Brazil's neglected biome: The South Brazilian Campos

The South Brazilian grasslands occupy some 13.7 million ha and support very high levels of biodiversity. This paper reviews the current state of ecological knowledge on South Brazilian Campos and of threats and challenges associated with their conservation. The principal factors shaping grassland physiognomy and diversity are discussed, and information is presented on diversity of plant species; best estimates suggest that 3000–4000 phanerophytes occur in the South Brazilian grasslands.It is argued that, despite their high species richness, Campos vegetation is not adequately protected under current conservation policies. In the past three decades, approximately 25% of the grassland area has been lost due to land use changes, and this trend continues. However, representation of Campos grasslands in conservation units is extremely low (less than 0.5%), and the management in most of these is inadequate to preserve the grasslands, as grazing and fire are important factors for their persistence. In conclusion, the following urgent needs are identified: (1) to create more conservation units in different regions, including different grassland types throughout southern Brazil, (2) to develop proper management strategies where grasslands are subject to shrub encroachment and forest expansion, (3) to conduct research on biodiversity and ecological processes in the Campos region and (4) to raise public awareness of the value and vulnerability of this vegetation type.     

The tadpole of Hypsiboas leptolineatus (Braun and Braun, 1977), a species in the Hypsiboas polytaenius clade (Anura; Hylidae).

The larval morphology of Hypsiboas leptolineatus was studied. The tadpole has an ovoid body in lateral view, wider than deep; snout rounded with dorsal reniform nostrils; spiracle sinistral with lateral wall attached to body; anal tube dextral; tail fins convex with acuminate tip; oral disc ventral; labial tooth row formula is 2(2)/3(1); moderately developed beaks with serrated jaw sheaths. These external oral features are compared with those of the known tadpoles in the Hypsiboas polytaenius clade. The oral cavity was studied using an electron microscope. Life history aspects are commented.     

Rotavirus neutralizing protein VP7: antigenic determinants investigated by sequence analysis and peptide synthesis.

The rotavirus neutralizing antigen, VP7, is a 37,000-molecular-weight glycoprotein which is a major component of the outer shell of the virion. The amino acid sequence of VP7 for strain S2 (human serotype 2) and Nebraska calf diarrhea virus (bovine serotype) has been inferred from the nucleic acid sequence of cloned copies of genomic segment nine. Comparison of the amino acid sequences of these two VP7 proteins with those already determined for other rotavirus strains reveals extensive sequence conservation between serotypes with clusters of amino acid differences sited predominantly in hydrophilic domains of the protein. Six peptides have been synthesized that span the hydrophilic regions of the molecule. Antisera to these peptides both recognize the respective homologous peptides in a solid-phase radioimmunoassay and bind to denatured VP7 in a Western blot. However, none of the antisera either recognize virus or exhibit significant neutralizing activity, indicating that these peptide sequences are not available on the surface of the virus.     

Improvement of solubility and stability of the antimicrobial peptide nisin by protein engineering.

Nisin is a 3.4-kDa antimicrobial peptide that, as a result of posttranslational modifications, contains unsaturated amino acids and lanthionine residues. It is applied as a preservative in various food products. The solubility and stability of nisin and nisin mutants have been studied. It is demonstrated that nisin mutants can be produced with improved functional properties. The solubility of nisin A is highest at low pH values and gradually decreases by almost 2 orders of magnitude when the pH of the solution exceeds a value of 7. At low pH, nisin Z exhibits a decreased solubility relative to that of nisin A; at neutral and higher pH values, the solubilities of both variants are comparable. Two mutants of nisin Z, which contain lysyl residues at positions 27 and 31, respectively, instead of Asn-27 and His-31, were produced with the aim of reaching higher solubility at neutral pH. Both mutants were purified to homogeneity, and their structures were confirmed by one- and two-dimensional 1H nuclear magnetic resonance. Their antimicrobial activities were found to be similar to that of nisin Z, whereas their solubilities at pH 7 increased by factors of 4 and 7, respectively. The chemical stability of nisin A was studied in the pH range of 2 to 8 and at a 20, 37, and 75 degrees C. Optimal stability was observed at pH 3.0. Nisin Z showed a behavior similar to that of nisin A. A mutant containing dehydrobutyrine at position 5 instead of dehydroalanine had lower activity but was significantly more resistant to acid-catalyzed chemical degradation than wild-type nisin Z.     

Effect of 5′-terminal structure and base composition on polyribonucleotide binding to ribosomes

Ribopolymers of variable base composition and 5′-terminal structure were synthesized with polynucleotide phosphorylase. Under primer-dependent conditions, m⁷GpppGmpC (m⁷G-cap)², its alkali-treated m⁷G ring-opened derivative, GpppGpC and ppGpC but not m⁷GpppGmpCp, m⁷GpppGm or GpppG were incorporated as 5′-termini. The ribopolymers were compared with reovirus mRNA, which contains m⁷G-cap, for their ability to form initiation complexes with wheat germ 40 S ribosomal subunits and 80 S ribosomes. The presence of 5′-terminal m⁷G was required for stable complex formation by some ribopolymers while for others binding was increased by two- to fourfold. The final level of binding observed was similar to that with reovirus mRNA. In addition to 5′-terminal m⁷G, the base composition of the ribopolymers markedly influenced binding. Some ribopolymers including m⁷G-cap (A)_n did not bind significantly; m⁷G-cap (U)_n formed 40 S complexes while m⁷G-cap (A,U)_n bound to 80 S ribosomes. The ribopolymer m⁷G-cap (A₂,U₂,G)_n directed protein synthesis as measured by amino acid incorporation into polypeptides, methionine tRNA association with 40 S complexes, and puromycin reactivity of 80 S-associated methionine and, like reovirus mRNA, its binding to ribosomes was inhibited by 7-methylguanosine 5′-monophosphate.     

A general strategy for cloning double-stranded RNA: nucleotide sequence of the Simian-11 rotavirus gene 8

Using Simian-11 rotavirus RNA, a strategy has been developed for the production of full length cloned copies of the genes of double-stranded (dsRNA) viruses. Genomic RNA segments were polyadenylated and reverse transcribed to yield a mixture of full length cDNA copies of both possible polarities. The cDNAs were annealed, filled in to complete any partial copies, tailed and inserted into the PstI site of pBR322 using dG/dC tailing. Cloned rotavirus cDNA gene copies were assigned to genomic RNA segments by Northern hybridization. The complete sequence of gene 8 which codes for NCVP3, a non-structural protein of SA11 rotavirus, was determined from a cloned gene copy. It is 1059 bases in length and has an open reading frame which could code for a protein containing 317 amino acids. The apparent 5' and 3' terminal non coding regions are 46 and 59 bases in length, respectively. The sequence ATGTGACCOH at the 3' end of the plus strand is conserved in four of the eleven genes examined. The cloning procedures used should be generally applicable to viruses with segmented dsRNA genomes.     

Conservation and variation in the hemagglutinins of Hong Kong subtype influenza viruses during antigenic drift.

The nucleotide sequence was determined for the hemagglutinin gene of the Hong Kong subtype influenza strain A/Bangkok/1/79. The amino acid sequence predicted from these data shows a total of 36 amino acid changes as compared with hemagglutinin for a 1968 Hong Kong strain, 11 more than had occurred in a 1975 strain. The distribution of these changes confirmed that there are conserved and highly variable regions in hemagglutinin as the viral gene evolves during antigenic drift in the Hong Kong subtype. Of the four variable regions found in this study, only two have been seen previously. Correlation of highly variable areas in the hemagglutinins of Hong Kong subtype field strains with sites of amino acid changes in antigenically distinct influenza variants enabled us to predict likely antigenic regions of the protein. The results support and extend similar predictions made recently, based on the three-dimensional arrangement of hemagglutinin from a 1968 influenza strain.     

A rapid and non-destructive method to assess leaf injury caused by the cassava green mite,Mononychellus tanajoa (Bondar) (Acarina: Tetranychidae)

A relative scale of Leaf Damage Indices (LDI) from 0 to 5 describes the visible injury to leaves of cassava, Manihot esculenta Crantz caused by the cassava green mite, Mononychellus tanajoa (Bondar). As the scale is ordinal and thus not quantitative, the observed LDIs are converted individually to relative loss of chlorophyll on a ratio scale before an average injury is determined. This calibration is required because the ordinal and the ratio scales are not linearly related. A calibration curve was established on the basis of laboratory experiments to determine the chlorophyll content, c, of leaves representing various leaf damage indices. Several monotonously decreasing functions were fitted to the experimental data yielding the following relation , where c _o is the chlorophyll content of unijured leaf tissue and a is a constant describing the steepness of the curvilinear relation. This means that LDIs could be converted to relative loss of chlorophyll, d, where . The photosynthetically active leaf area of plants can be estimated by combining the relative loss of chlorophyll with leaf area assessments and adds the effects of defoliation and suspended growth to the chlorophyll depletion. The difference in photosynthetically active area that arises between uninjured and injured plants over a period of time provides a measure of spider mite injury that can be related to growth and yield. The method integrates the injury inflicted over a period of time, allows successive observations of the same plants, and is rapid and reasonably precise considering the time savings.     

Phytoplankton and zooplankton (Cladocera,Copepoda) relationship in the eutrophicated River Danube (Danubialia Hungarica,CXI)

The seasonal variation in primary production, individual numbers, and biomass of phyto- and zooplankton was studied in the River Danube in 1981. The secondary production of two dominant zooplankton species (Bosmina longirostris and Acanthocyclops robustus) was also estimated. In the growing season (April–Sept.) individual numbers dry weights and chlorophyll a contents of phytoplankton ranged between 30–90 × 10⁶ individuals, l^–1, 3–12 mg l^–1, and 50–170 µg l^–1, respectively. Species of Thalassiosiraceae (Bacillariophyta) dominated in the phytoplankton with a subdominance of Chlorococcales in summer. Individual numbers and dry weights of crustacean zooplankton ranged between 1400–6500 individuals m^–3, and 1.2–12 mg m^–3, respectively. The daily mean gross primary production was 970 mg C m^–3 d^–1, and the net production was 660 mg C m^–3 d^–1. Acanthocyclops robustus populations produced 0.2 mg C m^–3 d^–1 as an average, and Bosmina longirostris populations 0.07 mg C m^–3 d^–1. The ecological efficiency between phytoplankton and crustacean zooplankton was 0.03%.