Anjum Nasim Sabri

Auxin production by plant associated bacteria: impact on endogenous IAA content and growth of Triticum aestivum L.

Aims:  The aim of this study was to investigate the potential of bacterial strains of Bacillus, Pseudomonas, Escherichia, Micrococcus and Staphylococcus genera associated with wild herbaceous flora to enhance endogenous indole‐3‐acetic acid (IAA) content and growth of Triticum aestivum var. Inqalab‐91.

Bacterial exopolysaccharide and biofilm formation stimulate chickpea growth and soil aggregation under salt stress

To compensate for stress imposed by salinity, biofilm formation and exopolysaccharide production are significant strategies of salt tolerant bacteria to assist metabolism. We hypothesized that two previously isolated salt-tolerant strains Halomonas variabilis (HT1) and Planococcus rifietoensis (RT4) have an ability to improve plant growth, These strains can form biofilm and accumulate exopolysacharides at increasing salt stress. These results showed that bacteria might be involved in developing microbial communities under salt stress and helpful in colonizing of bacterial strains to plant roots and soil particles. Eventually, it can add to the plant growth and soil structure. We investigated the comparative effect of exopolysacharide and biofilm formation in two bacterial strains Halomonas variabilis (HT1) and Planococcus rifietoensis (RT4) in response to varying salt stress. We found that biofilm formation and exopolysaccharide accumulation increased at higher salinity. To check the effect of bacterial inoculation on the plant (Cicer arietinum Var. CM-98) growth and soil aggregation, pot experiment was conducted by growing seedlings under salt stress. Inoculation of both strains increased plant growth at elevated salt stress. Weight of soil aggregates attached with roots and present in soil were added at higher salt concentrations compared to untreated controls. Soil aggregation was higher at plant roots under salinity. These results suggest the feasibility of using above strains in improving plant growth and soil fertility under salinity.

Growth stimulation of Triticum aestivum seedlings under Cr-stresses by non-rhizospheric pseudomonad strains

Four chromium-resistant non-rhizospheric strains SPCr-1, SPCr-2, SPCr-3 and SPCr-4 (Pseudomonads), which were isolated from the effluents of an ICI paint factory and could tolerate 2-3 mg ml(-1) chromium in a minimal medium and 40 mg ml(-1) in a rich medium, were used to inoculate seeds of Triticum aestivum. Both inoculated and non-inoculated seeds were germinated and grown under different concentrations of chromium salts (K2CrO4, 0, 100, 250, 500; CrCl3, 0, 250, 500, 1000 microg ml(-1)). Germination and growth parameters were severely affected by chromium-salts. K2CrO4 had more drastic effects than CrCl3 treatments. Seedlings had a hard and brittle texture and showed symptoms of hypertrophy. Brown spots on leaves and stems were visible and the tips of leaves were bifurcated and curled. The root system was also impaired, ranging from a browning of the tip to complete destruction of cortical tissues. Under chromium-stress conditions, inoculated plants had significantly better germination and growth as compared to non-inoculated treatments. Bacterial growth enhancement of seedlings was associated with reduced chromium-uptake, increased auxin content and the formation of stress specific proteins. With bacterial inoculations, symptoms of chromium toxicity were reversed, especially at lower concentrations of chromium salts.

Effect of Biocides on Biofilm Bacteria from Dental Unit Water Lines

Microbial biofilm formation in dental unit water lines (DUWL) is a phenomenon that has been recognized for nearly four decades. Water delivered by DUWL can harbor high numbers of bacteria, including opportunistic pathogens. Biofilms on tubing within DUWL may serve as a reservoir for these microorganisms and should therefore be controlled. In this study, the effects of eight biocides were monitored on DUWL biofilms individually and in combination by epifluorescence microscopy and total viable counts (TVC). The effects of sodium dodecyl sulphate (SDS), hydrogen peroxide (H2O2), sodium hypochlorite (NaOCl), phenol (Phe), Tween 20 (Tw 20), ethylenediaminetetraacetic acid (EDTA), chlorohexidine gluconate (CHX), and povidine iodine (PI) were tested on DUWL biofilms alone and in combination. PI was found to have negligible effects on biofilm removal either applied alone or in combined form with CHX. Applying all biocides simultaneously did not completely eliminate viable bacteria nor did they remove biofilm. Overall, when combined, the biocides performed better than singly applied products. The most effective biocides were NaOCl and Phe (both alone and in combination).

Analysis of cell wall constituents of biocide-resistant isolates from dental-unit water line biofilms.

In past years, the significance of microbial resistance to biocides has increased. Twenty biocide-resistant bacterial strains were isolated from dental-unit water line biofilm. All strains resisted high biocide concentrations (up to 100 μg ml−1): sodium dodecyl sulphate, hydrogen peroxide, sodium hypochlorite, phenol, Tween 20, ethylenediaminetetraacetic acid, chlorohexidine gluconate, and povidine iodine. Among bacteria, biocide sensitivity is based on permeability of biocides through the cell wall. Gram-positive bacteria are more permeable and susceptible to biocides, whereas Gram-negative bacteria have a more complex cell wall and are the least sensitive bacteria. The present study was designed to study the effect of biocides on the cell wall of biocide-resistant bacteria. Peptidoglycan (PG), diaminopimelic acid (DAP), and teichoic acid contents of the cell wall were determined in L-broth and L-broth supplemented with biocides at different temperatures (37°C and 45°C) and pH levels (7 and 9). In general and Gram staining–specific comparison, a significant increase (p < 0.05) in the DAP content of biocide-resistant bacteria was observed at pH 7 and at both temperatures. In tubing-specific comparison, a significant increase in the amount of teichoic acid in air water tubing (37°C at pH 9) and DAP in patient tubing (pH 7 at both temperatures) was observed. In main water pipe, a significant decrease (p > 0.05) in PG content was noticed at 45°C and pH 9. Overall, a significant increase in DAP content may be an important constituent in the manifestation of isolate resistance against various biocides.

Biofilm formation in moderately halophilic bacteria is influenced by varying salinity levels

Bacteria in a biofilm have a co‐dependent lifestyle resulting in a harmonized and complex coordination of the bacterial cells within an exopolysaccharide (EPS) matrix. We hypothesized that biofilm formation and EPS production in salt‐tolerant bacteria are helpful for plant growth improvement in saline soil, but that they are influenced differently. To investigate this hypothesis, we tested the effect of different salinity levels on the biofilm formation of the bacterial strains PAa6 (Halomonas meridiana), HT2 (Kushneria indalinina) and ST2 (Halomonas aquamarina) on different abiotic and biotic surfaces. Maximum biofilm formation was established at 1 M salt concentration. However, EPS production was maximal at 0–1 M NaCl stress. We also studied the effect of salt stress on EPS produced by the bacterial strains and confirmed the presence of EPS on Cicer arietinum var. CM 98 roots and in soil at different salinity levels, using Alcian blue staining. Overall, the strain PAa6 was more effective in biofilm formation and EPS production. Under saline and non‐saline conditions, this strain also colonized the plant roots more efficiently as compared to the other two strains. We conclude that the strain PAa6 has the potential of biofilm formation and EPS production at different salinity levels. The presence of EPS in the biofilm helped the bacterial strains to better colonize the roots. (© 2012 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)

Hydropericardium syndrome in Pakistan: a review

Hydropericardium syndrome has caused huge economic losses to the poultry industry in Pakistan. The disease was reported for the first time in 1987 at Angara Goth, a broiler producing area near Karachi and given the name Angara disease. The disease is prevalent in 3–7 week old broilers and characterized by its sudden onset, accumulation of a jelly like fluid in the pericardial sac, inflamed liver with basophilic intranuclear inclusion bodies, congested kidneys and up to 70% mortality rates. Fowl adenovirus 4 has been found to be the causative agent. In Pakistan, the status of scientific research on various aspects of hydropericardium syndrome is quite unsatisfactory. There is an extensive use of formalin inactivated liver homogenate vaccines for the prevention and control of disease. Moreover, the gross lesions seen in post mortem examination are only the criteria for the clinical diagnosis of disease. Keeping in view the economic significance of hydropericardium syndrome, the need for additional scientific research on proper diagnosis and control of disease is suggested.

Isolation and characterization of biocides resistant bacteria from dental unit water line biofilms.

Six biocides resistant isolates were isolated from dental unit water lines (DUWL) in Pakistan. All isolates could tolerate 150 μg ml–1 of biocides (5.25% sodium hypocholrite, 35% H2O2, 4% tween 20, 1% povidine iodine, 0.2% chlorohexidine gluconate, 1% ethylene di‐amino tetra acetic acid and 1% phenol) on l‐agar and 100 μg ml–1 in l‐broth. The growth rate of all isolates was determined by generating growth curves at 37 °C for 48 h. The isolates were found to differ in growth rates with lag phase varying from (4–6 h) in biocides supplemented media compared to 2–4 h in biocides free medium. They have wide temperatures (24–42 °C) and pH (5–9) ranges. Traditional ways of identification of bacteria by phenotypic characteristics were accomplished by phenotypic and biochemical characterization. Heavy metals and antimicrobial susceptibility tests indicated that all isolates examined were resistant to trimethoprim, chloramphenicol while sensitive to HgCl2 and Pb (NO3)2. Almost all isolates were opportunistic pathogens. The 16S rRNA‐encoding genes from these six isolates were sequenced to confirm the identity of these isolates. 5 different genera (Bacillus, Achromobacter, Pseudomonas and Klebsiella) of bacteria were identified by 16S rDNA genes amplified from genomic DNA of biocides resistant DUWL biofilm isolates. Analysis of 16S rDNA genes revealed a much more clear identification of microrganisms than culture methods. However, different species of the same genera can have the same 16S rRNA gene sequence but are different due to phenotypic differences or different clinical manifestations. (© 2009 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)

Effects of temperature and pH on conjugal transfer of zinc-resistant plasmids residing in Gram-negative bacteria isolated from industrial effluents

Two zinc (Zn)-resistant strains, AnZn-1 and AnZn-2, which were resistant to ZnSO4 up to 12.5 mg ml(-1) were isolated from industrial effluents. Both were Gram-negative with motile cells. They exhibited tolerance to Ba2+, Ni+, Co2+, Mn2+, Cu2+, Fe2+, Ni2+, Cd2+, kanamycin, chloramphenicol, ampillicin and tetracycline, but were sensitive to Hg2+ and streptomycin. For AnZn-1 and AnZn-2, the optimum pH for growth was 7. Both were facultative anaerobes and had cytochrome oxidase and urease enzymes, while catalase was present only in AnZn-2. Both strains had the ability to hydrolyse gelatin, reduce nitrate, and yield acid from arabinose and rhamnose. The two strains shared maximum characters with Vibrionaceae. Each strain carries a single Zn-resistant conjugative plasmid. The plasmid residing in AnZn-1 (pSH1211) displayed a lower level of resistance than the plasmid of AnZn-2 (pSH1212). Both required a minimum of 24 h for mating and showed highest transfer frequency at 25 degrees C. pSH1211 preferred pH 7 and pSH1212 pH 8.5 for their transfer. Both plasmids, when allowed to mate with Escherichia coli at 25 degrees C, alkaline pH values of 8-8.5 (pSH1211) of pH 7.5 (pSH1212), showed increased transfer frequency.

Chloroform extract of turmeric inhibits biofilm formation, EPS production and motility in antibiotic resistant bacteria

In the form of biofilms, bacteria exhibit more resistance to antibiotics. Biofilm formers can withstand severe conditions and the hosts defense system. Therefore, it is necessary to search for effective biofilm inhibitors. In this study, we investigated the effect of a chloroform extract of turmeric on biofilm formation against antibiotic resistant bacteria. The extract exhibited its antibiofilm effect by altering adherence, motility, extracellular polymeric substance (EPS) production and cell surface hydrophobicity; important attributes of biofilm formation. Cell attachment assays indicated that a chloroform extract resulted in a 38.9-60.2% inhibition of cell adherence to a polystyrene surface, and a 44.5-58.3% inhibition to a glass surface. Static biofilm formation assays indicated that a chloroform extract resulted in a 23-74.5% reduction in biofilm formation. The chloroform extract inhibited flagella-directed swarming and swimming motility and pilus-directed twitching motility in a dose-dependent manner. In addition to repression of motility, a chloroform extract also significantly (p < 0.05) altered the hydrophobic behavior, and bacterial strains such as K. pneumoniae and E. cloacae exhibited hydrophilic behavior after the addition of the extract, as compared with control cells. The presence of the extract also significantly (p < 0.05) increased the detachment of biofilms by a surfactant as compared with controls. Fourier transformed infrared spectroscopy (FTIR) had indicated a loss of vital functional groups of polysaccharides and proteins from the EPS of cells treated with a chloroform extract. Gas chromatography mass spectrometry (GC-MS) analysis indicated the presence of many phytochemical constituents, mainly sesquiterpenes and fatty acid groups. These results clearly suggested that turmeric could affect multiple cellular activities in biofilm formers exhibiting antibiotic resistance by modulating adherence, EPS production, motility and surface hydrophobicity.