Aims: The primary aim of the present study is to evaluate the effect of rearing substrates on the nutritional content of black soldier fly larvae (BSFL) by incorporating Cupriavidus necator cells containing intracellular polyhydroxybutyrate (PHB) in BSFL diet to further increase the protein content and simultaneously to biologically extract the polymer by utilizing the digestive system of BSFL. The potential application of BSFL as a biological PHB extraction agent was determined. Methodology and results: Two feeding strategies consists of a mixture of protein (P) to carbohydrate (C) with a ratio of P50:C50 food waste (control feeding) and feed with bacterial cells (modified feeding). A comparison on the proximate analysis between this research and two commercially available products were conducted. Feeding BSFL with P50:C50 food waste revealed the highest crude protein content of 81.3 ± 0.2%. Additional bacteria cells in the BSFL diet, however, showed a negligible decrease in crude protein content of 0.67% as compared to the control feeding. Howbeit, this results comparably higher in contrast to the commercial products, with increment of crude protein content by 12.1% and 40.8%, respectively. Conclusion, significance and impact of study Two desirable products were obtained from the feeding with cells: (1) high protein content of BSFL and (2) biologically extracted polymer. This is the first study to demonstrate the utilization of BSFL as a biological extraction agent to partially extract biopolymer and increase the protein content by feeding with cells.
Diptera ; Polyhydroxybutyrates ; Animal Feed

The physis of a long bone may get 'sandwiched' and crushed between the metaphysis and the epiphysis if it is traumatically loaded along its long axis. Such a physeal injury may lead to complications like angular deformities and growth restrictions and hence, management of such injuries requires adequate planning and attentive execution. Two patients with distal femoral physeal crush injury were treated using a ring fixator such that one ring had the wires passing through the epiphysis and the other through the femoral shaft. On table image intensifier controlled distraction of the crushed physis was done to bring the height of the physis similar to that of the opposite limb. Patients were followed up for more than two years clinically and radiologically. There was no clinical or radiological angular deformity of the operated limbs. MRI scans showed intact physes with no physeal bar formation in either of the two patients. The distraction obtained by the ring fixator appears to have provided ample 'breathing space' to the compressed physis and that the growth potential may have been re-gained by the procedure. However, two years is a relatively short duration of follow-up and further follow-up of longer duration and in greater number of patients is needed to gauge the actual effectiveness of the technique used by us.

Background: Unsafe injection practice can transmit various blood borne infections. The aim of this study was to assess the knowledge and practice of injection safety among injection providers, to obtain information about disposal of injectable devices, and to compare the knowledge and practices of urban and rural injection providers. Methods: The study was conducted with injection providers working at primary health care facilities within Kaski district, Nepal. Ninety-six health care workers from 69 primary health care facilities were studied and 132 injection events observed. A semi-structured checklist was used for observing injection practice and a questionnaire for the survey. Respondents were interviewed to complete the questionnaire and obtain possible explanations for certain observed behaviors. Results: All injection providers knew of at least one pathogen transmitted through use/re-use of unsterile syringes. Proportion of injection providers naming hepatitis/jaundice as one of the diseases transmitted by unsafe injection practice was significantly higher in urban (75.6%) than in rural (39.2%) area. However, compared to urban respondents (13.3%), a significantly higher proportion of rural respondents (37.3%) named Hepatitis B specifically as one of the diseases transmitted. Median (inter-quartile range) number of therapeutic injection and injectable vaccine administered per day by the injection providers were 2 (1) and 1 (1), respectively. Two handed recapping by injection providers was significantly higher in urban area (33.3%) than in rural areas (21.6%). Most providers were not aware of the post exposure prophylaxis guideline. Conclusion: The knowledge of the injection providers about safe injection practice was acceptable. The use of safe injection practice by providers in urban and rural health care facilities was almost similar. The deficiencies noted in the practice must be addressed.
Health Personnel ; Injections

Aims: The study was carried out to isolate and identify the spontaneously growing populations of bacteria and fungi on the surface of biologically recovered polyhydroxyalkanoate (PHA) copolymer, poly(3-hydroxybutyrate-co-3- hydroxyhexanoate) [P(3HB-co-3HHx)]. Methodology and results: Large-scale PHA biosynthesis was carried out using 300 L fermenter and a biological methodology developed in our laboratory was utilized for PHA recovery. Using standard microbiological and molecular biology techniques the naturally growing microbial populations on the surface of biologically recovered PHA were identified. Scanning electron microscopy (SEM) analysis showed that the identified bacterial (Bacillus cereus and Burkholderia cepacia) and fungal isolates (Aspergillus niger, Byssochlamys nivea, Penicillium citrinum and Penicillium griseofulvum) were able to grow on and degrade the P(3HB-co-3HHx) copolymer. Conclusion, significance and impact of study: This is the first report on biologically recovered PHA pellet addressing the occurrence of microorganisms that grew spontaneously on it during storage under laboratory conditions. Fungi appeared to be dominant over bacteria in their ability to colonize the biologically recovered PHA.
Bacteria ; Fungi

Aims: This study is focused on the isolation, characterization and screening of new bacterial strains isolated from soil and wastewater samples that are able to produce PHA utilizing waste glycerol as sole carbon source in order to create useful products from waste glycerol and at the same time reduce the PHA production cost. A new isolate, Burkholderia contaminans Kad1 strain was investigated for its ability to biosynthesize PHA copolymers containing 3HV monomers from waste glycerol and 3-hydroxyvalerate (3HV) precursors. Methodology and results: PHA producing bacteria were screened using Nile Red and 1% of Nile Blue method. The presence of PHA granules was detected using phase contrast and fluorescence microscopy. Burkholderia contaminans Kad1, one out of 23 positive samples, was selected for further study because of its ability to produce high PHA content (47 wt%) and dry cell weight (DCW), (4.2 g/L) when waste glycerol 2% (v/v) was used as the sole carbon source. The 16S rDNA and the PHA synthase gene were sequenced and the PHA produced was confirmed by NMR analysis. A mixture of waste glycerol and sodium valerate fed to the culture gave rise to poly(3-hydroxybutyrate-co-3- hydroxyvalerate) [P(3HB-co-3HV)]. The mole fraction of 3HV monomer in the co-polymer P(3HB-co-3HV) sample analyzed using 1H NMR was 23 mol%. Conclusion, significance and impact of study: This study demonstrated for the first time B. contaminans Kad1 was able to use waste glycerol for PHA biosynthesis including the P(3HB-co-3HV) copolymer using a mixture of waste glycerol with sodium valerate as the precursor.
bacteria

Petrochemical-derived plastics have become a source of pollution for decades, and finding alternative plastics that are environmentally friendly has become a matter of urgency. Polyhydroxyalkanoate (PHA), a biopolyester synthesized by microbial cells, has properties that make it suitable as a biodegradable plastic material. The diversity of PHA makes it applicable to a wide range of products, from packaging to biomedical devices. The main challenge in commercialization of PHA is the cost of production. Although many studies have been focused on obtaining high yields of PHA, up until now, there is no absolute definition of efficient production of PHA, as there are many factors that could contribute to the efficiency of a process. Efficiency in PHA recovery also contributes to the commercial viability of PHA production. This review focuses on the efficiency of PHA biosynthesis from several aspects relating to the criteria for efficient production. The development of new strategies for improved production, including utilization of low cost carbon sources, genetic modification of PHA-producing microbes, and fermentation strategies are discussed here. Advances in recovery of PHA, as well as the potential of biological recovery techniques, are also highlighted in this review.

Aims: This project was aimed to study the microbial diversity of the limestone soil and its correlation with the environment. Methodology and results: The study was carried out using samples obtained from Gunung Lang, Ipoh, Perak in August 2013. X-ray diffraction analysis of the rock structure confirmed that the samples were of limestone origin. Besides that, soil analysis revealed that this area was fertile and rich in nutrients. It therefore served as a suitable habitat for microorganismal diversity to flourish. This was proven by the 16S rDNA metagenomic analysis which targeted on 16S rDNA variable region V3-V5 using Illumina MiSeq sequencer. Using this approach, a variety of microorganisms was identified and many yet to be characterized microorganisms were detected from this area. Conclusion, significance and impact of study: This is the first study in Malaysia that aimed to study the microbial diversity of limestone soils through metagenomic approach. The study showed that limestone is rich in microbial diversity and it is a place worth looking for novel microbes and genes of interest in biotechnology.
Calcium Carbonate

STUDY DESIGN: Cross-sectional study. PURPOSE: The aim of the study was to determine relationship between the degrees of radiologically demonstrated anatomical lumbar canal stenosis using magnetic resonance imaging (MRI) and its correlation with the patient's disability level, using the Oswestry Disability Index (ODI). OVERVIEW OF LITERATURE: The relationship between the imaging studies and clinical symptoms has been uncertain in patients suffering from symptomatic lumbar canal stenosis. There is a limited number of studies which correlates the degree of stenosis with simple reproducible scoring methods. METHODS: Fifty patients were selected from 350 patients who fulfilled the inclusion criteria. The patients answered the national-language translated form of ODI. The ratio of disability was interpreted, and the patients were grouped accordingly. They were subjected to MRI; and the anteroposterior diameters of the lumbar intervertebral disc spaces and the thecal sac cross sectional area were measured. Comparison was performed between the subdivisions of the degree of lumbar canal stenosis, based on the following: anteroposterior diameter (three groups: normal, relative stenosis and absolute stenosis); subdivisions of the degree of central canal stenosis, based on the thecal sac cross-sectional area, measured on axial views (three groups: normal, moderately stenotic and severely stenotic); and the ODI outcome, which was also presented in 20 percentiles. RESULTS: No significant correlation was established between the radiologically depicted anatomical lumbar stenosis and the Oswestry Disability scores. CONCLUSIONS: Magnetic resonance imaging alone should not be considered in isolation when assessing and treating patients diagnosed with lumbar canal stenosis.
Constriction, Pathologic* ; Cross-Sectional Studies ; Humans ; Intervertebral Disc ; Low Back Pain ; Lumbar Vertebrae ; Magnetic Resonance Imaging* ; Methods ; Observational Study* ; Radiculopathy ; Research Design

Aims: Improper disposal of domestic wastes, such as waste cooking oil (WCO), contributes to the deterioration of the environment and may lead to health problems. In this study, we evaluated the potential of plant-based WCO as a carbon source for the commercial biosynthesis of the bio-plastics, poly(3-hydroxybutyrate) and poly(3-hydroxybutyrate-co-3-hydroxyhexanoate). The consumption of WCO for this purpose would mitigate their pollution of the environment at the same time. Methodology and Results: WCO collected from several cafeterias in USM was tested as the carbon source for polyhydroxyalkanoates (PHA) production. A selection of suitable nitrogen source was first conducted in order to obtain an acceptable number of dry cell weight (DCW) and PHA content. Urea was found to be a suitable nitrogen source for the two bacterial strains used in our study, Cupriavidus necator H16 and its transformed mutant, C. necator PHB¯4 harboring the PHA synthase gene of Aeromonas caviae (PHB¯4/pBBREE32d13). With WCO as the sole carbon source, C. necator H16 yielded a relatively good dry cell weight (DCW=25.4 g/L), with 71 wt% poly(3-hydroxybutyrate) P(3HB) content. In comparison, the DCW obtained with fresh cooking oil (FCO) was 24.8 g/L. The production of poly(3 hydroxybutyrate-co-3-hydroxyhexanoate) [P(3HB-co-3HHx)] from WCO by the transformant C. necator PHB¯4 was comparable, yielding a DCW of 22.3 g/L and P(3HB-co-3HHx) content of 85 wt%. Lipase activities for both bacterial strains reached a maximum after 72 h of cultivation when time profile was conducted. Conclusion, significance and impact of study: The use of WCO as a carbon source in the biosynthesis of the bioplastic, PHA, turns a polluting domestic waste into a value-added biodegradable product. This renewable source material can thus be exploited for the low cost production of PHA.

Aims: This study evaluates potentials of Cupriavidus necator PHB4 transformant harboring the highly active polyhydroxyalkanoate synthase gene (phaC) of a locally isolated Chromobacterium sp. USM2 for its ability to incorporate 3-hydroxyheptanoate (3HHp) monomer. Methodology and results: A mixture of fructose and sodium heptanoate fed to the culture gave rise to poly(3- hydroxybutyrate-co-3-hydroxyvalerate-co-3-hydroxyheptanoate), [P(3HB-co-3HV-co-3HHp)] terpolymer synthesis, with traces of 3HHp monomers confirmed through gas chromatography (GC), proton (1H) and carbon (13C) NMR spectra. Conclusion, significance and impact of study: This study has revealed that the PHA synthase of Chromobacterium sp. USM2 has a broad range of substrate specificity. The synthase is able to polymerize 3-hydroxyalkanoate monomers having 4–7 carbon atoms.