Characterization of halo-tolerant plant growth promoting endophytic Bacillus licheniformis MHN 12.

BACKGROUND: Endophytic bacteria overlay significant role in plant growth promotion, eliminating phyto-pathogens and combating stress-conditions. In the present study, we aimed to screen high salt tolerant bacteria and study their adaptive response to elevated salt concentrations. A total of 46 endophytic bacterial isolates from Vigna radiata were screened for salt tolerance. The high salt tolerant endophytic isolate was characterized for alteration in morphology, growth rate, protein profiling, and compatible solute concentrations. RESULTS: The isolate MHN12, based upon biochemical characterization and partial 16S rDNA sequencing identified as B. licheniformis (accession number MG273753) was able to tolerate up to 15% NaCl (Sodium Chloride) (2.6 M) concentration. The isolate possessed 1-aminocyclopropane-1-carboxylic acid deaminase (ACCD) activity along with indole acetic acid (IAA), siderophore, ammonia, organic acid and hydrogen cyanide (HCN) production. Accumulation of proline was apparent up to 7.5% NaCl concentration and declined afterwards. Ultrastructure analysis using TEM (transmission electron microscopy) revealed the morphological alteration from rods to filaments. CONCLUSION: Acclimatization to salt stress and plant growth promoting activities could contribute to utilization of this bacterium as bioinoculant to enhance the crop yield and discourage the application of chemical fertilizers.

Exploration of plant growth-promoting endophytic bacteria from Pisum sativum and Cicer arietinum from South-West Haryana.

In the present study, nonrhizobial endophytes were isolated from Pisum sativum and Cicer arietinum from Haryana, India. A total of 355 bacterial endophytes were screened for plant growth promoting traits. Out of all, 96 bacterial endophytes were selected based on morphological characters and multi-PGP traits, and their diversity analyzed by amplified ribosomal DNA restriction analysis. Based on their ARDRA profile, the 25 representative isolates (12 from P. sativum and 13 from C. arietinum), were selected and identified by 16S ribosomal DNA sequencing. Genetic relatedness based on BLAST analysis revealed the similarity of these isolates with members of three prominent phyla, that is, Proteobacteria, Firmicutes, and Actinobacteria. The dominant cluster, Firmicutes, constituted 60% of the isolates, assigned to four different genera, Bacillus, Staphylococcus, Ornithinibacillus, and Lysinibacillus. Phylum α-proteobacteria included two genera, namely Paenochrobactrum and Ochrobactrum and three genera in phylum γ-proteobacteria, namely Pseudomonas, Pantoea and Proteus. The phylum Actinobacteria was constituted of two genera, Microbacterium and Arthrobacter. Bacillus zhangzhouensis, Bacillus safensis, Arthrobacter enclensis from P. sativum and Bacillus haynesii, Paenochrobactrum sp. from C. arietinum are documented as plant growth promoting endophytic bacteria for the first time in the present study. The in vitro and in vivo assessment based on bonitur score revealed that the endophytic isolates Bacillus mojavensis PRN2, Pseudomonas chlororaphis PHN9, B. safensis PRER2, Pseudomonas sp. RCP1, Pseudomonas lini PRN1 and B. haynensii RCP3 from P. sativum and C. arietinum significantly enhanced the plant growth parameters. Therefore, these potential isolates can be further harnessed for preparation of bioformulations to enhance sustainable agriculture.

Endophytism: A Multidimensional Approach to Plant-Prokaryotic Microbe Interaction.

Plant growth and development are positively regulated by the endophytic microbiome via both direct and indirect perspectives. Endophytes use phytohormone production to promote plant health along with other added benefits such as nutrient acquisition, nitrogen fixation, and survival under abiotic and biotic stress conditions. The ability of endophytes to penetrate the plant tissues, reside and interact with the host in multiple ways makes them unique. The common assumption that these endophytes interact with plants in a similar manner as the rhizospheric bacteria is a deterring factor to go deeper into their study, and more focus was on symbiotic associations and plant-pathogen reactions. The current focus has shifted on the complexity of relationships between host plants and their endophytic counterparts. It would be gripping to inspect how endophytes influence host gene expression and can be utilized to climb the ladder of "Sustainable agriculture." Advancements in various molecular techniques have provided an impetus to elucidate the complexity of endophytic microbiome. The present review is focused on canvassing different aspects concerned with the multidimensional interaction of endophytes with plants along with their application.

Annular intubation with pigtail probe for canalicular lacerations.

PURPOSE: Pigtail probe as a procedure of choice for canalicular laceration. METHODS: Retrospective, consecutive interventional case series of patients with eyelid laceration involving the canaliculus undergoing repair. All patients were subjected to repair with pigtail probe as first choice and only if this was not possible, they were repaired by other method. Outcome was analyzed in terms of cosmetic, functional, and anatomic success. RESULTS: A total of 35 patients (mean age, 10.88 years) underwent eyelid and canaliculus repair by a single surgeon (RM). In all, 32 (91.42%) canalicular lacerations were repaired by annular intubation using a pigtail probe, while the remaining 3 (8.58%) lacerations in which pigtail probe intubation was not possible, were repaired by an alternative method. Upper canaliculus was involved in 6 (17.14%) and lower canaliculus in 29 (82.86%) eyes. Mean follow-up was 8.2 months (range 3-13 months). Intubation tubes were removed after at least 3 months (range 12-20 weeks). None of the patients had complaints of epiphora. All the patients had good cosmesis, anatomic alignment and functional success as assessed by dye disappearance test in younger children and lacrimal irrigation in older children and adults. CONCLUSION: Bicanalicular annular repair with pigtail probe achieved excellent functional and cosmetic results. The loop minimizes the chances of extrusion of the tube, maintains natural anatomic alignment of the cut ends of the canaliculus and thus retains the integrity of the delicate canalicular system. Pigtail probe intubation can be considered as the first choice in canalicular lacerations.

Isolation and characterization of ACC Deaminase Producing Endophytic Bacillus mojavensis PRN2 from Pisum sativum.

BACKGROUND: Endophytic bacteria reside inside healthy plant tissues and provide several benefits to their host, and help them to tolerate various stresses. Aminocyclopropane-1-carboxylate deaminase (ACCD) production is one of the mechanisms by which these bacteria help the plant to survive under ethylene stress. OBJECTIVES: The main focus of this study was to isolate endophytic bacteria and effectively screen them for ACCD production. The selected isolate was identified and assessed for plant growth-promoting potential under pot conditions. MATERIALS AND METHODS: Endophytic bacteria were isolated from root nodules of Pisum sativum plants, grown in northern India (Haryana state). ACCD activity was initially screened on DF minimal salt medium with ACC as a sole nitrogen source. To narrow down the number of the isolates, another screening method was adopted using a modified medium containing indicator dyes along with ACC. The strain producing ACCD as well as a significant amount of Indole 3 acetic acid (IAA) was identified using 16S rDNA gene sequencing and amplification of acdS gene. Its ability to promote plant growth was evaluated under pot culture conditions. RESULTS: Twenty-six endophytic bacteria were isolated from nodules of P. sativum plants. Sixteen isolates showed growth on DF minimal salts medium supplemented with ACC along with negative control. On the modified medium containing indicator dyes, two isolates, PJN13 and PJN17, showed zones of the color gradient. The ACC deaminase activity was further confirmed by enzymatic assay. The strains PJN13 and PJN17 produced 160 and 130 µM of α-ketobutyrate m.g-1 protein h-1, respectively. The IAA production in the strain PJN13 (79.04 ± 0.78 µg.mL -1) was significantly more than that in the strain PJN17 (38.36 ± 1.89 µg.mL-1). It could enhance pea plant growth parameters, including root and shoot length and fresh and dry weight from 1 to 4 times compare to the control (untreated pea plants) under pot conditions. The results of 16S rDNA amplification and sequencing showed that PJN13 has maximum similarity to Bacillus mojavensis, and the sequence submitted to GenBank under accession number MH298523. Also, a band about 800 bp was amplified for the acdS gene. CONCLUSIONS: Though Bacillus is known as a predominant non-rhizobial endophytic genus, however in the present study, a B. mojavensis Bacillus mojavensis PRN2 (MH298523) was reported for the first time as an endophyte from the nodules of pea plants. The isolated strain possesses ACC deaminase activity along with IAA production capability, and high potentials as PGPE (Plant growth-promoting endophyte) for plant growth, so it has potential to be used as biofertilizers in pea fields.

Combined endothelial keratoplasty and clear lens extraction for corneal decompensation in irido-corneal endothelial syndrome.

A 38-year-old woman presented with corneal decompensation in left eye secondary to irido-corneal endothelial (ICE) syndrome. She underwent simultaneous Descemet's stripping endothelial keratoplasty (DSEK) and clear lens extraction with posterior chamber intraocular lens implantation. The surgery was accomplished comfortably without rupture of peripheral anterior synechiae (PAS). 5 weeks postoperatively, the graft was attached, the cornea was clear and best-corrected visual acuity improved from 20/400 to 20/30. DSEK combined with clear lens extraction appears to be an effective measure to treat corneal decompensation in patients with ICE syndrome. Associated lens extraction in such cases increases the working space in anterior chamber for DSEK, which minimizes the intra-operative graft manipulation. This also avoids a future difficult cataract surgery in the presence of PAS and an endothelial graft, which may increase the chances of graft survival.

Thyroid associated orbitopathy.

Thyroid associated orbitopathy, also known as Graves' orbitopathy, is typically a self-limiting autoimmune process associated with dysthyroid states. The clinical presentation may vary from very mild disease to severe irreversible sight-threatening complications. Despite ongoing basic science and clinical research, the pathogenesis and highly effective therapeutic strategies remain elusive. The present article reviews the pathophysiology, clinical presentation, and management of this common, yet poorly understood disease, which remains a challenge to the ophthalmologist.

Tonometry and Care of Tonometers.

Intraocular pressure (IOP) remains the only modifiable risk factor in the management of glaucoma. Hence, IOP and its appropriate measurement deserve our ongoing interest. Over the years, not only has our understanding of glaucoma changed but also has changed our approach to the measurement of the IOP. This review is an attempt to elucidate the commonly techniques of tonometry, and critically evaluate each of them, in current glaucoma practice. How to cite this article: Maheshwari R, Choudhari NS, Singh MD. Tonometry and Care of Tonometers. J Current Glau Prac 2012;6(3):124-130.

Muller's muscle resection for ptosis and relationship with levator and Muller's muscle function.

PURPOSE: Results of Muller's muscle resection in patients with ptosis and its relationship with levator and Muller's muscle function. METHODS: Prospective review of medical records of all pateints who underwent open sky Mullerectomy for correction of upper eyelid ptosis. The study period was from January 2008 to July 2009. Levator function, MRD1, Phenylepherine test result and extent of ptosis correction were recorded. Excised muscle tissue in few cases were subjected to histopathology. RESULTS: In 11 patients who underwent 13 mullerectomy procedures for correction of upper eyelid ptosis, MRD1 increased an average by 2.54 mm. The amount of ptosis correction was 4.28 mm in pateints with either good levator function and/or Muller's action and 1mm in patients where both responses to phenylepherine and levator functions were poor. Histopathology revealed both straited and smooth muscle tissue in the excised specimens in all cases. CONCLUSION: Mullerectomy porbably involves plication of the posterior layer of the levator aponeurosis. Results of Muller's muscle resection is good in patients with good levator function and/or good Mullers action and poor if both Muller and levator function are poor.

Phacotrabeculectomy without mitomycin C in primary angle-closure and open-angle glaucoma.

PURPOSE: To compare the surgical outcomes of single site phacotrabeculectomy without mitomycin C (MMC) in primary angle-closure glaucoma (PACG) and primary open-angle glaucoma (POAG). METHODS: We retrospectively reviewed the records of all patients with a diagnosis of PACG and POAG, who underwent single site phacotrabeculectomy without MMC between January 2001 and December 2005 and had a minimum follow-up of 12 months. The primary outcome measure was cumulative success probability, defined as complete [intraocular pressure (IOP) ≤21 mm Hg without antiglaucoma medications or additional surgery] and qualified (IOP <21 mm Hg with medications). Secondary outcome measures were reduction of IOP, the number of antiglaucoma medications at last follow-up, and complication rates. RESULTS: Seventy-one eyes of 63 PACG patients (mean age 61.2 y) and 72 eyes of 57 POAG patients (mean age 64.0 y) were analyzed. Mean duration of follow-up was 38.7 and 41.7 months in the PACG and POAG groups, respectively. Complete success in PACG (72.1%) was more than the POAG group (56.1%), but the difference was not statistically significant (P=0.06). Qualified success in the PACG and POAG group was 87.4% and 92.8%, respectively (P=0.43). IOP reduction was greater (P=0.03) in the PACG group and PACG group required fewer antiglaucoma medications postoperatively (P=0.03) for IOP control. CONCLUSIONS: Survival probability of single site phacotrabeculectomy without MMC was not significantly different between the PACG and POAG groups. IOP reduction was greater and the need for antiglaucoma medications after surgery was lesser in the PACG group.

Bilateral simultaneous acute angle closure caused by sulphonamide derivatives: a case series.

The sulphonamide group of drugs is implicated in bilateral acute angle closure (AAC) due to an idiosyncratic response. We report a series of three cases with bilateral AAC caused by different sulphonamide derivatives, their presentation and management.

Review of orbital exenteration from an eye care centre in Western India.

PURPOSE: Indications for orbital exenteration performed over a 10-year period from a tertiary eye care center in Western India. METHOD: Retrospective review of medical records of all patients who underwent exenteration of orbit over a period from January 1999 to April 2009, at a tertiary eye care center in western India. RESULT: Out of total 236 major orbital surgeries performed, 15 (6%) underwent orbital exenteration. Age ranged from 19 years to 75 years. Of these 9 were male and 6 female. Except for one case all exenteration were for malignant neoplasm. Lesions originated from eyelid in 8 (54%) cases, orbit in 5 (33%) cases and bulbar conjunctiva in 2 (13%) cases. Histopathological classification of the neoplasm were sebaceous carcinoma (4 cases), basal cell carcinoma (3 cases), squamous cell carcinoma (3 cases), adenoid cystic carcinoma (1 case), transitional cell carcinoma (1 case), malignant fibrous histiocytoma (1 case), lymphoma (1 case) and mucormycosis (1 case). CONCLUSION: Majority of exenterations performed were for neglected ocular or adnexal malignant lesions that could have been avoided provided patients reported earlier for treatment. Exenterations can be reduced by aggressive patient education about eye malignancy.

Agreement among 3 methods of optic disc diameter measurement.

PURPOSE: To assess the agreement among clinical, Heidelberg Retina Tomograph (HRT), and optic disc photography in the optic disc diameter measurement. METHODS: Prospective, observational study of 55 eyes of 55 subjects who underwent horizontal and vertical optic disc diameter measurements with +90 diopter lens on slit-lamp biomicroscopy (clinical) and on HRT and optic disc photography. Measurements were compared and agreement among the methods was assessed using intraclass correlation coefficients and Bland and Altman plots. RESULTS: Mean vertical diameter measured by clinical (1.93±0.28 mm) and on disc photography (1.9±0.22 mm) were comparable (P=0.812) and both were significantly greater (P<0.0001) than that measured on HRT (1.69±0.20 mm). Mean horizontal diameter by clinical (1.82±0.19 mm) and on disc photography (1.79±0.22 mm) were comparable (P=0.789) and both were significantly greater (P<0.01) than that measured on HRT (1.66±0.16 mm). The 95% limits of agreement on Bland and Altman plots ranged from 0.54 mm (clinical and disc photography) to 0.93 mm (clinical and HRT) for vertical diameter measurements and from 0.51 mm (HRT and disc photography) to 0.63 mm (clinical and HRT) for horizontal diameter measurements. Proportional bias was detected in the agreement between clinical and HRT as well as clinical and disc photography in vertical disc diameter measurement. CONCLUSIONS: Agreement between clinical and disc photography for optic disc diameter is better than that between either of these 2 methods and HRT. With the increasing disc size, there is an increasing disagreement in the vertical disc diameter measurements between the clinical and HRT as well as clinical and disc photography.

Extramedullary pasmacytoma masquerading as chalazion.

Extramedullary plasmacytoma (EMP) is a rare entity that belongs to the category of non-Hodgkin lymphoma. EMP make up 4% of all plasma cell tumors and occur mainly in the upper aerodigestive tract. Primary plasmacytoma involving the orbit is rare. There is a relation between solitary plasmacytoma and subsequent development of multiple myeloma. As no predictors of progression have been identified patients need indefinite follow-up. The authors report a case of EMP involving the eyelid in an otherwise healthy adult female.

Acute dacryocystitis causing orbital cellulitis and abscess.

PURPOSE: Acute dacryocystitis commonly presents as preseptal cellulitis as lacrimal sac lies anterior to the orbital septum and hence infection localizes in the preseptal space. The purpose of this study is to describe presentation and management of cases of acute dacryocystitis presenting as orbital cellilitis and abscess secondary to acute dacryocystitis. METHOD: This retrospective, non-comparative, interventional case series included 6 patients presenting with orbital cellulitis and abscess secondary to acute dacryocystitis. All patients underwent drainage of abscess under general anesthesia. External dcaryocystorhinostomy was advised as the definitive treatment for all pateints. RESULT: Abscess was localized in the extraconal space in 4 patients and in the intraconal space in 2 patients. Patients with intraconal abscess had visual loss on presentation while vision could be preserved in patients with abscess in extraconal space. Both patients with intraconal abscess had history of acute dacryocystitis. CONCLUSION: Acute dacryocystitis is an ophthalmic emergency that can cause orbital cellulitis with abscess formation, and even vision loss, if left untreated.

Balloon catheter dilation for complex congenital nasolacrimal duct obstruction in older children.

PURPOSE: To evaluate the effectiveness of lacrimal balloon catheter dilation in the treatment of complex congenital nasolacrimal duct obstruction in older children. METHODS: In a retrospective study, the case records of 65 eyes of 59 children 2 to 6 years (mean: 37 months) who underwent probing for congenital nasolacrimal duct obstruction were evaluated. Complex obstruction was noted in 21 (32.3%) of the 65 children older than 2 years who underwent probing. Secondary balloon dacryoplasty was performed in 8 (38.09%) of these 21 lacrimal systems. Patient age ranged from 3 to 6 years (mean: 47 months). Balloon catheter dilation was performed according to a standard protocol. Clinical patency of the nasolacrimal system was the main outcome measure and was defined as complete resolution of signs and symptoms (crusting, discharge, and watering) on follow-up. The procedures were performed under general anesthesia. All dilations were performed by the same lacrimal surgeon. RESULTS: Secondary balloon dacryoplasty was successful in seven of the eight lacrimal systems (87.5%). Complex obstruction at the distal end of the nasolacrimal duct was noted in all children. All children were older than 3 years (mean: 37 months), and all underwent secondary procedures because previous probing was unsuccessful. CONCLUSION: Balloon catheter dilation is an effective procedure for complex nasolacrimal duct obstruction in older children. It can be an alternative to silicone intubation and dacryocystorhinostomy in children after unsuccessful probing.

Extranodal Rosai-Dorfman disease presenting as an isolated epibulbar mass.

Rosai-Dorfman disease is a rare idiopathic disorder characterized by painless lymphadenopathy with cervical involvement in more than 80% cases. We report a case of Rosai-Dorfman disease presenting as an isolated epibulbar mass in a healthy young adult male. Epibulbar involvement in Rosai-Dorfman disease is a rare presentation as can be seen from a review of all literature. The presentation, differential diagnosis and treatment are discussed.

Success rate and cause of failure for late probing for congenital nasolacrimal duct obstruction.

PURPOSE: To evaluate the outcome of late probing for congenital nasolacrimal duct obstruction in children 2 to 6 years old and to identify the cause of failure in these children. METHODS: Records of 65 nasolacrimal duct obstructions in 62 children (39 boys and 23 girls) aged 2 to 6 years who underwent primary probing for congenital nasolacrimal duct obstructions were evaluated. The main observations were the type of obstruction encountered during probing (complex/simple), age at the time of probing, and success rate depending on the type of obstruction. Success was the main outcome measure and was defined as complete resolution of all signs and symptoms at 1 week of follow-up. The Mann-Whitney test for statistical significance was used to compare the results between simple and complex nasolacrimal duct obstructions. RESULTS: Six children had bilateral obstruction. The average age at the time of probing was 37.71 months in children with complex obstruction and 37.27 months in children with simple obstruction. Twenty-one eyes (32.20%) had complex obstruction and 44 eyes (67.69%) had simple obstruction. The success rate was 33% in children with complex obstruction and 97.72% in children with simple obstruction, which was statically significant. The overall success rate of the whole cohort was 76.92%. CONCLUSION: Older children with simple obstruction undergoing late probing had an excellent success rate in this study (97.72%). Complex obstruction is the main risk factor for failure of probing in older children. Probing is a successful and viable option in older children.