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Genus Halomonas is reported from most of the saline environments regardless of their geographical location, including the marine environment, salterns, saline lakes, and soil ( Llamas etal., 2006). The MRB isolated from the coastal areas of Kuwait were identified as Alcanivorax borkumensis, Marinobacter hydrocarbonoclasticus, and Halomonas taeheungii ( Sorkhoh etal., 2010). The 16S rDNA analysis and phenotypic characteristics revealed that the heavy metal-resistant halophilic bacteria WQL9 belong to the genus Halomonas sp ( Abdel-Razik etal., 2020). The Marinobacter genus is comprised of widespread marine bacteria found exceptionally in marine or terrestrial environments rich in sodium salts, in the deep sea, coastal seawater, sediment, oceanic basalt, etc. ( Handley and Lloyd, 2013). Alcanivorax xenomutans are also rich in the saline environment as they are halophilic and favorable to living in the marine environment. A comparative analysis of the partial sequence of the 16S rDNA of the four strains revealed a high level of similarity to the corresponding sequence of environmental isolates. Detection of mercury reductase gene Centre for Ocean Science and Technology for Islands, National Institute of Ocean Technology, Ministry of Earth Sciences, Government of India, Port Blair, India The pathogenesis of HTN involves oxidative stress. Another mechanism involved is the decreased bioavailability of nitric oxide (NO) [ 38]. Physical exercise could be a potential lifestyle intervention to treat HTN due to its beneficial effects on endothelial function and oxidative stress [ 38]. Exercise exerts an anti-inflammatory action via the hypothalamic-pituitary-adrenal axis and via the sympathetic nervous system, thus affecting BP directly [ 9]. The physiologic effects of exercise are further divided into acute, post-exercise, and chronic [ 9]. Aerobic exercises like speed walking, jogging, running, cycling, dancing, and swimming have been shown to decrease resting BP and BP reactivity to stressors [ 9]. A study by Ozemek et al. revealed the following about how diverse types of exercises affect BP: (1) Aerobic exercise of 90 to 150 minutes per week with 65%-75% heart rate reserve has been shown to impact SBP by −5/8 mm Hg in hypertensive individuals and by −2/4 mm Hg in normotensive individuals. (2) Dynamic resistance exercise of 90 to 150 minutes per week with 50%-80% one rep maximum, six exercises, three sets/exercise, and ten repetitions/set has been shown to decrease SBP by 4 mm Hg in hypertensive individuals and 2 mm Hg in normotensive individuals. (3) Isometric resistance exercise of 4 × 2 min (hand grip), 1 min rest between exercises, 30%-40% maximum voluntary contraction, and three sessions per week for 8-10 weeks have been shown to lower SBP by 5 mm Hg in hypertensive individuals and 4 mm Hg in normotensive individuals [ 11].
According to the National Institute on Alcohol Abuse and Alcoholism (NIAAA), low-risk drinking is considered four drinks per day or less than fourteen drinks per week for men and less than or equal to three drinks per day and less than seven drinks per day for women [ 49]. Binge drinking is the consumption of five drinks by men and four drinks by women in a period of two hours [ 47, 50]. According to studies performed by O’Keefe et al. and Peng et al., it was shown that excessive consumption of alcohol has contributed to 16% of HTN cases worldwide [ 45, 51]. The pathophysiology of HTN in chronic alcohol consumption is by vagal inhibition and sympathetic stimulation [ 52]. The Mediterranean diet (Med Diet) has also helped lower BP. As per the Maine-Syracuse Longitudinal Study conducted in the United States in 2020 by a group of researchers who followed 851 US older adults, for every one unit increase in the Med Diet score in participants, it was found that there was a corresponding reduction of 0.69 units in SBP, a reduction of 0.33 in DBP, and a reduction of 0.45 on mean arterial pressure (MAP) [ 32]. Although this seems to be small, this change can have a noteworthy effect at the level of the population; that is, a decrease of 2 mm Hg in SBP can lead to a decrease of 10% when it comes to the population [ 32]. According to the observational studies conducted in Mediterranean countries, higher adherence to a Med Diet is associated with a decreased risk of cardiovascular disease, overall mortality as well as neoplastic disease [ 33, 34]. The Med Diet consists of higher consumption of extra virgin olive oil, vegetables, fruits, whole grains, nuts, cereals, as well as seeds; moderate consumption of fish, poultry, red wine, and dairy; and lower consumption of processed foods and red meat [ 35]. The health effects of alcohol intake are variable and are based on the amount of intake (low, moderate, or heavy) and intake pattern (acute, chronic, or binge) [ 45- 47]. In the US, the amount of pure alcohol in one standard drink is 14 g, which can be found in 12 oz of regular beer (about 5% alcohol), 5 oz of wine (about 12% alcohol), and 1.5 oz of distilled spirits (about 40% alcohol) [ 48]. Moderate alcohol intake recommendations according to the Dietary Guidelines for Americans are the consumption of two standard drinks per day for men and one standard drink per day for women [ 49]. Moderate alcoholic intake has been shown to reduce the risk of chronic disease [ 49]. Physicians play a key role in encouraging and helping patients achieve smoking cessation [ 56]. HTN has also been associated with second-hand smoke. According to research done by Bernabe-Ortiz et al. on 897 individuals in Peru in 2021 to assess the association of second-hand smoke with HTN and cardiovascular risk, 15% of adults reported second-hand smoke overall, and this emphasizes the necessity to keep places smoke-free to reduce the risk of cardiovascular disease [ 57].It can be observed that high Hg concentration causes structural deformities which may be due to its toxic nature. In our previous study, we reported that cultures accustomed to grow in the presence of HgCl 2 manifested major morphological abnormalities ( Joshi etal., 2021). We also suggested several structural asymmetries allied with the cell wall and cytoplasmic membrane, significantly affecting the cellular mechanism. Resultantly, there is a substantial delay at the beginning of growth and cell division in a higher concentration of Hg 2+. Metabolites formed during Hg (II) reduction Many studies suggest that mer operon is located on any one of either location such as mobile elements like plasmids, transposons, genomic or chromosomal DNA ( Liebert etal., 1999; Schelert etal., 2004; Zeng etal., 2010). The mer operon contains many functional genes along with the operator, promoter, and regulatory elements. All functional genes encrypt for the diverse proteins that contribute to the detection, transportation, and reduction of Hg ions ( Barkay etal., 2003; Dash and Das, 2012; Naguib etal., 2018). An extremely noteworthy positive correlation was found among the harboring of mer gene with the phenotypic resistance to Hg and concentration of the Hg in the source environment ( Osborn etal., 1997). In this study, we did not find any Hg from the ecosystem (detectable limit - 0.73 µg/L), but the amplification of mer genes and available literature suggested that these are extensively dispersed in a non-highly contaminated or contamination-free environment such as an open ocean, Antarctic sea-ice, high Arctic snow, sea ice brine etc. and plays a key role in the biogeochemical cycle of Hg ( Christakis etal., 2021). Thus, the Hg adaptation by these isolates is reasonably accepted. Confirmation of Hg 2+ reduction by the isolates
Weight loss is one of the most important non-pharmacological interventions to lower BP. Numerous interrelated pathophysiologic mechanisms stimulate higher BP in obesity [ 12]. In overweight/obese individuals, accelerated vascular aging can lead to HTN due to inflammation, oxidative stress, and insulin resistance [ 13]. Obese individuals also experience increased activity of the sympathetic nervous system and the renin-angiotensin-aldosterone system [ 14]. The combined effect results in increased sodium resorption by the kidney, impaired vasodilation, volume expansion, and decreased natriuresis, thus leading to elevated BP [ 15]. Ozemek et al. trials have shown that weight loss helped lower systolic BP in hypertensive individuals by 5 mm Hg and in normotensive individuals by 2 to 3 mm Hg. According to the Centers for Disease Control (CDC), a BMI of 25 to 29.9 is considered overweight, and a BMI of 30 or higher is considered obese. In overweight or obese individuals, achieving their ideal body weight is best, but it is good to aim for at least a 1 kg reduction in body weight. The study by Ozemek et al. also revealed that for every 1 kg reduction in body weight, we can expect about a 1 mm Hg reduction in blood pressure.
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The statistical analysis of Hg removal potential among the different species i.e., Alcanivorax xenomutans, Halomonas sp., Marinobacter hydrocarbonoclasticus and within the Halomonas spp. revealed that the Hg removal by NIOT-EQR_J251 was significantly higher (p< 0.01) than NIOT-EQR_J7 and NIOT-EQR_J258. There was a significant difference (p< 0.05) in the removal of Hg between NIOT-EQR_J248 vs. NIOT-EQR_J258 and NIOT-EQR_J248 vs. NIOT-EQR_J251. Al-Mailem etal. (2011) reported Haloferax sp. (HA1 and HA2), Halobacterium sp. HA3, and Halococcus sp. HA4 effectively volatilized (from 40 to 65%) the available 100 mg/L of Hg after 8 days. Many other isolates such as Bacillus sp., Pseudomonas stutzeri, Pseudomonas putida, Vibrio fluvialis could volatilize 60%-95%, 94%, 100%, 60% of Hg 2+, respectively, from culture ( Zhang etal., 2012; Dash etal., 2013; Giri etal., 2014; Saranya etal., 2017; Zheng etal., 2018). The GC-MS study was included to get insight into the metabolic changes in the MRB isolates under Hg stress. It revealed that the metabolic profiles of MRB isolates are diverse with different biological properties. GC-MS metabolic profiles revealed that the number of compounds increased in the presence of Hg compared to the control isolates, where some compounds were found to be different in Hg-treated samples. A total of 50 metabolites were identified in the absence of Hg, whereas with 50 mg/L of Hg, 64 compounds were identified. The minimum metabolites (6) were identified in NIOT-EQR_J258 without Hg, whereas the maximum metabolites (22) was identified in NIOT-EQR_J251 with Hg 2+. The NIOT-EQR_J7 metabolic profile was almost the same in both control and Hg-treated samples. A total of 14 metabolites were present in the control sample, whereas 15 metabolites were identified with Hg. In the case of NIOT-EQR_J248, NIOT-EQR_J251, and NIOT-EQR_J258, a total of 10, 20, and 6 compounds were identified in the control sample, whereas 16, 22, and 11 metabolites were present with Hg, correspondingly. GC-MS analysis was performed to characterize the bacterial response and the metabolomic changes leading to Hg tolerance. The solvent extraction method was used to extract the bacterial metabolites. In brief, the freshly inoculated and exponentially grown MRB and MMRB bacterial cells in ZMB medium (non-exposed and exposed to 50 mg/L of Hg) were freeze-dried using BENCHTOP lyophilizer (VIRTIS Instrument, Gardiner, NY). For extraction of compounds, 50 mg of lyophilized bacterial cells were suspended in ethyl acetate and chloroform (1:1; v/v) and homogenized. After homogenization, the solution (crude extract) containing the metabolites was transferred to the clean glass vial by pipetting. These steps were repeated two-three times to obtain a pure and ample amount of sample. The separated organic fractions (crude extract) were treated with anhydrous NaSO 4 (Sigma-Aldrich) to remove moisture, which was again concentrated on the rotary evaporator (BUCHI Rotavapor R-215/V advanced, Switzerland) at RT and stored at -80°C until further analysis. The concentrated crude extract was re-suspended in 1 mL of Dichloromethane (DCM) and 5 µL of the sample was injected into the GC-MS analyzer (Agilent Technologies Instrument 7890A GC System, 240 Ion Trap GC/MS, USA). The GC-MS analysis was carried out under external ionization mode using a fused silica column HP 5 MS column (30 m × 0.320 mm × 0.25 µm). High purity helium was used as a carrier gas at a constant flow rate of 1 mL/min. For analysis, the chromatographic conditions i.e. initial injector and detector temperature, were set at 250°C and 330°C, respectively. The temperature of the column was programmed from 50°C (hold for 2 min) to 320°C (2 min hold), with a constant 5°C increment per minute and 1 min hold at 330°C. A metabolic library of all the separated compounds found via GC-MS analysis of bacterial extract was created and identified using NIST mass spectral library match. The PubChem CID, structures, names, and molecular weight of those bioactive compounds were obtained from the PubChem database. In-silico analysis The following four randomized controlled trials have been conducted to show that exercise helped lower both systolic and diastolic BP in participants: In the meta-analyses of randomized controlled trials conducted by Fagard et al., 72 trials have been conducted with an average of 40 participants per trial, which involved a 16-week study of 40 minutes of exercise sessions three times/week with an average intensity of 65% of heart rate [ 39]. This study showed a decrease in SBP of 6.9 mm Hg and a lowering of DBP of 4.9 mm Hg [ 39]. In 27 randomized controlled trials by Lee et al. with 1842 participants, the exercise regimen involved walking for 26.5 min/day for 4.4 days/week for a mean of 19 weeks [ 40]. This study shows that there is a larger effect with more intense and frequent exercise regimens for a longer duration [ 40]. There was a mean decrease in SBP of 5.2 to 11 mm Hg and in DBP of 3.8 to 7.7 mm Hg [ 40]. In the meta-analysis of randomized control trials performed by Cornelissen et al., 15 trials with 633 participants involving the exercise of 30-60 min, two to five times/week, at 50% to 75% HR reserve for six to 52 weeks showed a daytime decrease in SBP of 3.2 mm Hg and in DBP of 2.7 mm Hg [ 41]. However, no blood pressure reduction was seen at night [ 42]. The meta-analysis of randomized controlled trials performed by Cornelissen and Smart included 105 trials with 3957 participants [ 42]. This study concluded that moderate aerobic exercise involving walking and jogging for 30 to 60 min/session three to five times/week for four to 52 weeks showed a reduction in SBP of 3.5 mm Hg and in DBP of 2.5 mm Hg [ 42]. According to the American College of Cardiology, a BP higher than 180/120 mm Hg is considered a hypertensive emergency or crisis. Patients with these blood pressures need emergency medical help. Untreated high BP may increase the risk of myocardial infarction, stroke, and other serious complications. Monitoring BP every two years, starting at age 18, is important to diagnose and treat hypertension timely to prevent complications. HTN is diagnosed by performing repeated careful measurements of blood pressure. Blood pressure is categorized as follows: Normal blood pressure, defined as systolic blood pressure (SBP) less than 120, and diastolic blood pressure (DBP) less than 80. An elevated BP is an SBP of 120 to 129 and a DBP of less than 80. HTN is defined as a systolic pressure more than or equal to 130 or a diastolic pressure more than or equal to 80.
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