Deciphering the mechanisms of zinc tolerance in the cyanobacterium Anabaena sphaerica and its zinc bioremediation potential.

Cyanobacteria adopt a variety of changes at proteomic and metabolic levels for surviving under harmful environmental conditions including heavy metal stress. The current study investigates the impact of zinc stress on the proteome of Anabaena sphaerica to get an insight into its molecular mechanisms of zinc tolerance. The study revealed three different aspects that were associated with the zinc tolerance in A. sphaerica: (i) the reduced expression of photosynthesis, nitrogen fixation, energy metabolism, respiratory, and transcriptional/translational proteins probably to conserve energy and utilizing it to sustain growth; (ii) the enhanced expression of metallothionein and ferritin domain protein All 3940 to chelate free zinc ions whereas upregulation of antioxidative proteins for detoxifying reactive oxygen species; and (iii) the expression of large numbers of hypothetical proteins to maintain the important cellular functions. Furthermore, over expressions of sulfate adenylyl transferase and cystathionine beta synthase along with the increased synthesis of peptidases and thiolated antioxidant proteins were also noticed which denoted cysteine synthesis under sulfur deprivation possibly by mobilizing the sulfur from dead cells and its channelization towards the production of thiolated antioxidants. Besides tolerating excess amount of zinc, A. sphaerica exhibited high zinc biosorption efficiency which confirmed its outstanding zinc bioremediation potential.

Degrading ability and robust antioxidative defence system led to SDS tolerance in cyanobacterium Fischerella sp. lmga1.

To test the tolerance and degradation potential of the cyanobacterium Fischerella sp. lmga1 for surfactant, sodium dodecyl sulfate (SDS), different doses of SDS (10, 30, 40, 50, 70 and 100 µM) were used for the growth. The lower doses of SDS supported the growth of cyanobacterium whereas the higher doses were found to be inhibitory but the cyanobacterium somehow managed its survival up to 100 µM SDS. However, a significant reduction was observed in the pigment and protein content. A substantial accumulation of carbohydrate at 70 µM SDS may act as an osmoprotectant for the survival of the cyanobacterium. The higher doses of SDS also triggered the ROS generation and lipid peroxidation which showed negative impact on the PSII efficiency. Simultaneously, an efficient ROS mitigation system (SOD and CAT activity) has also been worked up to 70 µM SDS while APX was enhanced only up to 50 µM SDS. Furthermore, the SDS degrading potential was investigated and almost 80% of the SDS was degraded after 6th days of treatment in the cyanobacterium. Hence, the results suggested that due to robust antioxidative defence system and ability to degrade the surfactant this cyanobacterium showed significant tolerance toward SDS.

Cyanobacterial bioactive compound EMTAHDCA recovers splenomegaly, affects protein profile of E. coli and spleen of lymphoma bearing mice.

The antibacterial and anticancerous properties of EMTAHDCA have already been reported in our previous study. However, mode of action of EMTAHDCA is still elusive. The present study was aimed to investigate the molecular targets in Escherichia coli and spleen of lymphoma-bearing mice in response to cyanocompound 9-ethyliminomethyl-12 (morpholin-4-ylmethoxy)-5, 8, 13, 16-tetraaza -hexacene-2, 3- dicarboxylic acid (EMTAHDCA) isolated from fresh water cyanobacterium Nostoc sp. MGL001. Differential expressions of proteins were observed in both E. coli and spleen of lymphoma-bearing mice after EMTAHDCA treatment. In continuation of our previous study, the present study revealed that the antibacterial agent, EMTAHDCA causes the drastic reduction in synthesis of proteins related to replication, transcription, translation and transportation in E. coli. Probably the direct or indirect interaction of this compound with these important metabolic processes led to the reduction in growth and cell death. Furthermore, the anticancerous property of the compound EMTAHDCA reflected as down regulation in proteins of cell cycle, cellular metabolism, signalling, transcription and transport together with up regulation of apoptosis, DNA damage and immunoprotection related proteins in spleen of lymphoma-bearing mice. In this study the EMTAHDCA induced modulations in expression of proteins of key metabolic pathways in E. coli and spleen cells of lymphoma bearing mice helped in understanding the mechanism underlying the antibacterial and anti-cancerous property.

Physiological mechanisms of aluminum (Al) toxicity tolerance in nitrogen-fixing aquatic macrophyte Azolla microphylla Kaulf: phytoremediation, metabolic rearrangements, and antioxidative enzyme responses.

To investigate the extent of aluminum toxicity tolerance of eco-friendly, fast-growing, fresh water, pteridophytic Azolla-Anabaena symbiotic association in terms of altered physiological signals; Azolla microphylla Kaulf was exposed to 0 (control), 100, 250, 500, and 750 µM AlCl3, at pH 4.5 for 6 days. The adversity of Al was increased in a dose-dependent manner and the highest was recorded at 750 µM AlCl3. Despite the significant loss in membrane integrity (80% electrolyte leakage) due to an enhanced generation of H2O2, A. microphylla reflected only 50% growth inhibition (fresh and dry weight) at 500 µM AlCl3 (LD50). However, the average root length of Azolla was drastically reduced at high concentration due to their direct contact with aluminum-containing growth medium. Contrary to this, the whole association maintained moderate chlorophyll, carbohydrate content, photosynthetic efficiency, nitrogen-fixing ability, and nitrogen content at high Al concentration. Probably, growth protection was pertained through significant detoxification of H2O2 by employing an efficient antioxidative defense system including antioxidative enzymes (SOD, APX, and CAT) and non-enzymatic antioxidant carotenoids. An enhanced level of phenolics and flavonoids in the root exudates possibly maintained a non-toxic level of aluminum inside the cell (195.8 µg Al/g FW) which makes A. microphylla a suitable pteridophytic plant to not only remove toxic Al from the contaminated sites but also to improve nitrogen status of those regions. Graphical abstract ᅟ.

Transcriptional regulation of acetyl CoA and lipid synthesis by PII protein in Synechococcus PCC 7942.

PII protein family is widespread in prokaryotes and plants. In this study, impacts of PII deficiency on the synthesis of acetyl CoA and acetyl CoA carboxylase enzyme (ACCase) was analyzed in the Synechococcus sp. PCC 7942 by evaluating the mRNA levels of pyruvate kinase (PK), pyruvate dehydrogenase (PDH), citrate synthase (CS), biotin synthase (BS), biotin carboxylase (BC), biotin carboxyl carrier protein (BCCP), carboxyl transferase (CT) α and ß subunits. The PII deficient Synechococcus sp. PCC 7942 showed upregulation of all the above-mentioned genes, except CS. Analyses of genes required for acetyl coA synthesis exhibited a substantial increase in the transcript levels of PK and PDH in the PII mutant strain. In addition, the PII mutant also displayed reduced acetyl CoA content, high ACCase activity, and increased lipid content. The lessening of acetyl CoA content was attributed to the rapid utilization of acetyl CoA in fatty acid synthesis as well as in the TCA cycle whereas the increased ACCase activity was ascribed to the rise in mRNA levels of BS, BC, BCCP, CT α, and ß genes. However, increased lipid content was correlated with the declined total protein content. Hence, the study suggested that PII protein regulates the synthesis of acetyl CoA and ACCase enzyme at the transcriptional level.

Sequential role of biosorption and biodegradation in rapid removal degradation and utilization of methyl parathion as a phosphate source by a new cyanobacterial isolate Scytonema sp. BHUS-5.

A new isolate of genus Scytonema distinct from its closest relative cyanobacterium, Scytonema hofmanni was found efficient in the removal and degradation of organophosphorus (OP) pesticide, methyl parathion (MP). The cyanobacterial isolate was also capable of utilizing the phosphorus present in the MP following its degradation, which was evident from the increase in growth (chlorophyll content), biomass, protein content, and total phosphorus in comparison to cyanobacterium grown in phosphate-deficient cultures. The rapid removal of MP by the cyanobacterium during initial 6 hours of incubation was defined by the pseudo-second-order biosorption kinetics model, which indicated the involvement of chemosorption in initial removal of pesticide. Further, degradation of MP was also confirmed by the appearance of p-nitrophenol in the medium after 24 hours of incubation. Thus, the cyanobacterial isolate of Scytonema sp. BHUS-5 seems to be a potential bioremediation agent for the removal of OP pesticide, MP from the habitat.

Modulation of fatty acids and hydrocarbons in Anabaena 7120 and its ntcA mutant under calcium.

Calcium being a signaling molecule and mediator of cell response, we examined the modulation in fatty acid and hydrocarbon profiles of wild type cyanobacterium Anabaena sp. PCC 7120 and its ntcA mutant under the influence of different calcium chloride concentrations (0-10 mM). Dynamic modifications in fatty acid and hydrocarbon profile were evident through GC-FID analysis of extracted lipids. In the wild type, increase in CaCl2 (10 mM) resulted in unsaturation of fatty acids (observed in terms of high MUFA/PUFA ratio) while hydrocarbon production was distinctly high in the mutant strain compared to wild type at all tested concentrations. The synthesis of short chain hydrocarbons (C5-C8) were dominated at inhibitory concentration (10 mM CaCl2) in mutant strain. Results suggest that the increase in MUFA/PUFA ratio at inhibitory concentration in wild type, and higher percentage of hydrocarbons in mutant strain, may be attributed to the survival and acclimation strategies under altered calcium environment. Our results also suggest the involvement of the ntcA gene (master regulator of N2 metabolism) in regulation of carbon metabolism; specifically fatty acid, hydrocarbon, and other metabolic compounds essential for maintenance and sustenance of growth under stress condition. Thus, our study outlines basic acclimation response along with possibilities of production of fatty acid and hydrocarbon derived biofuel and other bioactive compounds in Anabaena sp. PCC 7120 under altered calcium levels which could be of biotechnological interest.

Exopolysaccharide production in Anabaena sp. PCC 7120 under different CaCl2 regimes.

Influence of various levels of CaCl2 (0, 1, 10 and 100 mM) on exopolysaccharide production has been investigated in the cyanobacterium Anabaena 7120. At the concentration of 1 mM CaCl2, growth was found to be stimulatory while 100 mM was sub lethal for the cyanobacterial cells. Estimation of EPS content revealed that EPS production depends on the concentration of calcium ions in the immediate environment with maximum being at10 mM CaCl2. A possible involvement of alr2882 gene in the process of EPS production was also revealed through qRT-PCR. Further, FTIR-spectra marked the presence of aliphatic alkyl-group, primary amine-group, and polysaccharides along with shift in major absorption peaks suggesting that calcium levels in the external environment regulate the composition of EPS produced by Anabaena 7120. Thus, both quantity and composition of EPS is affected under different calcium chloride concentrations presenting possibilities of EPS with novel unexplored features that may offer biotechnological applications.

Profenofos induced modulation in physiological indices, genomic template stability and protein banding patterns of Anabaena sp. PCC 7120.

To understand the mechanism underlying organophosphate pesticide toxicity, cyanobacterium Anabaena PCC 7120 was subjected to varied concentrations (0, 5, 10, 20 and 30 mg L(-1)) of profenofos and the effects were investigated in terms of changes in cellular physiology, genomic template stability and protein expression pattern. The supplementation of profenofos reduced the growth, total pigment content and photosynthetic efficiency of the test organism in a dose dependent manner with maximum toxic effect at 30 mg L(-1). The high fluorescence intensity of 2', 7' -dichlorofluorescin diacetate and increased production of malondialdehyde confirmed the prevalence of acute oxidative stress condition inside the cells of the cyanobacterium. Rapid amplified polymorphic DNA (RAPD) fingerprinting and SDS-PAGE analyses showed a significant alteration in the banding patterns of DNA and proteins respectively. A marked increase in superoxide dismutase, catalase, peroxidase activity and a concomitant reduction in glutathione content indicated their possible role in supporting the growth of Anabaena 7120 up to 20 mg L(-1). These findings suggest that the uncontrolled use of profenofos in the agricultural fields may not only lead to the destruction of the cyanobacterial population, but it would also disturb the nutrient dynamics and energy flow.

Pulmonary tuberculosis: a public health problem amongst the Saharia, a primitive tribe of Madhya Pradesh, Central India.

OBJECTIVES: To estimate the prevalence of pulmonary tuberculosis (TB) disease amongst the Saharia, a 'primitive' tribe of Madhya Pradesh, Central India. METHODS: A community-based cross-sectional TB prevalence survey was undertaken in the Saharia, a 'primitive' tribal community of Madhya Pradesh. A representative random sample of villages predominated by tribal populations was chosen from the selected block of Sheopur District. All eligible individuals were screened for chest symptoms related to TB. Sputum samples were collected from all eligible individuals, transported to the laboratory, and examined by Ziehl-Neelsen smear microscopy and solid media culture methods. RESULTS: Of the 11,468 individuals eligible for screening, 11,116 (96.9%) were screened for symptoms. The overall prevalence of pulmonary TB disease was 1518 per 100,000 population. Prevalence increased with age and the trend was statistically significant (p<0.001). The prevalence of pulmonary TB was also significantly higher in males (2156/100,000) than females (933/100,000) (p<0.001). CONCLUSION: The findings suggest that TB disease remains a major public health problem in the Saharia 'primitive' tribal community of Madhya Pradesh, Central India.

Prevalence of pulmonary tuberculosis amongst the tribal population of Madhya Pradesh, central India.

BACKGROUND: This was a prevalence survey of pulmonary tuberculosis (PTB) disease in the tribal population of Madhya Pradesh state, central India. METHODS: A community-based cross-sectional tuberculosis (TB) disease prevalence survey was undertaken among adults aged > or = 15 years in the tribal population of Madhya Pradesh. A multistage stratified cluster sampling was adopted. A representative random sample of villages predominated by tribal populations was selected from 11 districts. All eligible individuals were questioned for chest symptoms relating to TB. Sputum samples were collected from all eligible individuals, transported to the laboratory, and examined by Ziehl-Neelsen (ZN) smear microscopy and solid media culture methods. RESULTS: Of the 23,411 individuals eligible for screening, 22,270 (95.1%) were screened for symptoms. The overall proportion of symptomatic individuals was 7.9%. Overall prevalence (culture and/or smear positive) of PTB was 387 [95% confidence interval (CI): 273-502] per 100,000 population. The prevalence increased with age and was also significantly higher among males (554/100,000; 95% CI: 415-693) as compared with females (233/100,000; 95% CI: 101-364) (P < 0.001). CONCLUSION: The findings suggest that the TB situation amongst the tribal population is not that different from the situation among the non-tribal population in the country. However, TB remains a major public health problem amongst the tribal population and there is a need to maintain and further strengthen TB control measures on a sustained and long-term basis.

Seroprevalence of sexually transmitted viruses in the tribal population of Central India.

OBJECTIVE: To determine the seroprevalence of human immunodeficiency virus (HIV), hepatitis B and C viruses (HBV, HCV), and herpes simplex virus type 2 (HSV-2) in the tribal population of central India. METHODS: A community-based cross-sectional survey was carried out in the tribal population of Jabalpur district. Blood samples were drawn from 326 patients with sexually transmitted infections (STIs) and 526 randomly selected adults. These were tested for HIV, HBV, HCV, and HSV-2 using commercial ELISA kits. RESULTS: The prevalence of IgG antibodies to HSV-2 was 20.8% in STI patients compared to 12.4% in the general population. The HBV carriage rate was 3.4% in STI patients against 2.9% in the general population. HCV prevalence was 3.9% in STI patients and 4.6% in the general population. No HIV infection was found in the study population. CONCLUSIONS: In view of the high prevalence of viral STIs in the tribal community of Central India, there is a need to strengthen the STI control program in this under-privileged group.