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Biomedical subjects

A Gorenflot

Publications and source records attributed to A Gorenflot.

At least 37 records · Page 2Linked to original sources
PubMed1996

Human babesial infections in Europe.

The tick transmission of Babesia from infected animals to human has been clearly demonstrated. Including the first case described by Skaraballo in 1957 in Yugoslavia, 22 cases of human babesiosis have been reported in Europe. B. divergens, which is the most common pathogen for cattle, was involved in 17/22 cases. France, British Isles and Ireland accounted for more than half of the cases, occurring especially between May and October, the period of maximal activity of Ixodes ricinus, considered as the major vector. Splenectomy was the main factor of risk which was found in 86% of the patients. Clinically, babesial infections appeared suddenly with a non periodic high fever, shaking chills, sweat, headache, myalgia and jaundice induced by intravascular hemolysis. The mortality rate was higher than 50%. Treatment should be promptly initiated with a massive blood exchange, followed by intravenous clindamycin (600 mg) three time daily. For prophylaxis, attenuated vaccines could provide effective protection in cattle, but no vaccine is available for human protection.

Animals↗
PubMed1996

Female Ixodes ricinus (Acari, Ixodidae) in cattle of western France: infestation level and seasonality.

Data collected from a longitudinal survey carried out over a 2 year period in four herds located in western France were used to assess the infestation level in cattle by female ixodes ricinus and to describe its seasonality. Farms were visited once a month. The population studied consisted of calves, heifers and dairy cows (of Holstein and Norman de breeds). The total number of animals monitored varied from 207 to 173 throughout the study, because of the culling of dairy cows and birth of calves. The infestation level by l ricinus varied from one herd to another. Heifers were found to have a greater level of infestation than dairy cows and calves. The infestation rate appeared to be low, ie, most animals (between 47 and 83%) were infested by only one tick in the axilla. The seasonal distribution of ticks showed two peaks, one in spring and a second lower one in autumn, with slight fluctuations between herds and years. The number of ticks on cattle and the percentage of infested animals showed a parallel seasonality.

Age Factors↗
PubMed1995

Large scale survey of bovine babesiosis due to Babesia divergens in France.

Data on the epidemiology of bovine babesiosis in France were collected from two sources: a questionnaire sent to large animal veterinary surgeons and blood samples collected from cattle treated for clinical babesiosis. Babesia divergens was common and widespread. B major was found in only one sample. The disease was particularly widespread in north west, south west and central France. The mean clinical incidence was 0.4 per cent. The bimodal seasonality of the clinical cases was correlated with that of the tick vector (Ixodes ricinus). Of the affected animals 72.6 per cent were more than three years old.

Age Distribution↗
PubMed1995

Seasonal incidence and ecology of the tick Ixodes ricinus (Acari: Ixodidae) on grazing pastures in western France.

A longitudinal survey was carried out during a 2 year period in Western France to assess the infestation level of grazing pastures by Ixodes ricinus ticks. Four farms were visited once a month and each of the grazing pastures was sampled in the centre and at the border using the blanket dragging method. A total of 3562 I. ricinus (34 adults, 900 nymphs and 2628 larvae) were collected and the infestation was significantly higher during the first year (p < 0.0001). The infestation level by I. ricinus varied between grazing pastures and farms. Grazing pastures in the vicinity of forest were more infested than the others, all through the study. The seasonal distribution of ticks showed peaks, with low fluctuations between farms, years and stages. Tick abundance could not be related to vegetation, but only to the vicinity of woods.

Animals↗
PubMed1995

Babesia divergens: an ELISA with soluble parasite antigen for monitoring the epidemiology of bovine babesiosis.

An enzyme linked immunosorbent assay (ELISA) for bovine babesiosis caused by Babesia divergens was developed to analyse the evolution of the serological status of cattle living in an enzootic area. The antigen used was a soluble extract of B. divergens obtained from in vitro culture. Specificity, evaluated with negative sera, was 96.6%. The ELISA was compared to indirect immunofluorescence analysis (IFA) on naturally or experimentally infected animals. It appeared that IFA was positive for at least seven or eight weeks; on the contrary, B. divergens-specific antibodies were only transitorily detected by ELISA. Furthermore, the ELISA was more sensitive than the IFA for the detection of post-infection antibody increases, particularly when infection-pressure was low. These results suggest that IFA and ELISA could be complementary tools in epidemiological surveys; indeed, this ELISA could be the most efficient tool to detect new infections in cohort monitoring.

Animals↗
PubMed1995

A 37-kilodalton glycoprotein of Babesia divergens is a major component of a protective fraction containing low-molecular-mass culture-derived exoantigens.

The supernatants of in vitro cultures of Babesia divergens Rouen 1987 in human erythrocytes, obtained by using a semidefined medium based on human high-density lipoproteins, were fractionated by gel filtration chromatography into four fractions, F1 to F4. The crude supernatant as well as each fraction adjuvanted with Quil-A protected gerbils from mortality due to a homologous infectious challenge. Analysis of the humoral response of the 10 protected gerbils with fraction F4, containing major proteins with molecular masses lower than 50 kDa, showed that a few antigens (from 50 to 17 kDa) could be important candidates for an improved vaccine against B. divergens babesiosis. As an immunodominant response was directed against the 37-kDa antigen (Bd37) in two different B. divergens strains tested, a polyclonal antibody directed against Bd37 was produced in a rabbit. In an immunofluorescence assay, the anti-Bd37 antiserum strongly labelled small internal vesicles of the merozoites and the cell surface was diffusely labelled after fixation, whereas on live merozoites, this labelling was not observed. [3H]glucosamine-radiolabelling experiments demonstrate that Bd37 is a glycoprotein. The Bd37 protein can also be labelled with [14C]palmitate but not with [3H]myristic acid. In Triton X-114 temperature phase partitioning of B. divergens-infected erythrocyte extracts, Bd37 was exclusively found into the detergent phase, indicating that the palmitoylated Bd37 protein was in the membrane fraction. In the in vitro supernatant, the glycoprotein Bd37 was found in a nonpalmitoylated form, indicating excretion and/or release of the glycoprotein from the merozoite.

Animals↗
PubMed1993

Babesia divergens: characterization of a 17-kDa merozoite membrane protein.

Large amounts of viable merozoites were purified from in vitro cultures of Babesia divergens by a two-step sieving procedure. A monoclonal antibody produced against B. divergens merozoites, mAb DG7, stained the merozoite plasma membrane and an intra-parasitic linear organelle in indirect immunofluorescence. Immunogold labeling in electron microscopy demonstrated that the antigen recognized by mAb DG7 was localized just beneath the merozoite plasma membrane. Immunoprecipitations of metabolically labeled ([35S]methionine) B. divergens antigens showed that the epitope recognized by mAb DG7 was present on a 17-kDa polypeptide (Bd17) and was shared in all B. divergens geographical isolates tested so far. Bd17 was always present in the in vitro culture supernatants of all these isolates. Furthermore, Triton X-114 phase separation of babesial antigens demonstrated the hydrophilic character of Bd17 which suggests that it is an extrinsic protein present on the cytosol side of the parasite membrane. When added to the culture medium, mAb DG7, purified from ascite fluids, drastically altered the growth of the parasite with concentrations inhibiting 50% of development (IC50) ranging between 16.6 and 26.1 micrograms/ml).

Animals↗
PubMed1993

Cellular and humoral immune responses induced in cattle by vaccination with Babesia divergens culture-derived exoantigens correlate with protection.

Previous results with the Babesia divergens gerbil vaccination model were extended in studies with cattle. Two calves were vaccinated with culture-derived B. divergens exoantigens, and two others were treated with control supernatant; both preparations were adjuvanted with Quil-A saponin. A parasite-specific humoral response was observed after the first vaccine injection and was boosted by two succeeding vaccine injections. Sera from the two vaccinated calves immunoprecipitated eight major parasitic proteins (with molecular masses ranging between 17 and 110 kDa) whose patterns were close to those observed in gerbil vaccine assays. The cellular immune response, monitored by lymphoproliferation assays, was slightly delayed in comparison with the humoral response; a significant proliferation occurred only after the second vaccine injection. Mononuclear cell proliferation was dose dependent in the presence of (i) lysates of B. divergens-parasitized erythrocytes, (ii) exoantigens of the whole supernatant, or (iii) protective exoantigens of two low-molecular-mass fractions obtained after supernatant gel filtration chromatography. An infectious challenge was administered 3 weeks after the third vaccine injection, with 3.6 x 10(10) B. divergens-parasitized erythrocytes. Erythrocyte count, rectal temperature, and parasitemia of the animals were monitored daily until they returned to initial values. All parameters indicated that the exoantigens induced protection from B. divergens infection for the two vaccinated calves.

Animals↗
PubMed1992

Human babesiosis in Europe.

Human babesiosis in Europe came to medical attention in 1957 and until now 19 cases have been reported, most of them due to Babesia divergens. The onset of the disease is characterized by hemoglobinuria, high fever and renal failure ensue rapidly. The patients were generally asplenic and resident in a rural area. Intraerythrocytic pleomorphic parasites (1-3 microns) observed in stained thin blood smears are essential for Genus diagnosis. Parasitemia varied from 5 to 80% of red blood cells. Massive blood exchange transfusion (2-3 blood volumes) followed by intravenous clindamycin (3-4 times daily) and oral quinine (600 mg base, 3 times daily) were successfully used in the treatment of three recent cases. Splenectomised individuals should be aware for prevention.

Acute Kidney Injury↗
PubMed1992

Babesia divergens vaccine.

A vaccine strategy against Babesia divergens bovine babesiosis was successfully developed after perfecting of an efficient in vitro culture. Crude supernatants and purified fractions were able to induce a vaccine protection in gerbils against B. divergens infection. More, supernatants induced an effective vaccine protection in cattle. The role of B. divergens exoantigens of 17, 37, 46, 70 and 90 kDa in the development of the immune response was clearly demonstrated in gerbils, cattle, and man.

Animals↗
PubMed1991

Role of cytoskeleton and surface lectins in Giardia duodenalis attachment to Caco2 cells.

An in vitro model of Giardia duodenalis and the Caco2 cell line enable the study of parameters that could play a part in trophozoite attachment. We explored the role of membranous lectins of G. duodenalis in attachment-inhibition studies using carbohydrates in solution. Attachment rates were reduced by 14% and 23% in the presence of 100 mmol/l mannose-6-phosphate and glucose, respectively, as compared with control values. No significant modification was observed after trophozoite trypsinization at room temperature or at 37 degrees C. The inhibitory effects of colchicine (35%) and nocodazole (70%) suggest a primordial role of the cytoskeleton; microtubules appear to be the principal effectors of trophozoite fixation. Scanning electron microscopy revealed circular imprints on the Caco2 brush border after trophozoite detachment. The mechanisms of attachment of G. duodenalis to intestinal enterocyte-like cells in culture are thus essentially of the mechanical or hydrodynamic type; surface lectins would appear to intervene in the specificity for duodenal cells.

Animals↗
PubMed1991

Cytological and immunological responses to Babesia divergens in different hosts: ox, gerbil, man.

A continuous in vitro culture system for Babesia divergens was initiated from a human isolate. It was maintained through 305 subcultures for 3 years using a low concentration of serum and a low haematocrit, with no decrease in the initial virulence. This in vitro system enabled the routine culture of all human and bovine B. divergens isolates thus far tested, with a mean parasitaemia level of 30%-40%. Different cytological aspects observed in the same isolate by optical and electron microscopy were described in parasitized ox, gerbil and human erythrocytes. The sequence of B. divergens antibody responses was determined in man and ox, enabling the precise identification of major B. divergens antigens as candidates for vaccines.

Animals↗
PubMed1991

Lipid trafficking between high density lipoproteins and Babesia divergens-infected human erythrocytes.

A two-fold increase in the amount of phospholipids was observed in Babesia divergens infected human red blood cells. In vitro incubation with [32P]-phosphorus and [3H]-glycerol demonstrated that B divergens has the ability to synthesize the phospholipid backbone. On the other hand, the low incorporation of [14C]acetate indicated the absence of a de novo fatty acid synthesis and suggested the necessity of an exogenous lipid source for the parasite. Several intra-erythrocytic growth cycles of B divergens could be achieved in vitro, using a serum-free medium supplemented only with fractions of human high density lipoproteins (HDL). At an HDL concentration of 0.5 mg/ml (protein concentration) and with a 1% starting parasitaemia, parasite growth was similar to that observed under standard culture conditions with 10% human serum, at least for the first 24 h, a time equivalent to three parasite erythrocytic life-cycles. Lipid transfer from HDL to the intra-erythrocytic parasites was demonstrated by uptake and exchange of fluorescent NBD-phosphatidylcholine (NBD-PC) loaded HDL at different temperatures. Kinetic experiments with [3H]-oleyl-PC-loaded HDL demonstrated a unidirectional transfer of lipids from radiolabelled HDL to the parasite; partial conversion of PC to phosphatidylethanolamine (PE) was also observed. In the semi-defined medium, the HDL fraction appeared to be the major source of lipids for the growth of B divergens in human erythrocytes.

Acetates↗
PubMed1991

Heat shock response of Babesia divergens and identification of the hsp70 as an immunodominant early antigen during ox, gerbil and human babesiosis.

Using antisera (alpha-R and alpha-C7Ag) directed against the conserved Gly-Gly-Met-Pro-epitope of the hsp70 family, a single antigen was identified in the human Babesia divergens Rouen 1987 isolate by Western immunoblotting and immunoprecipitation experiments. This B divergens hsp70 is highly conserved as shown by the analysis of five other geographical B divergens isolates from different hosts (human and bovine). Indirect immunofluorescence assay performed on the asexual intraerythrocytic stages showed that the hsp70 is mainly cytoplasmic and stage-independent. Heat-shock experiments, with 20 min incubation at 40 degrees C followed by a 10 to 50 min shift to 37 degrees C in the presence of [35S]-methionine, led to an increase of two hsp of 85 and 70 kDa while protein synthesis in general decreased within 10 min. Immunoprecipitations of [35S]-methionine radiolabelled proteins with human, ox and gerbil antisera raised against various B divergens isolates, showed the presence of a B divergens 70 kDa protein which was demonstrated to be a hsp70 by coupling immunoblotting assays with alpha-C7Ag serum on the same immunoprecipitated material. During human babesiosis, the B divergens hsp70 appears as an early antigen during the acute phase. These results are in agreement with the use of the B divergens hsp70 as an essential valence antigen in an anti-babesiosis vaccine.

Amino Acid Sequence↗
PubMed1991

Analysis of immune responses of different hosts to Babesia divergens isolates from different geographic areas and capacity of culture-derived exoantigens to induce efficient cross-protection.

The immunoprecipitation of [35S]methionine-radiolabelled antigens from different Babesia divergens isolates by using bovine, gerbil, and human immune sera has shown that many B. divergens proteins contain epitopes shared between isolates. The cross-protective capacity of culture-derived soluble immunogens from the B. divergens Rouen 1987 isolate was tested against different B. divergens isolates. Results showed complete protection against the 7107b French isolate and substantial protection against the Weybridge 8843 English isolate (80% protection) and the Munich 87 German isolate (60% protection). In order to explain these vaccination results and to assess both the common and variable antigenicity of B. divergens, the antigenic patterns of the challenge isolates (Rouen 1987, 7107b, Weybridge 8843, and Munich 87) were compared by immunoprecipitation, using gerbil antisera raised against the Rouen 1987 vaccine isolate. Differences in the antigenic patterns and in the cross-protection of gerbils in these heterologous challenges were examined by studying the virulence and the antigenic status of each isolate.

Animals↗
PubMed1991

[Giardia intestinalis: transmission electron microscopy study of in vitro excystation].

The in vitro excystation process in Giardia intestinalis was studied by transmission electron microscopy (TEM). Untreated cysts served as controls. The excystation process was monitored by examination of organisms after the in vitro induction and at several times during the incubation phase. The control cyts had a thick wall, made of microfibrils, that appeared not to contain any weak areas. The peritrophic space extended between the cyst wall and the organism peripherally, the space was delimited by a thin cytoplasmic layer, "the outer cytoplasmic envelope" that subtended the cyst wall. During the in vitro incubation, the trophozoite cytoplasm retracted from the wall; thus, the peritrophic space became progressively larger. The outer cytoplasmic envelope detached from the cyst wall, then broke up forming numerous small vesicles lodged between the wall and the organism. The tight arrangement of the wall microfibirils was lost. Electron-dense vacuoles appeared in the peripheral cytoplasm of the trophozoite. The organism emerged through the posterior end of the cyst, leaving behind the empty husk. Emergence was followed by cell division. The possible interrelationships of biochemical and mechanical factors affecting the process of excystation are discussed in light of the present TEM findings.

Animals↗
PubMed1990

Identification of major Babesia divergens polypeptides that induce protection against homologous challenge in gerbils.

[35S]methionine-radiolabeled proteins from the Babesia divergens Rouen 1987 isolate were immunoprecipitated with immune sera from three potential hosts: human, ox, and gerbil. The results showed a constant humoral response against major babesial antigens. Similarly, immunoprecipitation of radiolabeled in vitro culture supernatant demonstrated that the exoantigens of 37, 46, 70, and 90 kDA were the immunodominant polypeptides, whatever the host. The effects of vaccination with concentrated supernatant from B. divergens Rouen 1987 in vitro cultures (30 to 40% parasitemia) were examined in gerbils inoculated with the homologous B. divergens isolate. Gerbils having received two or three injections of a whole vaccine dose (1.5 ml of parasitized culture supernatant equivalent [PCSE]) or of a 1:5 diluted vaccine dose (0.3 ml of PCSE) showed 100% survival after intraperitoneal challenge with 10(6) B. divergens-infected gerbil erythrocytes. Moreover, two or three injections of a 1:25 diluted vaccine dose (0.06 ml of PCSE) or 9% NaCl or 1.5 ml of unparasitized culture supernatant equivalent resulted in a mortality rate of 80 to 90% of the infected gerbils. Immunoprecipitation and immunofluorescence assays performed with antisera from vaccinated and control gerbils demonstrated that a single vaccine injection induced a humoral response, which increased slightly after the second or third injection. After challenge, antibody levels increased significantly, although the immunoprecipitation did not display any modification of Babesia antigen patterns.

Animals↗