Influence of EDTA and magnesium on DNA extraction from blood samples and specificity of polymerase chain reaction

This study consisting of two trails conducted to examine the impact of initial EDTA level added to blood samples on quantity and quality of genomic DNA isolated from avian fresh blood and the influence ofinitial EDTA level with various levels of MgCl2 added to polymerase chain reaction (PCR) final volume on amplification pattern. EDTA level added to collected blood samples had no significant impact onquantity as well as quality of extracted genomic DNA. However, higher levels of EDTA increased the OD260 and enhanced the OD260/OD280 ratio beyond 1.8-1.9 which is broadly accepted as an indicator of high quality DNA. To avoid such an error, EDTA level in initial blood sample must not exceed 9 ìg/ìl blood. The initial amount of EDTA has no influence on PCR process if a valid DNA isolation protocol is used. Addition of MgCl2 from 1.0 to 2.4 ìl in a final volume of 25 ìl could support the amplification properly. Low levels of MgCl2 results in incomplete amplification but levels higher than 2.4 ìl impedes the amplification by negative interference to the whole reactions

Association of GSTP1 gene polymorphisms with performance traits in Deoni cattle

Heat stress directly and indirectly stimulate excessive intra/inter cellular toxicants such as malondialdehyde (MDA) and reactive oxygen species (ROS). Glutathione S-transferase Pi (GSTP1) plays the central role in the detoxification of ROS. In this investigation, we studied the genetic variation in GSTP1 gene using polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) technique in 70 Deoni breed cows. All the seven exons of GSTP1 gene were amplified by PCR using a total of six sets of primers. The genetic variants were determined by PCR-SSCP technique. Two unique SSCP patterns were observed in fragment 1, 3, 5 and 6 of GSTP1 gene. Sequence analysis with reference to GenBank Acc. no AC_000186.1 revealed polymorphisms at position 210, 746, 2438, 2439, 2443, 2507, 2695 and insertions between positions 707 and 708, 2700-2701 and 2775-2776. All the observed variations in coding regions were silent mutations. The cows with SSCP pattern B of fragment-5 had higher age at first calving while the cows with pattern A had higher lactation length and lactation yield as compared to pattern B (P≤0.05). The animals with pattern A of fragment 6 in GSTP1 gene had higher lactation length and lactation yield as compared to animals with pattern B. There was no significant difference in enzyme activity and calving interval in cows with different patterns in different SSCP fragments.Keywords: Deoni, GSTP1, polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP), single nucleotide polymorphism (SNP), heat stressAfrican Journal of Biotechnology Vol. 12(24), pp. 3768-377

Genetic variability of Indian yaks using random amplified polymorphic DNA markers

Indian yaks are categorized into common yaks, bisonian yaks and bareback yaks. Genetic similarities and divergence among them was analyzed by random amplified polymorphic DNA (RAPD) technique using ten decamer oligonuclotide primers. Only five primers (ILO 526, OPAV 15, ILO 1127, ILO 1065 and ILO 876) out of the ten primers tried produced consistant polymorphic fingerprints. Of the 76 fingerprints produced, 49 were present in all types, 21 were individual specific and 6 were polymorphic for different types. The pair wise comparison studied for different primers indicated the average bands sharing ranged from 78% in bareback to 88% in common type. There was no significant difference (P .0.05%) between the mean average percentage (MAPD) values observed between Indian yaks.Key words: Yak types, RAPD, DNA fingerprints

Influence of EDTA and magnesium on DNA extraction from blood samples and specificity of polymerase chain reaction

This study consisting of two trails conducted to examine the impact of initial EDTA level added to blood samples on quantity and quality of genomic DNA isolated from avian fresh blood and the influence of initial EDTA level with various levels of $MgCl_2$ added to polymerase chain reaction (PCR) final volume on amplification pattern. EDTA level added to collected blood samples had no significant impact on quantity as well as quality of extracted genomic DNA. However, higher levels of EDTA increased the $OD_{260}$ and enhanced the $OD_{260}/OD_{280}$ ratio beyond 1.8-1.9 which is broadly accepted as an indicator of high quality DNA. To avoid such an error, EDTA level in initial blood sample must not exceed $9 \mu g/ \mu l$ blood. The initial amount of EDTA has no influence on PCR process if a valid DNA isolation protocol is used. Addition of $MgCl_2$ from 1.0 to $2.4 \mu l$ in a final volume of $25 \mu l$ could support the amplification properly. Low levels of $MgCl_2$ results in incomplete amplification but levels higher than $2.4 \mu l$ impedes the amplification by negative interference to the whole reactions

Genetic variability of bovine GHR, IGF-1 and IGFBP-3 genes in Indian cattle and buffalo

The identification of genetic polymorphisms in the genes that play a crucial role in regulatiing growth and development of livestock enables us to evaluate the biological similarities and to acquire a better perspective of quantitative traits. The present study was undertaken to characterize genetic variability in the bovine growth hormone receptor (GHR), insulin-like growth factor 1 (IGF-1) and insulin-like growth factor binding protein 3 (IGFBP-3) genes among Bos indicus (Malnad Gidda, Khillar), Bos taurus (Holstein Friesian, Jersey) cattle and Asian water buffalo Bubalus bubalis (Murrah, Surti) using polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) analysis. These polymorphisms were confirmed by direct sequencing. The comparative gene sequence analysis in cattle and buffalo breeds revealed 18 single nucleotide polymorphisms (SNPs) across different loci. Eight SNPs were detected in the bovine growth hormone receptor (GHR) gene, of which four were found in the promoter region and four in the exon 4 region. In the IGF-1 gene, two SNPs were observed in the 5ˈUTR, three SNPs in the intron 3 region and two SNPs in the coding region of exon 4. Three SNPs were detected in the exon 2 region of the bovine IGFBP-3 gene. The frequency of rare alleles observed in the present study ranged from 0.04 to 0.16. The present results revealed high levels of genetic variability in the GHR, IGF-1 and IGFBP-3 genes in cattle and buffalo reared in India.Keywords: PCR-SSCP, genetic polymorphism, cattle, buffal

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Not AvailableThe major objectives of the present study were to estimate the incidence of hoof disorders and to identify the various risk factors associated with the hoof disorders in crossbred dairy cattle maintained under field conditions in the state of Karnataka, India. A total of 204 crossbred dairy cattle were investigated for hoof disorders. The overall incidence of hoof disorders was found to be 30.4%. The Holstein Friesian crossbred dairy cows (36.2%) were more vulnerable to hoof disorders than Jersey crossbred dairy cows (20.8%). The age of the animals greater than six years (60%), parity greater than three (44%), body weight greater than 410 kg (43%), animals having white-colored hooves (58.8%), injured hocks (51.7%) and animal having abnormal claw set (32.6%) were significant risk groups for higher incidence of hoof disorders Non-infectious lesions were more common than infectious lesions (94.7 vs 5.3%). Interdigital hyperplasia was the most commonly observed lesion (48.48%) followed by scissor claws (25%), horizontal cracks (18.95%), heel horn erosion (4.54%), white line disease (2.27%) and foot rot (0.76 %). Thus, our study establishes that non-infectious lesions like interdigital hyperplasia, scissor claws, horizontal cracks and white line diseases were the major hoof disorders in the study area compared to infectious lesions (heel horn erosion and foot rot). It can also be concluded that HF crossbred were more vulnerable to hoof disorder than Jersey crossbred and there was an increase in the incidence of hoof disorders with the advancement of age, higher body weight, light coloured hooves, injured hock conditions and abnormal claw set in crossbred dairy cattle.ICAR-NDR

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Not AvailableSmooth transition from pregnancy to lactation is important for high productive and reproductive performance during later postpartum period in dairy animals. On the other hand, the poor transition often leads to huge economic loss to dairy farmers due to compromised production and reproduction. Therefore, understanding the causes and consequence of metabolic changes during the transition period is very important for postpartum health management. In this review, metabolic changes with reference to negative energy balance in transition cow and its effect on health and reproduction during the later postpartum period in dairy animals are discussed besides the role of metabolic inflammation in postpartum performance in dairy animalsNot Availabl

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Not AvailableThis aim of this study was to assess the relationship, if any, of scrotal surface temperature gradient (SSTG), scrotal circumference (SC) and testicular covering thickness (TCT) with semen quality in Murrah buffalo breeding bulls. For this, buffalo breeding bulls (n = 130) were selected from four different semen centres. The ejaculates of each bull were evaluated for ejaculate volume (EV), mass motility (MM); concentration (SPC), motility (SM), viability and abnormalities (SA). The SSTG, SC and TCT of individual bulls were quantified using digital infrared thermography, measuring tape and ultrasonography, respectively. The bulls were divided into three groups on the basis of SSTG (≤ 4 °C, 4.1 to 6.4 °C and ≥ 6.5 °C), and SC (< 31, 31 to 35 and > 35 cm) and into two groups on the basis of TCT (5 to 7.2 and 7.4 to 10.4 mm). Results indicated the bulls with a larger temperature gradient and larger SC produced greater quality semen than those with a lesser temperature gradient. The MM (P < 0.01) and SPC (P < 0.05) varied among the groups along with SSTG. Among the SC groups the EV, MM, SPC and SM (P < 0.01), and viability (P < 0.05) varied as did the SC. The bulls with a lesser TCT had a lesser SA (P < 0.05) as compared to the group with the greater TCT. Buffalo bulls having a greater SSTG, SC and lesser TCT produced semen of greater quality and these variables may be used as criteria for breeding soundness evaluationNot Availabl

TRANSITION PERIOD INFLUENCES LACTATION PERFORMANCE IN ZEBU

Not AvailableWe evaluated the changes in the concentration of innate immune molecules (haptoglobin: Hp, serum amyloid A: SAA, IL-6, TNF-D, IL-1E, and IL-8), energy indicators [NEFA, dry matter intake (DMI) and body condition scoring (BCS)] during the transition period in dual-purpose Zebu (Deoni breed) cows in relation to milk yield. Blood collection was done at weekly intervals (–21±2, –14±1, –7±1, d pre-partum, day 0 (date of calving) and 3±1, 7±1, 14±1, 21±2 d postpartum period) for estimation of above plasma variables using commercially available bovine specific ELISA kits. We also recorded DMI and BCS during the corresponding period. Transition cows were classified based on their milk yield during the study period as high (6), medium (6) and low (6) yielding cows and data were analyzed by using Mixed-model repeated measure analysis. High yielding (HY) cows had significantly higher concentrations of SAA, TNF-D, and IL-6 during pre-partum and early postpartum period than low yielding (LY) cows. DMI was significantly higher in HY cows than MY (3rd and 7th d) or LY cows (21st d) while, BCS was significantly higher in HY than LY cows during pre-partum period (-7th d). LY cows had significantly higher concentration of NEFA during the postpartum period (14th and 21st d). It is concluded that the active functioning of the immune system and more dry matter intake in transition Deoni cows enabled to synthesise more milk during the postpartum period.ICAR-NDR