In vitro and in vivo effect of 2,6-Di-tert-butyl-4-methylphenol as an antibiofilm agent against quorum sensing mediated biofilm formation of Vibrio spp.

This study unveils the in vitro and in vivo antibiofilm potential of 2,6-Di-tert-butyl-4-methylphenol (DTBMP) from Chroococcus turgidus against Vibrio spp. In the preliminary study, cell free culture supernatant (CFCS) of C. turgidus inhibited the violacein production in biomarker strain Chromobacterium violaceum and its mutant strain CV026 in a dose dependent manner. The effective biofilm inhibitory concentration (BIC) of pure compound DTBMP from C. turgidus was identified as 250 µg/ml concentration in tested Vibrio species. Furthermore, DTBMP proved to effectively inhibit the bioluminescence production in V. harveyi and other biofilm related virulence traits such as exopolysaccharides (EPS) production, hydrophobicity index, swimming and swarming motility at its BIC concentration in three major pathogenic vibrios: V. harveyi, V. parahaemolyticus and V. vulnificus. The antibiofilm potential of DTBMP was validated through light, confocal laser scanning and scanning electron microscopic analyses. In addition, the non-bactericidal effect of DTBMP was determined through growth curve and 2,3-bis (2-methyloxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide (XTT) assay. Real-time PCR studies revealed the down-regulation of master quorum sensing (QS) regulator genes of V. harveyi such as luxR, luxS, luxP, luxQ and luxO on treatment with DTBMP. In vivo results confirmed that DTBMP augmented the survival rate of Litopenaeus vannamei larvae up to 75, 88 and 66% upon infection with V. harveyi, V. parahaemolyticus and V. vulnificus, respectively. The results of this study ascertain the promising effects of DTBMP as an antibiofilm agent, which could be positively explored to treat biofilm-associated vibrios infections in aquaculture.

Sargassum wightii-synthesized ZnO nanoparticles - from antibacterial and insecticidal activity to immunostimulatory effects on the green tiger shrimp Penaeus semisulcatus.

The green synthesis of metal nanoparticles using phytochemical from marine seaweeds is a fast-growing research field in nanotechnology. Here, the biosynthesis of zinc oxide nanoparticles was achieved using the hot water extract of Sargassum wightii. The hot water extract prepared from S. wightii (H Sw) and ZnO NPs were studied by UV-visible and FTIR spectroscopy, SEM and XRD. Then, both products were evaluated for antibiofilm activity towards aquatic pathogens. The nanoparticles' immunostimulating potential on green tiger prawns, Penaeus semisulcatus was studied through immersion and dietary administration. Shrimp immune parameters (i.e., total hemocytes count (THC), respiratory bursts (RBs), phenoloxidase (PO) and superoxide dismutase (SOD) activity) were significantly affected by exposure or ingestion of ZnO nanoparticles. In addition, the hot water extract and ZnO nanoparticles had high antibiofilm activity against Gram-positive (B. subtilis, S. aureus) and Gram-negative (S. sonnei, P. aeruginosa) microbial pathogens. It was accomplished that the ZnO nanoparticles can be used as the bacteriostatic and immunostimulant agents through immersion and dietary administration enhancing immunity of green tiger shrimp. Furthermore, the toxicity effects of ZnO nanoparticles were 100% at 24 h on Aedes aegypti 3 rd instar larvae at the concentration of 100 µg/mL and the greatest efficacy was accomplished by H Sw ZnO NPs against the Ae. aegypti after 24 h (LC50 49.22; LC90 86.96 mg/mL), if compared to the seaweed extract alone. Morphological and histological damages triggered by nanoexposure were investigated.

GFP tagged Vibrio parahaemolyticus Dahv2 infection and the protective effects of the probiotic Bacillus licheniformis Dahb1 on the growth, immune and antioxidant responses in Pangasius hypophthalmus.

In this study, the pathogenicity of GFP tagged Vibrio parahaemolyticus Dahv2 and the protective effect of the probiotic strain, Bacillus licheniformis Dahb1 was studied on the Asian catfish, Pangasius hypophthalmus. The experiment was carried out for 24 days with three groups and one group served as the control (without treatment). In the first group, P. hypophthalmus was orally infected with 1 mL of GFP tagged V. parahaemolyticus Dahv2 at two different doses (10(5) and 10(7) cfu mL(-1)). In the second group, P. hypophthalmus was orally administrated with 1 ml of the probiotic B. licheniformis Dahb1 at two different doses (10(5) and 10(7) cfu mL(-1)). In the third group, P. hypophthalmus was orally infected first with 1 mL of GFP tagged V. parahaemolyticus Dahv2 followed by the administration of 1 mL of B. licheniformis Dahb1 (combined treatment) at two different doses (10(5) and 10(7) cfu mL(-1)). The growth, immune (myeloperoxidase, respiratory burst, natural complement haemolytic and lysozyme activity) and antioxidant (glutathione-S-transferase, reduced glutathione and total glutathione) responses of P. hypophthalmus were reduced after post infection of GFP tagged V. parahaemolyticus Dahv2 compared to control. However, after administration with the probiotic B. licheniformis Dahb1 at 10(5) cfu mL(-1), P. hypophthalmus showed significant increase in the growth, immune and antioxidant responses compared to 10(7) cfu mL(-1). On the otherhand, the growth, immune and antioxidant responses of P. hypophthalmus infected and administrated with combined GFP tagged Vibrio + Bacillus at 10(5) cfu mL(-1) were relatively higher than that of GFP tagged V. parahaemolyticus Dahv2 and control groups but lower than that of probiotic B. licheniformis Dahb1 groups. The results of the present study conclude that the probiotic B. licheniformis Dahb1 at 10(5) cfu mL(-1) has the potential to protect the P. hypophthalmus against V. parahaemolyticus Dahv2 infection by enhancing the growth, immune and antioxidant responses. The probiotic B. licheniformis Dahb1 would be effectively used in the treatment of aquatic diseases for improvement of aquaculture industry.

Immune indices and identical functions of two prophenoloxidases from the haemolymph of green tiger shrimp Penaeus semisulcatus and its antibiofilm activity.

In the present study, we purified two prophenoloxidases (proPO) from haemolymph of green tiger shrimp, Penaeus semisulcatus by gel fermentation chromatography using blue Sepharose matrix. The two purified prophenoloxidase macromolecules are of about 76 and 75 kDa determined through SDS-PAGE and named as Penaeus semisulcatus prophenoloxidase I (PSproPO I) and Penaeus semisulcatus prophenoloxidase II (PSproPO II). It was further characterized by X-Ray Diffraction (XRD), Fourier Transform Infrared Spectroscopy (FTIR), Circular Dichroism (CD) and High Performance Liquid Chromatography (HPLC) analysis. The purified PSproPO I and PSproPO II showed the strongest agglutination titre against human erythrocytes compared to goat RBC. The PSproPO I and PSproPO II showed phagocytic activity against yeast Saccharomyces cerevisiae and encapsulation activity against Sepharose CL 6B beads compared to CM Sepharose and Sodium alginate beads. The functional analysis of purified PSproPO I and PSproPO II showed enhanced PO activity when added with the triggering molecules such as pathogen associated molecular patterns (PAMPs), metals and chemicals. In addition, eluted fraction containing PSproPO I and PSproPO II showed antibiofilm activity against Gram positive and Gram negative bacteria. The above results concluded that no significant differences were found between the purified PSproPO I and PSproPO II immune indices and functions. This study might provide a sensitive platform to understand more about the critical roles of PSproPO I and PSproPO II in crustacean immune system.

Survival of living donor renal transplant recipients in Sri Lanka: a single-center study.

Chronic kidney disease is one of the main public health concerns in Sri Lanka. In comparison with dialysis, successful kidney transplantation improves both patient survival and quality of life, relieves the burden of dialysis in patients suffering from end-stage renal disease and decreases the cost of healthcare to the society and government. The objective of this retrospective cohort study was to evaluate graft and patient survival rates in patients who were transplanted from living donors at the Nephrology Unit of the National Hospital of Sri Lanka from January 2005 to January 2011. Data were collected using an interviewer-administered questionnaire and through a review of past medical records. The Kaplan-Meier method was used to determine the survival rate, the log rank test was used to compare survival curves and the Cox proportional hazard model was used for multivariate analysis. Mean follow-up was 26.44±16.6 months. The five-year death-censored graft survival of kidney transplant recipients from living donors in our center was 93.5% and the five-year patient survival was 82.2%, which is comparable with other transplant programs around the world. The number of acute rejection episodes was an independent risk factor for graft survival. Delayed graft function, younger recipient age and unknown cause of end-stage renal disease were found to be risk factors for graft failure but after adjusting for confounding factors, and the difference was not apparent.

Unusual presentation of aortic dissection: post-coital acute paraplegia with renal failure.

We report the case of a 45-year-old chronic smoker who presented with acute paraplegia occurring during coitus and subsequently developed acute renal failure (ARF) requiring dialysis. He had absent peripheral pulses in the lower limbs with evidence of acute ischemia. Doppler study showed dissecting aneurysm of thoracic aorta, thrombotic occlusion of the distal aorta from L1 level up to bifurcation and occlusion of the right renal artery by a thrombus that was confirmed by magnetic resonance imaging of the spine. He was not subjected to any vascular intervention as his lower limbs were not salvageable due to delay in the diagnosis. Post-coital aortic dissection and aortic dissection presenting with acute paraplegia and ARF are very rare. This is probably the first case report with post-coital acute aortic dissection presenting with paraplegia and ARF. This case emphasizes the importance of a careful examination of peripheral pulses in patients presenting with ARF and paraplegia.

Reflex anuria following acute cardiac event.

BACKGROUND: Reflex anuria is an uncommon cause for acute renal failure, which occurs almost always after manipulation or irritation to kidneys, ureter, bladder or other pelvic organs. CASE PRESENTATION: Here we describe a case of acute renal failure due to reflex anuria following acute cardiac event. This patient had background history of urolithiasis. In the absence of other pre renal, renal or post- renal causes for acute kidney injury, we believe reflex anuria is the causative entity for acute renal failure in our patient. CONCLUSION: Acute renal failure due to reflex anuria is related to a reflex mechanism involving arteriolar vasoconstriction and urethral spasm. Patients with reflex anuria can be successfully managed with medical or surgical interventions. Our case suggests that reflex anuria should be considered as one of the differential diagnosis of acute renal failure following acute cardiac event, especially in patients with background urological problem.