Seasonal prevalence of malaria vectors and entomological inoculation rates in the rubber cultivated area of Niete, South Region of Cameroon.

BACKGROUND: Development of large scale agro-industries are subject to serious environmental modifications. In malaria endemic areas this would greatly impact on the transmission paradigm. Two cross-sectional entomological surveys to characterize the Anopheles fauna and their entomological inoculation rates were conducted during May 2010 (peak rainy season) and December 2010 (peak dry season) in the intense rubber cultivated area of Niete in southern forested Cameroon. METHODS: Mosquitoes were sampled by night collections on human volunteers, identified morphologically and members of the Anopheles gambiae complex further identified to species and molecular form. Parity status was determined following the dissection of the ovaries. Plasmodium falciparum circumsporozoite antigen indices were estimated after the identification of CS antigen by ELISA and the average entomological inoculation rates determined. RESULTS: A total of 1187 Anopheles was collected, 419 (35.3%) in the rainy season and 768 (64.7%) in the dry season. Species found were the M molecular form of An. gambiae s.s (66.8%), An. ziemanni (28.3%), An. paludis (4.7%), An. smithii (0.2%). An. gambiae M-form was the principal species in the dry (56.2%) and wet (86.2%) seasons. Average overall entomological inoculation rate for the malaria vectors varied between the dry season (1.09 ib/p/n) and the rainy season (2.30 ib/p/n). CONCLUSIONS: Malaria transmission in Niete occurs both in the dry and rainy season with the intensities peaking in the dry season. This is unlike previous studies in other areas of southern forested Cameroon where transmission generally peaks in the rainy season. Environmental modifications due to agro-industrial activities might have influenced vector distribution and the dynamics of malaria transmission in this area. This necessitates the possible implementation of control strategies that are related to the eco-geography of the area.

Expression and rapid one-step purification of biologically active His-tagged factor C by Ni(2+) affinity column chromatography.

Factor C is an unusual extracellular protein capable of inducing cytodifferentiation in certain Streptomyces strains. The protein is produced by Streptomyces griseus 45H at such a low amount that the study of its mode of action was hindered by the shortage of purified protein. We report here the expression of C-terminally hexa-His-tagged factor C in Streptomyces lividans and Escherichia coli. Expression in S. lividans is low while in E. coli it is relatively high, yielding about 5--10 mg of biologically fully active protein per liter culture.

Phenotypic heterogeneity of the progeny of Streptomyces griseus conidia.

In order to understand the complex ontogenetical processes, the development of Streptomyces (S.) griseus was applied as a model. The developmental cycle of S. griseus starts and ends as a conidium. In between, coenocytic mycelium develops which, if studied by cytomorphological or biochemical methods, exhibits conspicious heterogeneity. The hyphae develop into young, transient and old vegetative hyphae and different stages of reproductive forms. In developmentally blocked mutants these sequences of events appear mixed in all possible associations. It seems as if the program of development could be divided into several subprograms. The quantitative evaluation of the results show that the individual morphological markers exhibit certain independence from each other realized with a given probability. The conidia of S. griseus are also heterogeneous concerning all morphological and physiological traits examined so far (shape, size, light refraction, staining and shape of nucleoids with Feulgen, methyl green--pyronine, intensity and form of polysaccharide distribution, heat resistance, etc.). Kinetics of the survival curves of two S. griseus strains--a well-sporulating and its developmentally blocked mutant /24/--are different from each other, one has many more heat resistant conidia than the other but the kinetics of the survival curves of the two S. griseus strains indicate that spore populations of both react differently to heat treatment and heat resistance can be modeled by assuming the presence of two independent subpopulations of spores with different heat sensitivity. The emergence of two distinct subpopulations with (possibly) the same genetic make-up is designated: phenotypic segregation. Heat resistance is first of all species specific (genetically determined) but the epigenetic segregation seems to be characteristic of the developmental process. This process can in certain mutants be affected by environmental conditions and more importantly by the so-called autoregulators (A-factor and factor C). Factor C and A-factor are needed to normal development, if their quantity or the time of addition to the culture was not optimal, the quantity of spores decreased.

Amino acid sequence homology of factor C produced by Streptomyces griseus with regulatory proteins of zinc finger type.

Factor C is a regulatory protein produced by Streptomyces griseus 45H. Factor C-like antigen can be detected in the most diverse species examined. It is also present in human serum with an average of 219.4 U/ml of narrow dispersion. The level of factor C-like antigen shows an increase in the sera of patients with different hepatic disorders (341 U/ml), some values being 3-7 times higher than normal (600-1500 U/ml). Correlation was found between the elevated antigen levels and the amount of the bilirubin in the sera of patients with liver cirrhosis. Amino acid sequence homology was found between factor C and zinc finger motifs of several known DNA binding proteins. In fact, factor C was successfully bound to and eluted from a Zn2+ affinity column. Our data show that factor C is probably a zinc finger type DNA binding protein.

A short GC-rich sequence involved in deletion formation of cloned DNA in E. coli.

A spontaneous deletion probably caused by rec A independent recombination between short-direct repeat sequences was observed in pUC19 plasmid carrying a piece of Streptomyces genomic DNA after culturing in liquid medium. The deletion removed an unknown portion of the cloned DNA and the BamHI-EcoRI part of the multiple cloning site with an additional flanking 111 bp from the vector. At the junction a 13 bp GC-rich DNA sequence highly homologous to a known spontaneous deletion hotspot was found.

Detection and determination of factor C--a regulatory protein--in Streptomyces strains by antiserum and monoclonal antibody.

Rabbit antisera and monoclonal antibodies were raised against factor C, a regulatory protein of Streptomyces griseus. ELISA and immunoblotting techniques suitable to determine and characterize factor C antigen was detected in all the 23 Streptomyces strains and variants examined thus far and in one Bacillus subtilis too. Depending on the strain analysed it has a molecular mass of 34,000 or 70,000 in mycelial homogenates. Most of factor C was found excreted into the cultivation medium. The quantity of factor C antigen in different Streptomyces strains showed great variation. Amy+ strains were usually good producers of factor C while Amy- were not. This was consistent with our assumption that factor C was an inducer of reproductive phase in Streptomyces.

Acute non-dilating obstructive renal failure in a patient with AIDS.

A 30-year-old male who presented with acute renal failure was found to have acquired immunodeficiency syndrome (AIDS). Although sonography and computerized tomography did not show urinary tract dilatation, obstructive renal failure was demonstrated by retrograde pyelography. Relief of obstruction(s) due to encasement of the renal pelves and ureters with histiocytic lymphoma led to immediate return of normal renal function. Although the etiology of renal failure in this patient is highly unusual, the high incidence of lymphoma in patients with AIDS should make tumor-related renal disease a consideration in all such patients with renal dysfunction.

Protein patterns of Streptomyces griseus strains during development.

Protein patterns from mycelial extracts of Streptomyces griseus strains No. 45H and No. 52-1 were studied by one dimensional gradient PAGE with 20 cm run. The results are reproducible, the protein patterns are the same for the same developmental stage in a given strain. There are well-defined characteristic changes of the protein patterns during the life cycle. The proteins of spores show conspicuous differences compared to protein patterns of the old (72 h) mycelia in the same culture. There is no difference between protein patterns of spores from submerged and from solid media. The protein patterns of two closely related Streptomyces griseus strains significantly differ from each other. After pulse labelling with (14C)-labelled protein hydrolyzate for 40 min, the specific activities of the proteins show considerable differences. There are characteristic bands with low specific activity and others with high incorporation. The fluorograms of the 16, 40 and 64 h cultures show that different proteins are being synthesized intensively at different ages of the culture.

Comparison of protein patterns of Streptomyces griseus spores from surface and submerged cultures.

Protein patterns of conidia produced by a Streptomyces griseus strain in submerged cultures and on solid media were compared. Cell-free extracts (30 000 X g supernatant) were prepared and analyzed on gradient SDS polyacrylamide gels. The protein patterns of both kinds of conidia were found to be practically identical, and they differed from protein patterns of the old vegetative hyphae in a characteristic way.

A substance effecting differentiation in Streptomyces griseus. Purification and properties.

A conidium-producing variant of Streptomyces griseus, strain 45-H, produces a substance, factor C, which is capable of inducing conidium formation in the hyphae of a conidium-non-producing mutant, strain 52-1. Factor C can be determined quantitatively on the basis of this biological effect. The biologically active substance can be purified by ion-exchange chromatography on cellulose phosphate combined with affinity chromatography on DNA-agarose. The purified substance is concentrated at least 1700 times. The molecular weight of factor C, estimated by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulphate, is about 34500. On determining the amino acid composition of factor C 60% of the amino acids were found to be hydrophobic.

Detection and changes of nucleotide pyrophosphotransferase activity of Streptomyces griseus strains in submerged culture.

Nucleotide pyrophosphotransferase (NPT) activity of two Streptomyces griseus strains was studied in submerged culture during their life cycle. NPT activity could be detected only in the culture filtrate but not in the membrane fraction or in cell extract of the sporulating (streptomycin-non-producing) S. griseus No. 45-H. No enzyme could be detected in the non-sporulating (streptomycin-producing) S. griseus No 52--1 cultures.

Detection and changes of nucleotide pyrophosphotransferase activity in cultures of Streptomyces griseus strains on solid media.

The occurrence of nucleotide pyrophosphotransferase (ATP: nucleotide 5'-phosphate pyrophosphotransferase, EC 2.7.4) and the changes in its activity were studied during differentiation in cultures of two Streptomyces griseus strains grown on the surface of a solid medium. One of the strains does not sporulate and produce nucleotide pyro photransferase in submerged culture. Nucleotide pyrophosphotransferase could be detected, though in much differing amounts, in surface cultures of both strains.

Defective life cycle and low antibiotic production in submerged cultures of Streptomyces fradiae.

The life cycle of a Streptomyces fradiae strain producing high amounts of neomycin under industrial conditions has been investigated in liquid soybean medium where the production of antibiotic proved to be comparatively low. The changes occurring in the main macromolecular components and the enzyme activities of the mycelium during the life cycle and cytological observations proved that there was a block in the normal proecess of reproductive differentiation and a lack of exocellular alkaline phosphatase activity was found.

Inhibitory and mutagenic effects of sodium nitroprusside and its adenine complex on Escherichia coli.

Sodium nitroprusside and its adenine complex were found to decrease the growth rate of exponentially growing Escherichia coli cultures, and the adenine complex to exert in addition a bactericidal effect. In mutation experiments the latter compound failed to induce base-pair substitutions in the E. coli strains tested but the results do not allow to exclude other mutagenic mechanisms.