Isolation, culturing and characterization of feeder-independent amniotic fluid stem cells in buffalo (Bubalus bubalis)     

Dev K  Gautam SK  Giri SK  Kumar A  Yadav A  Verma V  Kumar P  Singh B 《Research in veterinary science》2012,93(2):743-748

Heterogeneous amniotic fluid contains various cell types. The aim of this study was to characterize and differentiate some of the key stemness attributes of the amniotic fluid-derived cells in buffalo (Bubalus bubalis). The amniotic fluid (AF) cells were cultured without feeder cells, in DMEM containing 15% FBS, 1% non-essential amino acids, 1% penicillin/streptomycin/ampicillin, 1% vitamin solution, and 1% l-glutamine in 5% CO(2) in humidified air at 38.5±0.5 °C. After 6 days of culture different types of cells viz., star shaped (62.7%), spherical without nucleus (1.9%), spherical with nucleus (26.4%), pentagonal (0.4%), and free floating/rounded cells (8.3%) were observed. Most of the cells started anchorage-dependent growth after day 7 of the culture. Expression of alkaline phosphatase (AP) and Oct-4, Nestin and FGF-5 were observed from the AF cells at different passages. Using species-specific primers, a PCR amplicon of 200, 296 and 210 bp were observed for Oct-4, Nestin and FGF-5, respectively. The cells were found to have a normal karyotype at different passages. These results may contribute towards establishing non-embryonic pluripotent stem cells for various therapeutic and reproductive biotechnological applications in the species.  相似文献   

An early gene of the flavonoid pathway, flavanone 3-hydroxylase, exhibits a positive relationship with the concentration of catechins in tea (Camellia sinensis)     

Singh K  Rani A  Kumar S  Sood P  Mahajan M  Yadav SK  Singh B  Ahuja PS 《Tree physiology》2008,28(9):1349-1356

Tea (Camellia sinensis (L.) O. Kuntze) leaves are a major source of flavonoids that mainly belong to the flavan 3-ols or catechins. Apart from being responsible for tea quality, these compounds have medicinal properties. Flavanone 3-hydroxylase (F3H) is an abundant enzyme in tea leaves that catalyzes the stereospecific hydroxylation of (2S)-naringenin to form (2R,3R)-dihydrokaempferol. We report a full-length cDNA sequence of F3H from tea (CsF3H Accession no. AY641730). CsF3H comprised 1365 bp with an open reading frame of 1107 nt (from 43 to 1149) encoding a polypeptide of 368 amino acids. Expression of CsF3H in an expression vector in Escherichia coli yielded a functional protein with a specific activity of 32 nmol min(-1) mg protein(-1). There was a positive correlation between the concentration of catechins and CsF3H expression in leaves of different developmental stages. CsF3H expression was down-regulated in response to drought, abscisic acid and gibberellic acid treatment, but up-regulated in response to wounding. The concentration of catechins paralleled the expression data. Exposure of tea shoots to 50-100 microM catechins led to down-regulation of CsF3H expression suggesting substrate mediated feedback regulation of the gene. The strong correlation between the concentration of catechins and CsF3H expression indicates a critical role of F3H in catechin biosynthesis.  相似文献   

Evaluation of multi-nutrient extractants for determination of available P,K, and micronutrient cations in soil     

Sanjay K. Sharma  Ankita Sharma  Surinder Rana  Naveen Kumar 《Journal of plant nutrition》2018,41(6):782-792

Sixty soil samples (0–0.15 m depth) collected randomly from different districts of Himachal Pradesh were used to evaluate ammonium bicarbonate-diethylenetriaminepenta acetic acid (AB-DTPA) and acid ammonium acetate-ethylene diamine tetra acetic acid (AAAc-EDTA) over standard methods for P, K, and micronutrient cations. Among Olsen P, Bray P1, AB-DTPA P, and AAAc-EDTA P, available P (kg ha^?1) varied from 7.8 to 44.3, 9.5 to 61.1, 6.5 to 38.8, 10.5 to 52.1; available K (kg ha^?1) among NH4OAc, AB-DTPA, and AAAc-EDTA ranged from 103.6 to 372.3, 86.6 to 364.9, and 74.6 to 362.5. In case of micronutrients cations, AB-DTPA in general, extracted more amounts as compared to DTPA alone. AAAc-EDTA correlated highly with the Olsen P. AB-DTPA-K and AAAc-EDTA-K correlated well with standard method (NH4OAc) however; AAAc-EDTA had higher correlation with AB-DTPA for K extractability. Among methods for micronutrients highest significant correlations were observed between AB-DTPA and DTPA for Cu, Fe, and Zn.  相似文献   

Ascertaining clonal fidelity of tissue culture raised plants of <Emphasis Type="Italic">Bambusa balcooa</Emphasis> Roxb. using inter simple sequence repeat markers     

Divya Negi  Sanjay Saxena 《New Forests》2010,40(1):1-8

One of the major applications of plant tissue culture technology is that the regenerants are genetically identical to the mother plant. Although, of the various methods of in vitro propagation, axillary branching method is regarded as the safest, the possibility of genetic variation (somaclonal variation) cannot be ruled out even in this method. In the present study, inter simple sequence repeat (ISSR) marker assay was employed to validate the clonal fidelity of in vitro raised Bambusa balcooa plantlets multiplied by enhanced axillary proliferation up to 33 passages. Fifteen ISSR primers generated a total of 99 amplicons among the tissue-cultured progenies. Analysis of ISSR patterns revealed that the bands were shared by both the parent clump and the in vitro raised plants confirming the genetic stability in the latter.  相似文献   

Replacement of live feed by formulated feed: effect on the growth and spawning performance of Siamese fighting fish (Betta splendens,Regan, 1910)     

Sagar C Mandal  Narottam P Sahu  Mahinder P Singh Kohli  Pronob Das  Sanjay K Gupta  Sukham Munilkumar 《Aquaculture Research》2010,41(11):1707-1716

Partial replacement of live feed (LF) with formulated feed (FF) was performed in Betta splendens to evaluate their growth, survival and reproductive performance. Three hundred B. splendens fry of uniform size (mean weight 0.19 g±0.01) were equally distributed in five treatment groups with each of three replicates in glass aquaria of 35 L capacity. Fish were offered diets of different ratio of LF and FF like T1 (control) – 100% LF; T2 – 75% LF, 25% FF; T3 – 50% LF, 50% FF; T4 – 25% LF, 75% FF and T5 – 100% FF. Highest (P<0.05) body weight gain (% BWG) (67.1±1.03) and specific growth rate (2.34±0.02) were observed in the T2 group. Best feed conversion ratio was also found in T2 (2.40±0.11), which was similar to T1 and T3. The highest survival rate (%) was recorded in T1, T2 and T3 groups (97±1.7), which was similar to T4 and the lowest in T5 (49±2.0). The highest gonadal weight (0.12 g±0.01) and gonadosomatic index (15.17±0.50) was observed in the T3 group during the first sampling after 63 days. No significant changes in spawning performance were observed in the second sampling after 21 days after first sampling. However, T4 group registered the highest fecundity and fertilized eggs at the end of 105 days of experiment. From the present study, it concludes that LF can be successfully replaced to the extent of 25% by FF without any detrimental effect on the growth, survival and spawning performance of B. splendens.  相似文献   

Developmental competence of HMC(TM) derived bovine cloned embryos obtained from somatic cell nuclear transfer of adult fibroblasts and granulosa cells     

Bhojwani S  Vajta G  Callesen H  Roschlau K  Kuwer A  Becker F  Alm H  Torner H  Kanitz W  Poehland R 《The Journal of reproduction and development》2005,51(4):465-475

To enable us to handle a large number of oocytes at a given time and to have an increased throughput of cloned embryos, we attempted the Handmade cloning (HMC) technique, a zona-free method of bovine somatic cell nuclear transfer. Our objective was to study the developmental competence of the HMC derived embryos obtained using different types of somatic cells. A total of 6,874 cumulus-oocyte-complexes were used with either 7th or 11th passage fibroblasts (1st and 2nd groups, respectively), which were prepared from male animals, or granulosa cells (3rd group) as nuclei donors. The average cleavage rate was 65%, accompanied by a blastocyst rate of just 2% for the cleaved products and 5% for the >8-cell embryos, and there was no significant difference between the three groups. Out of 27 blastocysts recovered, 22 blastocysts were transferred to 22 recipients, resulting in two pregnancies. One pregnancy was lost after the fourth week while the other progressed to full term with the birth of a male calf. This first successful cloning of a male calf with the HMC technique in Europe indicates the successful adoption and establishment of this technique in our laboratory, and that this technique can be successful in producing viable embryos.  相似文献   

Diagnostic investigation of emerging viruses of companion animals.     

Sanjay Kapil  Teresa Yeary  Bill Johnson 《Veterinary Clinics of North America: Small Animal Practice》2008,38(4):755-74, vii

In this article, the authors are specifically concerned with the timely and accurate detection of emerging diseases of small animals that are viral in origin. Veterinarians are bound to encounter emerging viruses in their practice. The problem is unavoidable, because viruses are highly mutagenic. Even the immune response dictates the nature of virus that evolves in a host. If the clinical signs and diagnostic methods fail to correlate, the veterinarian should work with the diagnostic laboratory to solve the diagnostic puzzle.  相似文献   

Variation in cardiac markers and electrocardiographic alterations in young calves naturally infected with bovine tropical theileriosis     

Kapil Kumar Gupta  Mukesh Srivastava  Vikrant Sudan  Shanker Kumar Singh  Soumen Choudhury  Daya Shanker 《Tropical animal health and production》2018,50(6):1227-1230

The present study was designed to assess the deleterious effects of bovine tropical theileriosis on the cardiovascular system and the consequent myocardial involvement in young calves. Myocardial effects in parasitic diseases are often neglected. Hemolytic anemia, associated secondary hypoxia, and vasculitis are cardinal features of bovine theileriosis. In the present study, electrocardiogram (ECG) alongside serum cardiac troponin I (cTnI) and creatinine phosphokinase-myocardial band (CPK-MB) concentrations were analyzed in infected, treated, and control groups of young calves. Non-significant alterations were noticed in ECG. However, certain signs like sinus tachycardia, first-degree AV block, atrial premature complex, left atrial hypertrophy, and right atrial hypertrophy were found on consistent basis in infected calves. A significant increase in the serum concentration levels of cTnI and CPK-MB was noticed in infected calves followed by significant fall in both these biomarkers post treatment. cTnI and CPK-MB can definitely be used as myocardial markers in theileriosis-affected animals.  相似文献   

Differential expression of endometrial toll-like receptors (TLRs) and antimicrobial peptides (AMPs) in the buffalo (Bubalus bubalis) with endometritis     

Kharayat  Nitish Singh  Sharma G  Chethan  Kumar  Gandham Ravi  Bisht  Deepika  Chaudhary  Gangaram  Singh  Sanjay Kumar  Das  Gautum Kumar  Garg  Anil Kumar  Kumar  Harendra  Krishnaswamy  Narayanan 《Veterinary research communications》2019,43(4):261-269

Veterinary Research Communications - Toll like receptors (TLRs) and β-defensins expressed in the endometrium are part of the innate uterine defense mechanism (UDM). In the present study,...  相似文献   

Clinical efficacy of intrauterine cephapirin benzathine administration on clearance of uterine bacteria and subclinical endometritis in postpartum buffaloes     

Dawoud A. Nehru  Gurcharan. S. Dhaliwal  Mustafa. H. Jan  Ranjna S. Cheema  Sanjay Kumar 《Reproduction in domestic animals》2019,54(2):317-324

This study examined the effect of single IU administration of cephapirin on clinical recovery, clearance of uterine bacteria and reproductive performance of postpartum buffaloes with subclinical endometritis (SCE). Buffaloes (n = 86) at 35 days postpartum (DPP) with >10% polymorphonuclear (PMN) cells in endometrial cytosmears were designated as positive (SCEP, n = 29), and buffaloes with ≤10% PMN cell were designated as negative (SCEN, n = 57) for SCE. Out of 29 positive buffaloes, 15 were administered a single intrauterine dose of cephapirin benzathine on 40 DPP (SCEP‐CB), while the remaining 14 animals were kept as untreated control (SCEP‐C). All animals were observed regularly for oestrous signs and were again subjected to cytobrush sampling on the first postpartum (FPP) oestrus. Buffaloes positive for SCE at 35 DPP were later considered “recovered” if their PMN cells dropped to ≤5% on the FPP oestrus. Presence of Escherichia coli, Arcanobacterium pyogenes and Fusobacterium necrophorum in uterus was detected based upon PCR amplification of genes related to bacteria‐specific virulence factors. A total of 66.7% of SCEP‐CB group buffaloes recovered as compared to 28.6% in SCEP‐C (χ² = 4.21; p < 0.05). Rate of bacterial clearance did not differ between treated (38.5%) and untreated buffaloes (8.3%) (χ² = 1.67; p > 0.05). The median days to first service did not differ significantly (p > 0.05) among the three groups, whereas cephapirin administration reduced (p < 0.05) the days open by 14 days in SCEP‐CB compared to SCEP‐C buffaloes. SCEP‐CB buffaloes were as likely to conceive as SCEN, whereas SCEP‐C had 0.28 hazard ratio for pregnancy. In conclusion, a single treatment with cephapirin benzathine at 40 DPP improved the reproductive performance of buffaloes with subclinical endometritis.  相似文献