Molecular characterization and serodiagnostic evaluation of the Echinococcus ortleppi recombinant glutaredoxin 1 protein for cystic echinococcosis in buffalo (Bubalus bubalis).

Cystic echinococcosis (CE), caused by the metacestode of Echinococcus granulosus sensu lato (s.l.), adversely affects the physiology of the vital organs in which they grow. Condemnation of meat causes substantial economic loss to the livestock industry. Conventionally the infection is detected by necropsy as serological diagnosis of the infection in livestock is ambiguous. Identification of specific diagnostic antigens would be a substitute for the cyst fluid antigens which lack adequate diagnostic sensitivity and specificity. BLAST analysis supported by the negligible pairwise nucleotide distance of the 389 nt COX1, 489 nt NAD1, and 425 nt ITS1 with the related sequences of E. ortleppi ascertained the association of E. ortleppi with CE in buffaloes. Given the extensive distribution of glutaredoxin 1 in every developmental stage of Echinococcus granulosus s.l that makes it an ideal serodiagnostic antigen for CE, we expressed the 14 kDa E. ortleppi glutaredoxin 1 (rEoGrx1) protein in E. coli BL21 (DE3) and tested a total of 225 sera samples, including 126 sera samples from the necropsy-positive buffalo, by the rEoGrx1 IgG-ELISA. The ELISA could detect a total of 82/126 sera samples as positive. The diagnostic sensitivity and specificity of the rEoGrx1 IgG-ELISA were 65.1 % and 51.5 %, respectively. The protein showed serological cross-reaction against Fasciola gigantica, Toxoplasma gondii, and Sarcocystis sp. The in silico bioinformatics analysis of the E. ortleppi, F. gigantica, and T. gondii glutaredoxin sequences revealed fully conserved amino acids at positions 11 and 21, the substitution of conserved amino acids at positions 14 and 6, and semi-conserved substitutions at positions 3 and 4, respectively. The findings partly explain the molecular basis of the serological cross-reactivity of the protein.

Chickpea (Cicer arietinum L.) as model legume for decoding the co-existence of Pseudomonas fluorescens and Mesorhizobium sp. as bio-fertilizer under diverse agro-climatic zones.

Microbial co-inoculation strategy utilizes a combination of microbes to stimulate plant growth concomitant with an increased phytopathogen tolerance. In the present study, 15 endophytic bacterial isolates from rhizosphere and roots of wild chickpea accessions (Cicer pinnatifidum, C. judiacum, C. bijugum and C. reticulatum) were characterized for morphological, biochemical and physiological traits. Two promising isolates were identified as Pseudomonas fluorescens strain LRE-2 (KR303708.1) and Pseudomonas argentinensis LPGPR-1 (JX239745.1) based on 16S rRNA gene sequencing. Biocompatibility of selected endophytes with Mesorhizobium sp. CH1233, a standard isolate used as a national check in All India Coordinated Research Project (AICRP) was assessed to develop functional combinations capable of producing Indole acetic acid, gibberellins, siderophores and improving seed vigour (in vitro). In vivo synergistic effect of promising combinations was further evaluated under national AICRP, (Chickpea) at two different agro-climatic zones [North-West plain (Ludhiana and Hisar) and Central zones (Sehore)] for three consecutive Rabi seasons (2015-18) to elucidate their effect on symbiotic, soil quality and yield parameters. On the pooled mean basis across locations over the years, combination of Mrh+LRE-2 significantly enhanced symbiotic, soil quality traits and grain yield over Mrh alone and highly positive correlation was obtained between the nodulation traits and grain yield. Superior B: C ratio (1.12) and additional income of Rs 6,505.18 ha-1 was obtained by application of Mrh+LRE-2 over Mrh alone and un-inoculated control. The results demonstrate that dual combination of Mrh and Pseudomonas sp. from wild Cicer relatives can be exploited as a potential bio-fertilizer for increasing soil fertility and improving chickpea productivity under sustainable agriculture.

Universal Hearing Screening in Newborns Using Otoacoustic Emissions and Brainstem Evoked Response in Eastern Uttar Pradesh.

The objectives were to determine the incidence of hearing impairment in a standardized population of neonates and to determine the significance of association of epidemiological and risk factors with neonatal hearing loss. A cohort of 600 newborns was selected for study and divided into two groups-525 in 'No Risk' group and remaining 75 in 'At Risk' group. The study protocol was carried out in three steps: (a) Screening of Hearing Loss with TOAE, done from 36 h after birth to 28 days of life, (b) Re-screening of hearing loss in newborns (of 4-12 weeks of age), who were tested positive for hearing loss in the first screening, done with DPOAE, (c) Confirmation of hearing loss with BERA, in those who were tested positive in both the first and second screening. In the study the incidence of hearing impairment in 600 infants screened was 6.67 per 1000 screened; 3.81 per 1000 screened in the Not at Risk group and 26.67 per 1000 screened in At Risk group. In At Risk group, admitted to the NICU, severe birth asphyxia and hyperbilirubinemia were found to be major risk factors. Loss to follow up was more in Not at Risk group and False Positive cases with TEOAE were more than DPOAE. BERA was found to be must for confirmation of hearing loss. Neonatal Hearing Screening of only At Risk population is likely to miss some hearing loss. Universal Hearing Screening should be the preferred strategy. Good follow up in the 'At Risk' group suggests that initial interventions in NICU had sensitized the parents for the possibility of hearing loss. This study recommends the introduction of two stage neonatal screening-rescreening protocol, using OAE and BERA, in the country in phased manner.

Effect of induced Fasciola gigantica infection during pre-patency on the performance of buffalo calves fed on different percentage of protein.

Thirty growing Murrah buffalo calves (8-12 months of age, 109.85 ± 2.43) were reared in parasite free conditions and randomly divided into three equal groups as per CRD. They were fed on iso-caloric (2.01 ME Mcal/Kg diet) diets containing standard protein (SP) diet at 100 %, 90 % of SP (medium protein, MP) and 80 % of SP (low protein, LP) of the protein requirements (Kearl 1982). After 21 days of feeding, each group was further subdivided into two sub-groups (A & B). Animals in sub-groups 'A' served as non-infected control, while in sub group 'B' were orally infected with Fasciola gigantica metacercarie (mc; 1,000 each). A metabolic trial of 40 days post infection was carried out in control and parasitized animals. Intake of digestible dry matter, organic matter and acid detergent fibre (ADF) was significantly higher (P < 0.05) in SP group compared to LP group. The digestibility of crude protein (CP) and ADF was significantly higher in SP group compared to MP and LP groups. The digestible crude protein (DCP) and total digestible nutrients (TDN) intakes (g/kgW(0.75)) were also significantly (P < 0.001) higher in SP than MP and LP groups. However, DCP intake was significantly (P < 0.001) lower in infected subgroups compared to control subgroups. Intake and balance (g/d) of nitrogen, calcium and phosphorus were significantly (P < 0.01) higher in SP than MP and LP groups. The average daily gain of buffalo calves fed on SP Uninfected (SPU), SP Infected (SPI), MPU, MPI and LPU, LPI groups was 333, 178, 356, 144, 222 and 144 g and was significantly (P < 0.01) lower in animals fed LP ration. The feed conversion ratio (FCR) was also significantly (P < 0.01) higher in infected sub-groups as compared to respective control groups. The results showed that a SP diet substantially improved the overall performance of buffalo calves in control and infected groups and reduced adverse effect of F. gigantica infection.

Regulation of alternative splicing of Bcl-x by BC200 contributes to breast cancer pathogenesis.

BC200 is a long non-coding RNA (lncRNA) that has been implicated in the regulation of protein synthesis, yet whether dysregulation of BC200 contributes to the pathogenesis of human diseases remains elusive. In this study, we show that BC200 is upregulated in breast cancer; among breast tumor specimens there is a higher level of BC200 in estrogen receptor (ER) positive than in ER-negative tumors. Further experiments show that activation of estrogen signaling induces expression of BC200. To determine the significance of ER-regulated BC200 expression, we knockout (KO) BC200 by CRISPR/Cas9. BC200 KO suppresses tumor cell growth in vitro and in vivo by expression of the pro-apoptotic Bcl-xS isoform. Mechanistically, BC200 contains a 17-nucleotide sequence complementary to Bcl-x pre-mRNA, which may facilitate its binding to Bcl-x pre-mRNA and recruitment of heterogeneous nuclear ribonucleoprotein (hnRNP) A2/B1, a known splicing factor. Consequently, hnRNP A2/B1 interferes with association of Bcl-x pre-mRNA with the Bcl-xS-promoting factor Sam68, leading to a blockade of Bcl-xS expression. Together, these results suggest that BC200 plays an oncogenic role in breast cancer. Thus, BC200 may serve as a prognostic marker and possible target for attenuating deregulated cell proliferation in estrogen-dependent breast cancer.

Regulation of breast tumorigenesis through acid sensors.

The low extracellular pH in the microenvironment has been shown to promote tumor growth and metastasis; however, the underlying mechanism is poorly understood. Particularly, little is known how the tumor cell senses the acidic signal to activate the acidosis-mediated signaling. In this study, we show that breast cancer cells express acid-sensing ion channel 1 (ASIC1), a proton-gated cation channel primarily expressed in the nervous system. RNA interference, knockout and rescue experiments demonstrate a critical role for ASIC1 in acidosis-induced reactive oxidative species and NF-κB activation, two key events for tumorigenesis. Mechanistically, ASIC1 is required for acidosis-mediated signaling through calcium influx. We show that as a cytoplasmic membrane protein, ASIC1 is also associated with mitochondria, suggesting that ASIC1 may regulate mitochondrial calcium influx. Importantly, interrogation of the Cancer Genome Atlas breast invasive carcinoma data set indicates that alterations of ASIC1 alone or combined with other 4 ASIC genes are significantly correlated with poor patient survival. Furthermore, ASIC1 inhibitors cause a significant reduction of tumor growth and tumor load. Together, these results suggest that ASIC1 contributes to breast cancer pathogenesis in response to acidic tumor microenvironments, and ASIC1 may serve as a prognostic marker and a therapeutic target for breast cancer.

Comparative screening of single nucleotide polymorphisms in ß-casein and κ-casein gene in different livestock breeds of India.

The most polymorphic milk protein gene is ß-casein; 13 protein variants are known in cattle. Milk protein genetic polymorphism has received considerable research interest in recent years because of possible associations between milk protein and economically important traits in livestock. The present study was undertaken to explore the genetic polymorphisms in exon 7 of ß-casein and exon 4 of κ-casein genes in Arunachali yaks (Bos grunniens), Sahiwal (Bos indicus) cattle, malpura sheep (Ovis aries) and Gaddi goat (Capra hircus). Results of the study revealed presence of 11 SNP variants in all livestock species. Four SNPs were observed in Bos indicus; two SNPs in Bos grunniens; three SNPs in Ovis aries and three SNPs in Capra hircus. These variations are found to be synonymous in nature as these variations do not result in their corresponding amino acids. A total of five polymorphic sites have been described at the κ-casein (CSN3) locus in the Indian domestic Gaddi goat (Capra hircus) when compared with exotic goat (X60763) while sequence analysis of κ-casein gene in sheep showed three novel nucleotide changes in malpura sheep when compared with the exotic sheep (AY237637). These results highlight the importance of taking into consideration the CSN3 SNPs when performing selection for milk composition in dairy livestock breeds.

Characterization of excretory-secretory antigens of adult Toxocara canis by western blotting.

Toxocara canis is one of the most common helminth worm of dogs which continues to stimulate both public health concern alongside the higher scientific interest. It may cause visceral and ocular damage in humans especially in children. The identification of specific antigens of T. canis is important so as to develop better diagnostic techniques. Excretory-secretory (ES) antigens were prepared by culturing the adult T. canis worms in RPMI 1640 medium without serum supplementation followed by ammonium sulphate precipitation. These antigens were separated using sodium dodecyl sulphate-electrophoresis (SDS-PAGE). Recovered proteins ranged from 30 to 384 kDa. The specific reactivity of the T. canis excretory-secretory (TC-ES) proteins was checked by western blotting. The immuno-reactivity of the naturally infected dog sera with the TC-ES antigens showed five bands at 43, 57,105, 139 and 175 kDa. The immuno-reactivity of the hyper immune serum raised in rabbits against TC-ES antigens was observed with ten polypeptides of 21, 25, 30, 37, 45, 50, 57, 69, 77 and 105 kDa. Common antigens band were observed at 57 and 105 KDa. These antigens merit further evaluation as candidate for use in diagnosis of toxocariasis in humans and adult dogs.

Pattern, severity and circumtances of injuries sustained in road traffic accidents: a tertiary care hospital-based study.

INTRODUCTION: Expansion in road network, motorization, and urbanization in the country has been accompanied by a rise in road accidents leading to road traffic injuries (RTIs). Today RTIs are one of the leading causes of deaths, disabilities, and hospitalizations with severe socioeconomic costs across the world. OBJECTIVES: The following study analyses the: Age and sex distribution of injured in road traffic accidents (RTAs).Circumstances leading to RTA.Pattern and severity of injuries sustained in RTAs cases. DESIGN: Retrospective record-based study. MATERIALS AND METHODS: The aim of this study was to audit retrospectively the circumstances, severity, and pattern of injury sustained by vehicle occupants presenting to the Saraswathi Institute of Medical Sciences (SIMS) hospital Hapur, for a period of one year. Data were collected using the case sheets of 347 patients from the medical records section of hospital and analyzed using SPSS computer software version 16.0. Results are interpreted in terms of percentage, mean, chi-square, and z-test. RESULTS: The pattern and severity of injuries sustained by 347 vehicle occupants admitted to the emergency department of SIMS, Hapur were retrospectively documented. Male victims 258 (74.35%) were more commonly involved than females 89 (25.65%) and majority of victims 141 (40.63%) were in age group of 20-30 years. Urban victims 222 (64.00%) outnumbered rural. The most frequently injured body regions were the extremities 499 (53.54%), followed by maxillofacial180(19.31%).. Out of total 802 external injuries, the most common type of injury was lacerations 307 (38.28%), abrasions 306 (38.15%)and followed by bruises 154 (19.20%). Multiple external injuries were more common on upper limb 216 (26.93%), lower limbs 210 (26.18%) and face 170 (21.20%), while crush injuries were more predominently seen in both the limbs. While laceration were common on face 120 (38.83%). Injuries to the chest 19 (2.36%), abdomen13 (1.61%), and spine 11 (1.36%) were seen in roughy equal proprotion of victims. The bones on right side 55 (55.55%) were more commonly fractured which is statistically significant. Skull injuries were mostly found on frontal 77 (47.53%), followed by parietal bone 33 (20.37%), mostly on right side. CONCLUSION: RTAs constitute a major public health problem in our setting. Urgent preventive measures targeting at reducing the occurrence of RTAs are necessary to reduce the morbidity and mortality resulting from these injuries.

Prevalence of canine toxocariasis in Bareilly, Uttar Pradesh, India.

Toxocara canis is one of the most common parasitic helminth worm of dogs and also a causative agent of zoonotic disease in humans. This pilot study was conducted to determine the presence of T. canis infection in dog population in and around Bareilly, Uttar Pradesh, India. A total of 558 faecal samples both from stray and owned dogs were screened and overall 24.3 % dogs were found positive for T. canis. A comparison between owned and stray dogs suggests that the higher prevalence was observed in the latter group. The age of the dogs had a considerable influence on prevalence, with a much higher proportion of younger dogs being infected. Among the stray dogs, the infection rate is much higher (62.79 %) in pups, as compared to 7.8 % in adult. Similarly, of the owned dogs screened 41.74 % pups were infected while the infection rate in adults was only 3.38 %. The higher rate of prevalence of this parasite in dogs could be the source of soil contamination for transmission of Toxocariasis which is of public health importance in this region.

Comparative sequence analysis in the exon 5 of growth hormone gene in the various livestock species of India.

The aim of the study was to identify genetic polymorphism in growth hormone (GH) gene locus of six different livestock species using PCR-Direct DNA sequencing method. In exon 5 of GH gene, 10 SNPs variants were identified in all livestock species studied, namely Bubalus bubalis, Bos indicus, Bos frontalis, Bos grunniens, Ovis aries, and Capra hircus. Four SNPs were observed in Bubalus bubalis, two SNPs in Bos indicus, one SNP in Ovis aries, and three SNPs in Capra hircus. No changes were observed in Bos grunniens and Bos frontalis when compared with the template sequence and the SNPs observed in the present investigation may be useful in the marker assisted selection.

Glyceraldehyde-3-phosphate dehydrogenase of the parasitic nematode Haemonchus contortus binds to complement C3 and inhibits its activity.

Haemonchus contortus is an economically important gastrointestinal parasite that infects primarily sheep and goats. To survive inside the host, the parasite must overcome the host immune response. In this study, we have identified and characterized a complement-C3-binding protein (H.c-C3BP) from this parasite employing biochemical and molecular biology tools. Initially, a truncated form of the protein was isolated from the excretory-secretory products of the parasite using C3-Sepharose column that facilitated its identification by mass spectroscopy. Subsequently, the parent molecule was generated in E. coli, and sequence analysis confirmed it as glyceraldehyde-3-phosphate dehydrogenase (GAPDH). GAPDH reacted with the antiserum raised against the truncated protein, and the truncated protein reacted with anti-GAPDH antiserum. The protein inhibited complement function as measured by haemolytic assay and membrane attack complex (MAC) formation. Sera from H. contortus-infected animals reacted with GAPDH as well as the truncated form of the protein, which further lend support to protein secretion. Thus, the C3-binding property of H. contortus GAPDH is a new function, and it represents a new entity of complement-binding protein. Identification and characterization of H.c-C3BP should facilitate development of new therapeutics considering a key role of this protein in immune modulation.

Evaluation of the effectiveness of diagnostic & management decision by teleophthalmology using indigenous equipment in comparison with in-clinic assessment of patients.

BACKGROUND & OBJECTIVES: There is a concern on the quality and the usefulness of teleophthalmology images, particularly those using indigenous equipment, in making a diagnosis and treatment decisions in ophthalmology. The present study was done to compare the level of agreement and sensitivity and specificity of diagnosis and management decisions of various eye diseases by teleophthalmology using indigenous equipment, compared to the in-clinic assessment. METHODS: Patients having different eye diseases were evaluated by two ophthalmologists - one ophthalmologist examined the patient in clinic setting while the other ophthalmologist made the diagnosis and management decision based on images sent by teleophthalmology. The images were taken by the ophthalmic technician using digital imaging system and fundus camera. The clinical findings and management decisions by the two ophthalmologists were masked to each others. RESULTS: In diagnosis of anterior segment eye diseases such as cataract and corneal diseases there was good to very good agreement (kappa values of 0.68 and 0.91 for cataract and corneal diseases respectively) between in-clinic assessment and assessment by teleophthalmology. There was moderate agreement (kappa values of 0.52 and 0.48 for glaucoma and retinal diseases respectively) between in-clinic assessment and assessment by teleophthalmology for the diagnosis of glaucoma and retinal diseases. For the management decisions of patients, there was moderate level of agreement in all groups of eye diseases. INTERPRETATION & CONCLUSIONS: Teleophthalmology, using indigenous equipment was found to be effective in diagnosis and management decision of anterior segment eye diseases such as cataract and cornea, and with some modification and continuous training to the technicians could become an effective tool for screening and referral of glaucoma and retinal diseases.

Lidar quantification of bank erosion in Blue Earth County, Minnesota.

Sediment and phosphorus (P) transport from the Minnesota River Basin to Lake Pepin on the upper Mississippi River has garnered much attention in recent years. However, there is lack of data on the extent of sediment and P contributions from riverbanks vis-à-vis uplands and ravines. Using two light detection and ranging (lidar) data sets taken in 2005 and 2009, a study was undertaken to quantify sediment and associated P losses from riverbanks in Blue Earth County, Minnesota. Volume change in river valleys as a result of bank erosion amounted to 1.71 million m over 4 yr. Volume change closely followed the trend: the Blue Earth River > the Minnesota River at the county's northern edge > the Le Sueur River > the Maple River > the Watonwan River > the Big Cobb River > Perch Creek > Little Cobb River. Using fine sediment content (silt + clay) and bulk density of 37 bank samples representing three parent materials, we estimate bank erosion contributions of 48 to 79% of the measured total suspended solids at the mouth of the Blue Earth and the Le Sueur rivers. Corresponding soluble P and total P contributions ranged from 0.13 to 0.20% and 40 to 49%, respectively. Although tall banks (>3 m high) accounted for 33% of the total length and 63% of the total area, they accounted for 75% of the volume change in river valleys. We conclude that multitemporal lidar data sets are useful in estimating bank erosion and associated P contributions over large scales, and for riverbanks that are not readily accessible for conventional surveying equipment.

Vaccination of buffaloes with Fasciola gigantica recombinant glutathione S-transferase and fatty acid binding protein.

Fasciola gigantica, causative agent of tropical fasciolosis, inflicts substantial economic losses on the livestock industry, affecting severely buffalo productivity in the tropical countries. Very few vaccination trials with different target antigens against F. gigantica infection have been conducted in this host. Present study describes a vaccination trial in buffaloes with F. gigantica recombinant glutathione S-transferase and fatty acid binding protein. The two recombinant proteins were expressed in Escherichia coli and evaluated for their immunoprophylactic potential in buffalo calves, using montanide 70 M-VG, a mineral oil-based adjuvant, for delivering the antigens. Buffalo calves were distributed in three groups, with group I, II and III calves immunized with recombinant glutathione S-transferase, fatty acid binding protein and a cocktail of these two antigens, respectively. Immunization of the calves evoked a mixed IgG1 and IgG2 antibody response. Present vaccination trial in these animals achieved a maximum protection level of 35%, when the two antigens were used in combination. Eosinophils were measured in both immunized and non-immunized challenge control animals, which showed a steady increase in their count in response to immunization with both the antigens and infection with F. gigantica, respectively.

Pentoxifylline therapy in the management of oral submucous fibrosis.

OBJECTIVES: Oral submucous fibrosis is a common premalignant condition in the Indian subcontinent and is caused by chewing areca nut and other irritants in various forms. Its medical treatment is not yet fully standardized. In this study we compared the efficacy of Pentoxifylline as compared to placebo. MATERIALS AND METHODS: 75 patients suffering from oral submucous fibrosis were randomly divided into two groups A and B. Group A patients received placebo, while Group B patients received 400 mg. Pentoxifylline for a period of 7 months. Treatment outcome was evaluated on the basis of improvement in symptom and sign scores. Student's t test was applied for comparing the results. RESULTS: The improvement in total (i.e. symptoms + sign) score was 25% in group A and 49.15% in group B. This difference was found to be statistically significant. (p < 0.05) CONCLUSION: Treatment regimen of group B was more effective. No significant side effects were seen. A follow up study is required to assess long term outcome of this therapy.

Effect of cigarette smoking on the periodontal health status: A comparative, cross sectional study.

CONTEXT: Survey. AIMS: The objective of the study was to evaluate the periodontal health status among cigarette smokers and non cigarette smokers, and oral hygiene measures. SETTINGS AND DESIGN: Cross sectional study. MATERIALS AND METHODS: The study included 400 male (200 cigarette smokers and 200 non smokers) aged 18-65 years. The subjects were randomly selected from the patients attending dental out-patient department of civil hospital and Himachal Dental College, Sundernagar. Community Periodontal Index (CPI) score was recorded for each patient and a questionnaire was completed by each patient. STATISTICAL ANALYSIS USED: Chi square and t-test. RESULTS: Periodontal condition as assessed by CPI score showed that there was statistically significant difference in the findings between cigarette smokers and non-smokers. CONCLUSIONS: Within the limits of this study, positive association was observed between periodontal disease and cigarette smoking. It was found that cigarette smoking was associated with lesser gingival bleeding and deeper pockets as compared to non-smokers.

Comparative potential of cultured skin fibroblast, cumulus, and granulosa cell to produce somatic cell nuclear transfer (SCNT) preimplantation embryos in buffaloes (Bubalus bubalis) in relation to gene expressions.

To improve the efficiency of somatic cell nuclear transfer (SCNT)-derived embryos in buffaloes (Bubalus bubalis), skin fibroblast, cumulus, and granulosa cells were cultured up to the 15th passage and cloned embryos were produced from each cell type. At the 15th passage the cumulative population doublings (CPDs) in cumulus cells was higher (60.78) than skin fibroblast (57.12) and granulosa (56.05) cell lines. Gene expression of chromatin remodelling proteins, that is, HDAC1, DNMT1, DNMT3a, and DNMT3b, were comparable at all five passages (P-3, P-6, P-9, P-12, and P-15) groups in cumulus cells but different in skin fibroblast and granulosa cells. Cleavage and blastocyst production rate in cumulus (65.9 and 27.4%)-derived embryos was higher than skin fibroblast (63.8 and 24.3%) and granulosa (62.5 and 22.3%)-derived embryos. Expressions of HDAC1, DNMT1, and DNMT3a mRNA in cumulus-derived blastocysts were similar to IVF blastocysts (control), whereas skin fibroblast and granulosa-derived blastocysts expression was significantly different (p < or = 0.05). DNMT3b mRNA expression in all the three donor cell types and IVF control were similar. The expression pattern of these genes showed the effect of donor cell type with different epigenetic reprogramming capabilities for SCNT embryo production rate. Overall, results indicated that cumulus cells are the best nuclear donor for SCNT.