-sorted semen. Sperm concentration needed for AI to obtain reasonable fertil...Low sperm numbers in artificial insemination (AI)-doses are being used widely to make the best use of high genetic value bulls as well as sex-sorted semen. Sperm concentration needed for AI to obtain reasonable fertility, taking genetic value of bull and numerous others components into consideration is one of the essential constituents for successful AI breeding program. However, low sperm concentrations in AI-doses lead to reducing post-thaw viability. The reduction in viability of low sperm doses may be affected by fresh semen volume, sperm number and seminal plasma level at final dilution. Reduction in quality and fertility of low sperm doses is one of the limitations for their use in successful AI programme. Sperm number per AI required to achieve optimum fertility is one of the main crucial things to AI industry, and numerous efforts have been made in this regard. Due to great variability among bulls, sperm number per AI could be a limiting factor in achieving acceptable fertility values. Fertility of low sperm doses may vary among bulls, and non-return rates (NRRs) with low sperm doses may be determined by fertility level of bull. On the basis of individual bulls, sperm numbers in AI doses needed to be adjusted to reduce the variations in NRRs among bulls. Utilizing high fertile bulls for low sperm doses with acceptable non-return rates (NRRs) may be a way to cover a large number of bovines under AI in countries like India. Deposition site within the uterine horn may alter non return rates following inseminations with low sperm doses. Following deep-uterine inseminations, acceptable pregnancies may be achieved with low sperm doses and even if ovulation side is unknown.展开更多
This article presents a review pertains the laser irradiation effects and its possible mechanisms of action on spermatozoa functions in domestic animals. To improve artificial insemination, laser is sensitive and cost...This article presents a review pertains the laser irradiation effects and its possible mechanisms of action on spermatozoa functions in domestic animals. To improve artificial insemination, laser is sensitive and cost effective technique, when compared to other conventional methods. Laser may have both positive and negative effects on spermatozoa functions. Since the effects of light are mediated by reactive oxygen species, and the levels of these reactive oxygen species following irradiating spermatozoa with laser may be responsible for determining the effects of laser on sperm. Dose of laser may be regarded as of great significance and this dosage of laser may be responsible for determining its effects on spermatozoa. Optimum dosage of laser for improving seminal attributes may vary among various species and this need to be standardized in each of them. The beneficial effects include improving sperm livability, acrosomal integrity, hypo-osmotic swelling response, mitochondrial function and computer-aided sperm analysis parameters. The increase in cytochrome c oxidase activity, ATP levels and mitochondrial membrane potential, in laser irradiated cells may be responsible for enhanced sperm quality parameters. Improving fertility with laser irradiated spermatozoa has been reported in few species like boar and need to be elaborated in other species. In conclusion laser may be regarded as an easy, cheap and time saving technology for improving artificial insemination;in addition, laser may have various potential applications in the field of reproductive biotechnology as well as in livestock farms and veterinary polyclinics.展开更多
Whey and toned milk was effectively used for the development of tea whitener. Cheese whey was concentrated to 12% solids in a vacuum evaporator, pasteurized at 72℃±2℃ for 15 s, diluted back to 11.5% solids and ...Whey and toned milk was effectively used for the development of tea whitener. Cheese whey was concentrated to 12% solids in a vacuum evaporator, pasteurized at 72℃±2℃ for 15 s, diluted back to 11.5% solids and was cold blended with toned milk of 11.5% solids in three ratios viz. 25:75, 50:50 and 75:25 and selected blends were homogenized at 1200-1500 psi. Tea was prepared by adding 80 mL whitener to 100 mL decoction and addition of sugar at 8% (w/v). The tea whitener was selected based on the sensory attributes of tea beverages. The colour of the beverages, tea whiteners and decoction were analyzed by reflectance meter and computer vision image analysis technique. The viscosity of the beverage was tested using a Brookfield viscometer. Coffee stability test was done to assess the stability of tea whitener under acidic conditions. Standard plate count, Coliform count and Presumptive Coliform test were done to assess the microbiological quality of the whitener. Homogenization had a positive impact on the viscosity and colour. The reflectance value of tea prepared from 25% homogenised whey milk blend and toned milk were in the range of 43-46 and 45-49, respectively. The viscosity of tea was found to increase with homogenization and whey levels. The sedimentation values of 25% whey blends and toned milk were 1.25 mL and 1 mL respectively. The shelf life of the blends was 2 days under refrigerated conditions. It can be concluded that a satisfactory tea whitener can be prepared by blending whey with toned milk in 25:75 proportion followed by homogenization.展开更多
Electrolyzed Oxidized Water (EOW) is produced by passing a diluted salt solution through an electrolytic cell, having anode and cathode electrodes. The anode and cathode are separated by a bipolar membrane. Negatively...Electrolyzed Oxidized Water (EOW) is produced by passing a diluted salt solution through an electrolytic cell, having anode and cathode electrodes. The anode and cathode are separated by a bipolar membrane. Negatively charged ions—chloride and hydroxide in the diluted salt solution move to anode to give up electrons and become gas (O2, Cl2) and hypochlorous acid and having redox potential of +700 to +800 mV with pH 4.0. It has a strong oxidation potential and a shortage of electrons giving it the ability to oxidize and sterilize. In microbial inactivation process, oxidized water damage cell membranes, create disruption in cell metabolic processes and essentially kill the cell. EOW, also a strong acid, is different to hydrochloric acid or sulfuric acid in that it is not corrosive to skin, mucous membrane, or organic material. It is easy to handle and suitable for the sanitation of the plant and decontamination of foods. Electrolyzed water has been tested and used as a disinfectant in the food industry and other applications.展开更多
文摘Dairy production plays an integral part in supporting smallholder farmers’ livelihoods. The desire to increase the number of dairy cattle is not feasible due to the reduced output of feed resources occasioned by climate change. Consequently, the need to increase productivity per cow is inevitable. Conventional protein supplements are costly;hence, the need to explore affordable nutrientdense alternative feed resources. Liquid brewers’ yeast (LBY), a by-product of the brewing industry, is a rich protein supplement in dairy production. This study aimed to assess the dairy farming conditions and utilization levels of LBY as a feed supplement in Githunguri Sub-county, Kiambu. Semi-structured questionnaires were administered to 457 dairy farmers in a cross-sectional survey. The findings revealed that most farmers (94.2%) fed their cattle on established forage/fodder and crop residues with supplementation. Even though 53.1% of the respondents were aware of the use of LBY, only 30.6% utilized it to supplement dairy cows, most of whom (96.0%) used it fresh without preservation. Membership in farmers’ organizations increased awareness of LBY (r = 0.732). Principal component analysis indicated that the benefits of using LBY outweigh the challenges involved with a loading matrix of 0.891 - 0.954 and 0.681 - 0.807, respectively. The low adoption and use levels of LBY as a source of protein supplements were due to low awareness. There is a need for concerted efforts by stakeholders in the industry to increase farmers’ knowledge base on the utilization and effectiveness of LBY in dairy production.
文摘Using probiotics as animal feed additives instead of antibiotics is gaining momentum to avert adverse negative effects on human health. Liquid brewers yeast (LBY) is an industrial by-product containing probiotic microorganisms and is also used as a protein supplement for dairy animals. Nevertheless, value chain actors lack of appropriate handling practices compromises the by-products quality and safety. This study aimed to determine the effect of variation in temperature on microbial diversity and probiotic effects during the storage time of LBY sampled from distributors and farmers from Githunguri sub-county of Kenya. The samples were stored at 20C, 25C and 30C, then tested on 0, 5, 10, 15 and 20 days. The studys parameters involved determining the pH levels, lactic acid bacteria (LAB), total coliform count (TCC), mould, and yeast in LBY. The rate (k) of the reaction kinetics model was used to extrapolate the expected probiotic shelf life. The LAB and yeast populations were reduced in a first-order reaction at all storage temperatures. The rate of reduction in the numbers of LAB reduced with an increase in temperature (k = 0.019 and 0.023) at 20C and 30C, respectively. Yeasts highest rate of growth reduction was 25C (k = 0.009) and least at 30C (k = 0.043). The minimum effective concentration for probiotics of 106 CFU/mL needed to observe the beneficial physiological impact on farm animals was achieved between 34.9 and 35.5 days at the tested storage temperatures. The study provides insight into the unexploited low-cost probiotic potential of LBY in dairy production. Conversely, handling practices and environmental microbial contamination along the value chain can compromise product quality and safety. There is a need to advocate its use in dairy for improved productivity and sensitize farmers to appropriate hygienic measures along the LBY value chain.
文摘Ladakh’s cropped and non-cropped areas suffer greatly from weedy invasion under subsistence agricultural system where weeding is not a priority but total crop + weed biomass together contribute to livestock feeding significantly. As agriculture along with livestock rearing is major activity of livelihood options, thereby contributing significantly to Ladakh economy and income generation for supporting resource poor farmers. Amongst various invasive weed species, the common reed (Phragmites australis) has become a serious ecological threat for agri-silvi-pastoral system biodiversity and accelerated its expansion in cropped lands including many wetland and salt-affected habitats of cold arid region due to its intermediate plant growth habit of C3 - C4 photosynthetic ecotypes. This character makes Phragmites a climate-resilient species, which is a major challenge for cold arid agriculture in the changing climatic patterns. Simultaneously, it is one of the best fodder substitute available under harsh climate. This paper deals with its unique characters and potential threats to the agrobiodiversity of Ladakh.
基金financial support provided under the Network Project of ICAR on ‘VTCC’ to carry the research further in this direction
文摘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.
基金supported by World Bank funded National Agricultural Innovation Project(C4/C30015)
文摘Background: The Y chromosome in mammal is paternally inherited and harbors genes related to male fertility and spermatogenesis. The unique intra-chromosomal recombination pattern of Y chromosome and morphological difference of this chromosome between Bos taurus and Bos indicus make it an ideal model for studying structural variation, especially in crossbred (Bos taurus x Bos indicus) bulls. Copy Number Variation (CNV) is a type of genomic structural variation that gives information complementary to SNP data. The purpose of this study was to find out copy number differences of four Y chromosomal spermatogenesis-related candidate genes in genomic DNA of crossbred and purebred Indicine bulls. Result: Four Y chromosomal candidate genes of spermatogenesis namely, sex determining gene on Ychromosome (SRY), DEAD box po/ypeptide 3-Y chromosome (DDX3 Y), Ubiquidn specific peptidase 9, Y-linked ( usPgY), testis-specific protein on Y chromosome (TSPY) were evaluated. Absolute copy numbers of Y chromosomal genes were determined by standard curve-based quantitative real time PCR. Copy numbers of SRYand TSPYgenes per unit amount of genomic DNA are higher in crossbred than Indicine bulls. However, no difference was observed in DDX3Yand usPgYgene copy numbers between two groups. Conclusion: The present study demonstrates that the structural organization of Y chromosomes differs between crossbred and Indicine bulls which are reproductively healthy as observed from analysis of semen attributes. The absolute copy numbers of SRY and TSPY genes in unit mass of genomic DNA of crossbred bulls are significantly higher than Indicine bulls. No alteration in absolute copies ofDDX3Yand usPgYgene was found between the genome of crossbred and Indicine bulls. This study suggests that the DDX3Yand USPgYare likely to be single copy genes in the genome of crossbred and Indicine bulls and variation in Y chromosome length between crossbred and Indicine bulls may be due to the copy number variation of SRY gene and TSPYarray.
基金a part of a PhD project of Sanjay Kumar that was supported by NDRI (ICAR) fellowshipNational Initiative on Climate Resilient Agriculture, India (NICRA) for providing partial support
文摘Direct-fed microbials(DFM), generally regarded as safe status, are successfully used in improving rumen ecology, gastro-intestinal health, feed efficiency, milk production and growth rate in ruminants. On the other hand, methanogenesis in rumen, which accounts for a significant loss of ruminant energy and increased greenhouse gas in environment, is of great concern, therefore, use of DFM for improving productivity without compromising the animal health and ecological sustainability is encouraged. The present study was conducted to investigate the methane reducing potential of bacteriocinogenic strain Pediococcus pentosaceus-34. Since, the culture showed no hemolysis on blood agar and DNase activity, hence, it was considered to be avirulent in nature, a prerequisite for any DFM. The culture also showed tolerance to pH 5.0 for 24 h with 0.5% organic acid mixture, whereas when given a shock for 2 h at different p H and organic acids concentrations, it showed growth at pH 3.0 and 4.0 with 0.1 and 1.0% organic acids, respectively, as having good animal probiotics attributes. The total gas production was significantly(P〈0.05) higher in live pedicoccal culture(LPC) and dead pedicoccal culture(DPC) both with wheat straw, when compared to the control. In sugarcane bagasse, gas production was significantly lower(P〈0.05) with LPC compared to the control and DPC both. Methane was reduced by the inclusion of LPC in sugarcane bagasse(0.07 mL CH4 mg–1 dry matter digestibility) with no effect on other rumen fermentation parameters. However, with wheat straw and LPC total gas, in vitro dry matter digestibility, total volatile fatty acids increased significantly but no reduction in methane production was observed in comparison to the control. Therefore, further research is warranted in this direction, if the bacteriocinogenic strains can be used as DFM for ruminants to improve the ruminant productivity.
文摘Study was conducted to assess the possible transfer of heavy metals from livestock feeds to animal products. In experiment-l, samples (n = 503) collected from conventional farms in Central Greece were analysed for heavy metal Copper (Cu), Zinc (Zn), Cadmium (Cd), Lead (Pb), Nickel (Ni) and Chromium (Cr) contents in muscle tissues, livers, kidneys, excremental and basic mixtures in livestock such as cows and sheep. Results showed that transfer of heavy metal contaminants from feed to animal products fluctuated below the permissible risk levels. In experiment 2, samples (n = 440) of feedstuffs used in different feeding regimens and seasons at different livestock farms [sheep (n = 140), dairy cows (n = 180), pigs (n = 120)] were collected. Results showed that relatively high concentration of Cu in the excrements of pigs (155 + 9.13) and Zn in the excrements of sheep (144.56 + 5.78) are explained by the use of Cu in the diet of developing piglets and sheep. Ovine showed higher concentration ofNi, Cr, and Pb in their excrements, while, the concentration of heavy metals in forages was under the allowed range to the EU legislation.
基金the financial support by the Open Project Program of State Key Laboratory of Dairy Biotechnology (No. SKLDB2018-002)National Natural Science Foundation of China (No. 31871805 & No. 31501437)+1 种基金Shanghai Municipal Education Commission (Plateau Discipline Construction Program)China Agriculture Research System (CARS-08-D2)。
文摘Components with strong adsorption capacity for cholates from buckwheat proteins were screened, separated and purified by several methods, and the effects of ultra-high-pressure(UHP) on the structure and function of buckwheat 13S globulin(BW13SG) were studied. Samples were treated by UHP at different p H(3.0 and 7.0) value(s) and at 100–500 MPa for 10–30 min. The results showed that the tertiary structure of BW13SG was partially denatured and aggregated. The decrease in the unordered structure indicated that UHP resulted in a looser secondary structure of BW13SG. UHP treatment also increased solubility, emulsion activity and stability, foaming capacity and stability. The samples treated at 500 MPa, p H 3.0 for 30 min had the most enhanced functionality. Moreover, under this condition, the sodium cholate and sodium deoxycholate adsorption capacities of BW13SG were both higher than 98% and the adsorption capacity of sodium taurocholate, which can be difficult to adsorb, was higher than 60%.
文摘Objective:To understand the level of progesterone (P4) in different quality of buffalo cumulus oocyte complexes (COCs) and further to evaluate the effect of exogenous P4 supplementation on maturation and subsequent developmental ability of poor quality brilliant cresyl blue (BCB-)COCs.Methods: Progesterone secreted by different quality of buffalo oocytes was estimated by enzyme linked immunosorbent assay and the concentration differences were translated into P4 doses to be incorporated in the maturation medium of BCB-ve COCs followed by expression analysis of genes involved in the cumulus expansion, extracellular matrix disintegration and progesterone receptor signalling. In addition, the study also evaluated the effect of exogenous P4 on sperm-cumulus interaction. Results: More than 10-fold up-regulated expression of progesterone receptor in P4 supplemented oocytes signified that P4 might be acting predominantly through this receptor. Also, exogenous P4 supplementation had significant effect on transcatheter arterial chemoembolization protease regulated by P4-progesterone receptor pathway which in turn had an important role in extracellular matrix disintegration. On the contrary, cumulus expansion genesHAS2,TNFAIP6,AREG were not altered upon P4 supplementation. Also, it was observed that P4 addition did facilitate passage of significantly more number of spermatozoa through P4 treated cumulus cells. Further, incorporation of different doses of P4 did not improve significantly the cleavage and blastocyst rates of BCB-ve COCs.Conclusions: Different qualities of buffalo COCs secrete substantially diverse levels of P4, and its supplementation has a role in oocyte maturation via modulation of cumulus characteristics but perhaps not fertilization.
文摘The well documented source for adult multipotent stem cells is spermatogonial stem cells (SSCs) of mammalian testis. It is foundation of spermatogenesis in the testis throughout adult life by balancing self-renewal and differentiation. SSCs isolation from mammalian testis is difficult because of their scarcity and the lack of well characterized cell surface markers. Thus, the isolation of SSCs is of great interest for exploration of spermatogonial physiology and therapeutic approaches for fertility preservation. CD9 is a surface marker expressed in mouse and rat male germline stem cells. In this study, CD9 positive SSCs were successfully isolated from the goat testis using enzymatic digestion followed by three step purification: Differential plating, Percoll discontinuous density gradient followed by Magnetic activated cell sorting (MACS). Percoll discontinuous density gradient showed significant differences in the percentage of CD9+ SSCs across individual fraction. The fraction 36% and 40% gave the highest percentage of CD9+ SSCs i.e. 82% ± 1.2 and 9.2% ± 1.3 respectively. Magnetic activated cell sorting of CD9+ cells in the magnetic fraction of goat testes was in the range of 15% - 18% which is upto threefolds. CD9+ SSCs were further recovered with appreciable efficiency after immunomagnetic isolation by using various bead: cells ratio in which 4:1 ratio gave the highest yield of 69.06 × 105 with 18% of CD9+ SSCs. Magnetic activated cell sorting using anti-CD9 antibodies provides an efficient and fast approach as compared to conventional approaches such as differential plating and percoll discontinuous density gradient for enrichment strategy for spermatogonial stem cells from goat testes for undertaking research on basic and applied reproductive biology.
文摘PH-20 (Spam-1) antigen appears to be a bi-functional sperm plasma membrane protein with hyaluronidase activity and its role is related to cumulus penetration as well as zona binding. (LDH-C4) is a key enzyme, distributed specifically in testis and is highly immunogenic, function of which relates to energy metabolism and capacitation of the sperm. Therefore, in this study, we observed the effect of purified native PH-20 and LDH-C4 antigens on sperm function and morphology in dog. Native PH-20 (46, 32 kDa) and LDH-C4 (36, 30, 28) sub units, purified from dog sperm SDS- extracts were used to test as immunocontraceptive candidates in stray dog. Dogs 1, 2, 3, 4, 5/6 and 7/8 were immunized with 46, 32, 36, 30/28, 46/32, 36/30/28 kDa sub units respectively by i/m route and semen was analysed at weekly intervals. High titre of 3200 - 6400 was achieved in all dogs after three boosters. Immune response in treated dogs was confirmed by increase in Ig level in blood serum and reaction of partially purified serum Ig of immunized dogs with respective antigens in double immune diffusion and immunoblots. Immunization of dogs either with individual PH-20/ LDHC sub units or pooled PH-20/LDHC4 sub units showed a significant effect on sperm count, percent motility, viability, HOST and sperm morphology. All the sperm parameters declined to a significant level (p < 0.05) between 45 - 115 DPI and remained low than that of the recommended values for fertile dog semen till 175 DPI. The incubation of spermatozoa in HOS solution in the presence of anti-PH-20 and anti-LDHC Ig significantly reduced the percent HOS positive spermatozoa. A significant decline (p ≤ 0.05) in acrylamide penetration assay was also observed in the presence of anti 46 and 30/28 kDa Ig. Since percent motility, sperm count, HOST and percent normal spermatozoa in immunized dogs were significantly less than that of recommended fertile dog semen, it can be concluded that PH-20 (46/32 kDa) and LDHC4 (36/30/28 kDa) sub units had an immunocontraceptive potential in dog.
文摘The production of cells capable of expressing gene(s) of interest is important for a variety of applications in biomedicine and biotechnology, including gene therapy and a novel method of stem cell therapy in the various diseases. Achieving high levels of transgene expression for the longer period of time, without adversely affecting cell viability and differentiation capacity of the cells, is crucial. In the present study, we investigated the efficiency of plasmid vector for the production of transgenic cMSCs and examined any functional change of cells after transfection. To do so first we have collected bone marrows from the adult goats and cultured them for isolation of mesenchymal stem cells (cBM-MSCs). These cells were characterized using MSC specific markers including differentiation into osteocytes and adipocytes. Transfection with plasmid vector did not adversely affect cBM-MSCs morphology, viability or differentiation potential, and transgene expression levels were unaffected beyond passage 12th. The results indicated that we have been able to generate transgenic caprine MSC (tcBM-MSC) and transfection of cBM-MSCs using plasmid vector resulted in very high and stable transfection efficiency. This finding may have considerable significance in improving the efficacy of MSC-based therapies and their tracking in animal model.
基金funded by "Niche area of excellence,Indian Council of Agricultural Research(ICAR),India"
文摘Background: An oviduct-specific glycoprotein, OVGP1, is synthesized and secreted by non-ciliated epithelial cells of the mammalian oviduct which provides an essential milieu for reproductive functions. The present study reports the effects of recombinant buffalo OVGP1 that lacks post-translational modifications, and native Buffalo OVGP1 isolated from oviductal tissue, on frozen-thawed sperm functions and in vitro embryo development.Results: The proportion of viable sperms was greater(P 〈 0.05) in the recombinant OVGP1-treated group compared to the native OVGP1-treated group at 2 h, 3 h, and 4 h of incubation. The proportion of motile sperms at3 h and 4 h of incubation; and membrane-intact sperms at 4 h was greater(P 〈 0.05) in the native OVGP1-treated group compared to the control and recombinant OVGP1-treated groups. The proportion of capacitated and acrosome-reacted sperms was greater(P 〈 0.05) in the native OVGP1-treated group compared to the recombinant OVGP1 group at 4 h. The rates of cleavage of embryos and their development to the blastocyst stage were greater(P 〈 0.05) in the presence of either native or recombinant OVGP1 in comparison to control at 10 μg/mL concentration as compared to 5 or 20 μg/mL.Conclusions: The study suggests that both native and recombinant OVGP1 impart a positive effect on various sperm features and in vitro embryo development. However, native OVGP1 was found to have a more pronounced effect in comparison to recombinant non-glycosylated OVGP1 on various sperm functions except viability. Hence,our current findings infer that glycosylation of OVGP1 might be essential in sustaining the sperm functions but not the in vitro embryo development.
基金funded by the National Agriculture Innovation Project Grant to Suresh Kumar Singla(C 2-1-(5)/2007)Manmohan Singh Chauhan(C-2067 and 075).
文摘Objective:To study the effect of L-ascorbic acid supplementation on the in vitro development of buffalo embryos and evaluate the relative mRNA abundance of some pro-apoptotic,anti-apoptotic,and embryonic development-related genes.Methods:In experiment 1,we evaluated the effect of the addition of 0(control),50,and 100μM L-ascorbic acid to the in vitro maturation medium on the developmental competence in terms of blastocyst rate and relative mRNA abundance of some pro-apoptotic(BAX,BID),anti-apoptotic(BCL-XL,MCL1),and embryonic development(GDF9,BMP15)related genes.Based on the results,we chose 50μM as the suitable dose of L-ascorbic acid for the subsequent experiments.We further evaluated the blastocyst rates following the addition of 50μM L-ascorbic acid to the in vitro culture medium(experiment 2),and in vitro maturation and in vitro culture media(experiment 3).In all three experiments,the maturation and culture media devoid of L-ascorbic acid served as the control group.Results:The blastocyst rate after adding 50μM L-ascorbic acid to the in vitro maturation medium was significantly higher than the control group(P<0.05),whereas 100μM L-ascorbic acid exhibited a negative effect on the blastocyst rate.The blastocyst rates for embryos cultured in 50μM L-ascorbic acid in the in vitro culture medium alone and both in vitro maturation and in vitro culture media were significantly higher than their corresponding control groups(P<0.05).The relative mRNA abundance of BAX significantly decreased in blastocysts produced after the addition of 50μM L-ascorbic acid as compared with the control group(P<0.05),whereas,for MCL1,it significantly decreased in blastocysts produced after the addition of 100μM L-ascorbic acid(P<0.05).Conclusions:The supplementation of 50μM L-ascorbic acid to in vitro maturation and in vitro culture media supports in vitro embryonic development in buffaloes by improving developmental competence and altering the expression of apoptosis-related genes.
基金The present study was carried out under the project“Molecular markers for improving reproduction in cattle and buffaloes”under the funding of Bill and Melinda Gates Foundation,USA and Indian Council of Agricultural Research-National Dairy Research Institute.
文摘Objective:To assess the stability of 10 candidate internal control genes(ICGs),namely GAPDH,ACTB,RPL23,RPS15A,ATPSF1,GLUT5,HMBS,ATP2B4,PPIA,and BRP to normalize the transcriptional data from testes samples of Zebu and crossbred bulls.Methods:Total RNA was isolated from testicular tissue of Zebu and crossbred bulls(n=6 each)between 2-8 years of age.cDNA was synthesized,and the quantitative real-time polymerase chain reaction(PCR)was performed.The cycle threshold values were used for the analysis of the stability of ICGs.Four different statistical algorithms:geNorm,Normfinder,BestKeeper,and RefFinder,were used to assess the stability of these genes.Results:ATPSF1,HMBS,PPIA,and RPS15A were the most reliable and stable ICGs for Zebu testes,and ATPSF1,RPL23,and PPIA for crossbred testes.Conclusions:A panel of stable ICGs(ATPSF1,HMBS,PPIA,RPS15A for Zebu and ATPSF1,RPL23,and PPIA for crossbred)for normalization of gene expression data in testes samples can be helpful for researchers to conduct functional genomics studies at the testicular level in cattle bulls.
文摘Quality semen production remains the main focus and objective of semen processing laboratories throughout the world. Bacterial and other microbial contaminants affect the semen quality and hence the fertility, and also lead to reproductive disorders as well as lower conception rates and increased embryonic mortality, abortion and other complications in females. Microbial contamination affects the semen adversely, by exerting direct spermicidal effect, formation of reactive oxygen species, toxin production, adherence with spermatozoa, deriving nutrients and oxygen from the medium and thus competing with spermatozoa for the factors of growth and normal functioning. Despite hygienic measures, several ubiquitous and opportunistic microbes find their ways into semen during collection, processing, and storage of semen, and survive even during freezing. Stringent sanitary precautions are therefore required at every step of collecting semen and its processing. Preputial cavity is considered as main source of semen contaminating microorganisms. Flushing the preputial cavity with normal saline or any suitable liquid combination with antimicrobial activity, prior to semen collection reduces the microbial load and thereby improves the semen quality.
文摘Low sperm numbers in artificial insemination (AI)-doses are being used widely to make the best use of high genetic value bulls as well as sex-sorted semen. Sperm concentration needed for AI to obtain reasonable fertility, taking genetic value of bull and numerous others components into consideration is one of the essential constituents for successful AI breeding program. However, low sperm concentrations in AI-doses lead to reducing post-thaw viability. The reduction in viability of low sperm doses may be affected by fresh semen volume, sperm number and seminal plasma level at final dilution. Reduction in quality and fertility of low sperm doses is one of the limitations for their use in successful AI programme. Sperm number per AI required to achieve optimum fertility is one of the main crucial things to AI industry, and numerous efforts have been made in this regard. Due to great variability among bulls, sperm number per AI could be a limiting factor in achieving acceptable fertility values. Fertility of low sperm doses may vary among bulls, and non-return rates (NRRs) with low sperm doses may be determined by fertility level of bull. On the basis of individual bulls, sperm numbers in AI doses needed to be adjusted to reduce the variations in NRRs among bulls. Utilizing high fertile bulls for low sperm doses with acceptable non-return rates (NRRs) may be a way to cover a large number of bovines under AI in countries like India. Deposition site within the uterine horn may alter non return rates following inseminations with low sperm doses. Following deep-uterine inseminations, acceptable pregnancies may be achieved with low sperm doses and even if ovulation side is unknown.
文摘This article presents a review pertains the laser irradiation effects and its possible mechanisms of action on spermatozoa functions in domestic animals. To improve artificial insemination, laser is sensitive and cost effective technique, when compared to other conventional methods. Laser may have both positive and negative effects on spermatozoa functions. Since the effects of light are mediated by reactive oxygen species, and the levels of these reactive oxygen species following irradiating spermatozoa with laser may be responsible for determining the effects of laser on sperm. Dose of laser may be regarded as of great significance and this dosage of laser may be responsible for determining its effects on spermatozoa. Optimum dosage of laser for improving seminal attributes may vary among various species and this need to be standardized in each of them. The beneficial effects include improving sperm livability, acrosomal integrity, hypo-osmotic swelling response, mitochondrial function and computer-aided sperm analysis parameters. The increase in cytochrome c oxidase activity, ATP levels and mitochondrial membrane potential, in laser irradiated cells may be responsible for enhanced sperm quality parameters. Improving fertility with laser irradiated spermatozoa has been reported in few species like boar and need to be elaborated in other species. In conclusion laser may be regarded as an easy, cheap and time saving technology for improving artificial insemination;in addition, laser may have various potential applications in the field of reproductive biotechnology as well as in livestock farms and veterinary polyclinics.
文摘Whey and toned milk was effectively used for the development of tea whitener. Cheese whey was concentrated to 12% solids in a vacuum evaporator, pasteurized at 72℃±2℃ for 15 s, diluted back to 11.5% solids and was cold blended with toned milk of 11.5% solids in three ratios viz. 25:75, 50:50 and 75:25 and selected blends were homogenized at 1200-1500 psi. Tea was prepared by adding 80 mL whitener to 100 mL decoction and addition of sugar at 8% (w/v). The tea whitener was selected based on the sensory attributes of tea beverages. The colour of the beverages, tea whiteners and decoction were analyzed by reflectance meter and computer vision image analysis technique. The viscosity of the beverage was tested using a Brookfield viscometer. Coffee stability test was done to assess the stability of tea whitener under acidic conditions. Standard plate count, Coliform count and Presumptive Coliform test were done to assess the microbiological quality of the whitener. Homogenization had a positive impact on the viscosity and colour. The reflectance value of tea prepared from 25% homogenised whey milk blend and toned milk were in the range of 43-46 and 45-49, respectively. The viscosity of tea was found to increase with homogenization and whey levels. The sedimentation values of 25% whey blends and toned milk were 1.25 mL and 1 mL respectively. The shelf life of the blends was 2 days under refrigerated conditions. It can be concluded that a satisfactory tea whitener can be prepared by blending whey with toned milk in 25:75 proportion followed by homogenization.
文摘Electrolyzed Oxidized Water (EOW) is produced by passing a diluted salt solution through an electrolytic cell, having anode and cathode electrodes. The anode and cathode are separated by a bipolar membrane. Negatively charged ions—chloride and hydroxide in the diluted salt solution move to anode to give up electrons and become gas (O2, Cl2) and hypochlorous acid and having redox potential of +700 to +800 mV with pH 4.0. It has a strong oxidation potential and a shortage of electrons giving it the ability to oxidize and sterilize. In microbial inactivation process, oxidized water damage cell membranes, create disruption in cell metabolic processes and essentially kill the cell. EOW, also a strong acid, is different to hydrochloric acid or sulfuric acid in that it is not corrosive to skin, mucous membrane, or organic material. It is easy to handle and suitable for the sanitation of the plant and decontamination of foods. Electrolyzed water has been tested and used as a disinfectant in the food industry and other applications.