A simple mathematical model of the dynamics of malaria invasion is proposed. The model is based on the hypothesis that replication slippage (slipped strand mispairing occurring in replicating DNAs), likely to take place within regular, low-complexity, regions of the plasmodium genome, is a significant mechanism of the ability for the parasite to escape the host immune defence. The model reconciles the conclusions of other formal approaches like periodic bursts of parasitaemia and, in particular, it predicts that antibody cross-reactivity affects the level and the time profile of malaria recrudescence. It also suggests that an efficient strategy for the parasite to escape host humoral immune defences is to express a single antigen. Linking local complexity to antigenic variability tentatively allows prediction of new potentially immunogenic sequences in the plasmodium genome.

Specific glycolipids (GLs) from Leishmania (Viannia) braziliensis promastigotes were isolated and purified. A monoclonal antibody directed to carbohydrate epitopes of these GLs was produced. mAb SST-1 recognizes a low molecular weight GL as established by solid-phase radioimmunoassay and HPTLC immunostaining, and does not cross-react with lipophosphoglycan isolated from L. (V.) braziliensis promastigotes. An indirect immunofluorescence study indicated that the antigenic GLs are present at the L. (V.) braziliensis promastigote surface. SST-1 reacted with promastigotes of L. (V.) naiffi and L. (V.) guyanensis, but not with species in the L. Leishmania subgenus i.e. L. (L.) amazonensis, L. (L.) chagasi, or L. (L.) major. All L. (V.) braziliensis serodemes tested were reactive with SST-1. These results indicate that SST-1 recognizes specific GLs expressed by species of the Viannia subgenus, and will be particularly useful for identification of L. (V.) braziliensis.

Co-treatment of mice infected with different strains of Leishmania donovani with a non-ionic surfactant vesicle formulation of buthionine sulfoximine (BSO-NIV), and sodium stibogluconate (SSG), did not alter indicators of Th1 or Th2 responses but did result in a significant strain-independent up-regulation of IL6 and nitrite levels by stimulated splenocytes from treated mice compared to controls. The efficacy of BSO-NIV/SSG treatment was dependent on the host being able to mount a respiratory burst indicating that macrophages are important in controlling the outcome of treatment. In vitro studies showed that SSG resistance was associated with a greater resistance to killing by activated macrophages, treatment with hydrogen peroxide or potassium antimony tartrate. Longitudinal studies showed that a SSG resistant (SSG-R) strain was more virulent than a SSG susceptible (SSG-S) strain, resulting in significantly higher parasite burdens by 4 months post-infection. These results indicate that SSG exposure may favour the emergence of more virulent strains.

The protozoan parasite Toxoplasma gondii produces a family of microneme proteins that are thought to play diverse roles in aiding the parasite's intracellular existence. Among these, TgMIC2 has a putative function in parasite adhesion to the host cell to initiate the invasion process. The invasion process may be localized and inhibited by monoclonal antibodies against the protein(s) involved. Here we report on the construction of a phage-displayed single-chain variable fragment (scFv) library from mice immunized with whole T. gondii parasites. The library was subsequently panned against recombinant TgMIC2 (rpTgMIC2) and 2 different groups of antibody clones were obtained, based on fingerprinting and sequencing data. The expressed recombinant scFv antibody was able to recognize rpTgMIC2 in a Western blot detection experiment. These results show that the phage display technology allows quick and effective production of monoclonal antibodies against parasite antigens. By panning the scFv-displayed library, we should be able to obtain a plethora of multi-functional scFv antibodies towards T. gondii proteins.

An immunochromatographic test (ICT), using recombinant truncated P50 (P50t), for the detection of antibodies to Babesia gibsoni was developed and evaluated. Whereas all sera from specific pathogen-free dogs were clearly negative, all sera from dogs experimentally infected with B. gibsoni were clearly positive in the ICT. In addition, the ICT detected no cross-reactivity with sera from dogs experimentally infected with closely related parasites, B. canis canis, B. canis vogeli, and B. canis rossi, or with Neospora caninum, and Leishmania infantum. Sequential sera from a dog experimentally infected with B. gibsoni were tested with the ICT; it was shown that the specific antibodies are detectable as early as 6 days post-infection (p.i.) and that strong antibody responses remained until the end of the experiment (144 days p.i.). To evaluate the clinical application of the ICT, a total of 54 serum samples collected from domestic dogs that had been identified as having signs of anaemia at veterinary hospitals in Japan, were tested with the ICT, the previously established enzyme-linked immunosorbent assay (ELISA) and with the indirect fluorescent antibody test (IFAT). Twenty-four of the tested samples (44·4%) were positive in both ICT and ELISA, and (51·8%) in IFAT. The concordance between ELISA and ICT was found to be 100%, and 85·7% with IFAT. Taken together, the results above suggest that the ICT using P50t is rapid, simple, accurate, and suitable for use at clinical sites for the diagnosis of B. gibsoni infection in dogs.

We produced a mAb against the Babesia caballi extracellular merozoite termed mAb 2H2 and used it to screen a cDNA expression library prepared from B. caballi merozoite mRNA for highly expressed proteins. The complete nucleotide sequence of the cloned gene had 1547 nucleotides and contained a 36-nucleotide intron. The 1398 nucleotide open reading frame predicts a 51 kDa protein showing similarity to protein disulfide isomerase (PDI) from other species. The PDI gene had a predicted N-terminal signal sequence of 19 amino acids and a C-terminal tetrapeptide sequence (His-Thr-Glu-Leu; HTEL) for retention in lumen of the endoplasmic reticulum (ER). The recombinant protein expressed in baculovirus showed an apparent mass of 51 kDa, identical to that the native B. caballi protein. Moreover, the ER retention signal site (HTEL) of the recombinant protein retained its function in ER of insect cells. This 51 kDa protein was strongly expressed by extracelluar B. caballi merozoites in indirect immunofluorescence antibody tests, and was not expressed in the early phase of trophozoite development. Interestingly, detailed observation showed that the reaction of anti-P51 antibody and mAb 2H2 against pear-shaped forms was very erratic, some displaying one or two brightly fluorescent patterns.

The virulence of Entamoeba histolytica is governed by adhesion/colonization in the gut which is mediated by a galactose specific lectin. Two morphologically identical but distinct species i.e. pathogenic E. histolytica and non-pathogenic E. dispar, can be differentiated by distinct epitopes in the lectin. Both species bind to colonic epithelial cells, but only E. histolytica infection induces an inflammatory response and subsequent pathogenesis. Thus, comparing the responses of the intestinal cells to pathogenic and non-pathogenic lectins is a point of interest. The pathogenic lectin causes cytolysis of epithelial and immune-competent cells. Our data (both qualitative and mRNA quantitation) indicate that the epithelial cells responded to E. histolytica lectin with an increased expression of pro-inflammatory IL-2, IL-6, IL-8, MIP-1α, MCP-1, RANTES, GROα and GMCSF as compared to E. dispar lectin. The pathogenic LCM induced a significant increase in intracellular calcium concentration, proliferative response and chemotaxis of lymphocytes from ALA patients as compared to non-pathogenic LCM. High RANTES and IL-6 were induced in patients' lymphocytes by pathogenic LCM, along with their receptors CCR5 and CD126 as compared to NP-LCM. The local release of such a complex network of cytokines/chemokines could explain the histopathology of E. histolytica infection. The comparative low levels of these chemokines/pro-inflammatory cytokines and high levels of anti-inflammatory IL-10 in response to non-pathogenic E. dispar could explain the absence of an acute inflammatory response and the disease process. The cytokines and chemokines may provide a mechanism for initiation, amplification or containment of inflammation during disease state.

Echinococcus granulosus antigen B (AgB) is encoded by a gene family and is involved in the evasion of the host immune response. E. granulosus exists as a number of strains (G1–G10) that differ in biological characteristics. We used PCR-SSCP followed by DNA sequencing to evaluate sequence variation and transcription profile of AgB in 5 E. granulosus strainsNucleotide sequence data reported in this paper are available in the GenBankTM database under the Accession numbers AY569341-AY569371, AY159484, AY159485, AY608606, AY608607, AY614001.. Twenty-four genomic sequences were isolated and clustered in 3 groups related to 2 of the 5 reported AgB genes. AgB4 genes were present in almost all strains, whereas AgB2 were present as functional genes exclusively in G1/G2 cluster, and as non-functional genes in G5 and the G6/G7 cluster, suggesting inter-strain variation. The AgB transcription patterns, analysed by RT-PCR, showed that AgB2 and AgB4 genes were transcribed in G1, while only the AgB4 gene was transcribed in G7 strain. Cysts from the same strain or cluster shared more genomic and cDNA variants than cysts from different strain or cluster. The level of nucleotide and deduced amino acid sequence variation observed is higher than that reported so far for coding genes of other helminths. Neutrality was rejected for AgB2 genes. These data show the genetic polymorphism of antigen-coding genes among genetically characterized strains of E. granulosus.

Although parasite communities have been studied extensively in recent years, spatial and temporal variation in factors affecting the communities has received less attention. This paper examined the similarity of parasite assemblages of perch (Perca fluviatilis) in 18 locations within a single lake in relation to geographical distance and temporal dynamics in the host and parasite populations. We expected that in the present study-scale where distinct but potentially interacting host subpopulations could occur, similarity of the assemblages could be affected by seasonal dynamics in host movements particularly during the spawning period. Parasite species showed differences in infection levels between the sampling locations and similarity of the assemblages of autogenic parasite species in winter, measured inversely as difference in parasite numbers, was negatively affected by geographical distance between the locations. However, no such relationship was observed in the allogenic species Ichthyocotylurus variegata. Furthermore, no relationship was found either in autogenic parasites when the locations were re-sampled in summer, after the spawning period of perch. We concluded that the effect of geographical distance on the similarity of parasite assemblages is a dynamic process, which is affected by seasonal dynamics in host and parasite populations.

We investigated variation in the abundance-prevalence relationships of fleas among 17 different flea-host associations as well as among different species of hosts and fleas in the Negev desert. We explored variation in the value of exponent of Taylor's power relationship with changes in flea community size and flea specialization (host specificity and seasonal pattern of activity). We tested if a simple epidemiological model can reproduce the pattern of the abundance-prevalence relationship. We confirmed aggregated distribution of fleas within a population of host species as well as across a whole host community and the existence of a positive relationship between local flea abundance and their prevalence. Prevalence, mean abundance and variance of abundance were significantly higher in host specific than host opportunistic fleas. When ecological specialization was considered, based on a seasonal pattern of activity, these parameters were higher in year-round-active than seasonal fleas. The degree of flea specialization and flea community richness affected the pattern of the relationship between mean abundance and its variance. Power law slopes decreased with increasing richness of flea community. A simple epidemiological model based on mean flea abundance and degree of aggregation, corrected for host sample size, can predict the observed pattern of prevalence. In some cases, observed flea prevalence was higher than that predicted from the epidemiological model. The discrepancy of the observed prevalence from that predicted by the model can be explained by either a relatively low negative effect of flea parasitism on a host (at least, in terms of pathology) or strong resistance of a host to flea parasitism or both.

We examined the relative merits of mitochondrial DNA loci and ribosomal DNA internal transcribed spacers for their use in prospecting for cryptic species of platyhelminth parasites. Sequence divergence at ITS1 and ITS2 was compared with divergence at 2 mtDNA loci (NADH dehydrogenase-1 and cytochrome c oxidase I) between closely related species of trematodes and cestodes. Both spacers accumulated substitutions substantially more slowly than mtDNA, which clearly shows a higher level of divergence among species relative to intra-specific variation. Besides a slow rate of substitution, other caveats that may be encountered when using ITS sequences as a prospecting marker are discussed. In particular, we note recent studies that suggest the existence of substantial levels of intra-individual variation in ITS sequences of flatworms. Because it is likely that closely related species share this phenomenon, it may confound the detection of cryptic species, especially if small sample sizes are studied. Although potential limitations of mtDNA are also recognized, the higher rate of evolution and smaller effective population size of this marker increases the probability of detecting diagnostic characters between cryptic species.

Thelazia callipaeda, commonly known as the ‘oriental eyeworm’, has been recently reported in Italy and other European countries. The insect/s that act as intermediate hosts and details of larval development inside the vector remain unclear. In order to (1) demonstrate the species of fly that may act as vector/s for T. callipaeda in southern Italy (Site A) and China (Site B) and (2) describe the larval development of the nematode in the body of flies, 847 Phortica (Drosophilidae) flies were collected from the above two sites, each with a history of human and/or canine thelaziosis. Flies were identified as Phortica variegata (245 – site A) and Phortica okadai (602 – site B), experimentally infected by 1st-stage larvae (L1), kept at different temperatures and dissected daily until day 180 post-infection (p.i.). Dead flies from site A were subjected to specific polymerase chain reaction (PCR) assay to detect T. callipaeda. To demonstrate the role of Phortica as vectors of T. callipaeda, 3rd-stage larvae (L3) recovered from the proboscis of flies were deposited onto the cornea of the eyes of dogs and rabbits. Following dissection, 3 (2·9%) of P. variegata in site A were found to be infected by L3 in the proboscis on days +14, +21 and +53 p.i., compared with 26 (18·4%) of Phortica flies recorded as being positive by PCR. Sequences from positive PCR products were 99% identical to sequences of the corresponding species available in GenBank (AY207464). At site B, 106 (17·6%) of 602 dissected P. okadai were found to be infected by T. callipaeda larvae (different stages) and in total 62 L3 were recovered from the proboscis of 34 (5·6%) flies. The shortest time in which L3 were found was at day +14, +17, +19, and +50 p.i. respectively, depending on the environmental temperatures. Of 30 flies overwintered for 6 months, 6 L3 were detected at day +180 p.i. in 3 flies (10%). The biology of larval development was reconstructed on the basis of the dissection of 602 P. okadai-infected flies and the morphology of larval stages in the insect body described. The present work provides evidence that P. variegata and P. okadai act as vectors for T. callipaeda in southern Europe and in China, respectively. The phenomenon of overwintering is described here for the first time for T. callipaeda and discussed. Finally, the relationship between T. callipaeda and its fly vector is considered in light of disease prophylaxis and to model its dissemination into habitats and environments favourable to Phortica flies.

Taeniosis/cysticercosis are important zoonotic infections which continue to be a significant problem in several parts of the world including Latin America, Asia and Africa. Control of these infections has always required the collaboration of both veterinary and medical public health services at a national level and these aspects have prompted the production of this publication jointly by the Food and Agriculture Organisation (FAO), the World Health Organisation (WHO) and the World Organisation for Animal Health (OIE). It has been produced to provide guidance on the design, implementation and standardization of control strategies against these zoonoses. It is a timely update on the previous guidelines produced in 1983 which were of significant importance in providing good overviews of the problems and strategies involved at the time.

Fourteen years ago I had the pleasure of reviewing the first edition of Mike Lehane's book. This new edition, long overdue, provides perhaps the most comprehensive, and a very readable, teaching text for medical entomology available today. The title truly reflects the content. Written from a biologist's viewpoint, we are treated to an overview of how blood-feeding insects function rather than the drier, phylogenetic-based treatment that is much more common. The scene is set by two chapters dealing with the importance and evolution of blood-feeding insects. We are then led, chapter-by-chapter through the blood-feeding process from host selection through to the digestion and utilization of a bloodmeal. The penultimate two chapters consider respectively the more intricate interactions of insects with their hosts and the parasites that they transmit. The classification of blood-feeding insects is covered in a final chapter.

Cyst fluid (CF) of Taenia solium metacestode (TsM) is an important source of serodiagnostic antigens. We have investigated the molecular characteristics of the 120 kDa protein complex in TsM CF purified by fast performance liquid chromatography. The structure of the purified protein was characterized by a variety of proteomic analyses. The protein was found to consist of 2 major components of 42–46 and 22–28 kDa, and shared 3 subunits of 14, 16 and 18 kDa. The 42–46 kDa component was determined to contain 3 additional subunits of 22, 28 and 38 kDa. These 6 subunits were shown to originate from either the 14 or 18 kDa precursor. We assessed the antibody reactivity of the native protein, its individual subunits and the recombinant 14 and 18 kDa proteins, and found that the 120 kDa protein, particularly 14 and 18 kDa subunits revealed high reliability for differentiation of active and mixed stage NC from chronic NC. The subunits of the 120 kDa protein complex identified herein represent some of the low-molecular weight glycoproteins which have been described in several previous studies. Recognizing and understanding the structural and immunological relationship of these proteins will facilitate the development of new serodiagnostic assays.

Nematology is divided, often arbitrarily, into several ‘subdisciplines’, amongst which the study of plant-parasitic nematodes is predicated on their considerable economic importance world-wide. Further subdivision of the discipline is rarely justified but, in the tropics and subtropics, control of nematode damage is often compromised by lack of knowledge and the cost of conventional management strategies. Thus, there is a need for a comprehensive, informative text on plant-parasitic nematodes of these regions. Plant Parasitic Nematodes in Subtropical and Tropical Agriculture is the second edition of a book whose aim is to meet these requirements.