Assessment of the influence of intrinsic environmental and geographical factors on the bacterial ecology of pit latrines

Improving the rate and extent of faecal decomposition in basic forms of sanitation such as pit latrines would benefit around 1.7 billion users worldwide, but to do so requires a major advance in our understanding of the biology of these systems. As a critical first step, bacterial diversity and composition was studied in 30 latrines in Tanzania and Vietnam using pyrosequencing of 16S rRNA genes, and correlated with a number of intrinsic environmental factors such as pH, temperature, organic matter content/composition and geographical factors. Clear differences were observed at the operational taxonomic unit, family and phylum level in terms of richness and community composition between latrines in Tanzania and Vietnam. The results also clearly show that environmental variables, particularly substrate type and availability, can exert a strong structuring influence on bacterial communities in latrines from both countries. The origins and significance of these environmental differences are discussed. This work describes the bacterial ecology of pit latrines in combination with inherent latrine characteristics at an unprecedented level of detail. As such, it provides useful baseline information for future studies that aim to understand the factors that affect decomposition rates in pit latrines.     

Higher helminth ova counts and incomplete decomposition in sand-enveloped latrine pits in a coastal sub-district of Bangladesh

Pit latrines are the most common latrine technology in rural Bangladesh, and untreated effluent from pits can directly contaminate surrounding aquifers. Sand barriers installed around the latrine pit can help reduce contamination but can also alter the decomposition of the fecal sludge and accelerate pit fill-up, which can counteract their benefits. We aimed to evaluate whether there was a difference in decomposition of fecal sludge and survival of soil-transmitted helminth (STH) ova among latrines where a 50-cm sand barrier was installed surrounding and at the bottom of the pit, compared to latrines without a sand barrier, in coastal Bangladesh. We assessed decomposition in latrine pits by measuring the carbon-nitrogen (C/N) ratio of fecal sludge. We enumerated Ascaris lumbricoides and Trichuris trichiura ova in the pit following 18 and 24 months of latrine use. We compared these outcomes between latrines with and without sand barriers using generalized linear models with robust standard errors to adjust for clustering at the village level. The C/N ratio in latrines with and without a sand barrier was 13.47 vs. 22.64 (mean difference: 9.16, 95% CI: 0.15, 18.18). Pits with sand barriers filled more quickly and were reportedly emptied three times more frequently than pits without; 27/34 latrines with sand barriers vs. 9/34 latrines without barriers were emptied in the previous six months. Most reported disposal methods were unsafe. Compared to latrines without sand barriers, latrines with sand barriers had significantly higher log₁₀ mean counts of non-larvated A. lumbricoides ova (log₁₀ mean difference: 0.35, 95% CI: 0.12, 0.58) and T. trichiura ova (log₁₀ mean difference: 0.47, 95% CI: 0.20, 0.73). Larvated ova counts were similar for the two types of latrines for both A. lumbricoides and T. trichiura. Our findings suggest that sand barriers help contain helminth ova within the pits but pits with barriers fill up more quickly, leading to more frequent emptying of insufficiently decomposed fecal sludge. Further research is required on latrine technologies that can both isolate pathogens from the environment and achieve rapid decomposition.     

Characteristics of Latrines in Central Tanzania and Their Relation to Fly Catches

The disposal of human excreta in latrines is an important step in reducing the transmission of diarrhoeal diseases. However, in latrines, flies can access the latrine contents and serve as a mechanical transmitter of diarrhoeal pathogens. Furthermore, the latrine contents can be used as a breeding site for flies, which may further contribute to disease transmission. Latrines do not all produce flies, and there are some which produce only a few, while others can produce thousands. In order to understand the role of the latrine in determining this productivity, a pilot study was conducted, in which fifty latrines were observed in and around Ifakara, Tanzania. The characteristics of the latrine superstructure, use of the latrine, and chemical characteristics of pit latrine contents were compared to the numbers of flies collected in an exit trap placed over the drop hole in the latrine. Absence of a roof was found to have a significant positive association (t=3.17, p=0.003) with the total number of flies collected, and temporary superstructures, particularly as opposed to brick superstructures (z=4.26, p<0.001), and increased total solids in pit latrines (z=2.57, p=0.01) were significantly associated with increased numbers of blowflies leaving the latrine. The number of larvae per gram was significantly associated with the village from which samples were taken, with the largest difference between two villages outside Ifakara (z=2.12, p=0.03). The effect of latrine superstructure (roof, walls) on fly production may indicate that improvements in latrine construction could result in decreases in fly populations in areas where they transmit diarrhoeal pathogens.     

Pollution de l’eau souterraine et risques sanitaires à Yaoundé au Cameroun

The aim of this article consists of developing an ecosystemic approach to collect information about groundwater pollution in order to determine its origin and infer the health risks in Mingoa's watershed through the study of 12 spontaneous settlements. From an environmental point of view, family interviews and direct observations allowed us to inventory springs, wells and latrines, and to collect information. So, we have selected, on the basis of predefined criteria, 21 wells and springs for physico-chemical and bacteriological laboratory analyses. Two hundred children aged less than 5 years were subjected to KOAP examination. A number of 1224 latrines were inventoried, for an average use of 15 people per latrine, producing 913.3 kg/year of excreta. Most of 91% of people use these latrines. The superficial aquifers, connected to the wells and springs, present high values of nitrogen, phosphorous pollutant and fecal contamination indicators (coliforms and fecal streptococcus), which make water inappropriate to human consumption. PCA analysis establishes that water pollution comes mainly from traditional pit latrines. The overall prevalence of diarrhea and parasite infestation was important (= 40%). The presence of cysts of Entamoeba histolytica, Ascaris limbricoides, and Entamoeba coli confirms this high prevalence and testify to very approximate hygiene and environment conditions.     

Effects of temperature and stocking density on intensive culture of Pacific white shrimp in freshwater

In aquaculture, the application of predictive techniques based on statistical-mathematical modeling allows not only to project and study individual growth trajectories, but also to evaluate the probable effect of external factors that would explain their behavior over time. This is the case of this work, which takes the above as a principle to demonstrate the effect of water temperature on the growth of the Pacific white shrimp Litopenaeus vannamei cultured in fresh water (0 mg L⁻¹), using densities of 90, 120, 180, 230, 280 and 330 shrimp m⁻². Shrimp were exposed to water temperature between 11.5 °C and 31.6 °C. Temperature effect was determined using a parameterized Gompertz growth model with experimental data from each initial culture density. The best shrimp productivity yield was obtained above 26 °C, and the least efficient was below 22 °C. Densities of 90–180 shrimp m⁻² and 230–330 shrimp m⁻² generated a maximum average size of 12.6 g and 8.8 g in 30 weeks, respectively. Here we present the implications of the effect of water temperature on the intensive culture of white shrimp with zero salinity (0 mg L⁻¹) using these techniques from a predictive analytical approach.     

Antibiotic resistance profiles of coagulase-positive and coagulase-negative staphylococci from pit latrine fecal sludge in a peri-urban South African community

The aim of this study was to assess pit latrine samples from a peri-urban community in KwaZulu-Natal (South Africa) for the presence of multidrug-resistant (MDR) Staphylococcus spp. Standard procedures were used to isolate Staphylococcus spp. from pit latrine fecal sludge samples, with confirmation at genus level by polymerase chain reaction (PCR). Sixty-eight randomly selected pit latrine Staphylococcus spp. isolates were further characterized by using established disk diffusion procedures. An average Staphylococcus spp. count of 2.1?×?10⁵ CFU per g fecal material was established using two randomly selected pit latrine samples. Of the 68-selected Staphylococcus spp. pit latrine isolates, 49% were identified as coagulase positive, 51% as coagulase negative and 65% (12 coagulase positive, 32 coagulase negative isolates) were categorized as MDR. The majority (66/68) of Staphylococcus spp. isolates displayed resistance to fusidic acid while only 5/68 isolates displayed resistance to chloramphenicol. The pit latrine samples analyzed in this study are a source of MDR Staphylococcus spp., highlighting the need for proper hygiene and sanitation regimes in rural communities using these facilities.     

Microcosm assays and Taguchi experimental design for treatment of oil sludge containing high concentration of hydrocarbons

Microcosm assays and Taguchi experimental design was used to assess the biodegradation of an oil sludge produced by a gas processing unit. The study showed that the biodegradation of the sludge sample is feasible despite the high level of pollutants and complexity involved in the sludge. The physicochemical and microbiological characterization of the sludge revealed a high concentration of hydrocarbons (334,766 ± 7001 mg kg⁻¹ dry matter, d.m.) containing a variety of compounds between 6 and 73 carbon atoms in their structure, whereas the concentration of Fe was 60,000 mg kg⁻¹ d.m. and 26,800 mg kg⁻¹ d.m. of sulfide. A Taguchi L₉ experimental design comprising 4 variables and 3 levels moisture, nitrogen source, surfactant concentration and oxidant agent was performed, proving that moisture and nitrogen source are the major variables that affect CO₂ production and total petroleum hydrocarbons (TPH) degradation. The best experimental treatment yielded a TPH removal of 56,092 mg kg⁻¹ d.m. The treatment was carried out under the following conditions: 70% moisture, no oxidant agent, 0.5% of surfactant and NH₄Cl as nitrogen source.     

Correlation of TCE cometabolism with growth characteristics on aromatic substrates in toluene-degrading bacteria

We have constructed a bacterial library consisting of 97 strains of toluene-degrading bacteria from soil and activated sludge samples for examining their physiological properties in terms of cometabolism of TCE. Large variation of TCE degradation ability was observed in Gram-positive and Gram-negative strains, as well as diverse patterns of availability of aromatics as a growth substrate. No clear correlation was observed between the number of available substrates for growth and TCE degradation ability. However, the growth on some of the aromatics showed positive or negative correlations with TCE degradation ability. Kendall correlation constants (tau) for the growth on cumene, m-xylene, p-xylene, and m-cresol with TCE degradation ability were statistically significant (P < 0.001): their values were −0.44, −0.31, −0.26, 0.37, respectively. Among 12 of aromatics, only m-cresol showed positive correlation with TCE degradation ability. These findings would be useful for enrichment and isolation of the microbes that have TCE-cometabolism ability, which is a selective disadvantage through the toxicity or competitive inhibition against growth substrates.     

Himalayan musk deer (Moshcus leucogaster) behavior at latrine sites and their implications in conservation

Elusive species often use latrines which also serves as communication and information hubs. Thus, studying behavior at latrines may provide critical insights into the species’ ecology and behavior. While it is established that musk deer use latrines for defecating, very little is known about the endangered Himalayan musk deer (Moschus leucogaster) and their latrines. We examined musk deer behavior from the various video clips lasting 238 min altogether, captured at latrine sites during both breeding and non‐breeding seasons in the Annapurna Conservation Area of Nepal. A total of 428 visits by musk deer and 479 behavioral events by them were captured. We constructed an ethogram to describe musk deer behavior and compared behavior across individuals and sex using parametric and non‐parametric tests. We found that musk deer are crepuscular and nocturnal animals. Both male and female musk deer repeatedly and independently visited shared latrine sites as well as exclusively used latrine sites. The proportion of male musk deer visited latrine sites were significantly higher than that of female musk deer. Hence, male musk deer were found more active (>2 times) than females during both seasons. The most frequently observed behavioral activities at the latrine sites were defecating, sniffing and browsing, followed by scrapping and covering, and ignoring the latrine sites. The defecating and sniffing activity were performed throughout breeding and non‐breeding seasons to establish communication among peers and to claim territory. Based on the behaviors observed at the latrine sites, we can presume that Himalayan musk deer likely use latrines to convey various messages, including personality, maturity, sexual status, and territory marking among conspecifics. These findings of this study can be used for the conservation of musk deer in its natural habitat and replicated in captivity to enhance breeding performance that improves long‐term conservation prospects for this species.     

Characterization of the spatial distribution of latrines in reintroduced mountain gazelles: do latrines demarcate female group home ranges?

Mountain gazelle Gazella gazella in Saudi Arabia are listed as 'vulnerable' by the IUCN. At present, the species' survival is secured by extensive captive-breeding programmes and reintroductions into protected areas. Two reintroduction attempts (Ibex Reserve and Uruq Bani Ma'Arid protected areas) in Saudi Arabia have been undertaken in the past two decades. Post-monitoring of released individuals is essential for the success of such reintroduction programmes; however, cryptic species like mountain gazelles are extremely difficult to observe directly. As radio-tracking is a cost-intensive and invasive post-monitoring technique, we asked: how can reintroduced or remnant pockets of natural gazelle populations be monitored indirectly ? Here, we propose the use of latrine mapping as an effective, cost-efficient and non-invasive tool to survey the social organization of reintroduced mountain gazelles as an indicator for repatriation success. In this study, we used released radio-collared animals to characterize the spatial distribution of latrines within female group home ranges. Distance to the next latrine, latrine size, as well as numbers of fresh faecal pellet groups per latrine or presence of urination marks were used as dependent variables for step-wise backward multiple regressions and were correlated with various ecological factors. Most dependent variables were correlated with distance or direction from the nearest tree, but not indicative of home-range cores. Only latrine densities were distinctly higher in core areas of female group home ranges, and no pattern of peripheral marking was detected. Hence, latrine density is a good indicator of home-range use in female group home ranges. Mapping latrines and determining latrine densities are therefore the methods of choice to survey mountain gazelle populations.     

Olfactory communication and counter-marking in brown brocket deer <Emphasis Type="Italic">Mazama gouazoubira</Emphasis>

Scent marking by defecation and urination in numerous small latrines may be related to resource defence in brown brocket deer (Mazama gouazoubira). Both males and females seem to be territorial, and both contribute to latrines where their ranges overlap. Latrines could thus potentially function as centres of information exchange and intrasexual competition. Counter-marking occurs when animals respond to invaders' marks with a greater number of marks. The objectives of this experimental study were to determine whether brown brocket deer distinguish dung of presumed invaders from their own and whether they counter-mark such faecal deposits. Two samples of dung (from unknown males or females and the experimental animal) were introduced near the latrines of 21 captive deer (13 males and eight females), and we observed their responses, including investigative (sniffing) and marking (urination and defecation) behaviours. Males investigated the introduced dung and their own latrine significantly more when the dung was from an unknown male than when it was their own. Females investigated unknown female dung significantly more than their own. Males counter-marked the introduced dung and their latrine significantly more when the dung was from an unknown male than when it was their own. Males marked with a shorter latency and at a greater frequency than females. Our data indicated that males counter-mark most intensively dung from male ‘intruders’ which may be related to intrasexual competition and resource defence. Females showed a non-significant tendency to counter-mark same sex intruders.     

Carbon demand,utilization, and metabolic diversity of bacterioplankton in the frontal regimes of the Indian sector of the Southern Ocean

Bacterial production, respiration and metabolic diversity were measured up to 120 m depth in the Sub-Antarctic Front (SAF) and Polar Fronts I and II (PFI and PFII) of the Indian Ocean sector of the Southern Ocean during 2010 Austral Summer. Prokaryotic cell count was maximum at PFI and PFII (~109 cells L−1) and minimum at SAF (~107 cells L−1). Furthermore, integrated bacterial production was higher at PFI (1.07 mg C m−2 h−1) and PFII (0.72 mg C m−2 h−1) compared to SAF (0.61 mg C m−2 h−1). At PFII, integrated bacterial growth efficiency was higher (8.96) compared to PFI (7.42) and SAF (7.17), signifying that the net contribution of PFII to the microbial loop could be relatively pronounced. Enhanced cell numbers and production at polar fronts indicate that the dissolved organic matter could be converted to secondary biomass through the microbial loop. However, integrated bacterial respiration rate at PFII (0.83 mg C m−2 h−1) was lower than that at PFI (1.84 mg C m−2 h−1) resulting in higher growth efficiency at PFII. Metabolic flexibility at SAF was clearly brought about by utilization of carboxylic acids like D-malic acid and itaconic acid, and carbohydrates like N-acetyl D-glucosamine, D-cellobiose and D-lactose. Utilization of amino acids like glycyl L-glutamic acid and L-threonine, and an amine, phenylethylamine, was critical in determining the metabolic variability at PFI. PFII hosted microbes that utilized phenolic compounds (2-hydroxy benzoic acid and 4-hydroxy benzoic acid) and polymers (like Tween 80). Utilization of polyols over carbohydrates in polar waters indicates a niche with lesser influence of the Antarctic melt waters on the bacterioplankton metabolism.     

Genomic annotation and validation of bacterial consortium NDMC-1 for enhanced degradation of sugarcane bagasse

This study aims at designing a consortium using rumen bacterial isolates for enhancing the hydrolysis of sugarcane bagasse (SB) for efficient biofuel formation. The microbial population was screened through biochemical and molecular tools along with enzymatic activity to obtain potential isolates for diverse cellulolytic and hemicellulolytic carbohydrate active enzyme (CAZyme). Five strains (Paenibacillus, Bacillus, Enterobacter, and Microbacterium) were selected for designing the consortium NDMC-1. The hydrolytic efficiency of NDMC-1 was determined based on cellulase production with simultaneous rise in monosaccharides, oligosaccharides, and soluble chemical oxygen demand (sCOD) concentration. Cellulolytic machinery of these isolates was further explored using genome sequencing. The isolates selected for consortia NDMC-1 interacted synergistically leading to enhanced cellulase production. Maximal endoglucanase (1.67 μmol ml−1 min−1), exoglucanase (0.69 μmol ml−1 min−1), and β-glucosidase (2.03 μmol ml−1 min−1) activity were achieved with SB as a sole carbon source after 48 h of incubation. Enhancement in SB hydrolysis employing NDMC-1 was evident by the increase in sCOD from 609 to 2589 mg/l and release of 1295 mg/l reducing sugar, comprising 59.8%, 8.23%, and 6.16% of glucose, cellobiose, and cellotriose, respectively, which resulted in 5.5-fold rise in biogas production. On genome annotation, total 472 contigs from glycoside hydrolase family: 84 from Microbacterium arborescens ND21, 72 from Enterobacter cloacae ND22, 61 from Bacillus subtilis ND23, 116 from Paenibacillus polymyxa ND24, and 140 from Paenibacillus polymyxa ND25 were identified. On further analysis, total 33 cellulases, 59 hemicellulases, and 48 esterases were annotated in the reported genomes. This work proposes the application of consortia-based bioprocessing systems over the conventionally favorable single organism approach for efficient hydrolysis of cellulosic substrates to fermentable sugars.     

Growth condition and bacterial community for maximum hydrolysis of suspended organic materials in anaerobic digestion of food waste-recycling wastewater

This paper reports the effects of changing pH (5–7) and temperature (T, 40–60 °C) on the efficiencies of bacterial hydrolysis of suspended organic matter (SOM) in wastewater from food waste recycling (FWR) and the changes in the bacterial community responsible for this hydrolysis. Maximum hydrolysis efficiency (i.e., 50.5% reduction of volatile suspended solids) was predicted to occur at pH 5.7 and T = 44.5 °C. Changes in short-chain volatile organic acid profiles and in acidogenic bacterial communities were investigated under these conditions. Propionic and butyric acids concentrations increased rapidly during the first 2 days of incubation. Several band sequences consistent with Clostridium spp. were detected using denaturing gel gradient electrophoresis. Clostridium thermopalmarium and Clostridium novyi seemed to contribute to butyric acid production during the first 1.5 days of acidification of FWR wastewater, and C. thermopalmarium was a major butyric acid producer afterward. C. novyi was an important propionic acid producer. These two species appear to be important contributors to hydrolysis of SOM in the wastewater. Other acidogenic anaerobes, Aeromonas sharmana, Bacillus coagulans, and Pseudomonas plecoglossicida, were also indentified.     

Production of surfactin and fengycin by Bacillus subtilis in a bubbleless membrane bioreactor

Surfactin and fengycin are lipopeptide biosurfactants produced by Bacillus subtilis. This work describes for the first time the use of bubbleless bioreactors for the production of these lipopeptides by B. subtilis ATCC 21332 with aeration by a hollow fiber membrane air–liquid contactor to prevent foam formation. Three different configurations were tested: external aeration module made from either polyethersulfone (reactor BB1) or polypropylene (reactor BB2) and a submerged module in polypropylene (reactor BB3). Bacterial growth, glucose consumption, lipopeptide production, and oxygen uptake rate were monitored during the culture in the bioreactors. For all the tested membranes, the bioreactors were of satisfactory bacterial growth and lipopeptide production. In the three configurations, surfactin production related to the culture volume was in the same range: 242, 230, and 188 mg l⁻¹ for BB1, BB2, and BB3, respectively. Interestingly, high differences were observed for fengycin production: 47 mg l⁻¹ for BB1, 207 mg l⁻¹ for BB2, and 393 mg l⁻¹ for BB3. A significant proportion of surfactin was adsorbed on the membranes and reduced the volumetric oxygen mass transfer coefficient. The degree of adsorption depended on both the material and the structure of the membrane and was higher with the submerged polypropylene membrane.     

木质纤维素分解复合菌系WSD-5组成菌株的分离及其产酶特性

复合菌系WSD-5具有高效的分解能力和产酶能力,以探明WSD-5的协同分解机理和优化高效组合为目的,通过纯培养分离手段,获得了11株细菌和3株真菌。16S rDNA比对结果表明,细菌分别为Pseudomonas sp.、Pseudomonas aeruginosa、Achromobacter sp.、Stenotrophomonas sp.、Bacillus fusiformis、Bacillus cereus、Brevundimonas sp.、Ochrobactrum sp.、Cytophaga sp.、Benzo(a)pyrene-degrading bacter、Flavobacterium sp.的近缘种;26S rDNA比对结果表明3株真菌分别为Pseudallescheria boydii、Coprinus cinereus的近缘种。分离菌株中有4株细菌和3株真菌能在CMC平板上产生透明圈,但以糖化力法测定酶活结果只有3株真菌具有产酶能力。3株真菌的酶活动态测定结果,酶活的高峰均出现在7?14 d,并且呈现多峰变化;3株真菌的酶活种类表现为,滤纸酶活性、纤维素内切酶活性和外切酶活性均以菌株F1最高,分别达到了1.05、5.53和0.56 U/mL,β-葡萄糖甘酶活性和木聚糖酶活性以菌株FC最高,分别达到0.44和58.95 U/mL,其木聚糖酶活为F1最高值的6倍。     

Production of lactic acid from paper sludge using acid-tolerant, thermophilic Bacillus coagulan strains

Production of lactic acid from paper sludge was studied using thermophilic Bacillus coagulan strains 36D1 and P4-102B. More than 80% of lactic acid yield and more than 87% of cellulose conversion were achieved using both strains without any pH control due to the buffering effect of CaCO₃ in paper sludge. The addition of CaCO₃ as the buffering reagent in rich medium increased lactic acid yield but had little effect on cellulose conversion; when lean medium was utilized, the addition of CaCO₃ had little effect on either cellulose conversion or lactic acid yield. Lowering the fermentation temperature lowered lactic acid yield but increased cellulose conversion. Semi-continuous simultaneous saccharification and co-fermentation (SSCF) using medium containing 100 g/L cellulose equivalent paper sludge without pH control was carried out in serum bottles for up to 1000 h. When rich medium was utilized, the average lactic acid concentrations in steady state for strains 36D1 and P4-102B were 92 g/L and 91.7 g/L, respectively, and lactic acid yields were 77% and 78%. The average lactic acid concentrations produced using semi-continuous SSCF with lean medium were 77.5 g/L and 77.0 g/L for strains 36D1 and P4-102B, respectively, and lactic acid yields were 72% and 75%. The productivities at steady state were 0.96 g/L/h and 0.82 g/L/h for both strains in rich medium and lean medium, respectively. Our data support that B. coagulan strains 36D1 and P4-102B are promising for converting paper sludge to lactic acid via SSCF.     

Efficient regeneration and antioxidant potential in regenerated tissues of <Emphasis Type="Italic">Piper nigrum</Emphasis> L.

The organogenic potential and antioxidant potential (1, 1-diphenyl-2-picrylhydrazyl-scavenging activity) of the medicinal plant Piper nigrum L. (black pepper) were investigated. Callus induction and shoot regeneration were induced from leaf explants of potted plants cultured on MS medium supplemented with different plant growth regulators. The best callogenic response was observed on explants cultured for 30 days on MS medium supplemented with either 0.5 or 1.5 mg l⁻¹ 6-benzyladenine (BA) + 1.0 mg l⁻¹ α-naphthaleneacetic acid. Subsequent transfer of the callogenic explants onto MS medium supplemented with 1.5 mg l⁻¹ BA + 1.0 mg l⁻¹ gibberellic acid (GA₃) achieved 85% shoot organogenesis after 30 days of culture. The maximum number (7.2) of shoots/explant was recorded for explants cultured in MS medium supplemented with 1.0 mg l⁻¹ BA. Following the transfer of shoots to an elongation medium, the longest shoots (5.4 cm) were observed on MS medium supplemented with 1.0 mg l⁻¹ BA + 1.0 mg l⁻¹ GA₃. The elongated shoots were rooted on MS medium supplemented with different concentrations of indole butyric acid. An assay of the antioxidant potential of the in vitro-grown tissues revealed that the antioxidant activity of the regenerated shoots was significantly higher than that of callus and the regenerated plantlets.     

Sequential anaerobic–aerobic degradation of munitions waste

A sequential anaerobic–aerobic biodegradation of hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX) was studied. The results demonstrated that: (i) a complete degradation of RDX was achieved within 20 days using a consortium of bacteria from a wastewater activated sludge, (ii) RDX degradation did not occur under aerobic conditions alone, (iii) RDX-degrading bacterial strain that was isolated from the activated sludge completely degraded RDX within 2 days, and (iv) RDX- induced protein expressions were observed in the RDX-degrading bacterial strain. Based on fatty acid composition and a confirmation with a 16S rRNA analysis, the RDX-degrading bacterial strain was identified as a Bacillus pumilus—GC subgroup B.