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Biomedical subjects

A Silvestre

Publications and source records attributed to A Silvestre.

At least 19 recordsLinked to original sources

Substitution of benzimidazole-resistant nematodes for susceptible nematodes in grazing lambs.

Multi-drug-resistant gastrointestinal nematode parasite populations are becoming more and more prevalent. Since anthelmintic treatments are of limited effectiveness, one solution could be to replace the anthelmintic-resistant population by a susceptible population, in order to re-establish the possibility of drug-based anthelmintic control. We investigated this substitution strategy in 4 paddocks of 0.7 ha, each of which was seeded with a benzimidazole-resistant Teladorsagia circumcincta population. The proportion of benzimidazole-resistant worms in these paddocks ranged from 20% to 89%. A 2-step replacement was performed: first, the paddocks were not grazed for 6 months (from December to July), and then the grass was cut to eliminate any residual infective larvae, before contaminating each of the paddocks with 10 seeder lambs experimentally infected with a benzimidazole-susceptible strain of T. circumcincta (from July to November). At the end of the experiment, all the populations on the 4 paddocks were phenotypically benzimidazole-susceptible, but genotyping indicated that 2 populations harboured 1% and 3% resistant worms respectively. This study demonstrates that nematode replacement is feasible in temperate areas, using semi-intensive stock management, even when the initial levels of benzimidazole-resistance are very high. Further research should next assess replacing the whole community to cope with the species diversity observed under field conditions.

Animals↗

The early drug selection of nematodes to anthelmintics: stochastic transmission and population in refuge.

We have developed an individual-based model to reflect the complexity of the early phase of drug resistance selection in a nematode/sheep model. The infection process consists of the stochastic ingestion of infective larvae spatially aggregated in clumps. Each clump corresponds to infective larvae, which are the offspring of the mature nematodes from a given sheep. We studied the dynamics of the parasitic population and the frequency of the recessive resistance alleles during selection by anthelmintic treatments. The interaction between genetic and demographic processes illustrated the trade-off between the control of the infection and the delay of resistance selection. We confirmed the importance of the number of treatments and their timing. The same treatment frequency may result in different outcomes on resistance selection in relation to the size of the refuge (infective larvae on pasture). Treatment applied during the summer (when the mortality of infective larvae on pasture was high), may lead to a rapid selection of drug resistance and a lack of control of sheep and pasture contamination. We showed that higher stocking rates were also a force in promoting the resistance allele selection.

Animals↗

The detection of anthelmintic resistance in nematodes of veterinary importance.

Before revised World Association for the Advancement of Veterinary Parasitology (WAAVP) guidelines on the detection of anthelmintic resistance can be produced, validation of modified and new methods is required in laboratories in different parts of the world. There is a great need for improved methods of detection of anthelmintic resistance particularly for the detection of macrocyclic lactone resistance and for the detection of resistant nematodes in cattle. Therefore, revised and new methods are provided here for the detection of anthelmintic resistance in nematodes of ruminants, horses and pigs as a basis for discussion and with the purpose that they are evaluated internationally to establish whether they could in the future be recommended by the WAAVP. The interpretation of the faecal egg count reduction test has been modified and suggestions given on its use with persistent anthelmintics and continuous release devices. An egg hatch test for benzimidazole (BZ) resistance is described. A microagar larval development test for the detection of benzimidazole and levamisole resistance provides third stage larvae for the identification of resistant worms. The sensitivity of these two tests can be increased by using discriminating doses rather than LD(50) values. Details are given of a PCR based test for the analysis of benzimidazole resistance in strongyles of sheep and goats, horses and cattle. Although promising for ruminant trichostrongyles, quantitative determination of gene frequency using real time PCR requires further development before PCR tests will be used in the field. Apart from faecal egg count reduction tests there are currently no satisfactory tests for macrocylic lactone resistance despite the great importance of this subject. Except for treatment and slaughter trials there are no validated tests for fasciolicide resistance or for the detection of resistance in cestodes.

Animals↗

Nematode parasites of animals are more prone to develop xenobiotic resistance than nematode parasites of plants.

In this paper, we concentrate on a comparison of plant and animal-parasitic nematodes, to gain insight into the factors that influence the acquisition of the drug resistance by nematodes. Comparing nematode parasite of domestic animals and cultivated plants, it appears that drug resistance threatens only domestic animal production. Does the paucity of report on nematicide field resistance reflect reality or, is nematicide resistance bypassed by other management practices, specific to cultivated plants (i.e. agricultural control)? First, it seems that selection pressure by treatments in plants is not as efficient as selection pressure in ruminants. Agronomic practices (i.e. sanitation, early planting, usage of nematodes resistant cultivar and crop rotation) are frequently used to control parasitic-plant nematodes. Although the efficiency of such measures is generally moderate to high, integrated approaches are developing successfully in parasitic-plant nematode models. Secondly, the majority of anthelmintic resistance cases recorded in animal-parasitic nematodes concern drug families that are not used in plant-parasitic nematodes control (i.e. benzimidazoles, avermectines and levamisole). Thirdly, particular life traits of parasitic-plant nematodes (low to moderate fecundity and reproductive strategy) are expected to reduce probability of appearance and transmission of drug resistance genes. It has been demonstrated that, for a large number of nematodes such as Meloidogyne spp., the mode of reproduction by mitotic parthenogenesis reduced genetic diversity of populations which may prevent a rapid drug resistance development. In conclusion, anthelmintic resistance develops in nematode parasite of animals as a consequence of an efficient selection pressure. Early detection of anthelmintic resistance is then crucial: it is not possible to avoid it, but only to delay its development in farm animal industry.

Agriculture↗

Diversity of benzimidazole-resistance alleles in populations of small ruminant parasites.

The resistance of gastro-intestinal nematodes of small ruminants (sheep and goat) to benzimidazole anthelmintic drugs seems to be linked primarily to a single mutation in the isotype 1 beta-tubulin gene. This study was carried out to investigate the origin and diversity of benzimidazole-resistance alleles in trichostrongylid nematodes. We sequenced a 550 bp fragment of the isotype 1 beta-tubulin gene from several benzimidazole-resistant Teladorsagia circumcincta populations isolated from dairy goat farms in the central and south-western France. We also sequenced the same beta-tubulin fragment from Trichostrongylus colubriformis and Haemonchus contortus populations in south-western France. We found eight benzimidazole-resistance alleles in all T. circumcincta populations studied, six in H. contortus populations, and only one in T. colubriformis populations. In most cases, only one benzimidazole-resistance allele was present in T. circumcincta and H. contortus populations, but two alleles were found in a fewer number of them. Some T. circumcincta populations shared the same benzimidazole-resistance allele whereas some others had a specific benzimidazole-resistance allele. Similar findings were obtained for H. contortus. As no parasites are introduced once the flock of dairy goat farms has been constituted, these data indicate for the three studied species that rare pre-existing benzimidazole-resistance alleles already present before the isolation of populations had been selected. On the other hand, the fact that some benzimidazole-resistance alleles were specific to one population of T. circumcincta or H. contortus, seems to be in agreement with the hypothesis of the selection of spontaneous mutations. Thus, the origin of benzimidazole-resistance alleles in trichostrongylid nematodes seems to involve primarily the selection of rare alleles and possibly of spontaneous mutations.

Alleles↗

Molecular approaches to studying benzimidazole resistance in trichostrongylid nematode parasites of small ruminants.

Molecular techniques are of growing importance in the study of anthelmintic resistance in trichostrongylid worm populations. A knowledge of the genetic determinants of benzimidazole (BZ) resistance has made it possible to construct a molecular tool for genotyping individual worms, in respect of mutation of the beta-tubulin gene responsible for BZ resistance. This tool offers new possibilities in the diagnosis of BZ resistance, and also in the study of anthelmintic use and other breeding management factors that can affect the selection of BZ-resistant alleles in worm populations. New molecular methods have also made it possible to study the origin and diversity of BZ-resistant alleles in trichostrongylid populations. The results demonstrate the value of a multidisciplinary approach to the study of anthelmintic resistance, combining molecular, ecological and epidemiological techniques.

Alleles↗

Prevalence of anthelmintic resistance in gastrointestinal nematodes of dairy goats under extensive management conditions in southwestern France.

The occurrence of benzimidazole (BZ) and levamisole resistance was investigated in 18 randomly selected dairy goat herds located in southwestern France and characterized by extensive management. On each of the 18 farms, 45 adult goats were randomly allocated into three groups of 15 animals each: an untreated control group, a group that was orally administered fenbendazole (10 mg kg(-1) body weight) and a group that received orally a levamisole drench (12 mg kg(-1) body weight). Individual faecal egg counts and pooled larval cultures were done 10 days after anthelmintic treatment. Naive lambs were infected with larvae obtained from control and fenbendazole treated groups and were necropsied 35 days after infection for worm recovery. Faecal egg count reductions (FERC) were calculated for fenbendazole and levamisole and, when less than 95 per 100, were considered as indicative of anthelmintic resistance. An in vitro egg hatch test (EHT) was conducted with thiabendazole on eggs isolated from pooled faeces of fenbendazole treated goats in nine farms. Faecal egg count reductions indicated the occurrence of benzimidazole resistance in 15 out of 18 farms. Among these farms, nine had EHT values above 0.1 microg thiabendazole ml(-1) confirming the benzimidazole resistance status. Levamisole resistance was detected in two farms through FECR. Based on necropsy results, the prevalence of benzimidazole resistance was higher in Trichostrongylus colubriformis, medium in Haemonchus contortus and lower in Teladorsagia circumcincta. In nine farms the benzimidazole resistance was monospecific whereas multispecific resistance was found in the six remaining farms. A negative relationship was found between FECR for fenbendazole and the average number of anthelmintic treatments given per year on the farm. Despite extensive management including a low number of treatments, the prevalence of benzimidazole resistance was very high suggesting that the repeated and sometimes exclusive use of benzimidazole drugs, even at low frequency, is probably the main cause in developing nematode resistance in dairy goat herds. The importance of other factors such as under-dosing or buying animals already carrying resistant nematodes are discussed.

Animals↗

Effect of benzimidazole under-dosing on the resistant allele frequency in Teladorsagia circumcincta (Nematoda).

This experiment was designed to determine the effects of under-dosing on the frequency of benzimidazole resistant allele in the nematode Teladorsagia circumcincta. Fenbendazole (FBZ) was tested at 1/32, 1/16, 1/8 and 1/4 of the recommended dose for sheep (5 mg/kg body weight). The fraction of the susceptible homozygote (SS), susceptible heterozygote (RS) and resistant homozygote (RR) genotypes were compared among FBZ dose groups to evaluate differences between SS and RS genotype selective advantage. Almost all SS genotype worms were eliminated by 1/4 of the FBZ recommended dose, whereas a significant fraction of the RS genotype worms survived treatment. The selective advantage was 4.5 times higher for the RS genotype. This selective advantage was determined at 1/4 of the manufacturer's recommended dose of FBZ. This value should be taken as an indictor of the selective advantage of RS over the SS genotype when lambs are under-dosed. A computer simulation was used to study the putative spread of anthelmintic resistance over a range of RS selective advantages (2, 4.5 and 10-fold), with two average sizes of individual host worm population (20 or 2000 worms/host) and two initial R allele frequencies (0.1%, or 1%). In all situations, the lowest selective advantage of the RS genotype over the SS genotype was sufficient to promote the spread of resistance in susceptible populations. When the RS genotype had no selective advantage over the SS genotype, genetic drift almost always led to the loss of the R allele, except in the largest populations (average size = 2000 worms).

Alleles↗

Transient osteoporosis.

Transient osteoporosis (TO) is an uncommon entity whose principal characteristic is to be a self-limited syndrome. Diagnosis is made upon clinical presentation and x-ray evidence of diffuse osteopenia around the affected joint followed by spontaneous healing after several months. When recurrent episodes occur at different times and locations it is called regional migratory osteoporosis. Magnetic resonance imaging and technetium-99 bone scan may be helpful in diagnosis during the early phase. Good results may be achieved with nonsteroidal antiinflammatory medication, protected weight bearing and physical therapy.

Diagnosis, Differential↗

Relationship between helminth species diversity, intensity of infection and breeding management in dairy goats.

Helminth infection, species diversity (proportion of each species in the community), species number, intensity of infection and anthelmintic resistance were investigated in 16 dairy-goat farms of south-western France. The aim of the study was to estimate which breeding management factors may affect diversity and intensity of infection. Anthelmintic resistance was investigated because this factor may modify composition of nematode communities. Farm history and breeding management were recorded by means of a questionnaire. Natural and experimental infections were compared to determine which strategy was the best to estimate diversity index and number of species from nematode community of a whole herd. A total of 17 species of helminths, among which 14 nematodes, one cestode (Moniezia spp.) and two trematodes (Paramphistomum daubneyi and Dicrocoelium lanceolatum) were recovered in the 26 necropsied culled goats. One culled goat worm burden represented conveniently species number but not species diversity (Shannon index) of parasite community harboured in the whole herd. Experimental sheep infection with larvae from pooled faeces of 15 goats was the best strategy to estimate species diversity in farms. Diversity could be predicted from the number of farms from which goats originated at constitution of the herd, the duration of goat winter withdrawal from pastures, and intensity of infection. Anthelmintic resistance was correlated with the number of farms of origin, area of permanent pastures and intensity of infection. The strategy adopted for the herd constitution was a critical step as species diversity and risk of anthelmintic resistance increased with the number of farms of origin. Intensity of infection was negatively correlated with species diversity. This suggested that more diverse communities were better controlled by the host, in agreement with the widespread statement that diversity would beget stability.

Abomasum↗

A molecular tool for species identification and benzimidazole resistance diagnosis in larval communities of small ruminant parasites.

This report describes a molecular method for determining in a first step the generic composition of a nematode community and in a second step, the resistance of each species to benzimidazole (BZ). We first established a polymerase chain reaction (PCR) linked to a restriction fragment length polymorphism strategy using the isotype 1 beta-tubulin gene. This method overcame the limitations of morphological identification of larval stages of trichostrongylid nematode species. Geographically distant isolates from the three main gastrointestinal species in temperate zones, Teladorsagia circumcincta, Haemonchus contortus, and Trichostrongylus colubriformis, were distinguished using this method. We then used an allele-specific PCR (AS-PCR) to detect mutations of residue 200 of the beta-tubulin, which is implicated in BZ resistance. The sequences of several samples confirmed the BZ-resistance genotype determined by AS-PCR. The ability to process large numbers of samples simultaneously makes this PCR-based strategy particularly suitable for epidemiological studies. It may also be useful for monitoring the emergence of resistant alleles in nematode communities.

Animals↗

Benefits of multiple methods for evaluating HIV counseling and testing sites in Pennsylvania.

In 1995-1996, an evaluation was conducted of publicly funded HIV counseling, testing, referral, and partner notification sites in the Commonwealth of Pennsylvania. A collaborative effort of public and private entities, this evaluation used multiple methods that proved to be more powerful in eliciting results than any of the methods used alone, and it did so with budget parameters determined by the public funding agency. This article examines the results of two methods: written mail surveys sent to all of the agencies in the population of publicly funded sites and interviews conducted at a random sample of sites. Results were found to be similar along lines of questions posed similarly by both approaches, and complementary in other areas. Benefits of triangulation in statewide evaluation of HIV/AIDS services are discussed.

AIDS Serodiagnosis↗

[Sanitary conditions of community dining halls in greater Buenos Aires, Argentina].

Because of the importance of food safety for public health, a study was done to determine the microbiologic quality of foods provided to patrons of community dining halls in greater Buenos Aires, Argentina. The study was descriptive and was conducted in three municipalities with intermediate or high poverty indices; a sample of 52 dining halls was selected randomly from those municipalities. Information was collected from April 1994 through April 1995 by means of surveys, direct observations, and microbiologic analyses of the water and food served. Water and food analysis followed the recommendations of the Argentine Food Code and the International Commission on Microbiologic Specifications for Foods, respectively. The results were interpreted according to the values for cooked foods agreed upon by the Working Group on Food Monitoring. The microbiologic analysis revealed that 28 (54%) of the dining halls had food or water of inadequate quality and 11 (21%) had inadequacies in both. Bacillus cereus was the most commonly found pathogenic bacterium in the food samples analyzed. The factors that were significantly associated with unsatisfactory microbiologic conditions in the food and water were lack of disinfection of the water (P = 0.009), lack of hygiene in the environment (P = 0.03), the presence of rodents and insects (P = 0.05), the supply system for raw materials (P = 0.01), and a relatively small number of users, i.e., fewer than 100 (P = 0.0008). It was concluded that water disinfection and general clean-up of the dining halls were measures that could be put in place by providing education in the areas of hygiene and food handling to supervisors and employees. The results obtained also indicate the need to include a food safety evaluation among the general evaluation criteria for food assistance programs.

Adolescent↗

[Dental-radicular cysts invading the maxillary sinus].

The authors report three cases of radicular cyst developing into the maxillary sinus which was partially or completely obscured. Looking for a thin rim of cortical bone should be undertaken in case of opacity of maxillary sinus. This rim correspond to the floor of the sinus, which is lifted up by the cyst. If the attenuation value of the intrasinusal process is near of that of water, the diagnosis of radicular cyst invading the maxillary sinus is very likely and may indicate intraoral surgical approach.

Adult↗

[Circumscribed myositis ossificans: bone changes detected by MRI].

The authors report a case of ossificans myositis, in which MRI showed inflammatory changes of the adjacent bone. T1 weighted fat saturation sequence with gadolinium injection showed enhancement of medullary and cortical bone. This potentially mistaking pattern must be known, to avoid misdiagnosing with malignant osseous tumor, specially before achievement of the characteristic pattern of zonal maturation and its calcified rim.

Adult↗