Variability in Colletotrichum infecting chilli plants of Andaman and Nicobar Islands, India

Aim: The aim of the present study was to understand the basic pathogen dynamics of Colletotrichum species infecting chilli crop in the Andaman and Nicobar Islands for disease management practices. Methodology: Twenty fungal isolates associated with chilli (Capsicum annuum) anthracnose disease from diverse locations of tropical Andaman, India were characterized using polyphasic approaches. Results: Upon morphological studies, out of twenty isolates, fourteen isolates were found chromogenic and eleven showed concentric ring pattern of mycelial growth. Based on conidial morphology, the isolates were grouped into two species (C. capsici and C. gloesporioides). The pathogenicity tests on three chilli varieties revealed difference in the pathogenic potential among the collected isolates. The sequence analysis using internal transcriber spacer (ITS) region revealed the presence of two other species C. siamense and C. plurivorum, in addition to C. gloesporioides and C. capsici. Interpretation: The present study revealed the presence of high level of genetic and pathogenic variability among the Colletotrichum isolates infecting chilli in Andaman Islands.

Study of Different Evapotranspiration Models under Polyhouse and Open Environment in Tarai Region of Uttarakhand

The average of mean monthly ET0 estimated under polyhouse by FAO PM (benchmark) model was 39.44 mm, but that of the FAO Penman, Hargreaves Stanghellini, Priestley-Taylor and FAO Radiation models were 38.37, 18.18, 37.80, 48.17 and 53.87 mm respectively. Whereas, the average of mean monthly ETo estimated under open environment by FAO PM (benchmark) model was 116.34 mm, but that of the FAO Penman, Hargreaves Stanghellini, Priestley-Taylor and FAO Radiation models were 119.33, 133, 126.41, 113.17 and 117.37 mm respectively. The FAO Penman and Hargreaves model are found to be most and least appropriate models for estimating daily ET0 under polyhouse. Whereas, FAO Radiation and Stanghellini model observed to be most and the least appropriate models in an open environment for estimating daily ET0 under polyhouse for the Pantnagar Tarai condition of Uttarakhand.

Bulk segregant analysis identifies SSR markers associated with Alternaria blight resistance in linseed (Linum usitatissimum L.)

Aim: Alternaria blight caused by Alternaria lini is one of the major diseases of linseed which severely affects the yield and productivity. Here, we utilizes F2 mapping population derived from a resistant (JRF-4) and a susceptible (Chambal) genotypes of linseed and SSRs to identify the markers associated with Alternaria blight resistance using bulk segregant analysis approach. Methodology: A population consisting of 154 F2 individuals was developed from the cross between JRF-4 (resistant) and Chambal (susceptible). All 154 F2 individuals were screened with 100 polymorphic SSRs to identify extreme phenotype. Two bulk of extremes phenotypes (disease resistant and disease susceptible) from F2 mapping population were used for the bulked segregant analysis. The SSR primers that distinguished the parental lines were used to amplify the DNA from two bulks and banding pattern was observed to identify the SSRs that can differentiate the resistant and susceptible phenotypes bulk for Alternaria blight. Markers validation was carried out by amplifying DNA from individual plants of each bulk. Results: Out of 100, only 10 markers showed polymorphism among the bulks and of which only three markers viz., LUSc 898_3_12, Lu 2472 and Lu 3078 were able to differentiate the disease resistant and susceptible individuals from F2 population. Further, single marker linear regression approach was used to validate the association of selected polymorphic markers with the disease. The markers LUSc 898_3_12 and Lu 2472 showed significant regression which confirmed their linkage with Alternaria blight resistance. Interpretation: The two markers having significant regression can be used for diseases resistance breeding during marker assisted selection.

Rapid detection of ethambutol-resistant Mycobacterium tuberculosis in clinical specimens by real-time polymerase chain reaction hybridisation probe method

Background: Early diagnosis of drug resistance (DR) to ethambutol (EMB) in tuberculosis (TB) remains a challenge. Simple and reliable method (s) are needed for rapid detection of DR Mycobacterium tuberculosis (MTB) in clinical specimens. Objectives: The aim of this study was to design fluorescence resonance energy transfer hybridisation probe-based real-time polymerase chain reaction (PCR) method for the early detection of EMB-resistant MTB direct from clinical sputa. Materials and Methods: Primers and probes were designed against 306 codon of embB gene which is commonly associated with EMB resistance. A comparative study was done between Lowenstein–Jenson (L–J) proportion and hybridisation probe-based real-time PCR method for susceptibility testing. DNA sequencing was used in nine representative isolates to validate the efficiency of real-time PCR method to detect emb306 mutation of MTB. Results: A total of 52 clinical sputum samples and corresponding culture isolates (from category II pulmonary TB cases) were included in this study. Out of 52 MTB isolates, 32 and 20 were resistant and susceptible to EMB, respectively, as determined by L–J proportion method. Real-time PCR showed 95% specificity, 75% sensitivity and 82.69% accuracy when compared with L–J proportion method. A 100% of concordance was observed by validating the real-time PCR results with DNA sequencing. Conclusions: Our real-time PCR hybridisation probe method promises for rapid detection of EMB-resistant MTB directly from clinical specimens. However, future studies and modifications of method by incorporating other potential loci along with targeted mutation (emb306) are still required to increase the sensitivity of method.

Tobacco use and cessation counseling in India-data from the Global Health Professions Students Survey, 2005-09.

Context: Tobacco use by health professionals reflects the failure of healthcare systems in protecting not only beneficiaries of the system but also those involved in health care delivery. Aim: The aim of this study was to report findings from the Global Health Professions Students Survey (GHPSS) conducted in medical, dental, nursing and pharmacy schools in India. Settings and Design: A cross-sectional survey was conducted in Indian dental and medical schools (in 2009), nursing (in 2007), and pharmacy (in 2008) schools. Materials and Methods: Anonymous, self-administered GHPSS questionnaire covering demographics, tobacco use prevalence, secondhand smoke (SHS) exposure, desire to quit smoking and training received to provide cessation counseling to patients was used. Statistical Analysis: Proportions and prevalence were computed using SUDAAN and SPSS 15.0. Results: Current cigarette smoking and other tobacco use ranged from 3.4-13.4% and 4.5-11.6% respectively, in the four health professional schools, with the highest numbers for medical schools and males. Enforcement of smoking ban in medical schools was low (53%) compared to nursing (86.4%), pharmacy (85.5%), and dental (90.8%) schools. Ninety percent students thought health professionals have a role in giving smoking cessation advice to their patients. Three out of five current smokers wanted to quit. However, one out of two reported receiving help/advice to quit. Although all expressed the need, 29.1-54.8% students received cessation training in their schools. Conclusion: Tobacco control policy, cessation training and initiatives to help students quit smoking should be undertaken.

Tobacco use, exposure to second-hand smoke, and cessation training among the third-year medical and dental students in selected Member States of South-East Asia region: A trend analysis on data from the Global Health Professions Student Survey, 2005-2011.

Background: The Medical and Dental Global Health Professions Student Surveys (GHPSS) are surveys based in schools that collect self-administered data from students on the prevalence of tobacco use, exposure to second-hand smoke, and tobacco cessation training, among the third-year medical and dental students. Materials and Methods: Two rounds of medical and dental GHPSS have been conducted in Bangladesh, India, Myanmar, Nepal, Sri Lanka, and Thailand, among the third-year medical and dental students, between 2005 and 2006 and 2009 and 2011. Results: The prevalence of any tobacco use among third-year male and female medical students did not change in Bangladesh, India, and Nepal between 2005 and 2006 and 2009 and 2011; however, it reduced significantly among females in Myanmar (3.3% in 2006 to 1.8% in 2009) and in Sri Lanka (2.5% in 2006 to 0.6% in 2011). The prevalence of any tobacco use among third-year male dental students did not change in Bangladesh, India, Nepal, and Thailand between 2005 and 2006 and 2009 and 2011; however, in Myanmar, the prevalence increased significantly (35.6% in 2006 to 49.5% in 2009). Among the third-year female students, a significant increase in prevalence was noticed in Bangladesh (4.0% in 2005 to 22.2% in 2009) and Thailand (0.7% in 2006 to 2.1% in 2011). It remained unchanged in the other three countries. Prevalence of exposure to second-hand smoke (SHS) both at home and in public places, among medical students, decreased significantly in Myanmar and Sri Lanka between 2006 and 2009 and in 2011. Among dental students, the prevalence of SHS exposure at home reduced significantly in Bangladesh, India, and Myanmar, and in public places in India. However, there was an increase of SHS exposure among dental students in Nepal, both at home and in public places, between 2005 and 2011. Medical students in Myanmar, Nepal, and Sri Lanka reported a declining trend in schools, with a smoking ban policy in place, between 2005 and 2006 and 2009 and 2011, while proportions of dental students reported that schools with a smoking ban policy have increased significantly in Bangladesh and Myanmar. Ever receiving cessation training increased significantly among medical students in Sri Lanka only, whereas, among dental students, it increased in India, Nepal, and Thailand. Conclusion: Trends of tobacco use and exposure to SHS among medical and dental students in most countries of the South-East Asia Region had changed only relatively between the two rounds of GHPSS (2005-2006 and 2009-2011). No significant improvement was observed in the trend in schools with a policy banning smoking in school buildings and clinics. Almost all countries in the SEA Region that participated in GHPSS showed no significant change in ever having received formal training on tobacco cessation among medical and dental students.

MPOWER and the Framework Convention on Tobacco Control implementation in the South-East Asia region.

The 11 member states of WHO's South-East Asia Region share common factors of high prevalence of tobacco use, practice of several forms of tobacco use, increasing prevalence of tobacco use among the youth and women, link of tobacco use with poverty, and influence of tobacco advertisements in propagating the use of tobacco, especially among young girls and women. The effects of tobacco use are many-fold, leading to high morbidity and mortality rates as well as loss of gross domestic product (GDP) to respective countries. The WHO Regional Office for South-East Asia has been actively involved in curbing this menace essentially by way of assisting member states in implementing the WHO Framework Convention on Tobacco Control (FCTC). This paper gives an overview of these activities and discusses the opportunities and challenges in implementing the FCTC and possible practical solutions.

Smokeless tobacco use in Nepal.

Tobacco use is widely prevalent in different forms in Nepal. These habits are deeply rooted among different age groups and gender. There is no information available on all aspects of smokeless tobacco (SLT) use and its implications in Nepal. To review the types of SLT products available in Nepal, prevalence of SLT use, correlates of SLT use, SLT-related harm, and existing gaps in information and policy implications. This review is mainly based on information from literature, and some WHO and other documents. The prevalence of smokeless tobacco use is high, particularly among males and disadvantaged groups. SLT users have multiple habits of tobacco chewing, smoking and drinking. Despite SLT products being manufactured in the unorganized sector, they are also largely imported from India. People have easy access to various SLT products. There is a general lack of information on the health hazards of SLT use to the population. This calls for research on the issue. In order to succeed in reducing SLT use in Nepal, addressing the challenges of enforcing the Tobacco Control Act through a multisectoral approach and developing capacity of sectors other than health is of vital importance.

Prevalence of smokeless tobacco use among adults in WHO South-East Asia.

Smokeless tobacco (SLT) use is an understudied problem in South-East Asia. Information on SLT use among the adult population was collected from various available sources. SLT use prevalence varies among countries in the region. The prevalence of SLT use is known for all countries at national level in the region with the exception of Bhutan and DPR Korea. For Bhutan, data pertains to Thimphu only. There is no available data on SLT use for DPR Korea. Using all available data from Bhutan, India, Myanmar, Nepal, and Sri Lanka, SLT use was found to be higher among males as compared to females; however, in Bangladesh, Indonesia, and Thailand, SLT use was higher among females as compared to males. Among males, prevalence of SLT use varied from 51.4% in Myanmar to 1.1% in Thailand. Among females, the prevalence of SLT use varied from 27.9% in Bangladesh to 1.9% in Timor-Leste. The prevalence also varies in different parts of countries. For instance, the prevalence of current use of SLT in India ranges from 48.7% in Bihar to 4.5% in Himachal Pradesh. In Thailand, prevalence of current use of tobacco use varies from 0.8% in Bangkok to over 4% in the northern (4.1%) and northeastern (4.7%) region. Among all SLT products, betel quid was the most commonly used product in most countries including Bangladesh (24.3%) and Thailand (1.8%). However, Khaini (11.6%) chewing was practiced most commonly in India. Nearly 5% of the adult population used tobacco as dentifrice in Bangladesh and India. SLT is more commonly used in rural areas and among disadvantaged groups. Questions from standard "Tobacco Questions for Surveys (TQS)" need to be integrated in routine health system surveys in respective countries to obtain standardized tobacco use data at regular intervals that will help in providing trends of SLT use in countries.

Effect of sulphur and phosphorus on yield, quality and nutrient status of pigeonpea (Cajanus cajan).

A field experiment was conducted to study the impact of Sulphur (S) and Phosphorus (P) on yield, nutrient status of soil and their contents in pigeonpea (Cajanus cajan) during the year 2008-2009. Seven treatments were studied in Factorial Randomized Block Design with three replications. The treatment combinations were derived from three levels of sulphur (0, 20 and 40 kg S ha-1) and four levels of phosphorus (0, 25, 50 and 75kg ha-1). The experimental soil was medium black, slightly calcareous, clay in texture and slightly alkaline in reaction. The results indicated a significant increase in grain yield (14.81 q ha-1) and straw yield (41.26 q ha-1) of pigeonpea after 20 kg S ha-1 and 50 kg P2O5 ha-1 treatment with common dose of nitrogen @ 30 kg ha-1. The increase in grain and straw yield was 102.77 and 52.87% as compare to higher over control. Maximum number of pods plant -1, maximum number of grains pod-1 and test weight by this treatment was also observed as compared to control. Application of S and P improved soil fertility status and S alone did not influence P availability. Hence, in order to maintain the fertility status of the soil at high level, combine application of 20 kg S ha-1 with 50 kg P2O5 ha-1 is essential. The residual fertility status of soil is advocated for rainfed pigeonpea crop grown on vertisol in Vidarbha region.

Impact of phosphorous on biochemical changes in Hordeum vulgare L. in mixed cropping with Chickpea.

Multiple cropping (i.e. intercropping or mixed cropping) plays an important role in agriculture because of the effective utilization of resources, significantly enhancing crop productivity compared with that of monocultured crops. The study was planed to assess the effect of various concentrations (00, 30, 60, 90 kg ha-1) of phosphorous on the biochemical composition of grains of Hordeum vulgare L. (NDB-1050) in mixed cropping system with Chickpea. Phosphorous is an essential ingredient for plants to convert atmospheric N (N2) into an ammonium (NH4) as a useable form. The available nitrogen content was found more in the year 2006 (131 kg ha-1) than year 2005 (105 kg ha-1). The results of available nitrogen content were showed that the mixed cropping system enhances N fixation process because phosphorous also influences nodule development through its basic functions in plants as an energy source. Reducing, non reducing and total sugar content of H. vulgare L. were influenced by changes in the phosphorous doses. Maximum protein (13.43 %) was obtained at 60 kg P2 O5 ha-1 during the year 2006. Lysine, tryptophan and methionine content were found maximum in year 2006, respectively. Total mineral content of grains of plant (0.99 g 100g-1) was found maximum by the application of 60 kg P2 O5 ha-1. It is possible that there was an increase in the soil N made available by the leguminous chickpea species, and this could be another reason why there was an increase in Hordeum vulgare L. shoot mass per plant with intercropping with chickpea.

Is Mycobacterium avium subsp. paratuberculosis, the cause of Johne’s disease in animals, a good candidate for Crohn’s disease in man.

Mycobacterium avium subspecies paratuberculosis (MAP) causes Johne’s disease or paratuberculosis, a gastro intestinal inflammatory condition in ruminants and other animals, which is similar to Crohn’s disease (CD) that occurs in man. The role of MAP in the causation of CD has been under intense investigation in the last few decades. This review summarizes the status of MAP in animals and the food chain and its association with CD in man.

Strain diversity within Mycobacterium avium subspecies paratuberculosis — A review.

Mycobacterium avium subspecies paratuberculosis (MAP), is the etiological agent of Johne’s disease (or paratuberculosis) in animals and has also been linked with Crohn’s disease of human beings. Extreme fastidious nature of the organism (MAP) has hampered studies on diversity within the organism. Studies based on phenotypic properties like growth rate, pigmentation, lipid profile etc., are unable to provide complete information on diversity of MAP organism in nature. However, with the advent of molecular assays (IS900 RFLP, PFGE, IS1311 PCR-REA, SSR typing, VNTR typing etc.) in last 2 decades, progress has been made to differentiate MAP strains. MAP isolates have been classified into various types and subtypes using these molecular tools. Optimization of these typing assays has led to generation of new information about MAP strains, subtypes, their comparative genomics, relative evolution, comparative virulence etc. Knowledge of strain diversity is important for better understanding of molecular and sero-epidemiology, infection and patho-biology, vaccine development and planning control strategies. The present review provides available information on MAP strains, host adaptations, their virulence, comparative genomics, relative genetic evolution and differentiation.

Effect of copper on growth, yield and concentration of Fe, Mn, Zn and Cu in wheat plants (Triticum aestivum L.).

A pot experiment was conducted at six graded levels of copper (Cu) viz., 0, 0.5, 1.0, 1.5, 2.0 and 2.5 mg kg-1 to test the response of wheat plants grown in a copper-responsive alluvial soil (entisol) under glass house conditions. The growth attributes like plant height, fresh and dry matter yield, percent dry matter enhanced with increasing Cu levels and was maximum at 1.5 mg kg-1 Cu while the number of tillers was minimum at this level. The grain yield at 1.5 mg kg-1 Cu was enhanced by 62.9% from the control. The increase in weight of 1000 grains ranged from 33.93 to 41.35 g in comparison to control (32.58 g). Harvest index (%) also increased and ranged from 39.42 to 47.73 in different treatments in comparison to control (35.92). Both 1000 grain weight and harvest index were maximum in the plants at 1.5 mg kg-1 copper. Cu concentrations in leaves, grain and straw enhanced with increasing levels of Cu application. The Fe concentration in leaves was significantly reduced by Cu application and the reduction was 10.3% at 2.5 mg kg-1 Cu and was not influenced in by Cu application in grain and straw. The Mn concentration was not affected by Cu application in any of the plant part studied. However, Zn concentration decreased significantly at higher levels of Cu (2.0 and 2.5 mg kg-1) in leaves and remained unaffected in the grain and straw.

Evaluation of four methods of DNA recovery from Mycobacterium avium subspecies paratuberculosis present in intestine tissue of goats and comparative sensitivity of IS900 PCR with respect to culture for diagnosis of Johne's disease.

Low sensitivity of PCR reaction for detection of Mycoobacterium avium subspecies paratuberculosis (MAP) in tissues and fecal samples is mainly attributed to false negative results. Present study was undertaken to compare four methods of DNA isolation from tissues of infected animals and to determine most sensitive protocol for the recovery of DNA, suitable for IS900 PCR based detection of Johne's disease infection. Method I, the traditional van Soolingen2 method of DNA isolation was adopted for the isolation of DNA from tissues. Method II was modification (hexadecyl pyridinium chloride-HPC treatment) of van Soolingen2 method. Method III was traditional tissue DNA isolation method based on tissue lysis buffer. Method IV was modification of method III (HPC treatment). Using four methods of DNA isolation from 25 intestinal tissues of clinically infected goats, DNA was isolated from 15 (60.0%), 18 (72.0%), 13 (52.0%) and 13 (52.0%) tissues using method I, II, III and IV, respectively. All isolated DNA preparations were positive for MAP in IS900 PCR. HPC treatment enhanced the recovery of DNA from tissues of infected animals using method II. Therefore, method II can improve the diagnosis MAP infection using IS900 PCR.

Mycobacterium avium subspecies paratuberculosis diagnosis and strain typing--present status and future developments.

Mycobacterium avium subspecies paratuberculosis (MAP) is the etiological agent of paratuberculosis, a chronic gastroenteritis of ruminants and has zoonotic importance. We present here a review of MAP with respect to--(i) present diagnostic techniques and important developments; and (ii) MAP strain-typing tools. A summary of the findings to date is presented, and advantages and disadvantages of each of the methods are compared and discussed.

Non-chemical method of DNA recovery and characterization of Mycobacterium avium subspecies paratuberculosis using IS 900 PCR.

In the present study, two methods of DNA isolation-routine, traditional and standard DNA isolation protocol for Mycobacteria (Method 1) and a new non-chemicals and non-enzymes (physical) method (Method 2) of DNA recovery have been compared and evaluated in IS900 PCR for the specific detection of pathogen. Using the new Method 2, DNA has been recovered from few (1 - 3 colonies), extremely minute and stunted colonies. DNA, thus, isolated from these colonies (colonies PCR) and cultured for the first time from the cases of Crohn's disease in human beings, dairy cattle, raw milk and pasteurized commercial milk samples has been characterized in the present study. It is the first report from India.