Morphometric, haematological and physio-biochemical characterization of Bactrian (Camelus bactrianus) camel at high altitude.

BACKGROUND: Biochemical and haematological parameters have not been determined in Bactrian camels kept at high altitude. Therefore, this study was undertaken to characterise different physiological, haematological, biochemical, and morphometric parameters of Bactrian camels of high altitude. For this, total fourteen high altitude healthy Bactrian camels were selected from Leh-Ladakh, India, a high altitude area, and thereafter divided into three age groups (N = 3 young; N = 6 adult; N = 5 old camels) to characterise for above parameters. All the results were compared with Lowlander Bactrian camels. RESULTS: Morphometric measurement showed significant difference in body height, body length, front-hump height and girth, back-hump height and girth, abdomen girth, neck length, and circumference of the shank in the young age group camels as compared to other age groups of Bactrian camels (p < 0.05). Furthermore, all the physiological and haematological parameters were similar in all the age groups of camels (p < 0.05). However, the leukocyte, erythrocyte, Hb, platelets, monocyte, and ESR level were towards the higher side of the normal reference range of Lowlander Bactrian camels. Whereas, the biochemical analysis revealed a significant increase in triglycerides and decrease in protein levels in the younger age group as compared to other age groups (p < 0.05). Although, albumin, aspartate aminotransferase, iron, magnesium, urea, and creatinine levels were insignificant among the different groups, but observed towards the higher side of the low altitude reference range. Interestingly, the glucose levels in all the groups were observed towards the lower side of the range, which showed metabolic adaptation to high altitude. CONCLUSION: These findings suggested there is morphometric and biochemical variation in Bactrian camel of high altitude. The results further helped in establishing novel reference ranges for these parameters in Highlander Bactrian camel. Hence, this study will be the basis of future research on a Bactrian camel from high-altitude cold desert and helpful for better camel husbandry and health management in high altitude.

A review on the biosynthesis of metal and metal salt nanoparticles by microbes.

Metal nanoparticles have received great attention from researchers across the world because of a plethora of applications in agriculture and the biomedical field as antioxidants and antimicrobial compounds. Over the past few years, green nanotechnology has emerged as a significant approach for the synthesis and fabrication of metal nanoparticles. This green route employs various reducing and stabilizing agents from biological resources for the synthesis of nanoparticles. The present article aims to review the progress made in recent years on nanoparticle biosynthesis by microbes. These microbial resources include bacteria, fungi, yeast, algae and viruses. This review mainly focuses on the biosynthesis of the most commonly studied metal and metal salt nanoparticles such as silver, gold, platinum, palladium, copper, cadmium, titanium oxide, zinc oxide and cadmium sulphide. These nanoparticles can be used in pharmaceutical products as antimicrobial and anti-biofilm agents, targeted delivery of anticancer drugs, water electrolysis, waste water treatment, biosensors, biocatalysis, crop protection against pathogens, degradation of dyes etc. This review will discuss in detail various microbial modes of nanoparticles synthesis and the mechanism of their synthesis by various bioreducing agents such as enzymes, peptides, proteins, electron shuttle quinones and exopolysaccharides. A thorough understanding of the molecular mechanism of biosynthesis is the need of the hour to develop a technology for large scale production of bio-mediated nanoparticles. The present review also discusses the advantages of various microbial approaches in nanoparticles synthesis and lacuna involved in such processes. This review also highlights the recent milestones achieved on large scale production and future perspectives of nanoparticles.

Valorization of waste glycerol for the production of poly (3-hydroxybutyrate) and poly (3-hydroxybutyrate-co-3-hydroxyvalerate) copolymer by Cupriavidus necator and extraction in a sustainable manner.

Glycerol is a by-product of many industrial processes and huge amounts of it are generated in the form of waste, thereby necessitating a search for the method of its disposal. An interesting solution is the valorization of crude glycerol into value added product such as polyhydroxyalkanoates (PHAs). The feasibility of producing PHAs by Cupriavidus necator was evaluated using crude glycerol (WG). Various cultivation strategies were designed for the production of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) copolymer by adding different organic acids as precursors at different concentrations levels. Batch cultivation of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) production showed accumulation of 6.76g/L biomass containing 4.84g/L copolymer on WG with a maximum 3-hydroxyvalerate content of 24.6mol%. PHAs extraction using a non-toxic and recyclable solvent, 1,2 propylene carbonate, showed the highest recovery yield (90%) and purity (93%) at 120°C temperature and 30min incubation. This is the first report on jatropha based glycerol valorization for poly(3-hydroxybutyrate-co-3-hydroxyvalerate) production coupled with extraction using non-toxic solvent.

Model-Based Nutrient Feeding Strategies for the Increased Production of Polyhydroxybutyrate (PHB) by Alcaligenes latus.

Polyhydroxyalkanoates (PHAs) are biodegradable polymers which are considered as an effective alternative for conventional plastics due to their mechanical properties similar to the latter. However, the widespread use of these polymers is still hampered due to their higher cost of production as compared to plastics. The production cost could be overcome by obtaining high yields and productivity. The goal of the present research was to enhance the yield of polyhydroxybutyrate (PHB) with the help of two simple fed-batch cultivation strategies. In the present study, average batch kinetic and substrate limitation/inhibition study data of Alcaligenes latus was used for the development of PHB model which was then adopted for designing various off-line nutrient feeding strategies to enhance PHB accumulation. The predictive ability of the model was validated by experimental implementation of two fed-batch strategies. One such dynamic strategy of fed-batch cultivation under pseudo-steady state with respect to nitrogen and simultaneous carbon feeding strategy resulted in significantly high biomass and PHB concentration of 39.17 g/L and 29.64 g/L, respectively. This feeding strategy demonstrated a high PHB productivity and PHB content of 0.6 g/L h and 75%, respectively, which were remarkably high in comparison to batch cultivation. The mathematical model can also be employed for designing various other nutrient feeding strategies.

Microbial glycolipoprotein-capped silver nanoparticles as emerging antibacterial agents against cholera.

BACKGROUND: With the increased number of cholera outbreaks and emergence of multidrug resistance in Vibrio cholerae strains it has become necessary for the scientific community to devise and develop novel therapeutic approaches against cholera. Recent studies have indicated plausibility of therapeutic application of metal nano-materials. Among these, silver nanoparticles (AgNPs) have emerged as a potential antimicrobial agent to combat infectious diseases. At present nanoparticles are mostly produced using physical or chemical techniques which are toxic and hazardous. Thus exploitation of microbial systems could be a green eco-friendly approach for the synthesis of nanoparticles having similar or even better antimicrobial activity and biocompatibility. Hence, it would be worth to explore the possibility of utilization of microbial silver nanoparticles and their conjugates as potential novel therapeutic agent against infectious diseases like cholera. RESULTS: The present study attempted utilization of Ochrobactrum rhizosphaerae for the production of AgNPs and focused on investigating their role as antimicrobial agents against cholera. Later the exopolymer, purified from the culture supernatant, was used for the synthesis of spherical shaped AgNPs of around 10 nm size. Further the exopolymer was characterized as glycolipoprotein (GLP). Antibacterial activity of the novel GLP-AgNPs conjugate was evaluated by minimum inhibitory concentration, XTT reduction assay, scanning electron microscopy (SEM) and growth curve analysis. SEM studies revealed that AgNPs treatment resulted in intracellular contents leakage and cell lysis. CONCLUSION: The potential of microbially synthesized nanoparticles, as novel therapeutic agents, is still relatively less explored. In fact, the present study first time demonstrated that a glycolipoprotein secreted by the O. rhizosphaerae strain can be exploited for production of AgNPs which can further be employed to treat infectious diseases. Although this type of polymer has been obtained earlier from marine fungi and bacteria, none of these reports have studied the role of this polymer in AgNPs synthesis and its application in cholera therapy. Interestingly, the microbial GLP-capped AgNPs exhibited antibacterial activity against V. cholerae comparable to ciprofloxacin. Thus the present study may open up new avenues for development of novel therapeutic agents for treatment of infectious diseases. Graphical abstract Development of novel therapeutic agents for treatment of cholera.

Development of a mathematical model for the growth associated Polyhydroxybutyrate fermentation by Azohydromonas australica and its use for the design of fed-batch cultivation strategies.

In the present investigation, batch cultivation of Azohydromonas australica DSM 1124 was carried out in a bioreactor for growth associated PHB production. The observed batch PHB production kinetics data was then used for the development of a mathematical model which adequately described the substrate limitation and inhibition during the cultivation. The statistical validity test demonstrated that the proposed mathematical model predictions were significant at 99% confidence level. The model was thereafter extrapolated to fed-batch to identify various nutrients feeding regimes during the bioreactor cultivation to improve the PHB accumulation. The distinct capability of the mathematical model to predict highly dynamic fed-batch cultivation strategies was demonstrated by experimental implementation of two fed-batch cultivation strategies. A significantly high PHB concentration of 22.65 g/L & an overall PHB content of 76% was achieved during constant feed rate fed-batch cultivation which is the highest PHB content reported so far using A. australica.

Use of NAD(P)H fluorescence measurement for on-line monitoring of metabolic state of Azohydromonas australica in poly(3-hydroxybutyrate) production.

Culture fluorescence measurement is an indirect and non-invasive method of biomass estimation to assess the metabolic state of the microorganism in a fermentation process. In the present investigation, NAD(P)H fluorescence has been used for on-line in situ characterization of metabolic changes occurring during different phases of batch cultivation of Azohydromonas australica in growth associated poly(3-hydroxybutyrate) or PHB production. A linear correlation between biomass concentration and net NAD(P)H fluorescence was obtained during early log phase (3-12 h) and late log phase (24-39 h) of PHB fermentation. After 12 h (mid log phase) cultivation PHB accumulation shot up and a drop in culture fluorescence was observed which synchronously exhibited continuous utilization of NAD(P)H for the synthesis of biomass and PHB formation simultaneously. A decrease in the observed net fluorescence value was observed again towards the end of fermentation (at 39 h) which corresponded very well with the culture starvation and substrate depletion towards the end of cultivation inside the bioreactor. It was therefore concluded that NAD(P)H fluorescence measurements could be used for indication of the time of fresh nutrient (substrate) feed during substrate limitation to further enhance the PHB production.

Estimation of fundamental kinetic parameters of polyhydroxybutyrate fermentation process of Azohydromonas australica using statistical approach of media optimization.

Polyhydroxybutyrate or PHB is a biodegradable and biocompatible thermoplastic with many interesting applications in medicine, food packaging, and tissue engineering materials. The present study deals with the enhanced production of PHB by Azohydromonas australica using sucrose and the estimation of fundamental kinetic parameters of PHB fermentation process. The preliminary culture growth inhibition studies were followed by statistical optimization of medium recipe using response surface methodology to increase the PHB production. Later on batch cultivation in a 7-L bioreactor was attempted using optimum concentration of medium components (process variables) obtained from statistical design to identify the batch growth and product kinetics parameters of PHB fermentation. A. australica exhibited a maximum biomass and PHB concentration of 8.71 and 6.24 g/L, respectively in bioreactor with an overall PHB production rate of 0.75 g/h. Bioreactor cultivation studies demonstrated that the specific biomass and PHB yield on sucrose was 0.37 and 0.29 g/g, respectively. The kinetic parameters obtained in the present investigation would be used in the development of a batch kinetic mathematical model for PHB production which will serve as launching pad for further process optimization studies, e.g., design of several bioreactor cultivation strategies to further enhance the biopolymer production.

Enhanced production of poly(3-hydroxybutyrate) in a novel airlift reactor with in situ cell retention using Azohydromonas australica.

Economic production of biodegradable plastics is a challenge particularly because of high substrate and energy cost inputs for its production. Research efforts are being directed towards innovations to minimize both of the above costs to economize polyhydroxybutyrate (PHB) production. A novel airlift reactor (ALR) with outer aeration and internal settling was utilized in this investigation. Although it featured no power consumption for agitation, it facilitated increased oxygen transfer rate and better cell retention than stirred tank reactor (STR), thereby resulting in enhanced PHB productivity. ALR with in situ cell retention demonstrated a significant improvement in biomass concentration and biopolymer accumulation. The total PHB production rate, specific biomass, and product yield in the ALR were observed to be 0.84 g/h, 0.43 g/g, and 0.32 g/g, respectively. The studies revealed that the volumetric oxygen mass transfer rate and mixing time for ALR were 0.016 s⁻¹ and 3.73 s, respectively, at 2.0 vvm as compared with corresponding values of 0.005 s⁻¹ and 4.95 s, respectively, in STR. This demonstrated that ALR has better oxygen mass transfer and mixing efficiency than STR. Hence, ALR with cell retention would serve as a better bioreactor design for economic biopolymer production than STR, particularly due to its lower cost of operation and simplicity along with its enhanced oxygen and heat transfer rates.