Characterization of a Coliphage AS1 isolated from sewage effluent in Pakistan

The emergence of multi-drug resistant (MDR) bacterial strains, which are posing a global health threat has developed the interest of scientists to use bacteriophages instead of conventional antibiotics therapy. In light of an increased interest in the use of phage as a bacterial control agent, the study aimed to isolate and characterize lytic phages from sewage effluent. During the current study, bacteriophage AS1 was isolated from sewage effluent against E.coli S2. The lytic activity of phageAS1 was limited to E.coli S2 strain showing monovalent behavior. The calculated phage titer was 3.5×109 pfu/ml. PhageAS1 was stable at a wide range of pH and temperature. The maximum stability was recorded at 37ºC and pH 7.0, while showing its normal lytic activity at temperature 60ºC and from pH 5.0 to11.0 respectively. At temperature 70ºC, phage activity was somewhat reduced whereas, further increase in temperature and decrease or increase in pH completely inactivated the phage. From the current study, it was concluded that waste water is a best source for finding bacteriophages against multi-drug resistant bacterial strains and can be used as bacterial control agent.

Quantitative Ecological Impact Assessments using Natural Abundance Carbon and Nitrogen Stable Isotope Signatures

<p>The use of delta15N and delta13C signatures to infer sources of enrichment in ecological systems relies on predictability in the transfer of delta15N and delta13C ratios. This thesis examines patterns of delta15N and delta13C change as pools of nitrogen and carbon move from a sewage effluent discharge into organisms in an adjacent coastal rocky reef community (Titahi Bay, New Zealand). These changes and their mechanisms are examined in the broader context of current research using carbon and nitrogen stable isotope ratios in marine ecology, with particular reference to impact assessment. Firstly this thesis examines the assimilation of nitrogen and carbon isotopes in Ulva sp. under varying light conditions and nitrogen source (e.g., nitrate or ammonium). In a field study, algae grown at depth and under lower light conditions showed comparatively lighter nitrogen isotope signatures relative to the predicted concentration of available 15N-enriched sewage nitrogen. In a complementary laboratory experiment, results from manipulated light availability and N source (either nitrate or ammonium, in equivalent molar concentrations) suggest that: 1) low-light conditions can produce algae with lighter nitrogen isotope signatures; and 2) this effect was more pronounced for ammonium (3.7 per mil difference between high light and low light treatments) than for nitrate (0.6 per mil difference between high light and low light treatments) sources. Stable carbon isotope ratios (delta13C) of Ulva sp.grown in conditions of low nitrogen availability were shown to be generally lower than those grown in nitrogen rich conditions in both field and laboratory studies. Where nitrogen supply was sufficient for growth, low light conditions also produced generally lower delta13C signatures than high light conditions. Experimental trials with a uniform dissolved inorganic carbon source and altered light and nitrogen enrichment levels produced delta 13C levels in Ulva sp. tissue that spanned the recorded delta13C ranges of many common algal species; -5.99 per mil (high light, with added ammonium and phosphate) to -17.61 per mil (high light without nutrient additions). Chapter 3 of this study examines the growth response of Ulva sp. to surplus nitrate and ammonium (the two most common forms of nitrogen available to plants in seawater), under light limited conditions. Ulva sp. experienced a temporary reduction in growth rate and nitrogen assimilation capacity (shown in tissue nitrogen indices) when grown on nitrate, relative to ammonium. The magnitude and the temporary nature of these results suggest that in natural populations the relative proportion of nitrate or ammonium available is unlikely to significantly affect the growth capacity of Ulva sp. In chapter 4, I use delta13C and delta15N signatures to separately trace the dissolved and particulate fractions of sewage effluent dispersal onto a rocky reef community. Delta15N signatures from tissue of the macroalga Carpophyllum maschalocarpum, and the herbivorous isopod Amphoroidea media tracked the distribution and signature of DIN from a sewage treatment plant that generated heavy delta15N signatures. Delta13C signatures from tissue of the filter-feeding half-crab Petrolisthes elongatus tracked the distribution and signature of suspended sewage particulate organic matter.</p>

Quantitative Ecological Impact Assessments using Natural Abundance Carbon and Nitrogen Stable Isotope Signatures

<p>The use of delta15N and delta13C signatures to infer sources of enrichment in ecological systems relies on predictability in the transfer of delta15N and delta13C ratios. This thesis examines patterns of delta15N and delta13C change as pools of nitrogen and carbon move from a sewage effluent discharge into organisms in an adjacent coastal rocky reef community (Titahi Bay, New Zealand). These changes and their mechanisms are examined in the broader context of current research using carbon and nitrogen stable isotope ratios in marine ecology, with particular reference to impact assessment. Firstly this thesis examines the assimilation of nitrogen and carbon isotopes in Ulva sp. under varying light conditions and nitrogen source (e.g., nitrate or ammonium). In a field study, algae grown at depth and under lower light conditions showed comparatively lighter nitrogen isotope signatures relative to the predicted concentration of available 15N-enriched sewage nitrogen. In a complementary laboratory experiment, results from manipulated light availability and N source (either nitrate or ammonium, in equivalent molar concentrations) suggest that: 1) low-light conditions can produce algae with lighter nitrogen isotope signatures; and 2) this effect was more pronounced for ammonium (3.7 per mil difference between high light and low light treatments) than for nitrate (0.6 per mil difference between high light and low light treatments) sources. Stable carbon isotope ratios (delta13C) of Ulva sp.grown in conditions of low nitrogen availability were shown to be generally lower than those grown in nitrogen rich conditions in both field and laboratory studies. Where nitrogen supply was sufficient for growth, low light conditions also produced generally lower delta13C signatures than high light conditions. Experimental trials with a uniform dissolved inorganic carbon source and altered light and nitrogen enrichment levels produced delta 13C levels in Ulva sp. tissue that spanned the recorded delta13C ranges of many common algal species; -5.99 per mil (high light, with added ammonium and phosphate) to -17.61 per mil (high light without nutrient additions). Chapter 3 of this study examines the growth response of Ulva sp. to surplus nitrate and ammonium (the two most common forms of nitrogen available to plants in seawater), under light limited conditions. Ulva sp. experienced a temporary reduction in growth rate and nitrogen assimilation capacity (shown in tissue nitrogen indices) when grown on nitrate, relative to ammonium. The magnitude and the temporary nature of these results suggest that in natural populations the relative proportion of nitrate or ammonium available is unlikely to significantly affect the growth capacity of Ulva sp. In chapter 4, I use delta13C and delta15N signatures to separately trace the dissolved and particulate fractions of sewage effluent dispersal onto a rocky reef community. Delta15N signatures from tissue of the macroalga Carpophyllum maschalocarpum, and the herbivorous isopod Amphoroidea media tracked the distribution and signature of DIN from a sewage treatment plant that generated heavy delta15N signatures. Delta13C signatures from tissue of the filter-feeding half-crab Petrolisthes elongatus tracked the distribution and signature of suspended sewage particulate organic matter.</p>

Quantitative Ecological Impact Assessments using Natural Abundance Carbon and Nitrogen Stable Isotope Signatures

<p>The use of delta15N and delta13C signatures to infer sources of enrichment in ecological systems relies on predictability in the transfer of delta15N and delta13C ratios. This thesis examines patterns of delta15N and delta13C change as pools of nitrogen and carbon move from a sewage effluent discharge into organisms in an adjacent coastal rocky reef community (Titahi Bay, New Zealand). These changes and their mechanisms are examined in the broader context of current research using carbon and nitrogen stable isotope ratios in marine ecology, with particular reference to impact assessment. Firstly this thesis examines the assimilation of nitrogen and carbon isotopes in Ulva sp. under varying light conditions and nitrogen source (e.g., nitrate or ammonium). In a field study, algae grown at depth and under lower light conditions showed comparatively lighter nitrogen isotope signatures relative to the predicted concentration of available 15N-enriched sewage nitrogen. In a complementary laboratory experiment, results from manipulated light availability and N source (either nitrate or ammonium, in equivalent molar concentrations) suggest that: 1) low-light conditions can produce algae with lighter nitrogen isotope signatures; and 2) this effect was more pronounced for ammonium (3.7 per mil difference between high light and low light treatments) than for nitrate (0.6 per mil difference between high light and low light treatments) sources. Stable carbon isotope ratios (delta13C) of Ulva sp.grown in conditions of low nitrogen availability were shown to be generally lower than those grown in nitrogen rich conditions in both field and laboratory studies. Where nitrogen supply was sufficient for growth, low light conditions also produced generally lower delta13C signatures than high light conditions. Experimental trials with a uniform dissolved inorganic carbon source and altered light and nitrogen enrichment levels produced delta 13C levels in Ulva sp. tissue that spanned the recorded delta13C ranges of many common algal species; -5.99 per mil (high light, with added ammonium and phosphate) to -17.61 per mil (high light without nutrient additions). Chapter 3 of this study examines the growth response of Ulva sp. to surplus nitrate and ammonium (the two most common forms of nitrogen available to plants in seawater), under light limited conditions. Ulva sp. experienced a temporary reduction in growth rate and nitrogen assimilation capacity (shown in tissue nitrogen indices) when grown on nitrate, relative to ammonium. The magnitude and the temporary nature of these results suggest that in natural populations the relative proportion of nitrate or ammonium available is unlikely to significantly affect the growth capacity of Ulva sp. In chapter 4, I use delta13C and delta15N signatures to separately trace the dissolved and particulate fractions of sewage effluent dispersal onto a rocky reef community. Delta15N signatures from tissue of the macroalga Carpophyllum maschalocarpum, and the herbivorous isopod Amphoroidea media tracked the distribution and signature of DIN from a sewage treatment plant that generated heavy delta15N signatures. Delta13C signatures from tissue of the filter-feeding half-crab Petrolisthes elongatus tracked the distribution and signature of suspended sewage particulate organic matter.</p>

Quantitative Ecological Impact Assessments using Natural Abundance Carbon and Nitrogen Stable Isotope Signatures

<p>The use of delta15N and delta13C signatures to infer sources of enrichment in ecological systems relies on predictability in the transfer of delta15N and delta13C ratios. This thesis examines patterns of delta15N and delta13C change as pools of nitrogen and carbon move from a sewage effluent discharge into organisms in an adjacent coastal rocky reef community (Titahi Bay, New Zealand). These changes and their mechanisms are examined in the broader context of current research using carbon and nitrogen stable isotope ratios in marine ecology, with particular reference to impact assessment. Firstly this thesis examines the assimilation of nitrogen and carbon isotopes in Ulva sp. under varying light conditions and nitrogen source (e.g., nitrate or ammonium). In a field study, algae grown at depth and under lower light conditions showed comparatively lighter nitrogen isotope signatures relative to the predicted concentration of available 15N-enriched sewage nitrogen. In a complementary laboratory experiment, results from manipulated light availability and N source (either nitrate or ammonium, in equivalent molar concentrations) suggest that: 1) low-light conditions can produce algae with lighter nitrogen isotope signatures; and 2) this effect was more pronounced for ammonium (3.7 per mil difference between high light and low light treatments) than for nitrate (0.6 per mil difference between high light and low light treatments) sources. Stable carbon isotope ratios (delta13C) of Ulva sp.grown in conditions of low nitrogen availability were shown to be generally lower than those grown in nitrogen rich conditions in both field and laboratory studies. Where nitrogen supply was sufficient for growth, low light conditions also produced generally lower delta13C signatures than high light conditions. Experimental trials with a uniform dissolved inorganic carbon source and altered light and nitrogen enrichment levels produced delta 13C levels in Ulva sp. tissue that spanned the recorded delta13C ranges of many common algal species; -5.99 per mil (high light, with added ammonium and phosphate) to -17.61 per mil (high light without nutrient additions). Chapter 3 of this study examines the growth response of Ulva sp. to surplus nitrate and ammonium (the two most common forms of nitrogen available to plants in seawater), under light limited conditions. Ulva sp. experienced a temporary reduction in growth rate and nitrogen assimilation capacity (shown in tissue nitrogen indices) when grown on nitrate, relative to ammonium. The magnitude and the temporary nature of these results suggest that in natural populations the relative proportion of nitrate or ammonium available is unlikely to significantly affect the growth capacity of Ulva sp. In chapter 4, I use delta13C and delta15N signatures to separately trace the dissolved and particulate fractions of sewage effluent dispersal onto a rocky reef community. Delta15N signatures from tissue of the macroalga Carpophyllum maschalocarpum, and the herbivorous isopod Amphoroidea media tracked the distribution and signature of DIN from a sewage treatment plant that generated heavy delta15N signatures. Delta13C signatures from tissue of the filter-feeding half-crab Petrolisthes elongatus tracked the distribution and signature of suspended sewage particulate organic matter.</p>

USING HISTORIC SITES IN DETERMINING PLANT-SOIL INTERACTIONS UNDER LONG TERM CONTAMINATED SOILS

The relationships between heavy metals in soils and crop tissues are critical in understanding the potential risk to crop yields from Zn, Cu and Ni (the principal phytotoxic elements of concern in sludge), and the possible effects on dietary Cd (the critical zootoxic element that is labile in sludge-treated soil and readily absorbed by plants. These relationships cannot be derived from the agronomic trials because the quantities of heavy metals applied to the soil in sludge are small. These are field sites have been treated with sludge for many years in operational practice and where the oncentrations of heavy metals have been significantly raised above background values, representing potentially a worse-case of soil contamination. The purpose of the study was to provide a surrogate for long-term sludge-treated agricultural soil by examining the effects on crops of heavy metals in soil irrigated with raw sewage effluent for periods of more than 80 years and containing significantly elevated concentrations of heavy metals. Methodology: Two surveys of the Gabal El Asfar Old Farm were undertaken to assess the long-term effects of heavy metals in sludge-treated soil on crop quality. In the first survey, the relationships between total and DTPA extractable heavy metals in soil and concentrations in citrus fruit were examined. Concentrations of heavy metals in leaves of citrus were measured in the second survey and related to total and DTPA extractable metals in soil. The heavy metal contents of citrus leaves and fruit (orange - eleven sampling sites; mandarin - four sampling sites), and total and DTPA extractable concentrations in soils were measured in samples collected from different areas of the Farm during two site surveys. Total and DTPA concentrations of heavy metals in the surveyed soils showed significant enrichment by long-term irrigation with sewage effluent. For example, the maximum total concentrations of Zn and Cu were 530 and 366 mg kg-1, respectively, representing a potential risk to crop yields The maximum Cd concentration detected was 9 mg kg-1 and may be a potential risk to the human food chain from uptake into staple crops grown at the farm. DTPA extractable metals were significantly (P<0.001) correlated with the total contents of Zn (r=0.91***), Cu (r=0.83***), Ni (r=0.63***) and Pb (r=0.85***) in soil when data from both surveys were pooled for statistical evaluation. There was also evidence of a weak relationship between DTPA extractable Cd and the total soil Cadmium is the only element of concern in terms of the risk to human health from uptake into food crops grown on sludge-treated soil. The total Cd concentration in soil was raised to a value 3 times the maximum EU limit for this element in sludge-treated agricultural soil. Despite the marked increase in soil Cd content, there was no detectable transfer into citrus leaves or fruit (Figure 1). The absence of Cd uptake into citrus fruit is to be expected because fruits are amongst the least sensitive plant parts to Cd accumulation. These data emphasise the minimal risk to the human diet from Cd in fruit crops grown on sludge-treated soil. . In all cases, leaf tissue concentrations were low and in some cases Cu status was below the deficiency threshold. The Cd content in leaves was small and generally <0.02 mg kg-1 DM.