Total Reducing Sugar Content Research Articles

Statistical Modeling And Optimization Of Pretreatment For Fermentable Sugars Production From Cotton Gin Waste

ABSTRACTBioethanol is produced from lignocellulosic materials, such as agricultural waste, forest residues, etc. In this study, cotton gin waste was utilized to produce fermentable sugars through an effective pretreatment method. Autoclaving, steam explosion, and liquid hot water pretreatments were compared for their effectiveness in disruption of the lignocellulose matrix. Autoclaving was proved to be an effective pretreatment method in terms of maximum reducing sugar production. The Box–Behnken design (BBD) was employed in the optimization of autoclaving pretreatment. Slurry density (X1, g/mL), temperature (X2, °C), and time (X3, min) were chosen as the independent variables and the total reducing sugar concentration (Y, mg/L) as the dependent variable (response). The total reducing sugar production from cotton gin waste was found to be 511.48 mg/L experimentally, at an optimum slurry density of 0.50 g/mL, a temperature of 138°C, and a time of 11 min.

Preservative Effect of Proteolytic Enzymes on Cane Molasses Storage

Molasses is an intermediate product obtained during sugar process. Usually, it has more than 850 Brix i.e. only 15% moisture. It contains more than 45% fermentable sugars. Basis composition, it is impossible to microorganism which could survive in this environment for too long. But microorganisms like Lactobacillus spp. and Gram –ve Coccobacilli possibly Pseudomonas spp. are able to survive in this medium and deteriorate the molasses. Therefore molasses preservation requires specific solutions which can stop deterioration and maintain total reducing sugar (TRS) value in molasses to produce alcohol. Therefore, two groups of proteolytic enzymes were developed which prevents the deterioration of molasses by preventing the microbial growth at 20ppm but also preserve the 3 – 5% of TRS content during storage of molasses up to six months or till application continued.

English

Sweet sorghum is considered one of the best sources of bioethanol due to its higher total reducing sugar content, which ferments completely to produce ethanol coupled with its adaptation to the changing climate. A study was carried out in Ethiopia, during the 2015/16 crop season, to determine the extent of phenotypic and genotypic relationships among 13 agronomic traits and six quality components of 28 sweet sorghum genotypes. Panicle weight and width, dry matter yield, thousand kernel weight and harvest index had significant positive correlation with grain yield and exerted favourable direct effects both at phenotypic and genotypic levels. Ethanol yield was also correlated with juice yield, sugar yield and fresh stalk yield. Therefore, these yield and quality components are suggested to receive due attention during sweet sorghum varietal selection. Moreover, days to maturity had negative correlation and imposed negative direct effect on grain yield, which may indicate the possibility to select high yielding, early maturing dual purpose varieties for dry environments where terminal drought is rampant. The studied genotypes were grouped into three clusters according to their D2 values, worthy of future breeding work considering the special merits in each cluster depending on the objectives of the breeding program. Some of the genotypes excelled as one of the two commercial sugarcane varieties used as controls for some quality traits. Therefore, considering their less water requirement, faster production cycle, and additional advantage of grain production over sugarcane, sweet sorghum stalks can serve as alternatives to sugarcane for use as feedstock in drier areas of the world under the changing climate. Key words: Bioethanol, correlation, juice, path-coefficient, sweet sorghum.

Elevated CO2 leads to carbon sequestration by modulating C4 photosynthesis pathway enzyme (PPDK) in Suaeda monoica and S. fruticosa

The C4 halophytic species Suaeda monoica and S. fruticosa, possess the C4 photosynthesis pathway without Kranz anatomy were grown at ambient (470ppm CO2) and elevated (850ppm CO2) atmospheric CO2 under control containment facility to study the plant response under CO2 stress condition. The relative growth of both Suaeda species was enhanced with atmospheric CO2 enrichment compared to control (ambient) condition. The photosynthesis rate was found 2.5μmolCO2m−2s−1 in both species under stress condition compared to about 1.9μmolCO2m−2s−1 under control conditions. About 0.3molH2Om−2s−1 conductance was detected under an unstressed condition which decreased significantly to ~0.07molH2Om−2s−1 on the 6th day of stress treatment. Similarly, transpiration rate was also decreased significantly from 4.4–5.2mmolH2Om−2s−1 to 1.7–1.9 under stress condition. In contrast, VpdL increased significantly from 1.9kPa to 2.5kPa under stress condition. A higher total chlorophyll content observed in S. monoica (56.36mgg−1 tissue) compared to S. fruticosa (33.12mgg−1 tissue) under unstressed (control) condition. A significant increase was found in the total chlorophyll content of S. fruticosa (45.47mgg−1 tissue) with stress treatment compared to control (33.12mgg−1 tissue). In contrast, the total chlorophyll decreased in S. monoica (51.58mgg−1 tissue) under similar stress condition compared to control plants (56.36mgg−1 tissue). About 6–6.8mg total sugar per gram tissue found under control condition which enhanced further (7.5 to 11mgg−1 tissue) under stress condition. Similarly, total reducing sugar (~2mgg−1 tissue) and total starch content (6.5–11mgg−1 tissue) increased under stress condition. About 6.5- and 3- fold higher expression of PPDK gene was observed for S. monoica and S. fruticosa, respectively under CO2 stress condition. PPDK (1.2- and 1.5- fold) and antioxidant enzymes; APX (12.7- and two-fold), CAT (2.2- and 6.4- fold) and SOD (4.6- and 94- fold) enhanced significantly in S. fruticosa and S. monoica, respectively under high CO2 stress condition compared to control plants. Overall, it was observed that PPDK enzyme plays a key role in C4 photosynthesis pathway and S. monoica is a potential candidate to be explored further for the saline agricultural and CO2 capture.

Synergistic action of an Aspergillus (hemi-)cellulolytic consortium on sugarcane bagasse saccharification

This study evaluated the synergistic efficiency of crude enzymatic extracts (CEE) produced by fungal strains Aspergillus fumigatus SCBM6 (AFE) and Aspergillus niger SCBM1 (ANE) in the hydrolysis of alkali pretreated sugarcane bagasse (2% NaOH (w v−1), ∼100 °C, 4 h and liquid-to-solid ratio of 25 (v w−1)). CEEs were produced by solid-state fermentation (SSF) and presented high concentration of β-glucosidase (AFE = 78.4 U g−1 and ANE = 43.02 U g−1), β-xylosidase (ANE = 78.0 U g−1) and xylanase (AFE = 1774.5 U g−1 and ANE = 3289.6 U g−1). A 24 full design of experiments was used to optimize the hydrolysis of pretreated bagasse as a function of time, temperature, amount of pretreated bagasse and AFE-to-ANE ratio. After hydrolysis optimization, the maximum concentration of total reducing sugars (TRS) was 21.68 g L−1. It was also noticed a significant production of xylose (135.59 mg) and arabinose (58.10 mg) per gram of hemicelluloses and a slight production of glucose (20.18 mg) per gram of cellulose. As fungi consortium were excellent producers of hemicellulases, it is suggested that they could be used for pentoses saccharification and production of ethanol via C-5 fermentation, xylitol, biopolymers, among other bioproducts.

Scrutinizing Efficacy of Ethyl Methane Sulphonate on Antioxidants of Sunflower

In current research the scrutiny of antioxidant potential was carried out. The sunflower seeds provide a good source of antioxidant potential. Different treatments of mutagenic substance, namely EMS, was given to the SF seeds and antioxidant potential along with other biochemical components was inquired. The total antioxidant’s activity was increased at 0.1%. The reducing power of seeds was elevated after the treatments of 0.5% and 0.2% as compared to the controls, where as the total phenolic and flavonoids contents were decreased after the treatments of 0.2% and 0.3% respectively. The total reducing sugar and total protein contents were boosted after the treatment at 0.3% and 0.1% respectively, where as total sugars were reduced.

Biochemical Defense Response: Characterizing the Plasticity of Source and Sink in Spring Wheat under Terminal Heat Stress.

Wheat is highly prone to terminal heat stress (HS) under late-sown conditions. Delayed- sowing is one of the preferred methods to screen the genotypes for thermotolerance under open field conditions. We investigated the effect of terminal HS on the thermotolerance of four popular genotypes of wheat i.e. WR544, HD2967, HD2932, and HD2285 under field condition. We observed significant variations in the biochemical parameters like protein content, antioxidant activity, proline and total reducing sugar content in leaf, stem, and spike under normal (26 ± 2°C) and terminal HS (36 ± 2°C) conditions. Maximum protein, sugars and proline was observed in HD2967, as compared to other cultivars under terminal HS. Wheat cv. HD2967 showed more adaptability to the terminal HS. Differential protein-profiling in leaves, stem and spike of HD2967 under normal (26 ± 2°C) and terminal HS (36 ± 2°C) showed expression of some unique protein spots. MALDI-TOF/MS analysis showed the DEPs as RuBisCO (Rub), RuBisCO activase (Rca), oxygen evolving enhancer protein (OEEP), hypothetical proteins, etc. Expression analysis of genes associated with photosynthesis (Rub and Rca) and starch biosynthesis pathway (AGPase, SSS and SBE) showed significant variations in the expression under terminal HS. HD2967 showed better performance, as compared to other cultivars under terminal HS. SSS activity observed in HD2967 showed more stability under terminal HS, as compared with other cultivars. Triggering of different biochemical parameters in response to terminal HS was observed to modulate the plasticity of carbon assimilatory pathway. The identified DEPs will enrich the proteomic resources of wheat and will provide a potential biochemical marker for screening wheat germplasm for thermotolerance. The model hypothesized will help the researchers to work in a more focused way to develop terminal heat tolerant wheat without compromising with the quality and quantity of grains.

Facile Strategy for Surface Functionalization of Corn Cob to Biocarbon and Its Catalytic Performance on Banana Peel Starch Hydrolysis

The sulphonated biocarbon (C-SO 3 H) has been prepared from carbonization of corn cob and followed by one-step hydrothermal treatment. This research focus on characterization of C-SO 3 H and evaluate the capability of C-SO 3 H for hydrolyzed banana peel starch, and also investigate the optimum concentrations of substrate and the reaction time for hydrolysis. The C-SO 3 H was produced from corn cob (±60 mm mesh); it was carbonized at 400 °C for 1 hour. The resulting carbon was sulphonated by hydroxy ethyl sulphonic acid at 180 °C for 4 hours, and then washed with 50% methanol and water, and then resulting biocarbon was dried at 80 °C for 6 hour in oven. Based on Scanning Electron Microscopy (SEM) observation, structure morphology of corn cob was changed due to carbonization process. The C-SO 3 H after sulphonation has random pores with diameter around 3 µm–10 µm. The X-Ray Diffraction (XRD) showed increased of crystalline index of corn cob about 133% after carbonization. Analysis of EDX result for S component on C-SO 3 H is 3.79% w/w and capacity of H + is 1.447 mmol/g. The apparent peaks of -SO 3 H functional groups (1207 cm -1 and 1720 cm -1 ) on C-SO 3 H were indicated the by Fourier Transform Infra Red (FTIR). Optimum hydrolysis concentration of banana peel starch is 7% w/v for 60 min at 100 o C which Total Reducing Sugar (TRS) concentration about 0.688 mg/mL. Reusability of C-SO 3 H for 4 repeated used which showed a good performance that only 6% of decreasing activity of C-SO 3 H. The utilization of corn cob can be enhanced of corn cob and also it can be utilized as heterogeneous catalyst.

Pretreatment conditions of palm oil mill effluent (POME) for thermophilic biohydrogen production by mixed culture

The availability of fermentable sugars in POME is one of the critical factors that determine the fermentative biohydrogen production yield. This study was carried out to determine the pretreatment conditions viz., temperature, hydrolysis time and acid concentration that can yield the highest monomeric sugars, to be utilized for production of biohydrogen from POME by mixed culture dark fermentation. Two different acids were used, nitric acid and phosphoric acid in the pretreatment of POME. Batch fermentation was performed to determine the potential of pretreated POME as a substrate for the production of biohydrogen under the tested pretreatment conditions. Higher hydrogen yield was successfully achieved using pretreated POME as compared to raw POME by mixed culture. Maximum hydrogen production was 0.181 (mmol/L/h), which corresponded to the yield of 1.24 mol H2/mol glucose achieved at 0.8% (w/v) phosphoric acid with initial total reducing sugar concentration of 18.47 g/L. Hence, the results implied that POME pretreated with 0.8% (w/v) phosphoric acid is a potential substrate for efficient biohydrogen fermentation yield that is 97% higher than untreated POME. While POME pretreated with 1% (w/v) of nitric acid showed 65% improvement in biohydrogen yield as compared to untreated POME. Therefore, the results implied that both pretreatment methods of POME showed significant increase in the biohydrogen production.

Evaluation of tropical seaweeds as feedstock for bioethanol production

Limited resources of freshwater and decreasing fossil fuel resources are two main reasons to consider the ocean as a huge resource for producing food, feed, fertilizer and feedstock for fuel. In this study, twenty-nine tropical seaweeds (11 green, 10 red and 8 brown seaweeds) collected in Malaysia were assessed as potential feedstock for bioethanol production. Total carbohydrate content ranged from 12.16 to 71.22% dry weight (DW) with total reducing sugar content ranging from 5.17 to 34.12% DW. During hydrolysis using dilute sulphuric acid, the dominant fermentation inhibitors were 5-hydroxymethylfurfural and phenolic compounds. Overliming was found to reduce the content of fermentation inhibitors by up to 79%. The red seaweeds, Kappaphycus alvarezii (Doty) Doty ex P.C.Silva and Gracilaria manilaensis Yamamoto and Trono, were selected for optimization of saccharification and fermentation of the hydrolysate, because they had the highest carbohydrate contents and are commercially cultivated. The most suitable dilute acid conditions obtained in present study was sulphuric acid (2.5%, w v−1) treatment at 121 °C for 40 min that produced 0.29 and 0.34 g g−1 DW reducing sugar for K. alvarezii and G. manilaensis, respectively. Fermentation of the hydrolysates with Saccharomyces cerevisiae produced bioethanol yields of 20.90 g L−1 (71.0% of theoretical yield) for K. alvarezii and 18.16 g L−1 (67.9% theoretical yield) for G. manilaensis.

Effect of Yeast and Acetic Fermentation on Phytochemical and Antioxidant Properties of Jackfruit Pulp(Artocarpus heterophyllus L.)

Jackfruit (Artocarpus heterophyllus L.) is one of the important tropical fruits in Malaysia, mainly used for fresh consumption because of its good source of energy, deliciously sweet in taste, proteins, dietary fiber, minerals and vitamins. To date, there is lack of fermentation study reported on jackfruit. Hence, the aim of this work is to evaluate the phytochemical content and antioxidant properties of fermented jackfruit which was subjected to acetic acid bacteria and yeast fermentation individually for a duration of 7 days. The changes in the pH value, brix value, total reducing sugar content, total phenolic and total flavonoid content were examined for both yeast and acetic acid fermented jackfruit samples. Few antioxidant activities such as ferric reducing antioxidant power (FRAP) value and 2,2-diphenyl -1- picrylhydrazyl (DPPH) free radical scavenging activity were also analyzed to determine the differences in the respective properties of jackfruit pulp after being subjected to yeast or acetic acid fermentation. There is a slight change on brix value and DPPH free radical scavenging activity (31.05-33.17% radical scavenging activity) for both yeast and acetic acid fermented samples. However, acetic acid fermented jackfruit exhibited higher total reducing sugar content (23.24 mg glucose/mL) and total phenolics content when compared to yeast fermented jackfruit sample. Both yeast and acetic acid fermented jackfruit samples showed higher FRAP value (0.435 -0.48 mg ascorbic acid/mL) than unfermented jackfruit sample (0.372 mg ascorbic acid/mL) except for total flavonoid content which exhibited a decreasing trend after fermentation.

Independent and interactive effects of waterlogging and salinity on carbohydrate metabolism and root anatomy in pigeonpea genotypes at different growth stages

Four pigeon pea genotypes (ICPH-2431, PARAS, UPAS-120, H09-33) were raised in polythene bags filled with half kg soil + FYM manure mixture. Waterlogging, salinity and waterlogging + salinity treatments were given for 8 days and observations were recorded 1 day after removal from treatment in 20 and 40 day old plants. Significant decline in percent survival was observed under waterlogging and combined treatment in 20 and 40 day old plants. Decline in total sugar content and non reducing sugar content was observed under all three treatments in 20 and 40 day old plant leaves. In roots, an initial increase in total soluble sugar content was observed with waterlogging and salinity treatment. Non reducing sugar content also decreased under all the three treatments. An increase in sucrose synthase and alcohol dehydrogenase activity under all three treatments and formation of aerenchyma in roots under waterlogging and combined treatment was also observed. Waterlogging and salinity in combination was more deleterious compared to alone treatments. Roots were more sensitive to all the stresses as compared to leaves. ICPH performed best among all the genotypes.

Influences of superheated steam roasting on changes in sugar, amino acid and flavour active components of cocoa bean (Theobroma cacao).

Roasting is one of the important unit operations in the cocoa-based industries in order to develop unique flavour in products. Cocoa beans were subjected to roasting at different temperatures and times using superheated steam. The influence of roasting temperature (150-250°C) and time (10-50 min) on sugars, free amino acids and volatile flavouring compounds were investigated. The concentration of total reducing sugars was reduced by up to 64.61, 77.22 and 82.52% with increased roasting temperature at 150, 200 and 250°C for 50 min, respectively. The hydrophobic amino acids were reduced up to 29.21, 36.41 and 48.87% with increased roasting temperature at 150, 200 and 250°C for 50 min, respectively. A number of pyrazines, esters, aldehydes, alcohols, ketones, carboxyl acids and hydrocarbons were detected in all the samples at different concentration range. Formation of the most flavour active compounds, pyrazines, were the highest concentration (2.96 mg kg-1 ) at 200°C for 10 min. The superheated steam roasting method achieves the optimum roasting condition within a short duration Therefore, the quality of cocoa beans can be improved using superheated steam during the roasting process. © 2017 Society of Chemical Industry.

Dielectric barrier discharge plasma pretreatment on hydrolysis of microcrystalline cellulose

Dielectric barrier discharge (DBD) plasma was used as a pretreatment method for downstream hydrolysis of microcrystalline cellulose (MCC). The degree of polymerization (DP) of MCC decreased after it was pretreated by DBD plasma under a carrier gas of air/argon. The effectiveness of depolymerization was found to be influenced by the crystallinity of MCC when under the pretreatment of DBD plasma. With the addition of tert-butyl alcohol in the treated MCC water suspension solution, depolymerization effectiveness of MCC was inhibited. When MCC was pretreated by DBD plasma for 30 min, the total reducing sugar concentration (TRSC) and liquefaction yield (LY) of pretreated-MCC (PMCC) increased by 82.98% and 34.18% respectively compared with those for raw MCC.

Cassava brown streak disease effects on leaf metabolites and pigment accumulation

Cassava Brown Streak Disease (CBSD) is a threat to productivity and product quality in East Africa. The objective of this study was to understand the effect of CBSD on the primary photosynthetic apparatus of cassava (Manihot esculenta Crantz). Three cassava varieties with varying levels of reaction to infection by CBSD were chosen and subjected to field disease pressure. Disease progression and the resulting effects on leaf morphology, metabolite and pigment accumulation were assessed over a period of five months, beginning from 3 months after planting. Slight increments in leaf dry matter were observed up to 4 MAP, and there after a drop was registered. A significant (P<0.05) reduction in photosynthetic pigments occured with a fall in the Chla:Chlb and Chla:Carotenoid ratio, indicative of specific reductions in chlorophyll a (-80%) compared to Chlorophyll b (-41 to -62%) and Carotenoid (-11 to -18%). Total reducing sugar and starch content also dropped significantly (-30 and -60%, respectively), much as NASE 14 maintained a relatively higher amount of carbohydrates. Leaf protein levels were significantly reduced at a rate of 0.07 ug g-1 leaf per month in diseased treatments. Significant reductions in primary metabolites show altered leaf photosynthetic and growth metabolism, resulting into a compromised plant system that cannot perform optimally.Keywords: Carotenoid, leaf metabolites, photosynthesis, protein

Biochemical response of salt-tolerant and salt-sensitive wheat cultivars to salinity

Two wheat (Triticum aestivum L.) varieties KRL 1–4 and UP 2338 were tested in pot culture under different salinity regimes for their biochemical response. Three different levels of salinity (100, 200 and 300 mM NaCl) were used. Untreated plants were kept as control. Samples were analyzed from 20 DAS till 60 DAS for various biochemical parameters viz., proline content, total reducing sugar content, total nitrogen content and total protein content. Protein profiling through SDS-PAGE for both the cultivars of wheat was performed. The amount of proline and reducing sugar increased with increasing salinity, the increase being more in tolerant than in sensitive cultivar. Total nitrogen and total protein content, however, decreased with increasing salt concentration and reduction being more in sensitive than in tolerant cultivar. The changes in protein synthesis were determined as new synthesis, complete loss, increase and decrease.

상황버섯균 접종 황기의 배양 중 이화학적 성분변화

This study analyzed the changes in physicochemical components of Astragalus membranaceus (AM) fermented with Phellinus linteus. Moisture content, pH, total acidity, total reducing sugar content, extraction yield, free sugar content, free amino acid and isoflavonoid (calycosin, formononetin) were investigated. The moisture content was increased during fermentation with Phellinus linteus. The pH level increased while the total acidity significantly decreased during fermentation. The reducing sugar content were in the range of 0.32~0.61%. The extraction yield using water was higher than that using 80% ethanol. The major free sugars were identified as glucose, fructose, sucrose and the content of free sugars decreased through fermentation. However, the glucose and sucrose contents of the water extracts were increased. In addition, the free amino acid increased significantly during fermentation. Finally, calycosin and formononetins contents in water extracts of after 30 days of AM fermentaion with Phellinus linteus were (3.91 mg/100 g) and (1.38 mg/100 g), respectively. These results suggest that fermentation with Phellinus linteus could be used to increase the bioactivity of AM. The mycelium-fermented AM could be a valuable source of functional material and edible resource for industry.

Prebiotic synthesis from rice using Aspergillus oryzae with solid state fermentation

Prebiotic synthesis from economical substrates (glutinous rice and rice bran) using Aspergillus oryzae TISTR 3102 and TISTR 3222 with solid state fermentation (SSF) was explored. The pH value, alpha-glucosidase activity and total reducing sugar (TRS) concentration were monitored during incubation with different substrate ratios of 1:1 and 1:2 weight per weight (w/w). After 7 d incubation under SSF in glutinous rice and rice bran at 1:2 w/w, the highest alpha-glucosidase activity (4.49 Unit/mL) and TRS concentration (16.84 g/L) were detected. The product mixtures consisted of several reducing sugars as prebiotic compounds, especially panose and isomaltotriose. The prebiotic properties were investigated for their growth stimulation on two probiotic strains (Lactobacillus plantarum and Lactobacillus acidophilus) and the inhibition of two pathogens (Escherichia coli and Salmonella paratyphi). These product mixtures under solid state fermentation (SSF) using A. oryzae in variable rice ratios displayed high efficiency with regard to prebiotic properties and were deemed appropriate for prebiotic production.

Chemical and volatile composition of jujube wines fermented by Saccharomyces cerevisiae with and without pulp contact and protease treatment

This study evaluated the chemical and volatile composition of jujube wines fermented with Saccharomyces cerevisiae A1.25 with and without pulp contact and protease treatment during fermentation. Yeast cell population, total reducing sugar and methanol contents had significant differences between nonextracted and extracted wine. The nonextracted wines had significantly higher concentrations of ethyl 9-hexadecenoate, ethyl palmitate and ethyl oleate than the extracted wines. Pulp contact also could enhance phenylethyl alcohol, furfuryl alcohol, ethyl palmitat and ethyl oleate. Furthermore, protease treatment can accelerate the release of fusel oils. The first principal component separated the wine from the extracted juice without protease from other samples based on the higher concentrations of medium-chain fatty acids and medium-chain ethyl esters. Sensory evaluation showed pulp contact and protease could improve the intensity and complexity of wine aroma due to the increase of the assimilable nitrogen.

Studies on fresh milt parameters and cellular changes during cryopreservation of spermatozoa of Deccan Mahseer, Tor Khudree (Sykes)

Studies on the physico-chemical characteristics of seminal plasma, along with ultrastructure and mitochondrial activity of fresh spermatozoa of the endangered Deccan mahseer Tor khudree were undertaken. The ultrastructure and mitochondrial activity of fresh spermatozoa were compared with those of cryopreserved-thawed spermatozoa to understand the nature and extent of cryo-damage. Physico-chemical analyses of fresh milt revealed sperm density of 3.93±0.11 x 107 spermatozoa ml-1, spermatocrit value of 67.08±1.22%, higher K+ concentration of 13.16±0.121 mg l-1, total reducing sugar and total protein concentration of 47.31±0.82 and 19.60±0.66 mg 100 ml-1 respectively. Ultrastructure of the fresh spermatozoa by both scanning (SEM) and transmission electron microscopy (TEM) revealed the spherical head without any acrosomal complex, small mid piece with mitochondria and a long tail. Cross section of tail by TEM revealed typical 9+2 doublet arrangement of the axoneme. Head measured 1.86±0.006 μ in dia with a mid-piece length of 0.53±0.012 μ and tail length of 33.53±0.220 μ. Ultrastructural damages to the spermatozoa following cryopreservation included, loosening of chromatin and disruption of the cytoplasmic membrane as compared to that of fresh spermatozoa. Nitroblue tetrazolium (NBT) assay revealed low levels of activity of the enzymes of the mitochondrial complex in cryopreserved-thawed spermatozoa when compared to fresh spermatozoa indicating damage to the functional integrity of the enzymes of the mitochondrial enzyme complex.