Biomedical subjects

J M Alunda

Publications and source records attributed to J M Alunda.

At least 37 records · Page 2Linked to original sources

PubMed1998

Infection-induced protection against Haemonchus contortus in merino and manchego sheep. Relationship to serum antibody response.

Resistance to primary and secondary infections with Haemonchus contortus was studied in 10-month-old Manchego and Merino sheep. No notable interbreed differences were observed after primary infections in the parameters determined (prepatency period, faecal egg output, abomasal worm burden). Previously infected sheep (200 L-3/kg live weight (lw)) from both breeds showed notable protection after challenge (400 L-3/kg lw), evidenced by lower eggs/g faeces (epg) values and worm burdens. A protective response in the Manchego breed was associated with arrested development of fourth stage larvae in the abomasal mucosa, whereas in the Merino breed a more rapid expulsion mechanism seems to be involved. Serum antibody levels (IgG, IgA) were infective dose-dependent and protection from re-infection was not clearly related to the parasite-specific IgG response estimated by enzyme-linked immunosorbent assay (ELISA) and Western blotting.

Abomasum↗

PubMed1996

Antibody response in dogs experimentally infected with Leishmania infantum: infection course antigen markers.

Five-month-old beagle dogs were experimentally infected with amastigotes of Leishmania infantum and kept for 14 months. Infection course was monitored by clinical examination, serum protein variations, and levels of specific antibodies against Leishmania estimated by indirect immunofluorescence test and Western blotting (WB). Infected animals developed notable changes in serum protein levels reaching maximum protein concentrations 2-3 months postinfection (p.i.) related to the gamma-globulin fraction. Specific antibody titers were in good agreement with the serum protein rise, reaching immunofluorescence values of over 1:800 3 months p.i. Serial Western blotting analysis with L. infantum promastigotes protein showed a strong response against immunodominant antigens of 50-57, 42, and 29 kDa during most of the studied period with immunofluorescence titers of over 1:100 and in addition the response was remarkably homogeneous among the infected dogs. Immunoreactivity patterns displayed time-related variations; the response against 29 and 50-57 kDa was seen very early, followed by the reaction around 42, 76, and 86 kDa. In addition the recognition of peptides around 34-35.4 and 26 kDa was restricted to the acute phase of the experimental infection. Preliminary results obtained in naturally infected dogs seem to support the predictive value of the WB.

Animals↗

PubMed1996

Identification and partial purification of a 26 kilodalton antigen of adult Haemonchus contortus.

Adult H. contortus soluble extracts were fractionated by means of gel filtration (S-200) and anion exchange chromatography (DEAE-Sephacel). Fractions from both analyses were checked by ELISA and western blotting (WB) with sera from lambs infected with H. contortus, monospecific heterologous sera (anti-Trichostrongylus colubriformis and anti-Teladorsagia circumcinta) and sera from naturally infected sheep with mixed trichostrongylid infections. High cross reactivity was seen between H. contortus and heterologous sera, particularly with the anti-T. colubriformis serum, when fractions from gel filtration were checked by ELISA. Individual fractions containing the highest positive/negative and positive/heterologous ratios were pooled and analysed by SDS-PAGE. One of the pools (A4) containing 2 regions around 48-55 and 25-27 kDa were strongly recognized by homologous sera in WB. Similar results were obtained with the first peak eluted in the DEAE-Sephacel chromatography with NaCl 0.1 M. The pooled fraction A4 from gel filtration was further fractionated by anion exchange chromatography and the peak obtained with the NaCl gradient contained a ca. 26 kDa antigen apparently specific for the diagnosis of H. contortus infections in lambs.

Animals↗

PubMed1996

Leishmania infantum: infection of macrophages in vitro with promastigotes.

Leishmania infantum promastigotes in axenic culture exhibit limited infectivity for mouse peritoneal macrophages (M phi) in vitro using standard culture conditions (37 degrees C; 95% air/5% CO2) compared to Leishmania donovani promastigotes which induce notable infections. The infectivity of logarithmic (log) and stationary (stat) phase promastigotes of L. infantum was enhanced by the addition of fresh homologous serum, but no amastigotes were observed after 4 days. Prolonged infections, including transformation and survival of intracellular amastigotes in BALB/c mouse and hamster resident peritoneal M phi and M phi cell line J774.G8 were obtained by incubating M phi for 48 h at 26 degrees C prior to standard culture. Enhanced infectivity was observed in a number of L. infantum strains subject to this transient thermal change.

Animals↗

PubMed1994

Polyamine content and drug sensitivities of different clonal lines of Leishmania infantum promastigotes.

Two isolates (skin and lymph node) of Leishmania infantum obtained from a naturally infected dog were cloned and the free pools of polyamines for both complete isolates and clones were determined. Putrescine (Put) and spermidine (Spd) levels were highly variable among the lines studied, ranging from 0.89 to 2.1 nmol Spd/10(7) promastigotes. The Put/Spd ratio was also variable (1.54-0.51) and correlated with the cell growth of the lines studied. There were important differences in the clones' sensitivities to difluoromethylornithine (DFMO) and Berenil, with some of the clones being almost refractory to the inhibitory effect of 50 microM DFMO, whereas the growth of others was reduced by 60%; similar findings were obtained with 50 microM Berenil. L. infantum sensitivities to DFMO and Berenil were correlated and apparently related to the values for the Put/Spd index.

Animals↗

PubMed1994

Cross antigenicity among ovine trichostrongyloidea. Preliminary report.

A preliminary trial on the extent of cross-antigenicity among the sheep strongylids Haemonchus contortus, Trichostrongylus colubriformis, Teladorsagia circumcincta and Nematodirus battus in 2.5- to 4-month-old lambs has been carried out using ELISA and Western blotting (WB). Cross antigenicity was tested using soluble extracts from adult and third stage larvae (L3) of H. contortus as antigenic source probed with sera from lambs with monospecific heterologous infections. There was cross-antigenicity between L3 of H. contortus and Trichostrongylus colubriformis in ELISA and WB. Immunodetection results with adult H. contortus antigen showed a closer relationship to Teladorsagia circumcincta. Certain heterologous sera reacted with H. contortus antigens more strongly than the homologous one, but sera from the H. contortus-infected animals had reactivity around the 25 kDa region from adult antigens which could have potential diagnostic use.

Animals↗

PubMed1993

Effect of subclinical coccidiosis in kids on subsequent trichostrongylid infection after weaning.

Seven-week-old female kids of the Murciana-Granadina breed naturally infected with coccidia were superinfected with a multispecific Eimeria inoculum (300,000 oocysts) or treated to control the coccidial infection with Amprolium (50 mg kg-1 liveweight day-1; 4 days every 14 days); 80 days later both animal groups received 2500 third-stage larvae of a sheep-derived mixture of Trichostrongylus colubriformis (50%), Teladorsagia circumcincta (40%) and Haemonchus contortus (10%). Kids experimentally superinfected with Eimeria showed reduced food intake at the early patency period, higher numbers of oocysts passed at weaning time and lower liveweight gain than the Amprolium-medicated kids. In the Eimeria superinfected kids the trichostrongyle infection caused a tendency to result in chronic coccidial infections. Further, these animals had higher numbers of nematode eggs in their faeces, lower dressed weights on slaughter and inferior meat quality.

Amprolium↗

PubMed1993

Lamb serum recognition of infective larvae and adult Haemonchus contortus antigens.

Sixteen- to eighteen-week-old lambs were infected with 2500 3rd stage larvae (L-3) of Haemonchus contortus or kept as uninfected controls. Two months later all animals were challenged with 5000 L-3 of this parasite. Soluble antigens of H. contortus L-3 and adult worms were analyzed by SDS-PAGE and Western blotting during experimental infection and challenge. Antigens from both sources, particularly of low molecular weight under reducing conditions, were recognised by the pooled sera of infected lambs. No single L-3 antigen was recognised by all infected lambs, whereas two peptides having around 25 and 26 kDa from adults were recognised by infected animals during the patency and could be of potential use in the diagnosis of lamb haemonchosis.

Animals↗

PubMed1992

A quantitative study of Eimeria infections of goats from central Spain.

A survey on coccidial infections in goats from central Spain was carried out. Fifty-five goat farms from 28 localities belonging to the three bioclimatic subregions of the area were visited. Individual samples (702) were obtained from the rectum of 130 young (under 1 year of age) and adult (572 over 1 year of age) goats. Coproscopical analyses showed a 100% prevalence and a moderate average intensity (7606.48 +/- 17918.78 Eimeria oocysts per gram of faeces), although wide variations were seen among herds and among individuals within each goat flock. Statistical analysis showed a strong age-related reduction in oocysts output in goats up to 4 years of age. The intensity of coccidial infections (young and adult goats, herds) was independent of the prevailing bioclimatic conditions. Heavier infections in young goats were found in bigger herds, whereas adult goats from these same big flocks (over 300 animals) showed the lowest oocyst output. Under our conditions, neither the female restocking rate nor the age structure seemed to play a significant role in the intensity of the Eimeria infections.

Age Factors↗

PubMed1991

Putrescine uptake regulation in response to alpha-difluoromethylornithine treatment in Leishmania infantum promastigotes.

The putrescine uptake/efflux regulation and their regulatory role on intracellular polyamine pools have been studied in the parasitic protozoa Leishmania infantum. Putrescine uptake was age-dependent with maximal values in logarithmic phase promastigotes and minimal in stationary phase. Moreover, putrescine uptake was activated in response to depletion of intracellular polyamines by alpha-difluoromethylornithine (DFMO)--a well known irreversible enzyme-activated inhibitor of ornithine decarboxylase. Kinetic studies of putrescine uptake induction showed a notable rise in Vmax without Km changes, suggesting a de novo synthesis of putrescine carriers. Putrescine uptake was able to replenish polyamine content and also to recover the proliferative rate in cells treated during 24 hours with DFMO.

Animals↗

PubMed1991

Leishmania infantum: polyamine biosynthesis and levels during the growth of promastigotes.

1. Decarboxylation of polyamine precursors: L-ornithine and L-methionine was determined along the growth curve of Leishmania infantum promastigotes in vivo, reaching maximum values on day 2 post-inoculum (mid-logarithmic phase). 2. Maximum values of L-ornithine and L-methionine decarboxylation were: 1.97 +/- 0.28 nmol CO2/hr/10(7) promastigotes and 3.18 +/- 0.34 nmol CO2/hr/10(7) promastigotes, respectively. 3. Total (free + conjugated) polyamine content was closely related with the proliferative stage of Leishmania infantum promastigotes. 4. D,L-alpha-difluoromethylornithine (DFMO) and Berenil depleted putrescine levels in a concentration-dependent manner. 5. Total and free putrescine/spermidine ratio varied significantly with the proliferative stage. Minimum values were found in late logarithmic phase (day 3 post-inoculum). 6. Small but detectable amounts of free spermine were detectable along the growth curve of Leishmania infantum promastigotes.

Animals↗

PubMed1991

Serum IgG response of Manchego lambs to infections with Haemonchus contortus and preliminary characterization of adult antigens.

Manchego lambs (16-18 weeks old) were infected with 2500 infective larvae (L3) of Haemonchus contortus and challenged 2 months later with 5000 L3. The serum IgG anti-Haemonchus response was estimated by enzyme-linked immunosorbent assay (ELISA) using soluble proteins from adults and L3. Previously infected Manchego lambs failed to mount a protective immune response against challenge, at least as assessed by faecal egg counts and pre-patency periods. Primary infection did not provoke any rise in specific anti-parasite serum antibodies, whereas a weak but significant rise was observed in challenged 6.5-month-old lambs which was very similar in both infected and non-infected lambs. The serum IgG anti-parasite response was higher against larval antigens than adult soluble proteins. Preliminary characterization of adult and larval soluble proteins by electrophoresis under reducing and denaturing conditions and Western blotting showed high cross-reactivity of both extracts. Immunoblots of adult H. contortus probed with infected and challenged lambs' sera did not yield conclusive results, although some low molecular weight peptides were recognized.

Animals↗

PubMed1989

Putrescine transport system in Leishmania infantum promastigotes.

A transport system for putrescine in Leishmania infantum (= L. donovani infantum) promastigotes has been identified and characterized by measuring the uptake of radioactively labelled putrescine. Putrescine uptake was time- and temperature-dependent without any accumulation taking place at 0 degrees C. Uptake of putrescine was maximal at pH values near neutrality. Putrescine uptake showed an apparent Km = 1.08 +/- 0.12 microM and a Vmax = 1.74 + 0.62 pmol min-1 (10(6) cells)-1. The effect of metabolic poisons and uncoupling agents suggests that the putrescine uptake was energy-dependent. The transport system seemed to be highly specific for putrescine as neither aminoacids nor related compounds tested showed any competition with putrescine uptake. The trypanocide Berenil inhibited almost completely the putrescine uptake with non-competitive kinetics and a value of Ki = 43 microM.

Amino Acids↗

PubMed1988

Preventive treatment of rabbit coccidiosis with alpha-difluoromethylornithine.

The effect of alpha-difluoromethylornithine (DFMO) administered in drinking water (0.2 and 0.5%) against liver and intestinal coccidioses in experimentally infected rabbits was studied. Zootechnical and clinical parameters were used to assess the efficacy of the compound. In both coccidioses the lower dose did not show any important effect. The higher dosage (0.5%) however, reduced the lesions induced by coccidia to an important extent. The animals infected and treated with 0.5% DFMO showed similar weight gain, food intake and relative liver weight figures to the non-infected controls. Serum transaminase levels in the rabbits infected with Eimeria stiedai and treated with 0.5% DFMO were close to those of the non-infected animals. Infected rabbits treated with this dosage also showed reductions (over 80%) in faecal oocyst output.

Animals↗

PubMed1987

The effect of the host's age on the pathology of Eimeria stiedai infection in rabbits.

The pathology of experimental Eimeria stiedai infection in 2- and 4-month-old rabbits raised coccidia-free was studied. Over 50 days of infection, the serum activities of aspartate (AST) and alanine (ALT) aminotransferases, alkaline phosphatase (AP) and gamma-glutamyltranspeptidase (GGTP) were analyzed as indicators of hepatic lesions and total serum globulins with respect to the immune response of the host; parasite development was followed by oocyst output; weight gain, food intake and relative liver weight were analyzed to control the performance of infected animals. The age of the host strongly affected parasite development and consequently the biochemical and zootechnical parameters differed more markedly in the younger animals; however, the enzyme ALT showed an increase independent of parasite development, and infection did not affect GGTP activity. The increase in age was responsible for resistance of rabbits to hepatic coccidiosis and older animals were less affected than young ones.

Age Factors↗

PubMed1987

Effects of methylglyoxal bis(guanylhydrazone) and two phenylated analogues on S-adenosylmethionine decarboxylase activity from Eimeria stiedai (Apicomplexa).

1. Activity of S-adenosylmethionine decarboxylase, one of the rate-limiting enzymes of polyamine biosynthesis, was determined in oocysts of Eimeria stiedai, a coccidian parasite of the rabbit. 2. Several properties of the enzyme were compared to the mammalian enzyme. It showed considerably less substrate affinity than the analog enzyme from the rabbit. 3. The E. stiedai enzyme showed a low sensitivity to methylglyoxal bis(guanylhydrazone), a frequently used inhibitor of the enzyme in mammals, and two phenylated derivatives. 4. Results with the inhibitors are discussed in view of their potential use in chemotherapy.

Adenosylmethionine Decarboxylase↗

PubMed1984

Some new molluscan hosts of Dicrocoelium dendriticum in Spain.

Twenty species of the family Helicidae (Gastropoda, Pulmonata) from the N.W. of Spain were tested as hosts for D. dendriticum in experimental conditions. The relationship between the egg hatching in the digestive tract and the development of sporocysts (108-110 days after egg ingestion) was studied. Three species were found to be new intermediate hosts of this trematode: Candidula intersecta , Helicella corderoi and H. jamuzensis . On the basis of the results obtained, the ecological value of the positive species in the life cycle of the parasite is discussed.

Animals↗

PubMed1984

Effect of some abiotic factors on the survival of dicrocoelium dendriticum eggs in laboratory conditions.

In the present work the results obtained from the study of the effect of temperature, RH, UV radiation and day-night thermal variations, in controlled laboratory conditions, on the survival of D. dendriticum eggs in sheep faeces are described. Mortality was seen to be maximum with a RH of 75-80% and temperatures higher than 18 degrees C. The results are compared with those obtained in natural conditions. It is suggested that the final cause of the mortality of the eggs both in laboratory and natural conditions could be the action of some microorganisms.

Animals↗