Current Research Trends in Biological Science Vol. 2

The Mangalitsa pig breed is one of the oldest breeds in Europe. The trend of an increasing n-3 polyunsaturated fatty acids concentration in animal tissue has continued, thus requiring an increase in using various types of oleaginous sources rich in n-3 fatty acid (FA). The objective of this study consist on investigate the influence of dietary linseed oil, as rich sources of the essential FA, particularly α-linolenic, on the plasma biochemical profile and the humoral immune response of the finishing Mangalitsa pigs. The implications of the addition of linseed oil on the FA composition of two types of muscles, longissimus dorsi and semitendinosus, were assessed as well. Twenty one Mangalitsa finishing barrows pigs, were assigned randomly to two groups fed: i) control diet based on classical ingredients; ii) linseed oil diet containing in addition vs. control diet, linseed oil produced by cold pressing 3.0%. Blood samples were aseptically collected by jugular venipuncture in order to determine biochemical and immunoglobulins profile. Samples of muscles were collected as well for establishing FAs composition by gas chromatography method. The use of linseed oil in the diets of barrow Mangalitsa pigs changed significantly most of the biochemical and immunological plasma parameters. The data show that the plasma constituents ranged within the physiological limits or slightly higher, but with no adverse effects on the health status of the animals. The meat from Mangalitsa pigs, improved nutritionally by the addition of linseed seed oil, as source rich in n-3 FA, brings several benefits beyond the controversies over the high-fat content: a higher content of α-linolenic FA; presence of the EPA, DPA, and DHA; a proper n-6:n-3 ratio, good for human health; a lower cholesterol level. These data are important because they can be used as a reference for biochemical determinations in humans due to the similarities existing between the human and pig organisms.

Aims: The study of strains belonging to local rhizobial population, concerning their diversity in the genetic, metabolic and symbiotic properties, and their prevalence in the microsymbiont population.

Methodology: 257 rhizobial isolates recovered from nodules of five pea (Pisum sativum cv. Ramrod) plants grown at one site were classified using PCR-RFLP analysis of 16-23S rRNA ITS. After that, for representative group of 55 strains, 16-23S rRNA ITS region was sequenced, nodA-F region was analyzed by PCR-RFLP and sequencing, metabolic capabilities were studied using Biolog`s and growth tests and symbiotic performance in plant tests were assayed.

Results: Individual plants were infected by numerous and diverse strains, however, in the entire sampled population of microsymbionts, only three large clusters of strains (one similar to Rhizobium pisi and two similar to Rhizobium leguminosarum) could be distinguished on the basis of PCR-RFLP and sequence analyses of 16S-23S rRNA ITS region. Rhizobium strains belonging to different groups varied in plasmid number and the amount of plasmid DNA, utilization of carbon and energy sources, growth on soil extract-based media and the ability for symbiotic plant growth promotion. The most numerous group of the isolates was characterized by the high plasmid DNA content, low number of utilized sugar substrates, and comprised numerous strains with low symbiotic efficiency.

Conclusion: Sampled population of pea microsymbionts had its own characteristic structure with clearly distinguishable sub-populations, composed of numerous strains - probably descendants of a few old lineages, which diversified in the lapse of time. These strains are still competing during root nodule colonization, resulting in the symbiosis of individual pea plants with broad spectrum of different Rhizobium strains

Aim: To analyze and compare the active sites of the multi-subunit (MSU) DNA dependent RNA polymerases (RNAPs)  of eubacteria and plant chloroplasts and find out the conserved motifs, metal-binding sites and catalytic regions and propose a plausible mechanism of action for the chloroplast MSU RNAPs using  Zea mays enzyme as a model enzyme.

Study Design: Bioinformatics, Biochemical, Site-directed mutagenesis and X-ray crystallographic data were analyzed.

Place and Duration of Study: School of Biotechnology, Madurai Kamaraj University, Madurai, India, between 2007-2013.

Methodology: Bioinformatics, Biochemical, Site-directed mutagenesis (SDM) and X-ray crystallographic data of these enzymes were analyzed. The advanced version of Clustal Omega was used for protein sequence analysis of the MSU DNA dependent RNAPs from various bacterial and chloroplast enzyme sources. Along with the conserved motifs identified by the bioinformatics analysis, the data already available by biochemical and SDM experiments and X-ray crystallographic analysis of these enzymes were used to confirm the possible amino acids involved in the active sites and catalysis.

Results:  Multiple sequence alignment (MSA) of RNAPs from both the sources showed many highly conserved motifs among them. The possible catalytic regions in the catalytic subunits β and β’ of eubacteria and their counterparts, viz. β, β’ and in  chloroplasts RNAPs consist of an absolutely conserved catalytic amino acid R, in contrast to a K as reported for DNA polymerases and single subunit (SSU) RNAPs. Besides, the invariant ‘gatekeeper/DNA template binding’ YG pair is also found to be absolutely conserved in the MSU RNAPs of chloroplasts, as reported in SSU, MSU RNAPs and DNA polymerases. The eubacterial β, the initiation subunit, is highly homologous to β subunit of chloroplast MSU RNAPs, i.e., the eubacterial and chloroplast β subunits exhibit very similar active site motifs, catalytic regions and distance conservations between the template binding YG pair and the catalytic R. However, the bacterial β’ elongation subunit is not completely similar to the  β’ elongation subunit of chloroplasts, but partly similar to the β’and β’’ subunits of chloroplast RNAPs. Interestingly, MSA analysis shows that the active sites are, in fact, shared  between β’ and β’’ in the MSU RNAPs of chloroplasts, i.e., the metal-binding site is found in the β’ subunit whereas the catalytic regions are located in β’’ subunit of chloroplast MSU RNAPs. Another interesting finding is, in the elongation subunits, i.e., in the eubacterial β’ and the chloroplast β’’ catalytic subunits, the proposed catalytic R is placed at double the distance, i.e., -16 amino acids downstream from the YG pair, in contrast to SSU RNAPs and DNA  polymerases where the distance is only ~8 amino acids downstream from the YG pair. An invariant Zn²⁺ binding motif reported in the eubacterial elongation subunit, viz., β’ is found in the β’’ subunits of chloroplasts. The catalytic R, along with the Zn binding motif is shifted towards the N-terminal in the elongation subunit of PEP.

Conclusions: MSA have shown that in both the MSU RNAPs of eubacteria and chloroplasts, the active sites, catalytic amino acids and metal-binding regions are absolutely conserved both in the initiation and elongation subunits. Therefore, it is suggested that the MSU RNAPs of chloroplasts may also follow very similar polymerization and proof-reading mechanisms as proposed for eubacteria. MSA data and the available experimental data show that both the eubacterial and chloroplast MSU RNAPs would have possibly evolved from a common ancestor.

Aims: Camelus dromedarius (dromedary or one-humped camels) are known to endure harsh conditions including extreme temperatures and high solar ultraviolet (UV) radiation in desert wilderness areas. This remarkable survival in the harsh desert conditions is attributed to distinctive bodily features enabling them to cope with this toxic environment. The present study hypothesized that the oil rendered from camel hump fat, consisting of saturated fatty acids with omega 3, 6, 9 and Vitamin E, has contributed to shield/protect/prevent UVA radiation damage.

Place and Duration of Study: This study was conducted in College of Veterinary Medicine, Baghdad University, Iraq, between June 2011 and July 2012.

Methodology: White BALB/c mice aged about 3 - 4 months weighing 24 - 31 gm were divided into four groups. Mice were shaved and three groups received different treatments of daily exposure to UVA radiation and one group was untreated as a control.

Results: Histopathological examinations of mice treated with camel oil prior to or following UVA radiation demonstrated that the camel oil acts as a protective agent, namely, protection of mice skin tissue from radiation-induced apoptosis. The mice treated with oil derived from cows and fat-tailed sheep demonstrated no improvement or worse results than untreated (control) mice. The results suggest that the camel oil protects the mice from UVA radiation injury and also acts as an injury-mitigator when applied following UVA exposure.

Conclusions: The major components in the camel hump fat including saturated fatty acids and noticeable values of omega 3, 6, 9 and Vitamin E have contributed to shield/protect/prevent UVA radiation damage, and may also have unique anti-tumor properties with novel dual radiation-protection and mitigation/healing properties.

Of the diverse hydrocarbonoclastic bacteria isolated from crude oil polluted IKO River estuarine and freshwater ecosystems, the bacterial isolate, ESW₁ and FSW₂ identified as Citrobacter amalonaticus strain – FSW₂ and Citrobacter amalonaticus strain - ESW₁.These efficiently degrade Bonny light crude oil sample recording 82.1 and 69.2% degradation respectively after 28 days of incubation. Biodegradability of the components of Bonny light crude oil was determined by gas chromatographic analysis. The chromatographic analysis after 28 days of incubation at 28°C also revealed that during the degradation of Bonny light crude oil, there was a decrease of the total hydrocarbon content (THC) from 10,906.9 mg L^-1 to 1,947.4 mg L^-1 and 3,357.9 mg L^-1 respectively by FSW₂ and ESW₁. These results suggest that Citrobacter amalonaticus is a good candidate for microbial seeding of Bonny light crude oil polluted aquatic ecosystem.

Aim: Mosquito are a major problem in almost all tropical and subtropical countries, as they are responsible for the transmission of pathogens which causes different diseases.

Study Design: The present study deals with the Comparison of Puntius conchonius and Gambusia affinis for feeding rate of mosquito larvae.

Place and Duration of Work: The mosquito larvae collected from badapur, Tal. Yeola Dist. Nashik between 2010 -2011.

Methodology: Fishes introduced into experimental jar individually and acclimatized for 1 hrs. Then 200 to 500 larvae introduced into jar for each fish. Readings were taken after 10 min, 20 min, 30 min initially and then after 1 hr, 3 hr, 6 hr, 9 hr, 12 hr and 24 hours, respectively. It determines feeding rate, food consumption and larvivorous potential of fishes.

Results: The comparative account of the feeding rate of two different species which came as Puntius conchonius > Gambisia affinis. Whereas, when the food consumption pattern was different i.e. Gambisia affinis > Puntius conchonius.

Conclusion: The length and weight increases, the feeding rate also increases. It was found that the larvivorous activity of the ornamental fishes was greater than the well-known Gambusia affinis.

A fish specimen of total length (5.5 cm) captured from Pharping was instantly photographed and preserved in 95% ethanol. Its morphological characters including morphometric and meristic data were recorded using digital calipers and backlit dissecting scope. The tissue removed from the caudal fin was placed in 95% ethanol and sent to a reputed lab in Kathmandu city for molecular analysis. The CO1 segment of mitochondrial DNA was amplified and sequenced using a cocktail of four fish specific primers. The DNA sequence (607 bp) was input in BLAST to find out highly similar sequences in GenBank. The sequence was uploaded in GenBank to obtain accession number. The external morphology mainly homodont teeth in jaws forming a crescent –shaped band on upper jaw and absence of post labial groove identified the fish specimen as Parachiloglanis hodgarti and it was found to be distantly related with other glyptosternoid fish based on multiple DNA sequence analysis. Based on the BLAST result the fish specimen was found to be homologous with Myersglanis blythi from Mugu, Nepal. The present study concluded that M. blythi from Mugu was most probably P. hodgarti which was misidentified, and further study on distribution and abundance of P. hodgarti and M. blythi was recommended as the peculiar water resources of Pharping was noteworthy for the study of rare species.

Seeds are a main dietary component for humans and animals. In this work, the physical, spectral, thermal, mineral and phenolic characteristics of seeds from 47 species belonging to 10 families (Arecaceae, Amaranthaceae, Anacardiaceae, Asteraceae, Fabaceae, Lamiaceae, Malvaceae, Rutaceae, Sapindaceae and Sapotaceae) are described. The seeds had moisture and ash contents in the 1.6–12.9% and 1.0–5.5% range, respectively, and the seed coat represented from 12 to 85% of the seed weight, depending on the species. The protein, starch and oil-associated vibrations in the seeds were characterized by infrared spectroscopy. The thermal behavior of the seeds, which involved an initial dehydration step, followed by free/bound moisture and volatile extractives release, and by protein, starch and oil decomposition, was studied by thermogravimetric (TG) and differential scanning calorimetry (DSC) techniques, evidencing noticeable differences between families. The total polyphenol and mineral contents widely varied from 185 to 3441 mg/100 g and from 831 to 7904 mg/100 g, respectively. Cluster analysis was used to categorize seed potentiality in terms of mineral contents. Variations in seed proximate parameters, spectral, thermal, phenolic and mineral characteristics as a function of the family to which the seeds belong to are also discussed.

The research was conducted to determine the anticarcinogenic properties of “rumput mutiara” (Hedyotis corymbosa (L.) Lam) and “pohpohan” (Pilea trinervia (Roxb.) Wight), by the microculture tetrazolium salt (MTT) assay on the human breast carcinoma dependent-hormone (MCF-7) cell lines. The preliminary results showed that the “rumput mutiara” extract displayed the cytotoxic effects against MCF-7 with IC₅₀-value of 22,67 µg/ml. However, the “pohpohan” extract did not show the IC₅₀-value against MCF-7 cell lines. The antioxidative activities of the extracts which could contribute to their cytotoxic properties were also studied. The “rumput mutiara” extract was found to have higher antioxidant activity compared with “pohpohan” extract. The strong cytotoxic properties of the “rumput mutiara” extract could be due to its high antioxidant activity. The combination effect of “rumput mutiara” and soursop were also studied.