Population genetic and phytochemical dataset of Saraca asoca: A traditionally important medicinal tree.

The data presented in this article is in support of the research paper "Genetic and phytochemical investigations for understanding population variability of the medicinally important tree Saraca asoca to help develop conservation strategies" Hegde et al., 2018. This article provides PCR based Inter-Simple Sequence Repeat (ISSR) and HPLC datasets of 106 individual samples of Saraca asoca collected from various geographical ranges of the Western Ghats of India. The ISSR data includes information on genetic diversity and images of population structures generated through amplified DNA products from samples of Saraca asoca leaf. Phytochemical data obtained from HPLC includes concentration (mg/g) of gallic acid (GA), catechin (CAT), and epicatechin (EPI). The data also presents information obtained from various statistical analysis viz. standard error of the mean values, distribution variables, prediction accuracy, and multiple logistic regression analysis.

Genetic and phytochemical investigations for understanding population variability of the medicinally important tree Saraca asoca to help develop conservation strategies.

Saraca asoca (Roxb.) De Wilde (Caesalpiniaceae) is a highly traded IUCN red listed tree species used in Ayurvedic medicines for the treatment of various disorders, especially gynaecological problems. However, information about the genetic variations between populations and corresponding variation in specialized metabolites of S. asoca remains unclear. To address this issue, we analysed 11 populations of S. asoca with 106 accessions collected from Western Ghats of India using ISSR markers along with selected phytocompounds using RP-HPLC. Twenty primers were screened, out of which seven were selected for further analysis based on generation of clear polymorphic banding patterns. These seven ISSR primers produced 74 polymorphic loci. AMOVA showed 43% genetic variation within populations and 57% among the populations of S. asoca. To estimate the genetic relationships among S. asoca populations, UPGMA and Bayesian Models were constructed, which revealed two clusters of similar grouping patterns. However, excluding minor deviations, UPGMA and dissimilarity analysis showed close association of genotypes according to their geographical locations. Catechin (CAT), epicatechin (EPI) and gallic acid (GA) were quantified from bark and leaf samples of corresponding genotypes collected from 106 accessions. ROC plots depicted the sensitivity and specificity of the concentrations of tested phytocompounds at various cut-off points. Although, multiple logistic regression analysis predicted some association between few loci with GA, EPI and CAT, but PCA for phytochemical data failed to distinguish the populations. Overall, there were no significant trends observed to distinguish the populations based on these phytocompounds. Furthermore, the study advocates the delineate provenance regions of S. asoca genotypes/chemotype snapshots for in-situ conservation and ex-situ cultivation.

Molecular identification of Saraca asoca from its substituents and adulterants.

ABSTRACT: Saraca asoca (Roxb.) De Wilde is an important medicinal plant from the Western Ghats of India, traditionally used in treatment of various gynecological disorders. Increasing commercial demand and decreasing numbers has resulted in this plant becoming endangered with crude drug materials being extensively substituted/adulterated with other plant species. The present study was undertaken with the objective of development and evaluation of multivariate cluster analysis of ISSR fingerprints against rbcL-based DNA barcodes as tool to understand the relationships and to differentiate common adulterants and substituents from S. asoca. ISSR-based Hierarchical Cluster Analysis was carried out on 41 samples of S. asoca and 5 each of the 5 common substituent/adulterant plants and the clustering patterns were evaluated against DNA-sequence-based barcoding of rbcL region of their plastids. Factorial analysis and Principal Coordinate Analysis revealed distinct groups of genetic pools of respective taxa thereby confirming the utility of ISSR fingerprinting as a useful tool for differentiation between the genuine and the adulterants/substituents. NCBI-BLAST search on DNA barcode rbcL region confirmed the results of ISSR assays. Therefore, our study demonstrated the utility of simple, cost-effective method of ISSR fingerprinting coupled with rbcL barcoding in differentiating this important medicinal plant from its common adulterants/substituents.

Determination of betulinic acid, oleanolic acid and ursolic acid from Achyranthes aspera L. using RP-UFLC-DAD analysis and evaluation of various parameters for their optimum yield.

Achyranthes aspera L. is a well known herb commonly used in traditional system of Indian medicine to treat various disorders, such as cough, dysentery, gonorrhea, piles, kidney stone, pneumonia, renal dropsy, skin eruptions, snake bite, etc. Here, we used RP-UFLC-DAD method for determining triterpenoids betulinic acid (BA), oleanolic acid (OA) and ursolic acid (UA) from A. aspera. Optimum yield of these compounds were studied and evaluated using parameters viz., method of extraction, time of extraction, age of plant and plant parts (leaves, stem and roots). Linear relationships in RP-UFLC-DAD analysis were obtained in the range 0.05-100 µg/mL with 0.035, 0.042 and 0.033 µg/mL LOD for BA, OA and UA, respectively. Of the variables tested, extraction method and parts used significantly affected content yield. Continuous shaking extraction (CSE) at ambient temperature gave better extraction efficiency than exposure to ultra sonic extraction (USE) or microwave assisted extraction (MAE) methods. The highest content of BA, OA and UA were determined individually in leaf, stem and root extracts with CSE. Collective yield of these triterpenoids were higher in leaf part exposed to 15 min USE method. To best of our knowledge, the study newly reports UA from A. aspera and the same was confirmed using ATR-FT-IR studies. This study explains the distribution pattern of these major triterpenoids and optimum extraction parameters in detail.

Vibrio cholerae O1 El Tor variant and emergence of Haitian ctxB variant in the strains isolated from South India.

Cholera still continues to be an important cause of human infection, especially in developing countries that lack access to safe drinking water and proper sanitation. In the present study, we report the emergence of new variant form of V. cholerae O1 El Tor biotype with a novel mutation in ctxB in strains isolated from various outbreaks during 2010-2014 in Belgaum situated in north-west Karnataka, India. A total of 14 occurrences of cholera were documented from Belgaum Division of North Karnataka during the 4-year period from 2010 to 2014. All the V. cholerae O1 isolates were subjected to DAMA PCR to detect the three different allelic subtypes of ctxB and PCR-based detection of virulent genes, and subsequently, 14 strains (one strain from each outbreak or sporadic case) were subjected to ctxB gene sequence and pulsed-field gel electrophoresis (PFGE) analysis. A total of 54 V. cholerae O1 strains were obtained of which 21 strains isolated during 2010-2011 had classical ctxB and remaining 33 strains isolated during 2012-2014 belonged to Haitian variant. In the cluster analysis, the PFGE profiles were divided into clades A with and B. Clade A contained eight strains with 94 % similarity and Haitian type of ctxB. Clade B contained six strains and had Haitian type of ctxB except one with classical ctxB. To the best of our knowledge, this is the first report of the Haitian variant of V. cholerae O1 Ogawa causing outbreaks and sporadic cases of cholera in South India.

Tribes in Karnataka: Status of health research.

The south Indian State of Karnataka, once part of several kingdoms and princely states of repute in the Deccan peninsula, is rich in its historic, cultural and anthropological heritage. The State is the home to 42,48,987 tribal people, of whom 50,870 belong to the primitive group. Although these people represent only 6.95 per cent of the population of the State, there are as many as 50 different tribes notified by the Government of India, living in Karnataka, of which 14 tribes including two primitive ones, are primarily natives of this State. Extreme poverty and neglect over generations have left them in poor state of health and nutrition. Unfortunately, despite efforts from the Government and non-Governmental organizations alike, literature that is available to assess the state of health of these tribes of the region remains scanty. It is however, interesting to note that most of these tribes who had been original natives of the forests of the Western Ghats have been privy to an enormous amount of knowledge about various medicinal plants and their use in traditional/folklore medicine and these practices have been the subject matter of various scientific studies. This article is an attempt to list and map the various tribes of the State of Karnataka and review the studies carried out on the health of these ethnic groups, and the information obtained about the traditional health practices from these people.

Evaluating Nothapodytes nimmoniana population from three localities of Western Ghats using camptothecin as phytochemical marker and selection of elites using a new-content range chart method.

BACKGROUND: Nothapodytes nimmoniana (Grah.) Mabb. is a high valued medicinal plant endemic to Western Ghats of India, distributed in fragmented populations. The plant is valued for potent anticancer drug camptothecin (CPT). OBJECTIVE: The study compares and expounds variation in CPT content from leaves and stems of N. nimmoniana obtained from three populations of Western Ghats, India. The study also describes a method for categorizing these populations using content range chart (CRC) method for percent yield of CPT. MATERIALS AND METHODS: A total of 60 samples were investigated including ten each of leaves and stems from three localities. Micro-extraction method was implemented to extract CPT. reversed phase ultra-performance liquid chromatography photo diode array technique was used to quantify CPT. RESULTS: Leaf samples of an individual collected from Joida, yielded lowest CPT content (0.002 ± 0.000 g/100 g), whereas a stem sample from Amgaon, yielded highest CPT content (0.123 ± 0.006 g/100 g). The findings suggest great variation in individuals producing and accumulating CPT. Using this data along with earlier published work, five categories of CPT yielding plants were made viz. I: Very low: <0.020, II: Low: 0.021-0.039, III: Moderate: 0.040-0.059, IV: High: 0.060-0.079 and V: Very high: >0.080. Based on CPT content in leaves, majority of individuals were under very low category (I(st)) and on the other hand stem samples were in 'II' category. Besides, very few individuals were observed in category 'V'. CONCLUSION: The study expounds use of CRC method for identifying elite population and suggests the need for its conservation.

Twin outbreak of cholera in rural North Karnataka, India.

BACKGROUND & OBJECTIVES: Successive outbreaks of acute watery diarrhoea occurred in Talikoti and Harnal, located in Bijapur District of the southern Indian s0 tate of Karnataka, in July and August 2012, respectively. These outbreaks were investigated to identify the aetiology and epidemiology. METHODS: Information was collected from the local population and health centres. Stool and water samples were collected from the admitted patients and their drinking water sources. Standard microbiological and PCR techniques were employed for isolation and characterization of the pathogen. RESULTS: While 101 people (0.38%) were affected in Talikoti, 200 (20.94%) were affected in Harnal which is a small remote village. All age groups were affected but no death occurred. While the outbreak was smaller, longer and apparently spread by person to person contact in Talikoti, it occurred as a single source flash outbreak at Harnal. A single clone of toxigenic Vibrio cholerae O1 Ogawa biotype El Tor was isolated from the two stool samples obtained from Talikoti and subsequently from three of five stool samples obtained from Harnal indicating village to village spread of the aetiological agent. Striking similarity in antibiotic resistance profiles of these isolates with a particular strain isolated from the city of Belgaum, 250 km away, in 2010, prompted tracking the lineage of the V. cholerae isolates by DNA fingerprinting. Random amplified polymorphic DNA (RAPD) fingerprinting assay helped confirm the origin of the incriminating strain to Belgaum. INTERPRETATION & CONCLUSIONS: Our study reported the first twin outbreak of cholera in two remote areas of Bijapur district, Karnataka, south India. It also indicated the need for immediate preparedness to deal with such emergencies.

Accumulation and trends in distribution of three triterpenoids in various parts of Achyranthes coynei determined using RP-UFLC analysis.

BACKGROUND: Achyranthes coynei Sant. (Family: Amaranthaceae) is a rare endemic medicinal plant used by local traditional practitioners to treat various diseases. The plant has been reported for promising antibacterial and antioxidant activities. However, the plant is not explored for its phytocompounds, especially triterpenoids. OBJECTIVE: TO STUDY THE ACCUMULATION AND TRENDS IN DISTRIBUTION OF TRITERPENOIDS: betulinic, oleanolic and ursolic acids (BA, OA and UA) in leaf, stem, root and inflorescence of A. coynei. MATERIALS AND METHODS: Extraction was achieved using refluxing and reversed phase-ultra flow liquid chromatographic (RP-UFLC) technique was employed for determination. Separation of triterpenoids was achieved on a Hibar 250-4.6 mm, 5 µ, Lichrospher 100, C18e column using methanol and water (90:10) as mobile phase (pH adjusted to 5.0 using GAA) in an isocratic mode. RESULTS: Oleanolic acid was higher in leaf (0.172 ± 0.009%) followed by stem (0.035 ± 0.002%) and root (0.028 ± 0.001%). Ursolic acid was accounted to be highest in the inflorescence (0.099 ± 0.005%). The contents of BA and UA were lower than OA in leaf and stem while it was remained undetected in roots of A. coynei. CONCLUSION: THE TRITERPENOIDS: BA, OA and UA were detected, quantified and reported for the first time from A. coynei. In the present study leaves were found to be the major source of BA and OA, whereas inflorescence was for UA.

Non-codified traditional medicine practices from Belgaum Region in Southern India: present scenario.

BACKGROUND: Traditional medicine in India can be classified into codified (Ayurveda, Unani, Siddha, Homeopathy) and non-codified (folk medicine) systems. Both the systems contributing equally to the primary healthcare in India. The present study is aimed to understand the current scenario of medicinal practices of non-codified system of traditional medicine in Belgaum region, India. METHODS: The study has been conducted as a basic survey of identified non-codified traditional practitioners by convenience sampling with semi structured, open ended interviews and discussions. The learning process, disease diagnosis, treatment, remuneration, sharing of knowledge and socio-demographic data was collected, analysed and discussed. RESULTS: One hundred and forty traditional practitioners were identified and interviewed for the present study. These practitioners are locally known as "Vaidya". The study revealed that the non-codified healthcare tradition is practiced mainly by elderly persons in the age group of 61 years and above (40%). 73% of the practitioners learnt the tradition from their forefathers, and 19% of practitioners developed their own practices through experimentation, reading and learning. 20% of the practitioners follow distinctive "Nadi Pariksha" (pulse examination) for disease diagnosis, while others follow bodily symptoms and complaints. 29% of the traditional practitioners do not charge anything, while 59% practitioners receive money as remuneration.Plant and animal materials are used as sources of medicines, with a variety of preparation methods. The preference ranking test revealed higher education and migration from villages are the main reasons for decreasing interest amongst the younger generation, while deforestation emerged as the main cause of medicinal plants depletion. CONCLUSION: Patrilineal transfer of the knowledge to younger generation was observed in Belgaum region. The observed resemblance in disease diagnosis, plant collection and processing between non-codified traditional system of medicine and Ayurveda require further methodical studies to establish the relationship between the two on a more objective basis. However, the practice appears to be at crossroads with threat of extinction, because of non-inheritance of the knowledge and non-availability of medicinal plants. Hence conservation strategies for both knowledge and resources at societal, scientific and legislative levels are urgently required to preserve the traditional wisdom.

Compound Specific Extraction of Camptothecin from Nothapodytes nimmoniana and Piperine from Piper nigrum Using Accelerated Solvent Extractor.

Effects of varying temperatures with constant pressure of solvent on extraction efficiency of two chemically different alkaloids were studied. Camptothecin (CPT) from stem of Nothapodytes nimmoniana (Grah.) Mabb. and piperine from the fruits of Piper nigrum L. were extracted using Accelerated Solvent Extractor (ASE). Three cycles of extraction for a particular sample cell at a given temperature assured complete extraction. CPT and piperine were determined and quantified by using a simple and efficient UFLC-PDA (245 and 343 nm) method. Temperature increased efficiency of extraction to yield higher amount of CPT, whereas temperature had diminutive effect on yield of piperine. Maximum yield for CPT was achieved at 80°C and for piperine at 40°C. Thus, the study determines compound specific extraction of CPT from N. nimmoniana and piperine from P. nigrum using ASE method. The present study indicates the use of this method for simple, fast, and accurate extraction of the compound of interest.

Chemical composition and antimicrobial activity of the essential oil of the leaves of Feronia elephantum (Rutaceae) from north west Karnataka.

The essential oil of the leaves of Feronia elephantum Corr. was analyzed by gas chromatography and gas chromatography/mass spectrometry. The main constituents were beta-pinene (28.4%), Z-anethole (22.1%), methyl chavicol (12.0%) and E-anethole (8.1%), among thirty-three identified compounds, which represented 92.6% of the total oil. The antimicrobial activity was tested against five Gram-positive and eight Gram-negative bacteria, and four fungi. The oil was active against Micrococcus luteus (Gram-positive bacterium), Proteus mirabilis (Gram-negative bacterium), Penicillium chrysogenum and Aspergillus niger (fungi) with MIC values of 0.31 +/- 0.06, 0.52 +/- 0.10, 0.20 +/- 0.50 and 0.26 +/- 0.52 mg/mL, respectively.

Effects of duration of diabetes on behavioural and cognitive parameters in streptozotocin-induced juvenile diabetic rats.

BACKGROUND: Diabetic encephalopathy is a recently recognised complication of early-onset type 1 diabetes in children. The abnormalities underlying diabetic encephalopathy are complex and poorly understood, and the impact of disease duration on behavioural and cognitive parameters also remains unclear. Hence, the present study was conducted to determine the effects of different durations of hyperglycaemia on behavioural and cognitive parameters in young streptozotocin-induced diabetic rats. METHODS: Diabetes was induced in young, weaned, age-matched rat pups by streptozotocin injection (50 mg/kg body weight, intraperitoneally). Diabetic status was confirmed on post-natal day 30. The rats were tested in the elevated plus maze 10 and 2o days after diabetes induction. RESULTS: Diabetic rats had significantly impaired behavioural and cognitive functions compared with age-matched controls. Increased anxiety levels and cognitive deficits were observed in rats that had been diabetic for 20 days compared with their 10-day counterparts. CONCLUSION: It is essential to diagnose and treat early-onset type 1 diabetes in young children to prevent irreversible cognitive dysfunction.

Chemical composition and antimicrobial activity of the essential oil of Anaphalis nubigena var. monocephala.

The essential oil composition of the aerial parts of Anaphalis nubigena DC. var. monocephala (DC.) C. B. Clarke collected from Pindari glacier at a height of 3300 m, was analyzed by using GC and GC/MS. Sixty components were identified, accounting for 95.9% of the total oil. The main constituents were alpha-guaiene (12.3%), gamma-muurolene (10.4%), gamma-cadinene (8.3%), alpha-muurolol (7.4%), alpha-gurjunene (6.0%) and alpha-bulnesene (5.8%). The oil was found to be rich in sesquiterpene hydrocarbons (60.1%). The oil was active against Escherichia coli (NCIM 2065) and Klebsiella pneumoniae (NCIM 2957), with MIC values of 125 microg/mL and 500 microg/mL, respectively.

Semen characteristics, cryopreservation, and successful artificial insemination in the Blue rock pigeon (Columba livia).

The present study was undertaken in the Blue rock pigeon (Columba livia) to evaluate the annual semen characteristics, to identify a suitable extender for semen short-term storage, to determine a protocol for cryopreservation of semen and finally to check whether intracloacal insemination would lead to the birth of a chick. Semen characteristics such as semen volume, sperm concentration, sperm motility, and percentage of normal spermatozoa were maximum during the monsoon season. TALP was observed to be the most suitable semen extender and the sperm survived best at 37 degrees C at a dilution of 1:100 in TALP. Further, cryopreservation studies on pigeon semen indicated that 8% DMSO with or without egg yolk (20%) proved to be a better cryoprotectant compared to glycerol and polyethylene glycol. In addition, the slow freezing protocol was better than the fast-freezing protocol and about 40% of the cryopreserved spermatozoa were motile following thawing. Computer-aided semen analysis indicated that pigeon spermatozoa were extremely active immediately after dilution in TALP and exhibited linear trajectories persisting up to 9h. But, with time there was a time-dependent decrease in the velocity parameters (VAP, VSL, and VCL). Cryopreserved spermatozoa following thawing also exhibited linear trajectories but had reduced velocity as evident from the significant decrease in VAP, VSL, and VCL. Further, artificial inseminations using fresh semen resulted in 45% fertilization and birth of a live chick.