Annual Research & Review in Biology

Aim: The study aimed at evaluating in vivo wound healing effect of herbal ointment formulated with 15% w/w Trichodesma zeylanicumm methanolic root extract, plant commonly used as traditional medicine.
Methodology: The wound healing potential of T. zeylanicumm was assessed using excision wound model and various biochemical parameters; L-hydroxyl, Hexose amine, Malondialdehyde and Ascorbic acid. Treatments were administered daily topically to three groups of albino rats: Ointment base only, 15% w/w T. zeylanicumm methanolic ointment and Neosporin ointment for 20 days.
Results: The results showed wound closure was slow in albino rats treated with 15% w/w T. zeylanicummmethanolic extract ointment but increased gradually to 76.95% on day 17, while there was steady increase for Neosporin ointment treated group (from 36.01% on day 5 to 92.89% on day 17). Wound contraction ability of the herbal ointment was significantly greater p<0.01 on 13^th day. The Neosporin ointment treated group had short epithelialization time (19.33±1.53) compared to herbal ointment treated group (21.33±3.06). The levels of mucopolysacharide content in the herbal ointment treated group were significantly decreasing P<0.05(from 2.03±0.11 to 1.17±0.13 on 4^th to 12^th day respectively), while collagen content levels were significantly increasing PË‚0.05 in the herbal ointment treated group (2.59±0.10 and 2.63±0.1 on 8^th and 12^th day). However, the levels of mucopolysacharide and collagen contents were significantly higher in the Neosporin ointment than in the herbal ointment treated group. For the blood plasma samples, the malondialdehyde levels were significantly much higher in the herbal ointment treated group (1.74±0.13) compared to Neosporin ointment treated group (1.55±0.08). The ascorbic content levels were significantly higher in the Neosporin ointment sample (0.92±0.26) compared to herbal ointment treated group (0.64±0.14).
Conclusion: The results showed that T. zeylanicumm methanolic root extract has wound healing potential; however, further clinical and toxicological experimentation is needed to scientifically validate its use as a topical ointment.

Aims: To find out a scientific validation for the traditional knowledge of tribals of Chittoor District, India for their usage of Caralluma umbellata Haw to cure stomach disorder and pain.
Methodology: Antibacterial activity of Caralluma umbellata Haw was studied on a few Gram positive and Gram negative bacteria. The dry roots and stems were extracted using hexane, benzene, diethyl ether, chloroform, acetone and methanol and were tested for their antibacterial activity.
Results: The root extracts were found to be effective against most of the organisms than the stem extracts. The extracts were highly effective against Bacillus subtillis, Bacillus cereus, Escherichia coli and Staphylococcus aureus. Chloroform extracts of both roots and stems exhibited good antibacterial activity against Gram positive and Gram negative bacteria except Pseudomonas aeruginosa.
Conclusion: The demonstration of antibacterial activity of C. umbellata against Gram positive (B. subtilisand B.cereus) and Gram negative bacteria (E.coli) provides the scientific basis for its use in the traditional treatment of stomach disorder.

Background: Fertility is declining in many countries and there has been substantial interest in the potential adverse effects of exposure to environmental hazardous chemicals, including pesticides on male reproduction, and it was evident that using pesticides play as the major reasons for sperm decline besides tested pesticides are used widely recently, that is why it was very important the investigate their draw back in fertilization. The objective of the present study focuses on the spermiotoxicity of some pesticides such as profenofos, chlorpyrifos, and lambda-cyhalothrin on male albino mice.
Study Design: To assess the effect of tested pesticides on sperm morphology of male albino mice treated for 30, 60 and 90 consecutive days with different doses of pesticides (1/10, 1/40 and ADI LD₅₀).
Place and Duration of Study: Institute of animal health, Ministry of Agriculture, Egypt, between May 2011 and March 2012.
Results: Data suggest a potential association between exposures to tested pesticides and decreased sperm quality and increased teratospermic (abnormal sperm morphology). Further support for testicular toxicity comes from studies in laboratory albino mice that showed associations between exposure tested pesticides and sperm shape abnormalities, as well as dose-response relationships between exposure and a decline in epididymal sperm count and motility and increased abnormal sperm.
Conclusion: Tested pesticides can cause male reproductive system abnormalities that include reduced sperm production. It is also possible that the genetic information of the sperm may potentially be altered prior to fertilization.

Jatropha curcas is an important plant belonging to the family Euphorbiaceae which is a potential candidate for biofuel production. Genetic transformation protocol for J. curcas callus mediated by Agrobacterium tumefaciens were optimized using a pCAMBIA1303 plasmid which carries green fluorescent protein (GFP) gene as a reporter. Results obtained were based on the highest percentage of GFP expression which was observed three days post-transformation. Immersion of callus into 1×10⁵ cfu ml^-1 (OD_600nm 0.6) of A. tumefaciens LBA4404 with addition of 300 µM of acetosyringone for 45 min, two days of pre-culture and three days of co-cultivation periods were determined to be ideal for J. curcas callus transformation. Putative transformants were selected in the presence of 25 mg/l hygromycin. Surviving calli were transferred into proliferation media (MS with 1 mg/l NAA and 1 mg/l BAP) to proliferate the callus for further molecular analyses and to confirm the presence of the target GFP transgene in the putative transformants. Polymerase chain reaction (PCR) was carried out using a 35S specific primer pair confirmed the presence of the 454 bp of 35S promoter region from the transformed callus. Quantitative real-time PCR (qRT-PCR) was carried out to demonstrate the integration and copy number of the 35S promoter in the putative tranformants. The 35S promoter gene (178 bp) as a target gene and J. curcas actin gene (179 bp) which functions as reference gene was designed to detect the positive transformants and control sample in real-time PCR reaction analysis. The results indicated that the actin specific PCR product was present in both the control and transformed calli, however the 35S PCR product was found only in the positive transformants. The similarity in CT values confirmed that both the genes were present as single copy thus confirming a single integration event.

An attempt was made to understand the genetic polymorphism and phylogenetic relationships among the eighteen silkworm races/breeds comprising six each of bivoltines (BVs), multivoltines (MVs) and autosomal mutants of the silkworm Bombyx mori belonging to both indigenous and exotic origins, which were drawn from the germplasm of the Department. The DNA was extracted from the whole moth and amplification was carried out with inter simple sequence repeat (ISSR) marker technique. Four ISSR primers namely UBC-812, UBC-813, UBC-826 and UBC-827 were utilized of which two primers UBC-812 and UBC-826 generated a total of 88 clear, distinct and reproducible bands ranging from 500bp to 2000bp corresponding to an average of 44 bands/primer. Among 88 bands 18 bands were monomorphic and remaining 70 were polymorphic. Thus, the percentage of polymorphism revealed for UBC-812 primer was 100%, whereas UBC-826 revealed 60.86% indicating the occurrence of genomic variation from the inbred population.
The unweight pair group method with arithmetic means (UPGMA) was constructed from the ISSR marker profiles have clearly discriminated the BVs, whereas MVs and an autosomal mutant namely pre and a BV race namely C₁₀₈ were genetically inter-linked together in cluster-I and II respectively. The significance of genetic polymorphism and phylogenetic relationships through the ISSR study are herein discussed.

Aims: The purpose of this research is to examine antimutagenicity and anticancer effect of Biebersteinia multifida.
Study Design: Currently cancer is considered as one of the main factors of mortality globally. Many chemicals in our environment can cause genetic mutations and are potentially responsible for millions of cancer-related deaths. Nowadays the scientists are looking for food materials which can potentially prevent the cancer occurrence.
Place and Duration of Study: Biebersteinia multifida DC. dried roots were purchased from local market in Tehran bazar, the center of Tehran province, Iran, between June 2012 and January 2013.
Methodology: In this study human leukemia pre B-cells (Nalm-6) were cultured in RPMI 1640 [Sigma], supplemented with 10% fetal bovine serum (FBS), penicillin -streptomycin and L-glutamine. The cultures were incubated at 37ºC, 5% CO₂ and then inhibitory effect of ethanolic extract on their proliferation was measured by MTT assay. The ethanolic extract was evaluated in terms of antimutagenicity properties by a standard reverse mutation assay (Ames Test). This was performed with histidine auxotroph strain of Salmonella typhimurium (TA100).Thus, it requires histidine from a foreign supply to ensure its growth. The aforementioned strain gives rise to reverted colonies when expose to carcinogen substance (Sodium Azide).
Results: The ethanolic extract prevented the reverted mutations and the hindrance percent 51.2% in antimutagenicity test. During MTT, human leukemia pre B-cells revealed to have a meaningful cell death when compared with controls (P = .01).
Conclusion: This study demonstrates the antimutagenicity effect of Biebersteinia multifida DC, and suggests that it may be potentially usefull as anticancer agent.

Soil samples collected from petrol bunks and workshops in Madurai were subjected to serial dilution and the development of fungal colonies in PDA plates. One of the colonies were selected and identified as Aspergillus sp. based on staining and cultural characteristics. Efficiency of this fungus on the degradation of 2.5, 5, 7.5 and 10% concentrations of petrol in minimal broth was studied for sixteen days. Decline in pH and increase in optical density and amount of CO2 released was noticed indicating the degradation of petrol by the fungus indirectly. It was also confirmed by the appearance of new peaks in HPLC analysis after sixteen days of treatment. Hence this strain can be used in cleaning oil polluted sites.

Aim: The aim of this study is to analyze the 34spores species by using SEM that grown in Northern parts of Iran as a contribution to the knowledge about the general morphology and assess if these characteristics could be useful for systematic purpose.
Methodology: For Scanning Electron Microscope, the material treated with hot 3% sodium carbonate, for 2 min, washed, dehydrated, suspended in 96% ethanol and then transferred to acetate plates and finally coated with gold. The shape, ornamentation, the equatorial and polar diameter, and the number of cells in annulus in 38 spores were studied.
Results: ornamentations of spores consist of rugulate, ornate, echinate or microechinate, verrucate, perforate and spinule or spinuluse. Based on our results the identification key was devised using spore characteristics.
Conclusion: Based on our results, the spore sculpture could be useful for systematic purpose, specially in Polypodiaceae, Thelypteridaceae and Hydrofilicinae ferns.

Aim: The aim of this study was to report the seasonal and altitudinal variation in herbaceous biomass along with the productivity of area.
Place and Duration of Study: This study was conducted in Nikyal valley located at altitudinal range of 1500-1900 m within the longitude 74o 04′ to 10′ east and latitude 33o 26′ to 29′ north during July 2012 to June 2013.
Methodology: The forage biomass was calculated by Harvest method using 1m2 quadrat. Five quadrats were selected from each community permanently. Above ground grasses and forbs were clipped up to 2cm and were placed in separate polythene bags. Both grasses and forbs were separately weighed and average was taken. The results were expressed in Kilo grams per hectares. Monthly sampling over one year was carried to determine its productivity.
Results: The average dry biomass production was 854 Kg/ha. The July and August were the most productive months (1387 Kg/ha and 1335 Kg/ha, respectively). The total dry biomass, biomass contributed by grasses and herbs generally increased from July to October and thereafter it progressively decreased till February and then again started increasing from March.
Conclusion: The variation in Biomass controlled by the amount and timing of precipitation and temperature inputs during the growing season.

Aims: Determine the chemical composition and ruminal degradation and fermentation of Kochia scopariaat three sowing dates (D1, D2, D3) and two cutting ages, 120 (C1) and 165 (C2) days post seeding.
Place and Duration of Study: Department of Animal Nutrition, between August 2011 and June 2013.
Methodology: Samples were analyzed using a randomized statistical design 3x2. Chemical composition, in vitro gas production (GP) 96 h and in sacco ruminal degradation at 24h was determined.
Results: Crude protein content was higher (P< .05) for dates D2 > D3 > D1, and for cutting interval, C1 > C2 (P<0.05). Interaction, D2C2 and D3C1 were higher in comparison with the rest of interactions. In vitro GP (ml gas/ g DM) was not different (P>0.05) for A, b, c and lag time, for the main effects of sowing date and cutting interval. In sacco rumen degradable protein [RDP], D2 was higher (P< .05) in comparison with the rest of the treatments. There were not differences (P>.05) among cuts.
Conclusion: Kochia scoparia could be used as a protein source for ruminant feeding with a higher rumen degradability, which will be influenced by the sowing establishment date and in lower magnitude for the cutting age.

Aim: The aim of this study was to assess the palatability and animal preference of 110 plant species of Nikyal rangeland during summer and spring season.
Place and Duration of Study: This study was conducted in Nikyal valley located at altitudinal range of 1500-1900 m within the longitude 74º04′ to 10′ east and latitude 33º26′ to 29′ north during July 2012 to June 2013.
Methodology: The palatability was assessed by collecting data through random sampling. The respondents (300) were randomly selected from different inhabitants of the area and were interviewed for this purpose. The respondents were those rearing the animal species considered for this study and they include: large scale farmers, domestic farmers, shepherds and house wives.
Results: There were 8 tree species (7.27%), 21 shrub species (19.09%) and 81herbaceous species (73.63%) which were grazed among four species of animals in different season. over all 60 species (55%) were palatable and 50 species (45%) were non-palatable. Among the palatable 10 species (16.66%) were highly palatable, 22 species (36.66%) mostly palatable, 19 species (31.66%) less palatable and 9 (15%) were rarely palatable species. Most acceptable plant parts by animals are leaves (42, 53%) while the least acceptable parts are flowers/fruits (14, 18 %). The goat browsed 52 species (33%), sheep 36 species (23%), cow 31 species (20 %) and buffalo 37 species (24%).
Conclusion: Along with the plant type, the palatability is dependent on different factor including animal type, seasonal type, area, habitat and weather.

Aim: Chinese hamster ovary (CHO) cells were used to identify the most likely apoptotic biochemical pathway induced by Methyl-α-D-glucopyranoside (MDG), D-fructofuranose-β(2→6)-methyl-α-D-glucopyranoside (DFMDG) and β-D-fructofuranosyl-(2→6)-α-D-glucopyranoside (DFDG) treatments isolated from wild garlic (Tulbaghia violacea).
Methodology: Assays, which detect different morphological and biochemical changes in the CHO cells were used to examine the apoptosis induction by individual agents, paying special attention to the mitochondria membrane potential (∆ψm) depolarization.
Results: The major events of apoptosis, namely, translocation of phosphatidylserine, mitochondria membrane potential (∆ψm) depolarization and activation of caspase-3 in CHO cells were initiated within 2 hours of treatments. The onset of the apoptosis induction was indistinguishable among the MDG, DFMDG and DFDG, and similar to that of staurosporine.
Conclusion: The study shows that CHO cells treated with MDG, DFMDG and DFDG undergo apoptotic death through the mitochondrial (intrinsic) pathway.

Aims: The purpose of this study is to evaluate the production of aflatoxin B1 and extracellular protein patterns produced by A .flavus. 
Study Design: Aspergillus flavus is among of fungi that wide dispersal and its massive contamination on feed thus, inevitable to inhibit the growth of this fungus and subsequently production of aflatoxin.
Place and Duration of Study: Department of Animal Science, Faculty of Agricultural Science, Malayer University, between August 2012 and March 2013.
Methodology: The using by acetonic, ethanolic, water, methanolic, chloroform and ethyl ether extracts of Aloe Vera fresh leaves, Antitoxin activity of the extracts was evaluated by HPLC. Also, effect of extract on extracellular proteins of Aspergillus flavus, analysed by SDS-PAGE technique has been investigated.
Results: The acetone extract of Aloe Vera were used to evaluate and study on results obtained from HPLC analysis revealed the inhibition of aflatoxin production in 2000µL in 50mL group for 40.94% and in 2µL in 50mL group for 18.14%. The SDS-PAGE results showed that with decrease in fungal mycelium growth, the proteins production rate was also decreased.
Conclusion: From this study it can be concluded that acetone extracts of Aloe Vera can be effective in reducing aflatoxin B1 and extracellular protein production rates by the fungus A. flavus.

A hallmark of crustacean physiology is the periodic shedding of their old exoskeleton achieved by moulting as an external manifestation of a discontinuous growth process. Crustacean metabolism, reproduction and behavior are all affected by the periodic shedding of the exoskeleton and the characteristics of moulting cycles. There are major gaps in our understanding of moulting patterns of commercially raised shrimp; hence, further investigations on the duration of each moult stage, in-moult cycle and the relationship between biochemical constituents and moult cycling are essentially needed. The present research work was aimed at describing characteristic features and biochemical changes that occur at various stages (A-D) during moulting cycle of the Indian green tiger shrimp Penaeus semisulcatus. Experimental animals (wet weight, 20 ± 2 g) were reared individually in aerated plastic aquaria under normal laboratory conditions (12L: 12D; 28.2ºC) and 50% of the water was replaced daily for one month. Animals were provided twice daily with commercial Magnum Scampi feed ad libitum and were observed regularly for moulting by setal development and biochemical analysis in conjunction with established parameters for morphological changes during the moulting cycle. Results showed that muscle protein content peaked during the post-moult stage A (51.23 ± 2.51 mg/g) and gradually declined through the inter-moult and pre-moult stages. Similarly in the hepatopancreas, distinct increase in the total protein content was observed during the post-moult stage B (15.78 ± 0.26 mg/g) and a steady decline was noticed thereafter (p < 0.05). Significantly higher levels of total sugars from muscle tissues were observed in late post-moult stage B (51.23 ± 2.65 mg/g) and minimum level of total sugars were observed in late pre-moult stage D_2-3 both in muscle tissue (28.43 ± 2.98 mg/g) and in hepatopancreas (18.79 ± 1.62 mg/g). A sharp fall in lipid content of muscle tissues was observed in inter-moult stage C (19.54 ± 1.45 mg/g) and a corresponding decline (48.21 ± 3.25 mg/g) was observed in the hepatopancreas. The present study documents and further expands our understanding of the physiological and biochemical changes occurring in P. semisulcatusduring four different moulting stages and will provide useful criteria for identifying different stages in the life cycle of this commercially farmed shrimp.

Studies have accumulated over the years to emphasize the importance of the magnetic fields, MF, used as a safe alternative choice to improve agricultural crops. The most important applications of a magnetic field are the treatments of irrigation water, dry seeds, wet seeds and seedlings. The studies also included the application of magnetic fields at different periods of times (minutes, hours and days) and different doses. The effect of different magnetic fields varies depending on the plant species and age during exposure, as well as the intensity of magnetic field and period of exposure. The variation of MF exposure intensities may transform the effect on the plants from a positive to stressful factor. In this review article we highlighted the influence of a magnetic field, and why it is one of the best methods to enhance a plant’s growth and productivity.