Biochemical pathways in prokaryotes can be traced backward through evolutionary time

For the first time, a credible prokaryotic phylogenetic tree is being assembled by Woese and others using quantitative sequence analysis of oligonucleotides in the highly conservative rRNA. This provides an evolutionary scale against which the evolutionary steps that led to the arrangement and regulation of contemporary biochemical pathways can be measured. This paper presents an emerging evolutionary picture of aromatic amino acid biosynthesis within a large superfamily assemblage of prokaryotes that is sufficiently developed to illustrate a new perspective that will be applicable to many other biochemical pathways.      

Genomic fingerprinting by arbitrarily primed polymerase chain reaction resolves Borrelia burgdorferi into three distinct phyletic groups.

The causative agent of Lyme disease, Borrelia burgdorferi, was first identified by Burgdorfer et al. in 1982 (W. Burgdorfer, A. G. Barbour, S. F. Hayes, J. L. Benach, E. Grunwaldt, and J. P. Davis, Science 216:1317-1319, 1982) and was isolated by Barbour et al. in 1983 (A. G. Barbour, W. Burgdorfer, S. E. Hayes, O. Peter, and A. Aeschlimann, Curr. Microbiol. 8:123-126, 1983). Since then, a large number of isolates have been collected, and there have been questions regarding the relationships among the various strains. Using genomic fingerprinting by an arbitrarily primed polymerase chain reaction, we resolved into three groups a collection of Eurasian and North American isolates of spirochetes that are generally categorized as B. burgdorferi. Group I strains have been identified in both North America and Eurasia, while strains belonging to Borrelia groups II and III have been found only in Eurasia. These same three groups have also been delineated by Baranton et al. (G. Baranton, D. Postic, I. Saint Girons, P. Boerlin, J.-C. Piffaretti, M. Assous, and P. A. D. Grimont, Int. J. Syst. Bacteriol. 42:370-375, 1992) by independent methods. Two isolates are distinct from all of the other strains in our collection but are clearly members of the genus Borrelia.     

Diets of sympatric flatfishes, Hippoglossoides platessoides, Pleuronectes ferrugineus, Pleuronectes americanus, from Sable Island Bank, Canada

Prey exploitation was documented for Hippoglossoides platessoides, Pleuronectes ferrugineus , and P. americanus collected from southeast Sable Island Bank in February and June 1989. Diets varied significantly by sample and with fish species and length. Of 239 species consumed by flatfishes, 66 were determined to be principal prey. Crustaceans, particularly amphipods, were the most frequently exploited prey of all three flatfish species. Small H. platessoides fed on suprabenthic fauna, while larger fish exploited epifauna and hyperiid suprafauna. P. ferrugineus exploited epibenthic fauna, consuming crustaceans and tunicates when small and polychaetes when larger. P. americanus preyed on epifauna; smaller fish exploited polychaetes while larger fish usually consumed crustaceans associated with ectoproct colonies. Feeding intensity was dramatically lower in February, food being found in stomachs of only 36% of H. platessoides and 49% of P. ferrugineus , and lacking in the stomachs of P. americanus . In June, stomachs of 86% of H. platessoides , 94% of P. ferrugineus and 96% of P. americanus contained food. Results indicate that prey resources for the three flatfish species are partitioned by specific habitat and prey type preferences.     

Growth factor stimulation of proliferating cell nuclear antigen (PCNA) in human breast epithelium in organ culture.

Normal human breast explants were maintained in serum-free culture for 7 days in the presence of either insulin hydrocortisone and cholera toxin (I/H/CT), epidermal growth factor (EGF) or transforming growth factor-alpha (TGF-alpha). Explants were labelled with [3H] thymidine, fixed in methacarn and processed for autoradiography. Parallel sections were immunolabelled with anti-PCNA antibody and analysed with a CAS 200 image analyser. Thymidine labelling index (TLI) and PCNA expression produced similar results with both indices increased in response to I/H/CT, EGF and TGF-alpha. In sections double labelled for PCNA and autoradiography the majority of labelled cells were positive for both markers.     

PCR-amplified length polymorphisms in tRNA intergenic spacers for categorizing staphylococci

The intergenic spacers between some adjacent tRNA genes were shown to be polymorphic in length when closely related Staphylococcus species were compared. A simple procedure was developed to detect and sequence these tRNA intergenic length polymorphisms (tRNA-ILPs). A comparison of homologous tRNA gene sequences flanking these ILPs in three Staphylococcus species was used to derive primers for high-stringency amplification of the ILPs by the polymerase chain reaction (PCR). The detection of tRNA-ILPs by PCR allowed the classification of virtually all strains from the five species of Staphylococcus that were examined. The procedure used to identify, sequence and derive primers for PCR detection of tRNA-ILPs in Staphylococcus should be applicable to many other genera of eubacteria. These primers could be used on uncultured material such as clinical samples.     

Phocanema decipiens: Growth,reproduction, and survival in seals

Captive harbor seals (Phoca vitulina) and gray seals (Halichoerus grypus) were fed infective larvae of Phocanema decipiens, an anisakine nematode from the flesh of Atlantic cod (Gadus morhus). Ova of P. decipiens were first detected in the feces of harbor seals 21(17–30) days after exposure; the patency period was 15 to 45 days. In gray seals, the prepatent period was 19(16–23) days; patency 20–60 days. By the sixth week of infection in harbor seals, mean body lengths of adult females and males of P. decipiens were 60.8(40.8–76.2) and 54.3(45.5–60.8) mm, respectively; mean fecundity of female nematodes was 156,000 ova. In infections of similar duration in gray seals, females and males of P. decipiens were 82.1(69.7–104.3) and 64.4(53.8–72.7) mm in length, respectively; mean fecundity of females was 366,000 ova. In sensitizing infections in harbor seals, 28% of P. decipiens survived to early patency (Days 25–30) while only 9% of the nematodes survived to midpatency (Days 35–45). In sensitizing infections in gray seals, 56% of P. decipiens survived to early patency (Days 20–30) and 48% survived to midpatency (Days 35–50). Seals with existing or recent P. decipiens infections resisted reinfection; <50% of the nematodes in challenge infections in gray seals survived to Day 3 and <10% survived to patency. Growth of the nematodes, however, was not retarded in the challenge infections and resistence to reinfection subsided when seals were maintained anisakinefree for 2–6 months after loss of prior natural or experimental infections. Natural anisakine infections were surveyed in 16 harbor and 53 gray seals from the Nova Scotia mainland. The mean incidence of P. decipiens was 62(5–177) in harbor seals and 577(11–1694) in gray seals; incidence varied seasonally and with age of host. Adult females of P. decipiens from harbor seals were 64.0(49.2–79.8) mm in length and contained 1.68(0.87–2.73) × 10⁵ ova; females from gray seals were 78.3(62.3–92.1) mm in length and contained 2.39(0.69–4.39) × 10⁵ ova.     

Restriction fragment fingerprint and genome sizes of Staphylococcus species using pulsed-field gel electrophoresis and infrequent cleaving enzymes.

Large restriction fragments of genomic DNA from Staphylococcus species were separated by pulsed-field gel electrophoresis (PFGE). Five different strains of S. aureus (ISP8, SAU3A, PS96, ATCC 6538, ATCC 15564) and three representative strains of S. haemolyticus SM102, S. warneri MCS4, S. cohnii LK478 from human hosts, and one strain of S. aureus (ATCC 8432) from an avian host were used in this study. Since Staphylococcus is A + T rich (approximately 67%), restriction fragments were obtained by digesting chromosomal DNA with endonucleases that recognize GC-rich sequences. Five enzymes Csp I, Sma I, Ecl XI, Ksp I, or Sac II were used for generation of few (7 to 16) distinctly separated fragments, with average sizes in the range of 200-300 kb. The size distribution of restriction fragments for each enzyme for each strain produced a strain-identifying fingerprint, and the genome size of each strain was determined from such restriction fragments separated by PFGE.     

Nitrogen loading from watersheds to estuaries: Verification of the Waquoit Bay Nitrogen Loading Model

World-wide eutrophication of estuaries has made accurate estimation ofland-derived nitrogen loads an important priority. In this paper we verifypredictions of nitrogen loads made by the Waquoit Bay Nitrogen LoadingModel (NLM). NLM is appropriate for watersheds with mixes of forested,agricultural, and residential land uses, and underlain by coarseunconsolidated sediments. NLM tracks the fate of nitrogen inputs byatmospheric deposition, fertilizer use, and wastewater disposal, and assignslosses of nitrogen from each source as the nitrogen is transported throughthe land use mosaic on the watershed surface, then through the underlyingsoils, vadose zones, and aquifers.We verified predictions of nitrogen loads by NLM in two independent ways.First, we compared NLM predictions to measured nitrogen loads in differentsubestuaries in the Waquoit Bay estuarine system. Nitrogen loads predictedby NLM were statistically indistinguishable from field-measured nitrogenloading rates. The fit of model predictions to measurements remained goodacross the wide range of nitrogen loads, and across a broad range in size(10–10,000 ha) of land parcels. NLM predictions were most precise whenspecific parcels were larger than 200 ha, and within factors of 2 for smallerparcels.Second, we used NLM to predict the percentage of nitrogen loads toestuaries contributed by wastewater, and compared this prediction to the¹⁵N signature distinguishable from N derived fromatmospheric or fertilizer sources. The greater the contribution ofwastewater, the heavier the ¹⁵N value in groundwater. Thesignificant linear relation between NLM predictions of percent wastewatercontributions and stable isotopic signature corroborated the conclusionthat model outputs provide a good match to empirical measurements. Thegood agreement obtained in both verification exercises suggests that NLMis an useful tool to address basic and applied questions about how land usepatterns alter the fate of nitrogen traversing land ecosystems, and thatNLM provides verified estimates of the land-derived nitrogen exports thattransform receiving aquatic ecosystems.