Identification and Characterization of P0 Protein as a Vaccine Candidate Against Babesia divergens, Blood Parasite of Veterinary and Zoonotic Importance

The molecular identification and antigenic characterization of P0 protein in Babesia divergens, a blood parasite of veterinary and zoonotic importance, were carried out in this study for use in developing subunit vaccines against B. divergens infection. Recombinant protein encoding P0 (BdP0) was developed in Escherichia coli, and its antiserum was generated in mice for further molecular characterization. Anti-rBdP0 serum had a specific interaction with the corresponding legitimate B. divergens protein, as confirmed by Western blotting and indirect fluorescent antibody tests. ELISA was used to assess the immunogenicity of BdP0 in a group of 68 bovine field samples, and significant immunological reactivity was found in 19 and 20 positive samples of rBdp0 and B. divergens lysate, respectively. The in vitro growth of B. divergens cultures treated with anti-rBdP0 serum was significantly inhibited (p < 0.05). Furthermore, after 6 h of incubation with 2 mg/ml anti-rBdP0 serum, the ability of pre-incubated free merozoites to invade bovine erythrocytes was reduced by 59.88%. The obtained data suggest the possible use of rBdP0 as diagnostic antigen and may serve as a vaccine candidate against babesiosis caused by B. divergens either in animal or human.

First Report of Haemoproteus (Haemosporida, Haemoproteidae) Megalomeronts in the Brain of an Avian Host, with Description of Megalomerogony of Haemoproteus Pastoris, the Blood Parasite of the Common Starling

Species of Haemoproteus (Haemoproteidae, Haemosporida) are common bird pathogens. Recent molecular studies combined with histopathology research have reported development of megalomeronts of these parasites in various organs, sometimes resulting in the death of the avian host. Five Common starlings (Sturnus vulgaris) were found naturally infected with Haemoproteus pastoris lineage hLAMPUR01. The parasite was identified using microscopic examination of blood films and DNA sequences. Infected bird organs were investigated histologically for (i) the presence of exo-erythrocytic stages and (ii) the patterns of development (morphology and localization) in different host individuals. For the first time, megalomeronts of Haemoproteus parasites were seen developing in the brain, while numerous others at different stages of maturation were found in the intestine, pancreas, kidneys, lungs, esophagus, spleen, gizzard, and trachea. Megalomeronts were predominantly roundish or oval, up to 800 μm, they were surrounded by a capsular-like wall and developed asynchronously in the same bird individual. After megalomeront maturation and rupture, a massive infiltration of blood cells occurred, indicating the hemorrhagic processes. Review of available data showed that different Haemoproteus species produce markedly different megalomeronts, morphology of which can probably be predicted using phylogenetic analysis based on partial sequences of cytochrome b gene.

Characterization of serum protein electrophoresis patterns and C-reactive protein in canine tick-borne diseases

Background and Aim: Canine tick-borne diseases are important diseases with a worldwide distribution. In Thailand, the most important canine tick-borne diseases are ehrlichiosis, babesiosis, and hepatozoonosis. This study aimed to determine the serum protein electrophoresis patterns (SPEPs) and C-reactive protein (CRP) levels associated with Ehrlichia canis, Babesia canis, or Hepatozoon canis single infections. Materials and Methods: A total of 650 canine blood samples were collected from animal hospitals and clinics in Bangkok and its vicinity to examine health status and blood parasite infection. Suspected blood parasite infections were examined by buffy coat thin blood smear and confirmed by polymerase chain reaction. Normal dog and positive E. canis, B. canis, and H. canis single infections and serum protein profiles were determined by agarose gel electrophoresis. CRP concentration was measured by fluorescent immunoassay. Results: In dogs infected with E. canis, B. canis, and H. canis single infections, albumin levels and A/G ratios significantly decreased, whereas β2-globulin levels increased (p<0.05). The γ-globulin level significantly increased in E. canis and H. canis infections (p<0.05). A monoclonal gammopathy pattern was observed in E. canis and B. canis single infections, whereas β-γ bridging patterns and increased β- and γ-globulin fractions were found in H. canis single infections. The CRP level increased in dogs with blood parasite single infections and may be related to the pathogenesis of the infection. Conclusion: SPEPs and CRP levels can be used to monitor health status and blood parasite problems in infected dogs.

High Granulocyte-Macrophage Colony Stimulating Factor to Interleukin 10 Ratio and Marked Antioxidant Enzyme Activities Predominate in Symptomatic Cattle Naturally Infected with Candidatus Mycoplasma haemobos, Theileria orientalis, Theileria sinensis and Trypanosoma evansi

The aim of this study was to measure the serum proinflammatory (IL-12, GM-CSF & IFN-γ) to anti-inflammatory (IL-10, IL-4) cytokine ratio, oxidant (MDA) level and antioxidant enzyme (SOD; GPx) activities after blood parasite infections. The blood and serum samples were obtained from 130 cattle and screened for identity of the infecting blood parasites by conventional PCR. The following blood parasite species were detected: Candidatus Mycoplasma haemobos (70/130); Theileria orientalis (65/130); Theileria sinensis (32/130); Anaplasma marginale (49/130); Anaplasma platys (7/130); and Trypanosoma evansi (4/130). The GM-CSF/IL-10 ratio showed significantly higher values in all the symptomatic blood parasite infected cattle groups except for symptomatic A. platys infected cattle groups. Anti-inflammatory cytokine immune responses were notable findings in symptomatic and asymptomatic cattle infected with C. M. haemobos and T. orientalis characterized by low serum IL-12:IL-10, IFN-γ:IL-10, IL-12:IL-4 and IFN-γ:IL-4 (p < 0.05). Therefore, high serum GM-CSF:IL:10 in the symptomatic blood parasite infected cattle, low serum IL-12:IL-10, IFN-γ:IL-10, IL-12:IL-4 and IFN-γ:IL-4 ratios in asymptomatic cattle, high MDA level, and increased antioxidant enzyme activities could be useful predictive tools for outcome of natural blood parasite infections in cattle.

Do blood parasites increase immature erythrocytes and mitosis in amphibians?

Introduction: In amphibians, blood may act as a hematopoietic tissue. However, the knowledge concerning hematological features is scarce, there is not much information that allows an analysis about the possible explanations of this physiological feature. Objective: This study aimed to evaluate the relationship between immature red blood cells (RBCs) mitosis and the presence of blood parasites in amphibians. Methods: We sampled 116 amphibians (31 species) in six Colombian localities. Blood was taken by cardiac puncture or maxillary vein puncture. Smears were prepared, fixed, and Giemsa stained for microscopical analysis. The variables analyzed were the percentage of immature RBCs, mitotic cells in peripheral blood, and blood parasite infection. Data were analyzed using Wilcoxon's rank test and exact Fisher statistical tests. Results: Sixty-two individuals showed mitosis in peripheral blood, and these mitotic RBCs shared morphological features with immature RBCs. Overall, parasite prevalence was 30.1 %, distributed as follows: Trypanosoma (24.1 %), Hepatozoon-like (6 %), Dactylosoma (4.3 %), Karyolysus-like (0.9 %), and Filarioidea (2.6 %). A positive association between the percentage of immature RBCs and the presence of mitotic RBCs was found, and also between the blood parasite infection and the percentage of immature RBCs. Conclusions: In this study, we found that the presence of blood parasites, immature RBCs, and RBCs mitosis are frequent events in amphibians' peripheral blood, and our analysis suggests an association between those features. Thus, the release of immature RBCs and the mitosis of those cells in peripheral blood may be a physiological response to blood parasite infection. Further studies characterizing hematology in amphibians and wildlife, in general, are desirable.

Spatially different annual cycles but similar haemosporidian infections in distant populations of collared sand martins

Background Populations of long-distance migratory birds experience different environments and are consequently exposed to different parasites throughout their annual cycles. Though, specific whereabouts and accompanied host-parasite interactions remain unknown for most migratory passerines. Collared sand martins (Riparia riparia) breeding in the western Palaearctic spend the nonbreeding period in Africa, but it is not yet clear whether specific populations differ in overwintering locations and whether these also result in varying infections with vector-transmitted endoparasites. Results Geolocator tracking revealed that collared sand martins from northern-central and central-eastern Europe migrate to distant nonbreeding sites in West Africa and the Lake Chad basin in central Africa, respectively. While the ranges of these populations were clearly separated throughout the year, they consistently spent up to 60% of the annual cycle in Africa. Ambient light recorded by geolocators further indicated unsheltered roosting during the nonbreeding season in Africa compared to the breeding season in Europe. We found 5–26% prevalence of haemosporidian parasites in three breeding populations and one migratory passage population that was only sampled but not tracked. In total, we identified seven Plasmodium and nine Haemoproteus lineages (incl. two and seven new lineages, respectively), the latter presumably typical for swallows (Hirundinae) hosts. 99.5% of infections had a low intensity, typical for chronic infection stages, whereas three individuals (0.5%) showed high parasitaemia typical for acute infections during spring migration and breeding. Conclusions Our study shows that blood parasite infections are common in several western Palaearctic breeding populations of collared sand martins who spent the nonbreeding season in West Africa and the lake Chad region. Due to long residency at the nonbreeding grounds blood parasite transmissions may mainly occur at host population-specific residences sites in Europe and Africa; the latter being likely facilitated by unsheltered roosting and thus high vulnerability to hematophagous insects. The rare cases of high parasitaemia during spring migration and breeding further indicates either relapses of chronic infection or primary infections which occurred shortly before migration and during breeding.

Characterisation of serum protein electrophoresis patterns and C-reactive protein in canine tick-borne diseases

Background Canine tick-borne diseases are important diseases with a worldwide distribution. In Thailand, the most important canine tick-borne diseases are ehrlichiosis, babesiosis and hepatozoonosis. The aim of this study was to determine the serum protein electrophoresis patterns and C-reactive protein levels associated with E. canis, B. canis or H. canis single infections. Results In dogs infected with E. canis, B. canis and H. canis single infections, the albumin levels and A/G ratios were significantly decreased, whereas β2-globulin levels were increased (P < 0.05). The γ-globulin level was significantly increased in E. canis and H. canis infections (P < 0.05). A monoclonal gammopathy pattern was observed in E. canis and B. canis single infections while β–γ bridging patterns and increased β- and γ-globulin fractions were found in H. canis single infections. The C-reactive protein level was increased in dogs with blood parasite single infections and may be related to the pathogenesis of the infection. Conclusions Serum protein electrophoresis patterns and C-reactive protein levels can be used to monitor health status and blood parasite problems in infected dogs.

Characterisation of serum protein electrophoresis patterns and C-reactive protein in canine tick-borne diseases

Background: Canine tick-borne diseases are important diseases with a worldwide distribution. In Thailand, the most important canine tick-borne diseases are ehrlichiosis, babesiosis and hepatozoonosis. The aim of this study was to determine the serum protein electrophoresis patterns and C-reactive protein levels associated with E. canis, B. canis or H. canis single infections.Results: In dogs infected with E. canis, B. canis and H. canis single infections, the albumin levels and A/G ratios were significantly decreased, whereas β2-globulin levels were increased (P < 0.05). The γ-globulin level was significantly increased in E. canis and H. canis infections (P < 0.05). A monoclonal gammopathy pattern was observed in E. canis and B. canis single infections while β–γ bridging patterns and increased β- and γ-globulin fractions were found in H. canis single infections. The C-reactive protein level was increased in dogs with blood parasite single infections and may be related to the pathogenesis of the infection.Conclusions: Serum protein electrophoresis patterns and C-reactive protein levels can be used to monitor health status and blood parasite problems in infected dogs.