Mutations affecting pigmentation and shape of the adult zebrafish

 Mutations causing a visible phenotype in the adult serve as valuable visible genetic markers in multicellular genetic model organisms such as Drosophila melanogaster, Caenorhabditis elegans and Arabidopsis thaliana. In a large scale screen for mutations affecting early development of the zebrafish, we identified a number of mutations that are homozygous viable or semiviable. Here we describe viable mutations which produce visible phenotypes in the adult fish. These predominantly affect the fins and pigmentation, but also the eyes and body length of the adult. A number of dominant mutations caused visible phenotypes in the adult fish. Mutations in three genes, long fin, another long fin and wanda affected fin formation in the adult. Four mutations were found to cause a dominant reduction of the overall body length in the adult. The adult pigment pattern was found to be changed by dominant mutations in wanda, asterix, obelix, leopard, salz and pfeffer. Among the recessive mutations producing visible phenotypes in the homozygous adult, a group of mutations that failed to produce melanin was assayed for tyrosinase activity. Mutations in sandy produced embryos that failed to express tyrosinase activity. These are potentially useful for using tyrosinase as a marker for the generation of transgenic lines of zebrafish. Received: 17 June 1996 / Accepted: 15 July 1996     

Identification of the mpl gene encoding UDP-N-acetylmuramate: L-alanyl-gamma-D-glutamyl-meso-diaminopimelate ligase in Escherichia coli and its role in recycling of cell wall peptidoglycan.

A gene, mpl, encoding UDP-N-acetylmuramate:L-alanyl-gamma-D-glutamyl-meso-diaminopimelat e ligase was recognized by its amino acid sequence homology with murC as the open reading frame yjfG present at 96 min on the Escherichia coli map. The existence of such an enzymatic activity was predicted from studies indicating that reutilization of the intact tripeptide L-alanyl-gamma-D-glutamyl-meso-diaminopimelate occurred and accounted for well over 30% of new cell wall synthesis. Murein tripeptide ligase activity could be demonstrated in crude extracts, and greatly increased activity was produced when the gene was cloned and expressed under control of the trc promoter. A null mutant totally lacked activity but was viable, showing that the enzyme is not essential for growth.     

Effects of nutrient and soil moisture on competition between shape Carex stricta,shape Phalaris arundinacea,and shape Typha latifolia

We investigated the importance of nutrients, soil moisture, arbuscular mycorrhizal fungi (AMF), and interspecific competition levels on the biomass allocation patterns of three wetland perennial plant species, Carex stricta Lam., Phalaris arundinacea L., and Typha latifolia L. A factorial experiment was conducted with high-low nutrient levels, high-low soil moisture levels, and with and without AMF inoculation. Under the experimental conditions, plant inoculation by AMF was too low to create a treatment and the AMF treatment was dropped from the total analysis. P. arundinacea and T. latifolia biomass were 73% and 77% higher, respectively, in the high nutrient treatment compared to the low nutrient treatment. Biomass allocation between shoots and roots remained relatively constant between environmental treatments, although shoot:root ratios of P. arundinacea declined in the low nutrient treatment. For C. stricta, the high nutrient and soil moisture treatments resulted in an increase in biomass of 50% and 15%, respectively. Shoot:root ratios were nearly constant among all environmental conditions. Biomass of T. latifolia and C. stricta was greatly decreased when grown with P. arundinacea. The rapid, initial height growth of P. arundinacea produced a spreading, horizontal canopy that overshadowed the vertical leaves of T. latifolia and C. stricta throughout the study. This pattern was repeated in both high and low nutrient and soil moisture treatments. When grown with P. arundinacea, C. stricta and T. latifolia significantly increased their mean shoot height, regardless of the nutrient or soil moisture level. The results of this experiment suggest that C. stricta and T. latifolia were light limited when growing with P. arundinacea and that canopy architecture is more important for biomass allocation than the other environmental conditions tested. The results also suggest that Phalaris arundinacea is an inherently better competitor (sensu Grime 1979) than C. stricta or T. latifolia.     

Distinct feeding behavior between sexes of shape Frankliniella occidentalis results in higher scar production and lower tospovirus transmission by females

Feeding behavior and scar production of male and female F. occidentalis (Pergande) (Thysanoptera: Thripidae) were studied in relation to transmission of tomato spotted wilt tospovirus (TSWV). Electrical penetration graph (EPG) analysis showed that females feed more frequently and intensively than males. The feeding intensity, reflected by silvery scar production and studied by an image analysis system, demonstrated that females induced more numerous scars than males. At the same time, males transmitted TSWV with a higher efficiency than females, indicating that TSWV transmission and scar production are not positively correlated. Furthermore, males produced significantly more local lesions of TSWV than females. These quantitative differences in scar production and transmission of TSWV can be explained by the lower mobility and higher consumption rate of females. The influence of the sex-ratio on crop damage and virus transmission, and thus to the spread of TSWV, is emphasized.     

ISOLATION AND CHARACTERIZATION OF PHASE-SPECIFIC COMPLEMENTARY DNAs FROM SPOROPHYTES AND GAMETOPHYTES OF PORPHYRA PURPUREA (RHODOPHYTA) USING SUBTRACTED COMPLEMENTARY DNA LIBRARIES1

The red alga Porphyra purpurea (Roth) C. Agardh has a life cycle that alternates between shell-boring, filamentous sporophytes and free-living, foliose gametophytes. The significant morphological differences between these two phases suggest that many genes should be developmentally regulated and expressed in a phase-specific manner. In this study, we prepared and screened subtracted complementary DNA (cDNA) libraries specific for the sporophyte and gametophyte of P. purpurea. This involved the construction of cDNA libraries from each phase, followed by the removal of common clones through subtractive hybridization. Sampling of the subtracted libraries indicated that 8–10% of the recombinant colonies in each library were specific for the appropriate phase. Of 20 putative phase-specific cDNAs selected from each subtracted library, eight unique clones were obtained for the sporophyte and seven for the gametophyte. After confirming their phase-specificities by hybridization to gametophyte and sporophyte messenger RNA, these 15 phase-specific cDNAs were sequenced, and the deduced amino acid sequences were used to search protein databanks. Two proteins encoded by the sporophyte-specific cDNAs and two by the gametophyte-specific cDNAs were identified by their similarity to databank entries.     

Ontogenetic development of the pineal organ,parapineal organ,and retina of the three-spined stickleback,Gasterosteus aculeatus L. (Teleostei)

The ontogenetic developments of the pineal organ, parapineal organ, and retina were studied by the use of light and electron microscopy in embryos and fry of the teleost, Gasterosteus aculeatus, from 60 to 168 h after fertilization. Sixty to 66 h after fertilization, the primordium of the pineal complex is discernible in the diencephalic roofplate; the parapineal anlage is located rostral to the pineal anlage. Photoreceptor cells endowed with outer segments are present in the embryonic pineal organ already after 72 h, whereas outer segments of retinal photoreceptors could not be demonstrated before 144 h (hatching occurs between 120-144 h). Furthermore, neuropil formations with synaptic specializations are present in the rostral part of the pineal organ 108 h after fertilization. At 72 h, the embryonic parapineal parenchyma is already differentiated into parapinealocytes, which give rise to the parapineal tract, and glia-resembling elements. Although parapinealocytes carry cilia (9 X 2 + 0), only a single outer segment of the photoreceptor type could be demonstrated in the parapineal organ of one adult stickleback. Photoreceptors present in the pineal organ of unhatched embryos are hardly involved in visual functions, but may already at this early developmental stage serve as photoneuroendocrine transducers.     

Fasciola hepatica: an antigen fraction derived from newly excysted juveniles, containing an immunoreactive 32-kDa protein, induces strong protective immunity in rats

Crude antigens of adult Fasciola hepatica and of newly excysted juveniles (NEJ) and a low-molecular-weight fraction of antigen from NEJs were tested for inducing protective immunity in rats. Two routes of vaccination were applied. The results showed that intraperitoneal vaccination induced significantly better protection (P <0.05) than intramuscular vaccination. Intraperitoneal vaccination with antigens from NEJs induced more effective protection: after challenge infection, rats that were so vaccinated had 92.6% (+/-2.5% SEM) fewer parasites in their liver and 57.3% (+/-13.3% SEM) fewer parasites penetrating the gut wall than control rats. Rats that were vaccinated with a low-molecular-weight fraction of antigen from NEJs were also highly protected against a challenge. F. hepatica antigens that are immunoreactive were identified on immunoblots, using sera collected from highly protected rats that had been vaccinated with NEJ antigens and also sera from cattle and rats that were experimentally infected with F. hepatica. The low-molecular-weight fraction of antigen from NEJs contained an immunodominant 32-kDa protein that was recognized by serum antibodies of vaccinated rats and immune cattle. This 32-kDa protein was not detected in partially purified antigens from adult flukes. We conclude that antigens of NEJs of F. hepatica, when injected intraperitoneally in rats, are highly protective. In particular, the 32-kDa protein contained in these antigens may be highly valuable for the development of an effective vaccine against F. hepatica.     

Harmonising the bioassessment of large rivers in the absence of near‐natural reference conditions – a case study of the Danube River

1. International river catchments pose challenges for effective water resource management. Catchment‐wide strategies are often complicated by differences in national bioassessment and quality classification. Intercalibration efforts aim to harmonise these differences, but rely on the consistent delineation of near‐natural reference sites that are almost unavailable in today’s landscape, especially for large rivers. 2. We introduce the concept of alternative benchmarking that is based on the notion of aquatic communities at similar (low) levels of impairment associated with least‐disturbed conditions (LDC) as defined by abiotic criteria. Using data acquired during the second Joint Danube Survey, we defined LDC sites based on a multivariate gradient of anthropogenic pressures, mostly related to morphological deterioration, that spans the entire navigable Danube. 3. The river was subdivided into four stretches, each featuring homogeneous biological assemblages. Indirect gradient analysis revealed relationships between the pressure gradient and selected features of the macroinvertebrate and macrophyte community but not for diatoms or phytoplankton. 4. We identified biological metrics suitable for the quality classification of individual stretches or the entire river. Impoundment is the major hydromorphological alteration on the Danube but various metrics still responded significantly to differences in the morphological condition of sites not affected by impoundment. 5. A comparison of macroinvertebrate sampling techniques (airlift versus kick‐and‐sweep) revealed differences in how the acquired data reflect the effects of anthropogenic pressure. Biological metrics based only on kick‐and‐sweep sample data were insensitive to habitat deterioration in the heavily modified Upper Danube. 6. This study exemplifies the empirical approach of alternative benchmarking in intercalibration and offers practical solutions to some of the challenges of large river bioassessment.     

Pneumococci in the African Meningitis Belt: Meningitis Incidence and Carriage Prevalence in Children and Adults

Background

The development of optimal vaccination strategies for pneumococcal conjugate vaccines requires serotype-specific data on disease incidence and carriage prevalence. This information is lacking for the African meningitis belt.

Methods

We conducted hospital-based surveillance of acute bacterial meningitis in an urban and rural population of Burkina Faso during 2007–09. Cerebrospinal fluid was evaluated by polymerase chain reaction for species and serotype. In 2008, nasopharyngeal swabs were obtained from a representative population sample (1 month to 39 years; N = 519) and additional oropharyngeal swabs from 145 participants. Swabs were evaluated by culture.

Results

Annual pneumococcal meningitis incidence rates were highest among <6-month-old (58/100,000) and 15- to 19-year-old persons (15/100,000). Annual serotype 1 incidence was around 5/100,000 in all age groups. Pneumococcal carriage prevalence in nasopharyngeal swabs was 63% among <5-year-old children and 22% among ≥5-year-old persons, but adding oropharyngeal to nasopharyngeal swabs increased the estimated carriage prevalence by 60%. Serotype 1 showed high propensity for invasive disease, particularly among persons aged ≥5 years.

Conclusions

Serotype 1 causes the majority of cases with a relatively constant age-specific incidence. Pneumococcal carriage is common in all age groups including adults. Vaccination programs in this region may need to include older target age groups for optimal impact on disease burden.