Risk assessment of heavy metal and metalloid toxicity through a contaminated vegetable (Cucurbita maxima) from wastewater irrigated area: A case study for a site-specific risk assessment in Jhang,Pakistan

The present research was conducted in district Jhang, Pakistan, to evaluate the concentration of metals/metalloids in soil and pumpkin (Cucurbita maxima) irrigated with domestic wastewater. Data revealed that the levels of metals and metalloids in soil samples from two different sites were below the safe limits except Cd, whereas, in the vegetable, the concentrations of As, Se, Ni, Mo, Pb, Mn, and Cu were above the safe limits. The levels of 12 metals and metalloids in the soil were ranged between 0.14 to 22.76 mg/kg at site-I and 0.16 to 22.13 mg/kg at site-II. The levels of these metals in the vegetable were found 0.35 to 61.13 mg/kg at site-I and 0.31 to 53.63 mg/kg at site-II. The transfer factor at both sites was highest for As and Co. The pollution load index recorded for Se, Cu, Cd, Mo, Pb, and Co was greater than 1. The daily intake of As, Mn, and Mo was above the oral reference dose, which reflects that the intake of pumpkin is not safe for the inhabitants of the selected sites. The control measures should be taken to phytoextract heavy metals and metalloids from polluted sites so as to reduce the health risks.     

Purification, N-terminal amino acid sequence and characterization of pH 2.5 optimum acid phosphatase (E.C. 3.1.3.2) from Aspergillus ficuum

An acid phosphatase from crude culture filtrate of Aspergillus ficuum was purified to homogeneity using three ion exchange chromatographic steps. SDS-PAGE of the purified enzyme gave a single stained band at approximately 68-KDa. The mobility of the native enzyme in gel filtration chromatography, however, indicated that the molecular mass to be about 130-KDa implying the active form to be a dimer. On the basis of a molecular mass of 68-KDa, the molar extinction coefficient of the enzyme at 280 nm was estimated to be 3.4 x 10(5) M-1 cm-1. The isoelectric point of the enzyme, as judged by chromatofocusing, was about 4.0. The purified enzyme is highly stable at 0 degree C. Thermal inactivation studies have indicated that the enzyme is unstable at 70 degrees C. The enzyme, however, exhibited a broad temperature optima with a maximum catalytic activity at 63 degrees C. The Km of the enzyme for p-nitrophenylphosphate is about 270 microM with an estimated turnover number of 2550 per sec. The enzyme is a glycoprotein as evidenced by the positive PAS staining; the sugar composition suggests the presence of N-linked high mannose-oligosaccharides. A partial N-terminal amino acid sequence up to the twenty-third residue was obtained. The enzyme was inhibited competitively by inorganic orthophosphate (Ki = 185 microM) and non-competitively by phosphomycin (Ki = 600 microM).     

Aspergillus ficuum phytase: partial primary structure, substrate selectivity, and kinetic characterization

Purified Aspergillus ficuum phytase's partial primary structure and amino acid and sugar composition were elucidated. Determination of kinetic parameters of the enzyme at different pH values and temperatures indicated no significant alteration of the Km for phytate while the Kcat was affected. The enzyme was able to release more than 51% of the total available Pi from phytate in a 3.0 hr assay at 58 degrees C, but the Kcat dropped to 15% of the initial rate. Substrate selectivity studies revealed phytate to be the preferred substrate. The pH optima of phytase was 5.0, 4.0, and 3.0 for phytate, ATP, and polyphosphate, respectively. The enzyme had varied sensitivity towards cations. While Ca++ and Fe++ produced no effect on the catalytic rate of the enzyme, Cu+, Cu++, Zn++, and Fe were found to be inhibitory. Mn++ was observed to enhance enzyme activity by 33% at 50 microM. Known inhibitors of acid phosphatases e.g. L (+)-tartrate, phosphomycin, and sodium fluoride had no effect on enzyme activity.     

Primary structure of the hemoglobin α-chain of rose-ringed parakeet (Psittacula krameri)

The structure of the hernoglobin α-chain of Rose-ringed Parakeet was determined by sequence degradations of the intact subunit, the CNBr fragments, and peptides obtained by digestion with staphylococcal Glu-specific protease and trypsin. Using this analysis, the complete α-chain structure of 21 avian species is known, permitting comparisons of the protein structure and of avian relationships. The structure exhibits differences from previously established avian α-chains at a total of 61 positions, five of which have residues unique to those of the parakeet (Ser-12, Gly-65, Ser-67, Ala-121, and Leu-134). The analysis defines hemoglobin variation within an additional avian order (Psittaciformes), demonstrates distant patterns for evaluation of relationships within other avian orders, and lends support to taxonomic conclusions from molecular data.     

The use of the probit model to predict pregnancy status of buffalo based on physio-chemical properties of estrual mucus

Improper timing of artificial insemination with respect to ovulation is one of the major factors hampering the conception rate in buffalo. The present study was an attempt to relate physio-chemical changes in estrual mucus to subsequent pregnancy status in order to find their optimal values for determining the time for artificial insemination (AI). Serum estradiol, total protein and dry matter contents of estrual mucus were evaluated to predict the subsequent pregnancy in 36 buffalo during October 1988 to February 1989. Serum estradiol was determined by radioimmunoassay (RIA); spinnbarkeit, dry matter and total protein were determined by standard methods. Multivariate probit analyses were carried out to relate these variables to subsequent pregnancy status. Elasticity and protein concentration were significantly related to prediction probability of pregnancy status, and they predicted the pregnancy status 86% of the times correctly (P < 0.05). The probability of pregnant animals being correctly classified was 0.76, whereas the corresponding value for non-pregnant animals was 0.95. The present study demonstrated the possibility of using such a statistical model on mucus characteristics for determining proper AI time for better conception rates in Nili-Ravi water buffalo.     

Positive identification of a lambda gt11 clone containing a region of fungal phytase gene by immunoprobe and sequence verification

Summary As the initial step in a project to provide a more cost-effective source of the phytase enzyme, this paper reports on the use of a polyclonal antibody raised to phytase purified from an isolate of Aspergillus niger (A. ficuum) to screen an A. niger lambda gt11 expression library and the use of amino acid sequencing to identify a clone containing part of the fungal phytase gene. The described use of amino acid sequence fragments to verify the cloning of a gene has potential applications in other cloning projects.Mention of a trademark or proprietary product does not constitute a guarantee or warranty by the U.S. Department of Agriculture and does not imply approval to the exclusion of other products that may also be suitable Offprint requests to: E. Mullaney     

Primary structure of hemoglobin β-chain fromColumba livia (gray wild pigeon)

Primary structure of -chain of pigeon is presented. It was determined by amino acid sequence analysis of intact -chain and its peptides obtained by the enzymatic and chemical cleavage. Comparison of amino acid sequence of the chain with other available data shows 14 Ile, 61 Lys, and 113 Ile as residues specific to pigeon. One important replacement at ₁₁ contact is 55 MetSer.     

Description of Dolichodorus cobbi n. sp. (Nematoda: Dolichodoridae) with Description of Dolichodorus cobbi n. sp. (Nematoda: Dolichodoridae) with Morphometrics and Lectotype Designation of D. heterocephalus Cobb, 1914

Dolichodorus cobbi n. sp. is described and illustrated from soil around roots of "woods" on the Stanley Farm, Scott County, Arkansas. This species is distinguishable from others of the genus by its short tail projection. It is most closely related to D. marylandicus but differs in having a short tail projection, longer stylet, greater body length, larger c value, and also in position of excretory pore. A limited number of Cobb''s original specimens and drawings of D. heterocephalus were examined. A lectotype and paralectotype were designated to establish the taxonomic base for the genus. Cobb''s original modified labeled drawing of D. heterocephalus is also included. Morphometric data on D. heterocephalus from the type locality and several other populations are given. Some variations in female tail shape, body length, and stylet length were noted.     

Isolation of the first trypsin inhibitor from the genusAspergillus

The fungusAspergillus flavipes was grown on a Czapeck sucrose medium; the biomass so obtained was treated with high concentration of sucrose to release intracellular metabolites. Sephadex G-75 chromatography of the latter yielded a pure protein having anti-trypsin activity in vitro.     

Rearing, reproductive behaviour and gamma sterilization of fruit fly, Dacus zonatus (Diptera: Tephritidae)

Investigations were made on rearing, reproductive behaviour and gamma sterilization of one-day old male adults of Dacus zonatus. The larvae were successfully reared on an artificial diet based on wheat shorts. Adult emergence ranged from 89–99% with a sex ratio of about 1:1. Mating occurred at dusk and its duration ranged from 8–13 hours. Males mated a second time with the same female but preferred mating if the already mated female was replaced with a sexually mature virgin female. The optimum dosage for inducing sterility amongst one-day old male adults was 12 kR.

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Zusammenfassung Zucht und Fortpflanzungsverhalten von Dacus zonatus (Saunders) wurde untersucht. Die Larven wurden vier Generationen lang an einer Diät aus Weizenkleie, Bierhefe, granuliertem Zucker, Agar, Nipagin, Salzsäure und Wasser gehalten. Die Arbeit gibt Daten über Verpuppungsprozentsatz (69,3%), Puppengewicht, Dauer der Ei + Larvenperiode, Schlüpfen der Adulten, Präovipositionszeit, Fruchtbarkeit, Fertilität und Lebensdauer der Adulten. Die Schlüpfrate der Adulten betrug 89–99%, das Geschlechtsverhältnis lag etwa bei 1:1. Die Kopulation findet während der Abenddämmerung statt, sie dauert 8–13 Stunden. Maximum der Kopulationen zwischen dem 10. und 15. Tag nach dem Schlüpfen. Männchen paarten sich ein zweites Mal mit dem gleichen Weibchen, bevorzugten jedoch geschlechtsreife jung-fräuliche Weibchen. Die Eiablage begann am 2.–7. Tag nach der Paarung, die Eizahl betrug bei gepaarten Weibchen 91–564.Die optimale Dosis zur Erzeugung von 99,3% Sterilität bei Bestrahlung von einem Tag alten Männchen war 12 kR. Die Lebensdauer der Bestrahlten war vermindert.