Direct isolation of Francisella spp. from environmental samples

Aims:  To develop a selective medium for isolation of F. tularensis, F. novicida and F. philomiragia from environmental samples.
Methods and Results:  A selective media, cysteine heart agar with 9% chocolatized sheep blood, containing polymyxin B, amphotericin B, cyclohexamide, cefepime and vancomycin (CHAB-PACCV) was developed and evaluated for growth of Francisella spp. No differences were observed in recovered colony forming units (CFUs) for F. tularensis , F. novicida and F. philomiragia on CHAB-PACCV vs nonselective CHAB. Growth of non- Francisella species was inhibited on CHAB-PACCV. When environmental samples were cultured on CHAB and CHAB-PACCV, only CHAB-PACCV allowed isolation of Francisella spp. Three new Francisella strains were isolated directly from seawater and seaweed samples by culture on CHAB-PACCV.
Conclusions:  CHAB-PACCV can be used for direct isolation of Francisella spp from environmental samples.
Significance and Impact of the Study:  Francisella spp. show a close association with environmental sources. Future utilization of CHAB-PACCV for isolation of Francisella spp. directly from environmental samples should prove valuable for investigating outbreaks and human infections attributed to environmental exposure.     

Purification and characterization of a novel, oligomeric, plasminogen kringle 4 binding protein from human plasma: tetranectin

Purification of alpha 2-plasmin inhibitor (alpha 2PI) from human plasma by affinity chromatography on plasminogen-Sepharose resulted in copurification of a contaminating protein with Mr 17,000 as judged by sodium dodecyl sulphate/polyacrylamide gel electrophoresis. This contaminating protein could not be removed from the purified alpha 2-PI preparation by several types of gel chromatography applied. The use of the kringle 1-3 part of plasminogen, K(1 + 2 + 3), bound to Sepharose for affinity chromatography, instead of plasminogen-Sepharose, resulted in an alpha 2PI preparation without this contaminant. The contaminating protein was found to interact specifically with the kringle 4 part of plasminogen (K4) and not with K(1 + 2 + 3) or miniplasminogen. The K4-binding protein was purified by ammonium sulphate precipitation, affinity chromatography on K4-Sepharose, ion-exchange chromatography and gel filtration on AcA 34. The relative molecular mass of the protein (Mr 68 000) was estimated by gel filtration. This suggests a tetrameric protein composed of four subunits (Mr 17,000), that are dissociated by 1% sodium dodecyl sulphate. Dissociation into subunits was also demonstrated by gel filtration in the presence of 6 M guanidine hydrochloride. A specific antibody was raised in rabbits against the purified protein and this antibody was shown not to react with any known fibrinolytic components. The pI of the K4-binding protein was found to be 5.8. The first three N-terminal amino acids were determined to be Glu-Pro-Pro. The concentration of the protein in plasma was estimated to be 0.20 +/- 0.03 microM (15 +/- 2 mg/l). The electrophoretic mobility of the K4-binding protein was shown by crossed immunoelectrophoresis to be influenced by the presence of Ca2+, EDTA and heparin. The protein was found to enhance plasminogen activation catalyzed by tissue-type plasminogen activator (t-PA) in the presence of poly(D-lysine). The protein appeared to be a novel plasma protein tentatively called 'tetranectin'.     

Mannopeptimycin esters and carbonates, potent antibiotic agents against drug-resistant bacteria

A series of ester and carbonate derivatives of the glycopeptide mannopeptimycin alpha (1) with potent activity against G+ bacteria, including the methicillin-resistant staphylococci and vancomycin-resistant enterococci, was synthesized. The SAR data obtained from natural and semisynthetic compounds demonstrated the importance of a hydrophobic group in the terminal mannosyl moiety for antibacterial activity.     

Specific protection from phenocopy induction by heat shock

Mild heat treatments applied to whole animals or cell cultures of Drosophila prior to lethal heat shocks result in increased survival and protection against phenocopy induction. The optimal condition for the preliminary mild heat treatment is that which induces the synthesis of heat-shock proteins but does not turn off the protein synthesis that is in progress. Recovery of protein synthesis but not RNA synthesis following a drastic heat shock is much enhanced by the pretreatments. The results suggest that the protection for survival and against phenocopy induction is due to storage of messenger RNA.     

Factors controlling nitrification and denitrification: A laboratory study with gel-stabilized liquid cattle manure

Nitrification and denitrification were studied in a millimeterscale microenvironment using a two-phase system with a liquid manure-saturated layer. Samples consisted of liquid cattle manure and air-dried soil stabilized with silica gel, placed between two aerobic soil phases with a water content near field capacity. A high potential for NH₄ ⁺ oxidation developed within 0–2 mm distance from the interface, and NH₄ ⁺ diffused only 10–20 mm into the soil. Some NH₄ ⁺ was probably immobilized by microorganisms in the soil between 0 and 4 days, after which nitrification was the only sink for NH₄ ⁺. A potential for denitrification developed within the manure-saturated zone. Maximum rates of both potential and actual denitrification were recorded by Day 4, but denitrification continued for at least 2–3 weeks. The potential for nitrification peaked after 14 days. When the pH of the manure was adjusted to 5.5, nitrification was reduced close to the interface, and NH₄ ⁺ penetrated further into the soil before it was oxidized. The pH adjustment had an inhibitory effect on denitrification: Both potential and actual rates of denitrification were almost eliminated for several days. The size of the manure-saturated layer strongly affected denitrification losses. With layers of 8 and 16 mm thickness, losses equivalent to 33 and 40% of the original NH₄ ⁺ pool, respectively, were estimated. When manure corresponding to a 12 mm layer was homogeneously mixed with the soil, only 0.3% was lost.Offprint requests to: S. O. Petersen.     

Selecting Biological Meaningful Environmental Dimensions of Low Discrepancy among Ranges to Predict Potential Distribution of Bean Plataspid Invasion

Background

The Bean plataspid (Megacopta cribraria) (Hemiptera: Pentatomidae), native to Asia, is becoming an invasive species in North America; its potential spread to soybean producing areas in the US is of great concern. Ecological niche modelling (ENM) has been used increasingly in predicting invasive species'' potential distribution; however, poor niche model transferability was sometimes reported, leading to the artifactual conclusion of niche differentiation during species'' invasion.

Methodology/Principals

We aim to improve the geographical transferability of ENM via environmental variable selection to predict the potential distribution of Bean plataspid invasion. Sixteen environmental dimensions between native and introduced Bean plataspid populations were compared, and classified into two datasets with different degrees of discrepancy by the interquartile range (IQR) overlap in boxplot. Niche models based on these two datasets were compared in native model prediction and invading model projection. Classical niche model approaches (i.e., model calibrated on native range and transferred outside) were used to anticipate the potential distribution of Bean plataspid invasion.

Conclusions/Significance

Niche models based on the two datasets showed little difference in native model predictions; however, when projecting onto the introduced area, models based on the environmental datasets showing low discrepancy among ranges recovered good model transferability in predicting the newly established population of Bean plataspid in the US. Recommendations were made for selecting biological meaningful environmental dimensions of low discrepancy among ranges to improve niche model transferability among these geographically separated areas. Outside of its native range, areas with invasion potential include the southeastern US in North America, southwestern Europe, southeastern South America, southern Africa, and the eastern coastal Australia.     

Early stress predicts age at menarche and first birth,adult attachment,and expected lifespan

Life history theory suggests that in risky and uncertain environments the optimal reproductive strategy is to reproduce early in order to maximize the probability of leaving any descendants at all. The fact that early menarche facilitates early reproduction provides an adaptationist rationale for our first two hypotheses: that women who experience more risky and uncertain environments early in life would have (1) earlier menarche and (2) earlier first births than women who experience less stress at an early age. Attachment theory and research provide the rationale for our second two hypotheses: that the subjective early experience of risky and uncertain environments (insecurity) is (3) part of an evolved mechanism for entraining alternative reproductive strategies contingent on environmental risk and uncertainty and (4) reflected in expected lifespan. Evidence from our pilot study of 100 women attending antenatal clinics at a large metropolitan hospital is consistent with all four hypotheses: Women reporting more troubled family relations early in life had earlier menarche, earlier first birth, were more likely to identify with insecure adult attachment styles, and expected shorter lifespans. Multivariate analyses show that early stress directly affected age at menarche and first birth, affected adult attachment in interaction with expected lifespan, but had no effect on expected lifespan, where its original effect was taken over by interactions between age at menarche and adult attachment as well as age at first birth and adult attachment. We discuss our results in terms of the need to combine evolutionary and developmental perspectives and the relation between early stress in general and father absence in particular. This work was supported by The University of Melbourne Department of Obstetrics and Gynaecology. James S. Chisholm is Professor in the School of Anatomy and Human Biology at the University of Western Australia. He is an anthropologist whose interests lie in the fields of human behavioral biology, evolutionary ecology, life history theory, and parental investment theory, where he focuses on infant social-emotional development, the development of reproductive strategies, and the integration of evolutionary, developmental, and cultural psychology and public health. Julie A. Quinlivan is Associate Professor in Obstetrics and Gynaecology at the University of Melbourne and Head of the Maternity Care Program at the Royal Women’s Hospital, Melbourne. Her interests are teenage pregnancy, domestic violence, child abuse prevention, and high-risk pregnancy. Rodney W. Petersen is Senior Lecturer in Obstetrics and Gynaecology at the University of Melbourne and Senior Consultant in Obstetrics and Gynaecology at the Royal Women’s Hospital and Sunshine Hospital in Melbourne. His interests are in psychosocial aspects of women’s health and cancer. David A. Coall is a Ph.D. student in the School of Anatomy and Human Biology at the University of Western Australia. His main interest lies in the application of evolutionary theory within an epidemiological framework. He is currently working on the synthesis of life history theory, parental investment theory, and parent-offspring conflict theory in exploring factors that influence variation in human birth weight and placental weight.