Identification and sequence comparison of a cuticular collagen of Brugia pahangi

The cuticle of filarial nematodes is a specialized extracellular matrix that covers the parasite and protects it from adverse conditions of the environment. As a surface structure it is in direct contact with the host defence mechanisms and therefore plays an important role in the molecular host-parasite relationship. Using polyclonal antisera raised against the insoluble components of the cuticle of the adult filarial parasite Brugia pahangi, we have isolated cDNA clones encoding collagen molecules of the cuticle. The protein domain structure of cDNA clone Bpcol-1 was compared with the known structures of cuticular collagens of the nematodes Brugia malayi, Caenorhabditis elegans, Ascaris suum and Haemonchus contortus, confirming interspecies similarities. Using affinity-purified anti-Bpcol-1 antibodies we identified Bpcol-1 antigenic determinants in different nematode extracts, and determined the localization of such epitopes within the cuticle of B. pahang

Sequence differences between histones of procyclic Trypanosoma brucei brucei and higher eukaryotes

Four histones, a, b, c, d from procyclic Trypanosoma brucei brucei, which show similarities with the amino acid composition of the core histones H3, H2A, H2B and H4, were isolated and cleaved with Endoproteinase Glu-C. The fragments were separated by FPLC reversed phase chromatography and a subset of the fragments (a5, a9, b6, c8, d3, d9, d11) was subjected to sequence analysis. A 54-71% identity was found in the sequences of the fragment c8 and the C-terminal half of H2B and of three fragments of protein d covering the N-terminal half as well as the C-terminal region of H4. The amino acid sequence of the fragment a9 showed a 57 and 54% identity with H3 sequences of Saccharomyces cerevisiae and Xenopus laevis. Neither the a5 nor the b6 sequence could be aligned with histone sequences of other eukaryotes. The significant differences of 21-48% between the T. b. brucei, histone sequences and those of calf thymus histones, which are more pronounced than the differences of Tetrahymena pyriformis and the higher eukaryote, resulted partially from replacements of amino acids with different properties and indicate specific patterns of histone-histone and/or histone-DNA contact sites in the nucleosome of T. b. brucei. These differences, together with the lack of a functional histone H1, may be sufficient to explain the lack of a salt-dependent formation of the nucleosome filament into the 30 nm fibre, which reflects alternative methods of organizing and processing the genetic information in the nucleus of the protozoan parasite and which may be of chemotherapeutic significanc

Trypanosoma brucei brucei: differences in the nuclear chromatin of bloodstream forms and procyclic culture forms

Nucleosome filaments of two stages of the life-cycle of Trypanosoma brucei brucei, namely bloodstream forms and procyclic culture forms, were investigated by electron microscopy. Chromatin of bloodstream forms showed a salt-dependent condensation. The level of condensation was higher than that shown by chromatin from procyclic culture forms, but 30 nm fibres as formed in rat liver chromatin preparations were not found. Analysis of histones provided new evidence for the existence of H1-like proteins, which comigrated in the region of the core histones in SDS-PAGE and in front of the core histones in Triton acid urea gels. Differences were found between the H1-like proteins of the two trypanosome stages as well as between the core histones in their amount, number of bands and banding pattern. It can be concluded that T. b. brucei contains a full set of histones, including H1-like proteins, and that the poor condensation of its chromatin is not due to the absence of H1, but most probably due to histone-DNA interaction being weak. It is obvious that structural and functional differences of the chromatin exist not only between T. b. brucei and higher eukaryotes, but also between various stages of the life-cycle of the parasite. It is therefore not adequate to investigate the chromatin only of the procyclic culture forms as a model for all stages of the life-cycle of T. b. bruce

Determination of Acaricide Resistance in Rhipicephalus (Boophilus) microplus (Acari: Ixodidae) Field Populations of Argentina, South Africa, and Australia With the Larval Tarsal Test

Infestations with ticks have an important economic impact on the cattle industry worldwide and resistance to acaricides has become a widespread phenomenon. To optimize their treatment strategy, farmers need to know if and against which classes potential acaricide-resistance does occur. Bioassays are used to assess the resistance level and pattern of Rhipicephalus (Boophilus) microplus populations. The objective of the current study was to assess the susceptibility of field populations originating from Argentina (8), South Africa (3), and Australia (2) using the Larval Tarsal Test. Nine acaricidal compounds from five major classes were tested: organosphosphates, synthetic pyrethroids (SP), macrocyclic lactones, phenylpyrazols, and amidines. The resistance ratios at concentrations inducing 50 and 90% mortality were used to detect established and emerging resistance. This study confirmed the newly reported presence of amitraz resistance in populations from Argentina. In addition, resistance to SP appeared to be widespread (88%) in the Argentinean farms, which had been selected based on the observation of lack of treatment efficacy by farmers. In South Africa one of the three populations was found to be resistant to SP and to a phenylpyrazol compound (pyriprol). Furthermore, resistance to organosphosphates and SP was observed in Australia. Finally, the Larval Tarsal Test proved to be a suitable test to evaluate the susceptibility of R. microplus field populations to the most relevant acaricidal classe

1+1+2 Electromagnetic perturbations on non-vacuum LRS class II space-times: Decoupling scalar and 2-vector harmonic amplitudes

We use the covariant and gauge-invariant 1+1+2 formalism of Clarkson and Barrett \cite{Clarkson2003} to analyze electromagnetic (EM) perturbations on non-vacuum {\it locally rotationally symmetric} (LRS) class II space-times. Ultimately, we show how to derive six real decoupled equations governing the total of six EM scalar and 2-vector harmonic amplitudes. Four of these are new, and result from expanding the complex EM 2-vector which we defined in \cite{Burston2007} in terms of EM 2-vector harmonic amplitudes. We are then able to show that there are four precise combinations of the amplitudes that decouple, two of these are polar perturbations whereas the remaining two are axial. The remaining two decoupled equations are the generalized Regge-Wheeler equations which were developed previously in \cite{Betschart2004}, and these govern the two EM scalar harmonic amplitudes. However, our analysis generalizes this by including a full description and classification of energy-momentum sources, such as charges and currents.Comment: 9 page

Genetically discrete populations of Trypanosoma congolense from livestock on the Kenyan coast

Twenty-seven stocks of Nannomonas trypanosomes isolated from livestock in 1982 on a ranch at Kilifi on the Kenyan coast were characterized by isoenzyme electrophoresis and by the abilities of the parasite's DNA to hybridize to two repetitive sequence DNA probes. Allthe Kilifi stocks which were examined had isoenzyme patterns which were markedly different from the 75 patterns previously described from 78 stocks of Trypanosoma congolense. On average only 15% of the enzyme bands present in the Kilifi stocks were present in those stocks of T. congolense which had previously been surveyed for isoenzymes. The DNA from all the Kilifi stocks which had been examined for isoenzymes hybridized with only the repetitive sequence probe isolated from a clone of a Kilifi stock. In contrast, the DNA from all 27 Kilifi stocks failed to hybridize with a repetitive sequence probe isolated from a clone from a different stock of T. congolense. Thus, the trypanosomes in all the Kilifi stocks examined were both phenotypically and genotypically discrete. These genetically discrete trypanosomes have also been detected in 2 stocks isolated from livestock from another location on the Kenyan coast. The results show that there is a wide range of genetic heterogeneity within the trypanosomes currently classified as T. congolense. We suggest that the limits of this genetic heterogeneity could represent incipient speciatio

Detecting malaria sporozoites in live, field-collected mosquitoes

A method is described for identifying malaria-infected mosquitoes, without killing them or hampering their fitness. Individual mosquitoes were induced to salivate on coverslips, and sporozoites, deposited on the glass surface, were visualized by Giemsa staining. Of 21 mosquitoes found to contain sporozoites by salivary gland dissection, 13 had delivered sporozoites on coverslips. A positive correlation was found between the amount of saliva expelled and ejection of sporozoites, indicating that the sensitivity of the method may be increased by improving the probing behaviour of the mosquitoes. The procedure described may be suitable for selecting infected mosquitoes which are able to eject sporozoites during probing. Being applicable to wild Anopheles and to large numbers of mosquitoes, the method lends itself for use in field studies on malari

Detecting malaria sporozoites in live, field-collected mosquitoes

A method is described for identifying malaria-infected mosquitoes, without killing them or hampering their fitness. Individual mosquitoes were induced to salivate on coverslips, and sporozoites, deposited on the glass surface, were visualized by Giemsa staining. Of 21 mosquitoes found to contain sporozoites by salivary gland dissection, 13 had delivered sporozoites on coverslips. A positive correlation was found between the amount of saliva expelled and ejection of sporozoites, indicating that the sensitivity of the method may be increased by improving the probing behaviour of the mosquitoes. The procedure described may be suitable for selecting infected mosquitoes which are able to eject sporozoites during probing. Being applicable to wild Anopheles and to large numbers of mosquitoes, the method lends itself for use in field studies on malari

1+1+2 Electromagnetic perturbations on general LRS space-times: Regge-Wheeler and Bardeen-Press equations

We use the, covariant and gauge-invariant, 1+1+2 formalism developed by Clarkson and Barrett, and develop new techniques, to decouple electromagnetic (EM) perturbations on arbitrary locally rotationally symmetric (LRS) space-times. Ultimately, we derive 3 decoupled complex equations governing 3 complex scalars. One of these is a new Regge-Wheeler (RW) equation generalized for LRS space-times, whereas the remaining two are new generalizations of the Bardeen-Press (BP) equations. This is achieved by first using linear algebra techniques to rewrite the first-order Maxwell equations in a new complex 1+1+2 form which is conducive to decoupling. This new complex system immediately yields the generalized RW equation, and furthermore, we also derive a decoupled equation governing a newly defined complex EM 2-vector. Subsequently, a further decomposition of the 1+1+2 formalism into a 1+1+1+1 formalism is developed, allowing us to decompose the complex EM 2-vector, and its governing equations, into spin-weighted scalars, giving rise to the generalized BP equations