D R Ranade

National Centre For Cell Science, Pune, Poona, Mahārāshtra, India

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Publications (35)58.52 Total impact

  • [show abstract] [hide abstract]
    ABSTRACT: Two coccoid, non-motile, obligate anaerobic, Gram stain negative bacteria mainly single or in pairs, with average size 1.4 - 2.5 μm; designated as NMBHI-10T and BLPYG-7 were isolated from faeces of two different healthy human volunteers aged 26 and 56 years respectively. Both the strains were affiliated to the sub-branch Sporomucosa of the class Clostridia as revealed by 16S rRNA gene sequence analysis. The isolates NMBHI-10T and BLPYG-7 showed 99.1 and 99.2 % 16S rRNA gene sequence similarity respectively with Megasphaera elsdenii JCM 1772T. DNA-DNA hybridization and phenotypic analysis showed that both the strains were distinct from their closest relative, M. elsdenii JCM 1772T (42 and 53% DNA-DNA relatedness respectively) but belong to the same species (DNA-DNA relatedness of 80.9% between the isolates). According to DNA-DNA hybridization results, the coccoid strains belong to the same genospecies, neither of which was related to any of the known species of Megasphaera. Strains NMBHI-10T and BLPYG-7 grow in PYG broth at temperatures between 15 and 40oC (optimum 37oC) but not at 45oC. The strains utilize a range of carbohydrates as sources of carbon and energy, including glucose, lactose, cellobiose, rhamnose, galactose and sucrose. Glucose fermentation resulted in formation of volatile fatty acids mainly caproic acid and organic acids such as succinic acid. Phylogenetic analysis and specific phenotypic characteristics and/or DNA G+C content also differentiated the strains from each other and from their closest relatives. The G+C content of strain NMBHI-10T and BLPYG-7 is 57.7 and 54.9 mol% respectively. Major fatty acids were 12:0 FAME and 17:0 CYC FAME. On the basis of this data, we propose strains NMBHI-10T and BLPYG-7 be classified as representing a novel species, Megspahaera indica sp. nov. The type strain is NMBHI-10T = (DSM 25563T ).
    INTERNATIONAL JOURNAL OF SYSTEMATIC AND EVOLUTIONARY MICROBIOLOGY 04/2014; · 2.11 Impact Factor
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    ABSTRACT: With increasing number of novel bacteria being isolated from the human gut ecosystem, there is a greater need to study their role in the gut ecosystem and their effect on the host health. In the present study, we carried out in silico genome-wide analysis of two novel Megasphaera sp. isolates NM10 (DSM25563) and BL7 (DSM25562), isolated from feces of two healthy individuals and validated the key features by in vitro studies. The analysis revealed the general metabolic potential, adaptive features and the potential effects of these isolates on the host. The comparative genome analysis of the two human gut isolates NM10 and BL7 with ruminal isolate Megasphaera elsdenii (DSM20460) highlighted the differential adaptive features for their survival in human gut. The key findings include features like bile resistance, presence of various sensory and regulatory systems, stress response systems, membrane transporters and resistance to antibiotics. Comparison of the "glycobiome" based on the genomes of the ruminal isolate with the human gut isolates NM10 and BL revealed the presence of diverse and unique sets of Carbohydrate-Active enzymes (CAZymes) amongst these isolates, with a higher collection of CAZymes in the human gut isolates. This could be attributed to the difference in host diet and thereby the environment, consequently suggesting host specific adaptation in these isolates. In silico analysis of metabolic potential predicted the ability of these isolates to produce important metabolites like short chain fatty acids (butyrate, acetate, formate, and caproate), vitamins and essential amino acids, which was further validated by in vitro experiments. The ability of these isolates to produce important metabolites advocates for a potential healthy influence on the host. Further in vivo studies including transcriptomic and proteomic analysis will be required for better understanding the role and impact of these Megasphaera sp. isolates NM10 and BL7 on the human host.
    PLoS ONE 01/2013; 8(11):e79353. · 3.73 Impact Factor
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    ABSTRACT: Glycerol is a promising carbon source which is used by number of organisms as a sole source of carbon under anaerobic conditions. 1,3 Propanediol (1,3 PDO) is the principle product obtained through glycerol fermentation. In the present investigation, optimization of fermentation pathway in the production of 1,3 PDO was carried out using a newly isolated culture BA11 from soil. The isolate was characterized morphologically, biochemically and phylogenetically (16S gene sequence analysis) and determined to be a strain of Klebsiella pneumoniae. Glycerol utilization and 1,3 PDO production by the organism was studied under aerobic and anaerobic conditions using pure glycerol as a sole source of carbon. Also, the effect of static and shaking conditions of incubation were investigated. The organism could utilize glycerol completely when grown under aerobic shaking condition with 8.3 g/l of 1,3 PDO after 72 h of incubation and a molar yield of 0.44 moles/moleglycerol utilized for initial glycerol concentration of 20 g/l. But, when grown under anaerobic shaking conditions, 8.1 g/l of 1,3 PDO was obtained with molar yield of 0.65 moles/moleglycerol utilized. Anaerobic shaking condition was found to be more efficient, as 8.03 g/l of 1,3 PDO was obtained within 24 h of fermentation. There was no increase in the molar yield with the increase in glycerol concentration. Shaking incubation condition and anaerobic mode of fermentation were effective for higher 1,3 PDO production. The isolate BA11 was found to be fast growing with a specific growth rate of 0.59 h-1 and a doubling time of 1.17 h. It could also tolerate a wide range of pH from 6-8.5 and also gave high yield of 1,3 PDO. Further, BA11 could show high glycerol utilization up to a temperature of 40 OC with 1,3 PDO yield of 0.53 moles/moleglycerol utilized.
    The 5th International Conference on Industrial Bioprocesses, Taipei; 10/2012
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    ABSTRACT: BACKGROUND: The gut micro flora plays vital role in health status of the host. The majority of microbes residing in the gut have a profound influence on human physiology and nutrition. Different human ethnic groups vary in genetic makeup as well as the environmental conditions they live in. The gut flora changes with genetic makeup and environmental factors and hence it is necessary to understand the composition of gut flora of different ethnic groups. Indian population is different in physiology from western population (YY paradox) and thus the gut flora in Indian population is likely to differ from the extensively studied gut flora in western population. In this study we have investigated the gut flora of two Indian families, each with three individuals belonging to successive generations and living under the same roof. RESULTS: Denaturation gradient gel electrophoresis analysis showed age-dependant variation in gut microflora amongst the individuals within a family. Different bacterial genera were dominant in the individual of varying age in clone library analysis. Obligate anaerobes isolated from individuals within a family showed age related differences in isolation pattern, with 27% (6 out of 22) of the isolates being potential novel species based on 16S rRNA gene sequence. In qPCR a consistent decrease in Firmicutes number and increase in Bacteroidetes number with increasing age was observed in our subjects, this pattern of change in Firmicutes / Bacteroidetes ratio with age is different than previously reported in European population. CONCLUSION: There is change in gut flora with age amongst the individuals within a family. The isolation of high percent of novel bacterial species and the pattern of change in Firmicutes /Bacteroidetes ratio with age suggests that the composition of gut flora in Indian individuals may be different than the western population. Thus, further extensive study is needed to define the gut flora in Indian population.
    BMC Microbiology 09/2012; 12(1):222. · 3.10 Impact Factor
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    ABSTRACT: Obesity is a consequence of a complex interplay between the host genome and the prevalent obesogenic factors among the modern communities. The role of gut microbiota in the pathogenesis of the disorder was recently discovered; however, 16S-rRNA-based surveys revealed compelling but community-specific data. Considering this, despite unique diets, dietary habits and an uprising trend in obesity, the Indian counterparts are poorly studied. Here, we report a comparative analysis and quantification of dominant gut microbiota of lean, normal, obese and surgically treated obese individuals of Indian origin. Representative gut microbial diversity was assessed by sequencing fecal 16S rRNA libraries for each group (n=5) with a total of over 3000 sequences. We detected no evident trend in the distribution of the predominant bacterial phyla, Bacteroidetes and Firmicutes. At the genus level, the bacteria of genus Bacteroides were prominent among the obese individuals, which was further confirmed by qPCR (P less than 0.05). In addition, a remarkably high archaeal density with elevated fecal SCFA levels was also noted in the obese group. On the contrary, the treated-obese individuals exhibited comparatively reduced Bacteroides and archaeal counts along with reduced fecal SCFAs. In conclusion, the study successfully identified a representative microbial diversity in the Indian subjects and demonstrated the prominence of certain bacterial groups in obese individuals; nevertheless, further studies are essential to understand their role in obesity.
    Journal of Biosciences 09/2012; 37(4):647-57. · 1.76 Impact Factor
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    ABSTRACT: A simple and easy sol–gel approach has been developed to directly synthesize in situ three-dimensionally interconnected uniform ordered bimodal porous silica (BPS) incorporating both the macroporosity and mesoporosity in the lattice without extra synthesis process performed in previous work. Multimodal porous carbon (MPC) was fabricated through the inverse replication of the BPS. The unique structural characteristics such as well-developed 3-D interconnected ordered macropore framework with open mesopores embedded in the macropore walls, large surface area (1120m2g−1) and mesopore volume (1.95cm3g−1) make MPC very attractive as an anode catalyst support in polymer exchange membrane fuel cell. The MPC-supported Pt-Ru alloy catalyst has demonstrated much higher power density toward hydrogen oxidation than the commercial carbon black Vulcan XC-72-supported ones.
    Electrochimica Acta - ELECTROCHIM ACTA. 01/2010; 55(26):7628-7633.
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    ABSTRACT: A mesophilic high hydrogen producing strain DMHC-10 was isolated from a lab scale anaerobic reactor being operated on distillery wastewater for hydrogen production. DMHC-10 was identified as Clostridium sp. on the basis of 16S rRNA gene sequencing. Various medium components (carbon and nitrogen sources) and environmental factors (initial pH, temperature of incubation) were optimized for hydrogen production by Clostridium sp. DMHC-10. The strain, in late exponential growth phase, showed maximum hydrogen production (3.35mol-H2mol−1 glucose utilized) at 37°C, pH 5.0 in a medium supplemented with organic nitrogen source. Butyric acid to acetic acid ratio was ca. 2.3. Hydrogen production declined when organic nitrogen was replaced with inorganic nitrogen.
    International Journal of Hydrogen Energy - INT J HYDROGEN ENERG. 01/2010; 35(19):10639-10644.
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    ABSTRACT: The removal of AOX from bleach plant effluent of pulp and paper industry was studied using upflow anaerobic filter. In this paper biodegradation of AOX at different concentrations and effect of electron donors like acetate and glucose thereon in an upflow anaerobic filter at 20 d HRT is described. Results showed significant improvement in AOX degradation when electron donors such as acetate and glucose were supplemented to the influent. AOX degradation was 88% at 28 mg AOX L(-1) and 28% at 42 mg AOX L(-1). The percent degradation efficiency was enhanced to 90.7, 90.2, and 93.0 at 28 mg AOX L(-1) when the influent was supplemented with glucose, acetate and both glucose and acetate, respectively. Similarly, the efficiency was 57, 56.6 and 79.6 at 42 mg AOX L(-1) when the influent was supplemented with glucose, acetate and both glucose and acetate, respectively. The GC-MS analysis data indicated that supplementation of the influent with electron donor increased the biodegradability of number of chlorinated organic compounds.
    Chemosphere 04/2009; 75(9):1179-85. · 3.14 Impact Factor
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    ABSTRACT: The methanogen community in biogas reactor running on cattle dung was investigated in two different seasons; summer (April, 36 degrees C) and winter (December, 24 degrees C), in the year 2004 by a culture-independent approach. Community structure was determined by phylogenetic analyses of 343 and 278 mcrA clones belonging to summer and winter month libraries, respectively. In summer month's library, 41.7% clones were affiliated to Methanomicrobiales, 30% to Methanosarcinales, 19% to Methanobacteriales, 5% to Methanococcales and a total of 4.3% clones belonged to unclassified euryarchaeotal lineages. In winter month's library, Methanomicrobiales encompassed 98.6% clones, and Methanobacteriales included 1.4% of total clone diversity. Biogas plant performance data collected during the winter month indicated significant reduction in daily biogas produced as compared to summer month because of lowering in ambient temperature and associated shift in microbial community. Results from this molecular study showed the existence of highly diverse and complex methanogens communities present in biogas plant.
    Bioresource Technology 10/2008; 99(13):5317-26. · 4.75 Impact Factor
  • S.R. Hajarnis, D.R. Ranade
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    ABSTRACT: Growth and methanogenesis by Methanobacterium formicicum were maximum at 0.5 gl-1 of ammonia and were completely inhibited by 70 g l-1. Revival following exposure to ammonia was inversely proportional to ammonia concentration and duration of exposure.
    Letters in Applied Microbiology 06/2008; 18(5):254 - 256. · 1.63 Impact Factor
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    ABSTRACT: Methanogen communities were characterized in cattle dung of different ages by using a culture-independent approach. Community structures were determined by the phylogenetic analyses of methyl-coenzyme M reductase A (mcrA) clones of fresh, 8-month-old, and 24-month-old-dry dung samples. The clones in the mcrA libraries of fresh and 8-month old dung samples were identified as belonging to Methanomicrobiales, Methanobacteriales, and Methanosarcinales. However, clones in the library of 24-month-old dung were not affiliated to Methanomicrobiales. Anaerobic digestion of 2-month-old dung produced only 15% less methane compared to fresh dung which indicated the possibility of using dry dung to fuel the biogas plants in areas where unavailability of fresh dung hinders their continuous functioning. Our results first time showed the presence of viable methanogens in dry cattle dung stored for prolonged periods of time.
    World Journal of Microbiology and Biotechnology 01/2008; 24(12):2973-2979. · 1.26 Impact Factor
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    ABSTRACT: The diversity of methanogenic archaea in enrichment cultures established from the sediments of Lonar Lake (India), a soda lake having pH approximately 10, was investigated using 16S rDNA molecular phylogenetic approach. Methanogenic enrichment cultures were developed in a medium that simulated conditions of soda lake with three different substrates viz., H(2):CO(2), sodium acetate, and trimethylamine (TMA), at alkaline pH. Archaeal 16S rRNA clone libraries were generated from enrichment cultures and 13 RFLP groups were obtained. Representative sequence analysis of each RFLP group indicated that the majority of the 16S rRNA gene sequences were phylogenetically affiliated with uncultured Archaea. Some of the groups may belong to new archaeal genera or families. Three RFLP groups were related to Methanoculleus sp, while two related to Methanocalculus sp. 16S rRNA gene sequences found in Lonar Lake were different from sequences reported from other soda lakes and more similar to those of oil reservoirs, palm oil waste treatment digesters, and paddy fields. In culture-based studies, three isolates were obtained. Two of these were related to Methanoculleus sp. IIE1 and one to Methanocalculus sp. 01F97C. These results clearly show that the Lonar Lake ecosystem harbors unexplored methanogens.
    Microbial Ecology 12/2007; 54(4):697-704. · 3.28 Impact Factor
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    ABSTRACT: The present study for the first time investigated the diversity of methanotrophs in a biogas reactor running on cattle dung by sequence analysis of the "functional" gene, particulate methane monooxygenase A (pmoA). Community structure was determined by operational taxonomic unit (OTU) phylogenetic analyses of 396 clones belonging to pmoA library. On the basis of 95% peptide identity, 396 inferred PmoA peptide sequences were clustered in 22 phylotypes. These OTUS were phylogenetically affiliated to alpha-proteobacteria (5.30% clones) and gamma-proteobacteria (94.7% clones) and were only distantly related to those of known methanotrophs, indicating the existence of unknown methanotrophs involved in aerobic methanotrophy inside the biogas plant. Good's coverage indicated that the present library covered 96.21% of the dominant species that could be cloned from the biogas reactor. The molecular approaches used in this study provided useful description of the microbial community involved in aerobic oxidation of methane.
    Microbiological Research 07/2007; 164(5):536-44. · 1.99 Impact Factor
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    ABSTRACT: The prokaryotic diversity associated with an Indian soda lake (Lonar Crater Lake) located in a basaltic soil area was investigated using a culture-independent approach. Community DNA was extracted directly from four sediment samples obtained by coring to depths of 10-20 cm. Small subunit rRNA genes (16S rDNA) were amplified by PCR using primers specific to the domains Bacteria and Archaea. The PCR products were cloned and sequenced. For the bacterial rDNA clone library, 500 clones were randomly selected for further analysis. After restriction fragment length polymorphism (RFLP) analysis and subsequent sequencing, a total of 44 unique phylotypes were obtained. These phylotypes spanned a wide range within the domain Bacteria, occupying eight major lineages/phyla. 34% of the clones were classified as firmicutes. The other clones were grouped into proteobacteria (29.5%), actinobacteria (6.8%), deinococcus-thermus (4.5%), cytophages-flavobacterium-bacteroidetes (13.3%), planctomycetes (6.8%), cyanobacteria (4.5%) and spirochetes (2.27%). In the case of the archaeal 16S rDNA library, analysis of 250 randomly selected clones revealed the presence of 13 distinct phylotypes; 5 phylotypes were associated with Crenarchaeota and 8 with Euryarchaeota. Most of the euryarchaeota sequences were related to methanogens. Findings from this molecular study of a site investigated for the first time have revealed the presence of a highly diverse bacterial population and a comparatively less diverse archaeal population. The majority ( approximately 80%) of the cloned sequences show little affiliation with known taxa (<97% sequence similarity) and may represent novel taxa/sequences and organisms specifically adapted to this basaltic soda lake environment. Diversity analyses demonstrate greater diversity and evenness of bacterial species compared to a skewed representation of species for Archaea.
    Research in Microbiology 12/2006; 157(10):928-37. · 2.89 Impact Factor
  • D V Savant, R Abdul-Rahman, D R Ranade
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    ABSTRACT: Adsorbable organic halides (AOX) are generated in the pulp and paper industry during the bleaching process. These compounds are formed as a result of reaction between residual lignin from wood fibres and chlorine/chlorine compounds used for bleaching. Many of these compounds are recalcitrant and have long half-life periods. Some of them show a tendency to bioaccumulate while some are proven carcinogens and mutagens. Hence, it is necessary to remove or degrade these compounds from wastewater. Physical, chemical and electrochemical methods reported to remove AOX compounds are not economically viable. Different types of aerobic, anaerobic and combined biological treatment processes have been developed for treatment of pulp and paper industry wastewater. Maximum dechlorination is found to occur under anaerobic conditions. However, as these processes are designed specifically for reducing COD and BOD of wastewater, they do not ensure complete removal of AOX. This paper reviews the anaerobic biological treatments developed for pulp and paper industry wastewater and also reviews the specific micro-organisms reported to degrade AOX compounds under anaerobic conditions, their nutritional and biochemical requirements. It is imperative to consider these specific micro-organisms while designing an anaerobic treatment for efficient removal of AOX.
    Bioresource Technology 07/2006; 97(9):1092-104. · 4.75 Impact Factor
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    ABSTRACT: In the present study, isolation of anaerobic bacteria from 24 different eco-niches was carried out. A total number of 300 bacterial isolates, including 230 obligate and 70 facultative anaerobes were obtained using anaerobic techniques. All the isolates were initially screened for succinic acid production by Fluorescein test and TLC method. During screening, 10 isolates found to produce succinic acid were further examined by HPLC and then finally confirmed for succinic acid by LC-MS analysis. Amongst 10 isolates, isolate SAP, a facultative anaerobe isolated from buffalo rumen fluid, showed maximum yield of 2.1 g/l of succinic acid from 10 g of glucose in 24 hr under anaerobic condition. This isolate was identified as Klebsiella pneumoniae strain SAP by 16S rDNA sequence and signature sequence analysis. Mouse lethality test for the strain SAP showed LD50 value of 3.3 x 10(8) CFU/ml, which shows non-virulent nature of the strain. This strain may become a candidate strain for succinic acid production because of its osmotolerant nature and higher succinate:acetate ratio.
    Indian journal of experimental biology 03/2006; 44(2):142-50. · 1.20 Impact Factor
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    ABSTRACT: Abstract 5-Aminolevulinic acid dehydratase from the archaebacterium Methanosarcina barken resembles the mammalian and yeast enzymes in its activation by Zn2+, whereas its activation by K+ resembles the characteristic of bacterial enzymes. This enzyme is activated with Ni2+ which is a component of F430, a cofactor present mainly in methanogens. The Mr of 280000 for the native enzyme and 30 000 ± 2000 for the individual subunit suggest that the enzyme is composed of eight apparently indentical subunits similar to mammalian and yeast enzymes. The enzyme has two pH optima, at 8.5 and 9.4. Higher levels of 5-aminolevulinic acid dehydratase in acetate-grown cells suggest the possibility that regulation and control of this enzyme could be different on various growth substrates.
    FEMS Microbiology Letters 01/2006; 127(1‐2):151 - 155. · 2.05 Impact Factor
  • Thakker C, Bhosale S, Ranade D
    Journal of Microbiology and Biotechnology 01/2006; 16(6):870-879. · 1.40 Impact Factor
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    ABSTRACT: The phylogeny of the genus Methanobrevibacter was established almost 25 years ago on the basis of the similarities of the 16S rRNA oligonucleotide catalogs. Since then, many 16S rRNA gene sequences of newly isolated strains or clones representing the genus Methanobrevibacter have been deposited. We tried to reorganize the 16S rRNA gene sequences of this genus and revise the taxonomic affiliation of the isolates and clones representing the genus Methanobrevibacter. The phylogenetic analysis of the genus based on 786 bp aligned region from fifty-four representative sequences of the 120 available sequences for the genus revealed seven multi-member groups namely, Ruminantium, Smithii, Woesei, Curvatus, Arboriphilicus, Filiformis, and the Termite gut symbionts along with three separate lineages represented by Mbr. wolinii, Mbr. acididurans, and termite gut flagellate symbiont LHD12. The cophenetic correlation coefficient, a test for the ultrametric properties of the 16S rRNA gene sequences used for the tree was found to be 0.913 indicating the high degree of goodness of fit of the tree topology. A significant relationship was found between the 16S rRNA sequence similarity (S) and the extent of DNA hybridization (D) for the genus with the correlation coefficient (r) for logD and logS, and for [ln(-lnD) and ln(-lnS)] being 0.73 and 0.796 respectively. Our analysis revealed that for this genus, when S = 0.984, D would be <70% at least 99% of the times, and with 70% D as the species "cutoff", any 16S rRNA gene sequence showing <98% sequence similarity can be considered as a separate species. In addition, we deduced group specific signature positions that have remained conserved in evolution of the genus. A very significant relationship between D and S was found to exist for the genus Methanobrevibacter, implying that it is possible to predict D from S with a known precision for the genus. We propose to include the termite gut flagellate symbiont LHD12, the methanogenic endosymbionts of the ciliate Nyctotherus ovalis, and rat feces isolate RT reported earlier, as separate species of the genus Methanobrevibacter.
    BMC Microbiology 06/2004; 4:20. · 3.10 Impact Factor
  • D P Kunte, T Y Yeole, D R Ranade
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    ABSTRACT: Anaerobic digestion offers a good alternative for human waste treatment. However, the fate of enteric bacterial pathogens present in human night soil (HNS) remains a major concern for hygienic safety of the process. A two-stage anaerobic digestion process, consisting of separate acidogenic and methanogenic digesters, was designed and its efficacy in the inactivation of Salmonella typhi was compared to a single-stage digestion process. In a single-stage digestion, complete pathogen inactivation was achieved only in the digesters with high levels of volatile fatty acids (VFA approximately equal to 18,000 mg/l) and acidic pH (approximately equal to 6.0). These digesters, however, showed drastic reduction in methane yield. In the two-stage digestion process, S. typhi was completely inactivated in the acidogenic digester and the methanogenic digester was free from the pathogen even after receiving a daily dose of the pathogen. The process also achieved complete inactivation of other enteric pathogens, viz., Shigella dysenteriae and Vibrio cholerae. The two-stage process was efficient in biogas generation from HNS. Thus, the two-stage process ensures complete hygienic safety in anaerobic digestion of human night soil.
    Water Science & Technology 01/2004; 50(6):103-8. · 1.10 Impact Factor