The prevalence of anthelmintic resistance in nematode parasites of sheep in southern Latin America: Brazil.

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Echevarria, F; Borba, M F; Pinheiro, A C; Waller, P J; Hansen, J W

1996-04-01

This survey was conducted in the southern Brazilian state of Rio Grande do Sul and involved 182 farms located in 26 counties. In addition to the three major broad-spectrum anthelmintic groups (viz. benzimidazole, levamisole and ivermectin) the combination benzimidazole and levamisole and the H. contortus specific anthelmintic, closantel, were tested by the faecal egg count reduction method for the prevalence of anthelmintic resistance. Resistance was found to be 90%, 84%, 13%, 73% and 20%, respectively. This is a crisis situation. Immediate, drastic action needs to be implemented, otherwise the sheep industry in this region (approx. 10 million head) will soon face a lack of any effective anthelmintics with the inevitable consequences of major restructuring or abandonment.

Gastrointestinal nematode control practices on lowland sheep farms in Ireland with reference to selection for anthelmintic resistance

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Patten Thomas

2011-03-01

Full Text Available Abstract Gastrointestinal parasitism is a widely recognised problem in sheep production, particularly for lambs. While anthelmintics have a pivotal role in controlling the effects of parasites, there is a paucity of data on how farmers use anthelmintics. A representative sample of Irish lowland farmers were surveyed regarding their parasite control practices and risk factors that may contribute to the development of anthelmintic resistance. Questionnaires were distributed to 166 lowland Irish sheep producers. The vast majority of respondents treated their sheep with anthelmintics. Lambs were the cohort treated most frequently, the majority of farmers followed a set programme as opposed to treating at sign of disease. A substantial proportion (61% administered four or more treatments to lambs in a 'normal' year. Departures from best practice in anthelmintic administration that would encourage the development of anthelmintic resistance were observed. In conclusion, in the light of anthelmintic resistance, there is a need for a greater awareness of the principles that underpin the sustainable use of anthelmintics and practices that preserve anthelmintic efficacy should be given a very high priority in the design of helminth control programmes on each farm. To this end, given that veterinary practitioners and agricultural advisors were considered to be the farmer's most popular information resource, the capacity of these professions to communicate information relating to best practice in parasite control should be targeted.

The development of anthelmintic resistance with best practice control of nematodes on commercial sheep farms in the UK.

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Learmount, Jane; Stephens, Nathalie; Boughtflower, Valerie; Barrecheguren, Alba; Rickell, Kayleigh

2016-10-15

increased sensitivity both to detect and to measure changes in response to anthelmintics with the dose-response assay compared to the dd and this is important particularly when allele frequencies are low as might be the case when novel compounds are released to the market. Anthelmintic use across years 5-7 was significantly lower for the farms in the SCOPS group compared to the TRADITIONAL group and farmers in the SCOPS group had selected products from the benzimidazole group less often than farmers in the TRADITIONAL group. Both groups had made minimal use of the imidazothiazole anthelmintic classes and the majority of ewe treatments were selected from the macrocyclic lactone class. Further research is required to determine the effect of these anthelmintic choices on the development of resistance to the macrocyclic lactones. Crown Copyright © 2016. Published by Elsevier B.V. All rights reserved.

Biotransformation of albendazole and activities of selected detoxification enzymes in Haemonchus contortus strains susceptible and resistant to anthelmintics.

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Vokřál, Ivan; Jirásko, Robert; Stuchlíková, Lucie; Bártíková, Hana; Szotáková, Barbora; Lamka, Jiří; Várady, Marián; Skálová, Lenka

2013-09-23

The increased activity of drug-metabolizing enzymes can protect helminths against the toxic effect of anthelmintics. The aim of this study was to compare the metabolism of the anthelmintic drug albendazole (ABZ) and the activities of selected biotransformation and antioxidant enzymes in three different strains of Haemonchus contortus: the ISE strain (susceptible to common anthelmintics), the BR strain (resistant to benzimidazole anthelmintics) and the WR strain (multi-resistant). H. contortus adults were collected from the abomasum of experimentally infected lambs. In vitro (subcellular fractions of H. contortus homogenate) as well as ex vivo (living nematodes cultivated in flasks with medium) experiments were performed. HPLC with spectrofluorimetric and mass-spectrometric detection was used in the analysis of ABZ metabolites. The in vitro activities of oxidation/antioxidation and conjugation enzymes toward model substrates were also assayed. The in vitro data showed significant differences between the susceptible (ISE) and resistant (BR, WR) strains regarding the activities of peroxidases, catalase and UDP-glucosyltransferases. S-oxidation of ABZ was significantly lower in BR than in the ISE strain. Ex vivo, four ABZ metabolites were identified: ABZ sulphoxide and three ABZ glucosides. In the resistant strains BR and WR, the ex vivo formation of all ABZ glucosides was significantly higher than in the susceptible ISE strain. The altered activities of certain detoxifying enzymes might partly protect the parasites against the toxic effect of the drugs as well as contribute to drug-resistance in these parasites. Copyright © 2013 Elsevier B.V. All rights reserved.

Anthelmintic resistant nematodes in goats in the Netherlands

NARCIS (Netherlands)

Borgsteede, F.H.M.; Pekelder, J.J.; Dercksen, D.P.

1996-01-01

A suspected case of anthelmintic resistance on a farm with Angora and Anglo-Nubian goats was confirmed in a controlled test. Twelve lambs of sheep were infected with larvae cultured from faeces of the goats. The lambs were allocated to four groups: untreated controls and lambs treated 21 days after

Prevalence and anthelmintic resistance of strongyle parasites in ...

African Journals Online (AJOL)

Prevalence of infection and level of anthelmintic resistance (AR) of strongyle nematodes to ivermectin (IV), albendazole (AB) and levamisole (LV) in Dorper lambs were determined. The overall prevalence was 67.0% and mean eggs per gram (EPG) of faeces was 357. Infection was light in 92.5%, moderate in 4.5% and high ...

Anthelmintic resistance and associated management practices in local horses in Sokoto metropolis, Nigeria

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Abubakar Musa Mayaki

2018-03-01

Full Text Available This study was carried out to assess the management practices used in the control of gastrointestinal (GI nematodes of horses and to determine the efficacy of three anthelmintics commonly used in Sokoto metropolis. A questionnaire was administered on management practices, while an anthelmintic efficacy test was carried out using 15 horses. The 15 horses were divided into three groups (A, B and C comprising of 5 each and treated with albendazole, ivermectin and fenbendazole, respectively. The faecal egg count reduction test (FECRT was used to determine the efficacy and faecal culture was used to determine the parasite species. Majority of the respondents (80% claimed to have worm control strategies, but only 32.5% used anthelmintics for the control of GI parasites. 62.5% of respondents designed their deworming plan, while only 25% relied on veterinarians. Most of the treatments were done by the horse owners and/or handlers and they largely depended on visual judgement in dosage determination. Their selection of anthelmintics was based on familiarity and 52.5% of the respondents dewormed their horses six times a year using a particular class of anthelmintic or herbal remedies. Resistance against albendazole as well as suspected resistance against fenbendazole by the GI nematodes identified was observed, while ivermectin demonstrated high efficacy against all nematodes isolated. In conclusion, a single dose of subcutaneous injection of ivermectin was highly effective against gastrointestinal parasites in horses, while the worm control strategies employed by respondents enhanced the selection of nematode resistance to albendazole and fenbendazole.

Managing anthelmintic resistance-Variability in the dose of drug reaching the target worms influences selection for resistance?

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Leathwick, Dave M; Luo, Dongwen

2017-08-30

The concentration profile of anthelmintic reaching the target worms in the host can vary between animals even when administered doses are tailored to individual liveweight at the manufacturer's recommended rate. Factors contributing to variation in drug concentration include weather, breed of animal, formulation and the route by which drugs are administered. The implications of this variability for the development of anthelmintic resistance was investigated using Monte-Carlo simulation. A model framework was established where 100 animals each received a single drug treatment. The 'dose' of drug allocated to each animal (i.e. the concentration-time profile of drug reaching the target worms) was sampled at random from a distribution of doses with mean m and standard deviation s. For each animal the dose of drug was used in conjunction with pre-determined dose-response relationships, representing single and poly-genetic inheritance, to calculate efficacy against susceptible and resistant genotypes. These data were then used to calculate the overall change in resistance gene frequency for the worm population as a result of the treatment. Values for m and s were varied to reflect differences in both mean dose and the variability in dose, and for each combination of these 100,000 simulations were run. The resistance gene frequency in the population after treatment increased as m decreased and as s increased. This occurred for both single and poly-gene models and for different levels of dominance (survival under treatment) of the heterozygote genotype(s). The results indicate that factors which result in lower and/or more variable concentrations of active reaching the target worms are more likely to select for resistance. The potential of different routes of anthelmintic administration to play a role in the development of anthelmintic resistance is discussed. Copyright © 2017 Elsevier B.V. All rights reserved.

First report of multiple anthelmintic resistance in nematodes of sheep in Colombia

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Carlos M.B. Gárcia

2016-03-01

Full Text Available This study aimed to report the presence of parasites resistant to the most used anthelmintic drugs in sheep in Colombia. Four farms (denominated farm 1, 2, 3 and 4 were selected where the animals were not treated with anthelmintics for two months before the trial. Animals with faecal egg count (FEC above 150 and of different ages were allocated into six groups, each consisting of at least 5 animals. The drugs and dosages used were: ivermectin 1% (0.2 mg/kg, albendazole 25% (5 mg/kg, fenbendazole 10% (5 mg/kg, levamisole 10% (5 mg/kg, and moxidectin 1% (0.2 mg/kg. Anthelmintic efficacy was determined by the FEC reduction test (FECRT with a second sampling 14 days post-treatment. The efficacy of albendazole and fenbendazole at farm 1 was above 95%, which was different from the others farms. The FECRT indicated the presence of multidrug resistance in the other farms where no tested drugs showed activity higher than 79% (albendazole: 0 to 55%, fenbendazole: 51.4 to 76.6%, ivermectin: 67.3 to 93.1%, levamisole: 0 to 78.1%, and moxidectin: 49.2 to 64.1%.Haemonchus contortus was the predominant (96% species, followed by a small presence of Trichostrongylus sp. (3% andCooperia sp. (1%. Therefore, we report for the first time the existence of multiple anthelmintic resistance in gastrointestinal nematodes of sheep in Colombia.

Managing anthelmintic resistance in small ruminant livestock of resource-poor farmers in South Africa : review article

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A.F. Vatta

2006-06-01

Full Text Available Gastrointestinal parasitism is one of the most important disease complexes of sheep and goats impacting on the resource-poor livestock farmer. Of the responsible nematodes, Haemonchus contortus, a blood-sucking worm of the abomasum, poses possibly the greatest threat. Over the past several decades, the worm has been controlled through the use of anthelmintics, but the emergence of anthelmintic resistance has threatened this chemotherapeutic approach. In Africa, the overall prevalence of anthelmintic resistance has not been extensively investigated, particularly within the resource-poor farming sector, but resistance has been reported from at least 14 countries with most of the reports emanating from Kenya and South Africa and the majority concerning H. contortus. While levels of resistance under commercial sheep farming systems in South Africa is considered to be amongst the worst in the world, resistance has also been reported from the resource-poor farming sector. Increases in productivity and reproduction of livestock and the development of markets for sale of animals are seen by international funding bodies as a way out of poverty for communities that keep livestock. This must lead to the greater need for parasite control. At such times, the risk of levels of anthelmintic resistance escalating is much greater and there is therefore a need to look at alternatives to their use. Proposed strategies include the appropriate, but judicious use of anthelmintics by application of the FAMACHA(c system and the use of alternatives to anthelmintics such as strategic nutrient supplementation. It is also very clear that there is a strong demand for knowledge about animal diseases, including helminthosis, and their effective management in the resource-poor livestock farming communities. This is an important challenge to meet.

A novel high throughput assay for anthelmintic drug screening and resistance diagnosis by real-time monitoring of parasite motility.

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Michael J Smout

Full Text Available BACKGROUND: Helminth parasites cause untold morbidity and mortality to billions of people and livestock. Anthelmintic drugs are available but resistance is a problem in livestock parasites, and is a looming threat for human helminths. Testing the efficacy of available anthelmintic drugs and development of new drugs is hindered by the lack of objective high-throughput screening methods. Currently, drug effect is assessed by observing motility or development of parasites using laborious, subjective, low-throughput methods. METHODOLOGY/PRINCIPAL FINDINGS: Here we describe a novel application for a real-time cell monitoring device (xCELLigence that can simply and objectively assess anthelmintic effects by measuring parasite motility in real time in a fully automated high-throughput fashion. We quantitatively assessed motility and determined real time IC(50 values of different anthelmintic drugs against several developmental stages of major helminth pathogens of humans and livestock, including larval Haemonchus contortus and Strongyloides ratti, and adult hookworms and blood flukes. The assay enabled quantification of the onset of egg hatching in real time, and the impact of drugs on hatch rate, as well as discriminating between the effects of drugs on motility of drug-susceptible and -resistant isolates of H. contortus. CONCLUSIONS/SIGNIFICANCE: Our findings indicate that this technique will be suitable for discovery and development of new anthelmintic drugs as well as for detection of phenotypic resistance to existing drugs for the majority of helminths and other pathogens where motility is a measure of pathogen viability. The method is also amenable to use for other purposes where motility is assessed, such as gene silencing or antibody-mediated killing.

Anthelmintic resistance of nematodes in communally grazed goats in a semi-arid area of South Africa

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F.R. Bakunzi

2003-07-01

Full Text Available A survey was conducted on the occurrence of anthelmintic resistance of nematodes in communally grazed goats in a semi-arid area in SouthAfrica. In herds belonging to 10 smallholder goat farmers, the efficacies of fenbendazole, levamisole and rafoxanide were tested by faecal egg count reduction (FECR tests. Efficacies of 80 % were considered a threshold for anthelmintic resistance. The FECR tests showed that all drugs tested more than 80 % effective in most instances, but there were notable exceptions. In 1 case, rafoxanide was only 31 % effective and in another case fenbendazole was only 47 % effective. The occurrence of anthelmintic resistance in this farming sector is of concern. Steps should be taken to prevent its further spread and to avoid the development of a situation as onnumerous commercial sheep farms in South Africa where resistance is very common.

Anthelmintic metabolism in parasitic helminths: proteomic insights.

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Brophy, Peter M; MacKintosh, Neil; Morphew, Russell M

2012-08-01

Anthelmintics are the cornerstone of parasitic helminth control. Surprisingly, understanding of the biochemical pathways used by parasitic helminths to detoxify anthelmintics is fragmented, despite the increasing global threat of anthelmintic resistance within the ruminant and equine industries. Reductionist biochemistry has likely over-estimated the enzymatic role of glutathione transferases in anthelmintic metabolism and neglected the potential role of the cytochrome P-450 superfamily (CYPs). Proteomic technologies offers the opportunity to support genomics, reverse genetics and pharmacokinetics, and provide an integrated insight into both the cellular mechanisms underpinning response to anthelmintics and also the identification of biomarker panels for monitoring the development of anthelmintic resistance. To date, there have been limited attempts to include proteomics in anthelmintic metabolism studies. Optimisations of membrane, post-translational modification and interaction proteomic technologies in helminths are needed to especially study Phase I CYPs and Phase III ABC transporter pumps for anthelmintics and their metabolites.

Comparison of two in vitro methods for the detection of ivermectin resistance in Haemonchus contortus in sheep

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Urda Dolinská M.

2016-06-01

Full Text Available Gastrointestinal parasitic nematodes in sheep cause severe economic losses. Anthelmintics are the most commonly used drugs for prophylaxis and therapy against parasitic helminths. The problem of drug resistance has developed for all commercially available anthelmintics in several genera and classes of helminths. In vitro and in vivo tests are used to detect anthelmintic resistance. Two in vitro methods (larval migration inhibition test and micromotility test for the detection of ivermectin (IVM resistance were compared using IVM-resistant and IVM-susceptible isolates of Haemonchus contortus. The degree of resistance for each test was expressed as a resistance factor (RF. The micromotility test was more sensitive for quantitatively measuring the degree of resistance between susceptible and resistant isolates. The RFs for this test for IVM and eprinomectin ranged from 1.00 to 108.05 and from 3.87 to 32.32, respectively.

Anthelmintic resistance in gastrointestinal nematodes of beef cattle in the state of Rio Grande do Sul, Brazil.

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Ramos, Fernanda; Portella, Luiza Pires; Rodrigues, Fernando de Souza; Reginato, Caroline Zamperete; Pötter, Luciana; Cezar, Alfredo Skrebsky; Sangioni, Luís Antônio; Vogel, Fernanda Silveira Flores

2016-04-01

Gastrointestinal nematodes resistant to anthelmintics have been reported in several regions of Brazil, and they may be associated with economic losses for the cattle industry. This study aimed to evaluate the resistance status of gastrointestinal nematodes from naturally infected beef cattle to several commercially available anthelmintics, as well as to test the efficacy of combinations of anthelmintics against multi-resistant gastrointestinal nematodes. Ten farms located in Rio Grande do Sul state were selected by: farmers' consent; extensive raising system; availability of calves aged from 7 to 9 months naturally infected by gastrointestinal nematodes; absence of anthelmintic treatment for 60 days before the study; and presence of 70-100 calves or more of both genders with ≥ 200 eggs per gram of feces (EPG) (sensitivity of 50 EPG). These calves were distributed into 10 groups (of 7-10 animals) per farm and treated with ivermectin, doramectin, eprinomectin, fenbendazole, closantel, nitroxynil, disophenol, levamisole, albendazole, or moxidectin. Feces were collected 2 days before treatment and 14 days after treatment. Additional groups of 7-10 calves were used to test six different two-drug combinations at four of the studied farms. In general terms, fenbendazole was the most effective drug, followed by levamisole, disophenol, and moxidectin. However, parasite resistance to multiple drugs was found in all herds, especially in the genera Cooperia spp., Trichostrongylus spp., and Haemonchus spp.. Some of the two-drug combinations were effective against nematode populations identified as resistant to the same compounds when used as single drugs. The most effective combinations were moxidectin + levamisole, doramectin + fenbendazole, and levamisole + closantel. In this study, parasites resistant to the main commercially available anthelmintics were found in all herds, and some combinations of two active components belonging to different chemical groups were effective

Anthelmintic resistance in gastrointestinal nematodes of beef cattle in the state of Rio Grande do Sul, Brazil

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Ramos, Fernanda; Portella, Luiza Pires; Rodrigues, Fernando de Souza; Reginato, Caroline Zamperete; Pötter, Luciana; Cezar, Alfredo Skrebsky; Sangioni, Luís Antônio; Vogel, Fernanda Silveira Flores

2016-01-01

Gastrointestinal nematodes resistant to anthelmintics have been reported in several regions of Brazil, and they may be associated with economic losses for the cattle industry. This study aimed to evaluate the resistance status of gastrointestinal nematodes from naturally infected beef cattle to several commercially available anthelmintics, as well as to test the efficacy of combinations of anthelmintics against multi-resistant gastrointestinal nematodes. Ten farms located in Rio Grande do Sul state were selected by: farmers' consent; extensive raising system; availability of calves aged from 7 to 9 months naturally infected by gastrointestinal nematodes; absence of anthelmintic treatment for 60 days before the study; and presence of 70–100 calves or more of both genders with ≥200 eggs per gram of feces (EPG) (sensitivity of 50 EPG). These calves were distributed into 10 groups (of 7–10 animals) per farm and treated with ivermectin, doramectin, eprinomectin, fenbendazole, closantel, nitroxynil, disophenol, levamisole, albendazole, or moxidectin. Feces were collected 2 days before treatment and 14 days after treatment. Additional groups of 7–10 calves were used to test six different two-drug combinations at four of the studied farms. In general terms, fenbendazole was the most effective drug, followed by levamisole, disophenol, and moxidectin. However, parasite resistance to multiple drugs was found in all herds, especially in the genera Cooperia spp., Trichostrongylus spp., and Haemonchus spp.. Some of the two-drug combinations were effective against nematode populations identified as resistant to the same compounds when used as single drugs. The most effective combinations were moxidectin + levamisole, doramectin + fenbendazole, and levamisole + closantel. In this study, parasites resistant to the main commercially available anthelmintics were found in all herds, and some combinations of two active components belonging to different chemical groups

Worm-control practices and prevalence of anthelmintic resistance using in vivo FECRTs on smallholder sheep farms in Lithuania

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Kupčinskas T.

2016-03-01

Full Text Available This study determined the prevalence of anthelmintic resistance (AR in parasitic nematodes on smallholder sheep farms in Lithuania from April to November 2014. Faecal samples were collected from two groups of 10-15 sheep treated with fenbendazole (FBZ or ivermectin (IVM on 18 sheep farms. Two samples were collected from each group: on day zero (T1 and 10-14 days after treatment. Faecal egg counts (eggs per gramme, EPG were determined using a modified McMaster technique. Animals with < 140 EPG on day zero were removed from the analysis. The prevalence of AR was estimated using the in vivo faecal egg count reduction test. AR to FBZ was detected on three of 15 farms where FBZ was used (20 % and was suspected on one farm (6.7 %. AR to IVM was detected on two of 16 farms where IVM was used (12.5 %. The main species of resistant gastrointestinal nematodes (GINs identified after treatment were Teladorsagia spp. and Trichostrongylus spp. A questionnaire surveying 71 sheep farmers estimated that 71.8 % of sheep farmers used anthelmintics against GINs. IVM was the most frequently (68.6 % applied anthelmintic, and 62.7 % of the respondents reported treating their animals twice a year. This study confirmed the presence of AR to GIN infections on sheep farms in Lithuania. Future studies should assess the prevalence of AR to GIN infection using in vitro methods.

Utilization of composite fecal samples for detection of anthelmintic resistance in gastrointestinal nematodes of cattle.

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George, Melissa M; Paras, Kelsey L; Howell, Sue B; Kaplan, Ray M

2017-06-15

Recent reports indicate that anthelmintic resistance in gastrointestinal nematodes of cattle is becoming increasingly prevalent worldwide. Presently, the fecal egg count reduction test (FECRT) is the only means available for detection of resistance to anthelmintics in cattle herds at the farm level. However, the FECRT is labor and cost intensive, and consequently is only rarely performed on cattle farms unless for research purposes. If costs could be reduced, cattle producers might be more likely to pursue drug resistance testing on their farms. One approach to reducing the cost of the FECRT, is the use of composite fecal samples for performing fecal egg counts (FEC), rather than conducting FEC on fecal samples from 15 to 20 individual animals. In this study FECRT were performed on 14 groups of cattle using both individual and composite FEC methods To measure how well the results of composite sampling reproduce those of individual sampling, Lin's Concordance Correlation Coefficient was utilized to describe both the linear relationship between methods and the slope and y-intercept of the line relating the data sets. There was little difference between the approaches with 98% agreement in mean FEC found between methods Mean FEC based on individual counts ranged between 0 and 670.6 eggs per gram of feces, indicating that the results of this study are applicable to a wide range of FEC levels. Standard error of the mean FEC and range of FEC are reported for each group prior to and following treatment to describe the variability of the data set. There was greater than 95% agreement in drug efficacy between individual and composite sampling methods, demonstrating composite sampling is appropriate to evaluate drug efficacy. Notably, for all groups tested the efficacy calculated by composite sampling was within the 95% confidence interval for efficacy calculated using individual sampling. The use of composite samples was shown to reduce the number of FEC required by 79

Anthelmintic resistance in gastrointestinal nematodes of beef cattle in the state of Rio Grande do Sul, Brazil

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Fernanda Ramos

2016-04-01

Full Text Available Gastrointestinal nematodes resistant to anthelmintics have been reported in several regions of Brazil, and they may be associated with economic losses for the cattle industry. This study aimed to evaluate the resistance status of gastrointestinal nematodes from naturally infected beef cattle to several commercially available anthelmintics, as well as to test the efficacy of combinations of anthelmintics against multi-resistant gastrointestinal nematodes. Ten farms located in Rio Grande do Sul state were selected by: farmers' consent; extensive raising system; availability of calves aged from 7 to 9 months naturally infected by gastrointestinal nematodes; absence of anthelmintic treatment for 60 days before the study; and presence of 70–100 calves or more of both genders with ≥200 eggs per gram of feces (EPG (sensitivity of 50 EPG. These calves were distributed into 10 groups (of 7–10 animals per farm and treated with ivermectin, doramectin, eprinomectin, fenbendazole, closantel, nitroxynil, disophenol, levamisole, albendazole, or moxidectin. Feces were collected 2 days before treatment and 14 days after treatment. Additional groups of 7–10 calves were used to test six different two-drug combinations at four of the studied farms. In general terms, fenbendazole was the most effective drug, followed by levamisole, disophenol, and moxidectin. However, parasite resistance to multiple drugs was found in all herds, especially in the genera Cooperia spp., Trichostrongylus spp., and Haemonchus spp.. Some of the two-drug combinations were effective against nematode populations identified as resistant to the same compounds when used as single drugs. The most effective combinations were moxidectin + levamisole, doramectin + fenbendazole, and levamisole + closantel. In this study, parasites resistant to the main commercially available anthelmintics were found in all herds, and some combinations of two active components belonging to different

Influence of sustained deworming pressure on the anthelmintic resistance status in strongyles of sheep under field conditions.

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Vijayasarathi, M K; Sreekumar, C; Venkataramanan, R; Raman, M

2016-10-01

Anthelmintic resistance (AR) status in Madras Red sheep from selected field flocks of a government funded scheme, covered by regular, sustained anthelmintic treatment for more than 10 years was determined. Parameters such as fecal egg count reduction test (FECRT), larval paralysis assay (LPA), and allele-specific-PCR (AS-PCR) were used to test the efficacy of fenbendazole, tetramisole, and ivermectin at recommended doses, in two seasons. Sheep belonging to non-beneficiary farmers were used as controls. Mean FECRT values of beneficiary group during winter and summer seasons were 77.77 and 76.04, 93.65 and 92.12, and 95.37 and 98.06 %, respectively, for fenbendazole, tetramisole, and ivermectin. In the non-beneficiary groups, the corresponding values were 74.82 and 81.09 %, 96.05 and 97.40 %, and 97.26 and 98.23 %, respectively. The results revealed resistance to fenbendazole, suspect resistance to tetramisole and susceptibility to ivermectin in beneficiary flock. In non-beneficiary flock, while resistance was noticed against fenbendazole, both tetramisole and ivermectin were effective. FECR values were found to be significantly different between beneficiary and non-beneficiary groups against tetramisole. The results of LPA confirmed this finding, as 50 % of the Haemonchus contortus larvae were paralyzed at the concentration of 0.0156 μg/ml in the beneficiary group, while those of non-beneficiary groups required lower concentrations of 0.0078 μg/ml. AS-PCR revealed the predominance of heterozygous susceptible population of H. contortus in the beneficiary group. In this study, resistance to fenbendazole was confirmed in both the beneficiary and non-beneficiary groups and this could be attributed to frequent use of benzimidazoles as seen from the deworming records. Emergence of tetramisole resistance was detected in the beneficiary group, where the drug was used continuously for 4 years. Ivermectin was found to be effective in all the flocks. It is

Decline in faecal worm egg counts in lambs suckling ewes treated with lipophilic anthelmintics: implications for hastening development of anthelmintic resistance.

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Dever, M L; Kahn, L P

2015-04-30

The aim for this experiment was to look for evidence of milk transfer of anthelmintic actives from ewes to their suckling lambs by reference to lambs' faecal worm egg count (WEC). The hypothesis was that WEC will decline in lambs suckling ewes treated with anthelmintics known to be lipophilic. One group of lactating Border Leicester×Merino ewes were treated (TX) with a combination of short (2.5mg/kg monepantel) and long-acting (1mg/kg moxidectin long-acting injection and a sustained release of 4.62g albendazole over 100 days) anthelmintics to remove gastrointestinal nematode (GIN) burden on day 0. The other group of lactating ewes (UTX) and all lambs (White Suffolk sires) were not treated. Ewes and lambs grazed as a single group and were exposed to GIN (predominately Haemonchus contortus) infection from pasture. Measurements were taken on days 0 and 7. WEC of lambs suckling UTX ewes increased from 6441 to 10,341 eggs per gram (epg) between days 0 and 7, while there was a 51% reduction in WEC for lambs suckling TX ewes. Packed cell volume (PCV) was significantly higher for lambs suckling TX ewes on day 7 compared to lambs suckling UTX ewes (28.5% vs. 24.9%, p=0.039). These results suggest that lambs suckling ewes treated with lipophilic anthelmintics received a sub-therapeutic dose via milk which would increase selection within the GIN (H. contortus) population for anthelmintic resistance. Copyright © 2015 Elsevier B.V. All rights reserved.

Anthelmintic resistance and multidrug resistance in sheep gastro-intestinal nematodes in France, Greece and Italy.

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Geurden, Thomas; Hoste, Herve; Jacquiet, Philippe; Traversa, Donato; Sotiraki, Smaragda; Frangipane di Regalbono, Antonio; Tzanidakis, Nikolaos; Kostopoulou, Despoina; Gaillac, Christie; Privat, Simon; Giangaspero, Annunziata; Zanardello, Claudia; Noé, Laura; Vanimisetti, Bindu; Bartram, David

2014-03-17

Anthelmintic resistance (AR) in ovine gastro-intestinal nematodes has been reported to affect the health and productivity of sheep globally. The objective of the present study was to evaluate the efficacy of commonly used oral drenches in sheep in France, Greece and Italy. In each country, 10 farms were selected. On each farm, 50 animals were blocked based on the pre-treatment faecal egg count (FEC). Within each block, animals were randomly allocated to one of 5 treatment groups. In addition to an untreated control group, there were 4 groups treated per oral route: moxidectin (MOX) and ivermectin (IVM), both at 0.2mg/kg bodyweight, levamisole (LEV; at 7.5mg/kg bodyweight) and a benzimidazole (BZ; at 3.75-5mg/kg bodyweight). In France, animals were not treated with LEV, but with netobimin (NET; at 7.5mg/kg bodyweight). The FEC was monitored using a modified McMaster technique. Two weeks after treatment, individual faecal samples were taken from all animals and efficacy was calculated as the difference between arithmetic mean FEC of the control group versus each respective treatment group. The results of the present study indicate the high efficacy of treatment with oral formulations of MOX (99-100%) and IVM (98-100%) on all farms, except on 1 farm in Greece. On this farm, multi drug resistance (MDR) was identified involving 4 anthelmintics (efficacy MOX: 91%; IVM: 0%; BZ: 58% and LEV: 87%). In Greece and Italy, AR against LEV and BZ was observed on some farms, with MDR involving both anthelmintics on 3 farms in Greece and on 2 farms in Italy. In France, AR against BZ and NET was observed on all 10 farms included. In all countries, Teladorsagia sp. was the most common nematode larva identified after treatment, followed by Haemonchus sp. and Trichostrongylus sp., with differences among farms and treatments. The current study confirms the high efficacy of oral treatments with MOX and IVM, even on farms with worm populations resistant to BZ, LEV or NET. This study also

Evaluation of Anthelmintic Resistance and Exhaust Air Dust PCR as a Diagnostic Tool in Mice Enzootically Infected with Aspiculuris tetraptera

Science.gov (United States)

Kapoor, Pratibha; Hayes, Yumiko O; Jarrell, Leslie T; Bellinger, Dwight A; Thomas, Rhiannon D; Lawson, Gregory W; Arkema, Jaclyn D; Fletcher, Craig A; Nielsen, Judith N

2017-01-01

The entry of infectious agents in rodent colonies occurs despite robust sentinel monitoring programs, strict quarantine measures, and stringent biosecurity practices. In light of several outbreaks with Aspiculuris tetraptera in our facilities, we investigated the presence of anthelmintic resistance and the use of exhaust air dust (EAD) PCR for early detection of A. tetraptera infection. To determine anthelmintic resistance, C57BL/6, DBA/2, and NCr nude mice were experimentally inoculated with embryonated A. tetraptera ova harvested from enzootically infected mice, followed by treatment with 150 ppm fenbendazole in feed, 150 ppm fenbendazole plus 5 ppm piperazine in feed, or 2.1 mg/mL piperazine in water for 4 or 8 wk. Regardless of the mouse strain or treatment, no A. tetraptera were recovered at necropsy, indicating the lack of resistance in the worms to anthelmintic treatment. In addition, 10 of 12 DBA/2 positive-control mice cleared the A. tetraptera infection without treatment. To evaluate the feasibility of EAD PCR for A. tetraptera, 69 cages of breeder mice enzootically infected with A. tetraptera were housed on a Tecniplast IVC rack as a field study. On day 0, 56% to 58% of the cages on this rack tested positive for A. tetraptera by PCR and fecal centrifugation flotation (FCF). PCR from EAD swabs became positive for A. tetraptera DNA within 1 wk of placing the above cages on the rack. When these mice were treated with 150 ppm fenbendazole in feed, EAD PCR reverted to pinworm-negative after 1 mo of treatment and remained negative for an additional 8 wk. The ability of EAD PCR to detect few A. tetraptera positive mice was investigated by housing only 6 infected mice on another IVC rack as a field study. The EAD PCR from this rack was positive for A. tetraptera DNA within 1 wk of placing the positive mice on it. These findings demonstrate that fenbendazole is still an effective anthelmintic and that EAD PCR is a rapid, noninvasive assay that may be a useful

Repurposing salicylanilide anthelmintic drugs to combat drug resistant Staphylococcus aureus.

Science.gov (United States)

Rajamuthiah, Rajmohan; Fuchs, Beth Burgwyn; Conery, Annie L; Kim, Wooseong; Jayamani, Elamparithi; Kwon, Bumsup; Ausubel, Frederick M; Mylonakis, Eleftherios

2015-01-01

Staphylococcus aureus is a Gram-positive bacterium that has become the leading cause of hospital acquired infections in the US. Repurposing Food and Drug Administration (FDA) approved drugs for antimicrobial therapy involves lower risks and costs compared to de novo development of novel antimicrobial agents. In this study, we examined the antimicrobial properties of two commercially available anthelmintic drugs. The FDA approved drug niclosamide and the veterinary drug oxyclozanide displayed strong in vivo and in vitro activity against methicillin resistant S. aureus (minimum inhibitory concentration (MIC): 0.125 and 0.5 μg/ml respectively; minimum effective concentration: ≤ 0.78 μg/ml for both drugs). The two drugs were also effective against another Gram-positive bacteria Enterococcus faecium (MIC 0.25 and 2 μg/ml respectively), but not against the Gram-negative species Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa and Enterobacter aerogenes. The in vitro antimicrobial activity of niclosamide and oxyclozanide were determined against methicillin, vancomycin, linezolid or daptomycin resistant S. aureus clinical isolates, with MICs at 0.0625-0.5 and 0.125-2 μg/ml for niclosamide and oxyclozanide respectively. A time-kill study demonstrated that niclosamide is bacteriostatic, whereas oxyclozanide is bactericidal. Interestingly, oxyclozanide permeabilized the bacterial membrane but neither of the anthelmintic drugs exhibited demonstrable toxicity to sheep erythrocytes. Oxyclozanide was non-toxic to HepG2 human liver carcinoma cells within the range of its in vitro MICs but niclosamide displayed toxicity even at low concentrations. These data show that the salicylanilide anthelmintic drugs niclosamide and oxyclozanide are suitable candidates for mechanism of action studies and further clinical evaluation for treatment of staphylococcal infections.

A field study on the anthelmintic resistance of Parascaris spp. in Arab foals in the Riyadh region, Saudi Arabia.

Science.gov (United States)

Alanazi, Abdullah D; Mukbel, Rami M; Alyousif, Mohamed S; AlShehri, Zafer S; Alanazi, Ibrahim O; Al-Mohammed, Hamdan I

2017-12-01

In the last decade, Parascaris spp. resistance to anthelmintics has been recorded in many countries. In Saudi Arabia, there are limited data available on Parascaris spp. resistance to anthelmintics. To determine the current status of ivermectin, abamectin and praziquantel combined, and fenbendazole resistance to Parascaris spp. in horses in Saudi Arabia. Three hundred and forty-one foals from eleven different farms were examined by faecal egg count (FEC). The foals were all Arab horses aged 17.2 ± 4.5 (SD) months. Ivermectin (n = 46 foals), abamectin and praziquantel combined (n = 46), and fenbendazole (n = 46) were administered on day 0 and faeces were collected on day 14. The study comprised 41 untreated foals as controls. Animals that have FEC of ≥100 eggs per gram (EPG) were used to measure anthelmintic efficacy. Parascaris spp. populations were considered susceptible when faecal egg count reduction (FECR) was ≥95% associated with a lower 95% confidence limit (LCL) >90%, suspected resistant when FECR ≤90% or LCL Saudi Arabia.

Overview of anthelmintic resistance of gastrointestinal nematodes of small ruminants in Brazil

Directory of Open Access Journals (Sweden)

Jordana Andrioli Salgado

2016-03-01

Full Text Available Abstract Frequent and inappropriate use of all classes of antiparasitic drugs in small ruminants has led to failures in their effectiveness, culminating in a global problem of anthelmintic resistance. Brazil stands out as one of the world’s leaders in publications about anthelmintic resistance, and for having the most numerous reports of this resistance in small ruminants in the Americas. These studies have involved mainly the fecal egg count reduction test (FECRT and its correlation with field management practices. In vivoeffectiveness testing is conducted in areas where livestock is of greater economic significance, e.g., in the South (sheep and Northeast (goats, or is important for research and economic centers, such as the Southeast (sheep. The most widely studied species is sheep, for which the widest range of drugs is also evaluated. Despite significant advances achieved in molecular research, laboratory analyses should include knowledge about the reality in the field so that they can become feasible for the producer. Moreover, molecular studies can be underpinned by the analysis of field studies, such as the maintenance of antiparasitic effectiveness over time and the mechanisms involved in this process.

A single amino acid substitution in isozyme GST mu in Triclabendazole resistant Fasciola hepatica (Sligo strain) can substantially influence the manifestation of anthelmintic resistance.

Science.gov (United States)

Fernández, V; Estein, S; Ortiz, P; Luchessi, P; Solana, V; Solana, H

2015-12-01

The helminth parasite Fasciola hepatica causes fascioliasis in human and domestic ruminants. Economic losses due to this infection are estimated in U$S 2000-3000 million yearly. The most common method of control is the use of anthelmintic drugs. However, there is an increased concern about the growing appearance of F. hepatica resistance to Triclabendazole (TCBZ), an anthelmintic with activity over adult and young flukes. F. hepatica has eight Glutathione S-Transferase (GST) isozymes, which are enzymes involved in the detoxification of a wide range of substrates through chemical conjugation with glutathione. In the present work we identified and characterized the GST mu gene isolated from the TCBZ-susceptible and TCBZ-resistant F. hepatica strains. Total RNA was transcribed into cDNA by reverse transcription and a 657 bp amplicon corresponding to the GST mu gene was obtained. The comparative genetic analysis of the GST mu gene of the TCBZ susceptible strain (Cullompton) and TCBZ resistant strain (Sligo) showed three nucleotide changes and one amino acid change at position 143 in the GST mu isozyme of the TCBZ-resistant strain. These results have potential relevance as they contribute better understand the mechanisms that generate resistance to anthelmintics. Copyright © 2015 Elsevier Inc. All rights reserved.

Anthelmintic Resistance of Strongyle Nematodes to Ivermectin and Fenbendazole on Cart Horses in Gondar, Northwest Ethiopia

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Zewdu Seyoum

2017-01-01

Full Text Available A study was conducted from November 2015 to April 2016 to determine fenbendazole and ivermectin resistance status of intestinal nematodes of cart horses in Gondar, Northwest Ethiopia. Forty-five strongyle infected animals were used for this study. The animals were randomly allocated into three groups (15 horses per group. Group I was treated with fenbendazole and Group II with ivermectin and Group III was left untreated. Faecal samples were collected from each cart horse before and after treatment. Accordingly, the reduction in the mean fecal egg count at fourteen days of treatment for ivermectin and fenbendazole was 97.25% and 79.4%, respectively. It was significantly different in net egg count between treatment and control groups after treatment. From the study, resistance level was determined for fenbendazole and suspected for ivermectin. In addition, a questionnaire survey was also conducted on 90 selected cart owners to assess their perception on anthelmintics. In the survey, the most available drugs in the study area used by the owners were fenbendazole and ivermectin. Most respondents have no knowledge about drug management techniques. Hence, animal health extension services to create awareness regarding anthelmintic management that plays a key role in reducing the anthelmintic resistance parasites.

Anthelmintic Resistance of Strongyle Nematodes to Ivermectin and Fenbendazole on Cart Horses in Gondar, Northwest Ethiopia.

Science.gov (United States)

Seyoum, Zewdu; Zewdu, Alemu; Dagnachew, Shimelis; Bogale, Basazinew

2017-01-01

A study was conducted from November 2015 to April 2016 to determine fenbendazole and ivermectin resistance status of intestinal nematodes of cart horses in Gondar, Northwest Ethiopia. Forty-five strongyle infected animals were used for this study. The animals were randomly allocated into three groups (15 horses per group). Group I was treated with fenbendazole and Group II with ivermectin and Group III was left untreated. Faecal samples were collected from each cart horse before and after treatment. Accordingly, the reduction in the mean fecal egg count at fourteen days of treatment for ivermectin and fenbendazole was 97.25% and 79.4%, respectively. It was significantly different in net egg count between treatment and control groups after treatment. From the study, resistance level was determined for fenbendazole and suspected for ivermectin. In addition, a questionnaire survey was also conducted on 90 selected cart owners to assess their perception on anthelmintics. In the survey, the most available drugs in the study area used by the owners were fenbendazole and ivermectin. Most respondents have no knowledge about drug management techniques. Hence, animal health extension services to create awareness regarding anthelmintic management that plays a key role in reducing the anthelmintic resistance parasites.

Effect of Anthelmintic Treatment on Insulin Resistance: A Cluster-Randomized, Placebo-Controlled Trial in Indonesia

NARCIS (Netherlands)

Tahapary, D.L.; Ruiter, K. de; Martin, I.; Brienen, E.A.T.; Lieshout, L. van; Cobbaert, C.M.; Soewondo, P.; Djuardi, Y.; Wiria, A.E.; Houwing-Duistermaat, J.J.; Sartono, E.; Smit, J.W.A.; Yazdanbakhsh, M.; Supali, T.

2017-01-01

Background: Emerging evidence suggests that helminth infections are associated with lower insulin resistance (IR). Current deworming programs might remove this helminth-associated protective effect. Therefore, we evaluated the anthelmintic treatment effect on changes in IR. Methods: We conducted a

Restrictions of anthelmintic usage

DEFF Research Database (Denmark)

Nielsen, Martin Krarup

2009-01-01

are based on parasitic knowledge. Despite recommendations given for the past two decades, strategies based on the selective therapy principle have not yet been implemented on a larger scale in equine establishments. In contrast, treatment regimens appear to be derived from recommendations originally given......Given the increasing levels of anthelmintic resistance in equine parasites, parasitologists now recommend traditional treatment approaches to be abandoned and replaced by more sustainable strategies. It is of crucial importance to facilitate veterinary involvement to ensure that treatment decisions...... that Danish veterinarians are now deeply involved with parasite management in equine establishments. However, little is known about the impact on levels of anthelmintic resistance and the risk of parasitic disease under these circumstances. In addition, the legislation makes huge demands on diagnosis...

Efficacy of anthelmintics on South American camelid (llama and alpaca) farms in Georgia.

Science.gov (United States)

Gillespie, Rose-Ann M; Williamson, Lisa H; Terrill, Thomas H; Kaplan, Ray M

2010-08-27

The number of South American camelid (SAC; llama and alpaca) farms is growing in the southeastern United States, and infection with gastrointestinal nematodes (GIN) is a major health concern in this region. There is widespread resistance to anthelmintic remedies in small ruminants (sheep and goats), but a paucity of information on llamas and alpacas. Anthelmintic resistance was evaluated on three SAC farms (two llama; one alpaca) in Georgia in the southern United States using fecal egg count reduction (FECR) tests. For each farm, animals were randomly assigned to 1 of 5 treatment groups based on initial fecal egg count (FEC) and number of animals available (2-5 groups, n=9-11 per treatment). Ivermectin (IVM, subcutaneous injection; 0.3mg/kg body weight (BW)) and a control group were tested on an alpaca farm, and fenbendazole (FBZ, oral; 10mg/kg BW; two farms), moxidectin (MOX oral; 0.2mg/kg BW; two farms), and levamisole (LEV, oral; 8 mg/kg BW; one farm) were added for the llama farms. Anthelmintic efficacy was determined by comparing FEC of treatment and control animals 14 days post-treatment, with resistance evaluated using the World Association for the Advancement of Veterinary Parasitology (WAAVP) guidelines. Based upon these guidelines, there was GIN resistance to IVM in both llamas and alpacas in Georgia and to FBZ on both llama farms where this drug was tested. There was MOX resistance on one llama farm using the FECR test, while there was no resistance to LEV detected in this study. These data demonstrate a serious emerging problem in the United States of llama and alpaca GIN resistant to drugs from two of the three major anthelmintic classes. (c) 2010 Elsevier B.V. All rights reserved.

Anthelmintic effect of Psidium guajava and Tagetes erecta on wild-type and Levamisole-resistant Caenorhabditis elegans strains.

Science.gov (United States)

Piña-Vázquez, Denia M; Mayoral-Peña, Zyanya; Gómez-Sánchez, Maricela; Salazar-Olivo, Luis A; Arellano-Carbajal, Fausto

2017-04-18

Psidium guajava and Tagetes erecta have been used traditionally to treat gastrointestinal parasites, but their active metabolites and mechanisms of action remain largely unknown. To evaluate the anthelmintic potential of Psidium guajava and Tagetes erecta extracts on Levamisole-sensitive and Levamisole-resistant strains of the model nematode Caenorhabditis elegans. Aqueous extracts of Psidium guajava (PGE) and Tagetes erecta (TEE) were assayed on locomotion and egg-laying behaviors of the wild-type (N2) and Levamisole-resistant (CB193) strains of Caenorhabditis elegans. Both extracts paralyzed wild-type and Levamisole-resistant nematodes in a dose-dependent manner. In wild-type worms, TEE 25mg/mL induced a 75% paralysis after 8h of treatment and PGE 25mg/mL induced a 100% paralysis after 4h of treatment. PGE exerted a similar paralyzing effect on N2 wild-type and CB193 Levamisole-resistant worms, while TEE only partially paralyzed CB193 worms. TEE 25mg/mL decreased N2 egg-laying by 65% with respect to the untreated control, while PGE did it by 40%. Psidium guajava leaves and Tagetes erecta flower-heads possess hydrosoluble compounds that block the motility of Caenorhabditis elegans by a mechanism different to that of the anthelmintic drug Levamisole. Effects are also observable on oviposition, which was diminished in the wild-type worms. The strong anthelmintic effects in crude extracts of these plants warrants future work to identify their active compounds and to elucidate their molecular mechanisms of action. Copyright © 2017 Elsevier Ireland Ltd. All rights reserved.

Biotransformation of selected anthelmintics in sheep tapeworm (Moniezia expansa)

OpenAIRE

Bečanová, Aneta

2013-01-01

Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Aneta Bečanová Supervisor: Mgr. Hana Bártíková, Ph.D. Title of diploma thesis: Biotransformation of selected anthelmintics in sheep tapeworm (Moniezia expansa) Biotransformation of anthelmintics is a process that prevents the parasite from adverse effects of xenobiotics. Therefore, it is the main factor that causes the reduction of an anthelmintic effect, resistance development and...

Efficacy of a combined oral formulation of derquantel-abamectin against the adult and larval stages of nematodes in sheep, including anthelmintic-resistant strains.

Science.gov (United States)

Little, Peter R; Hodge, Andrew; Maeder, Steven J; Wirtherle, Nicole C; Nicholas, David R; Cox, George G; Conder, George A

2011-09-27

Derquantel (DQL), a semi-synthetic member of a novel anthelmintic class, the spiroindoles, in combination with abamectin (ABA) [as the combination product STARTECT(®)] is a new entry for the treatment and control of parasites in sheep. The 19 studies reported herein were conducted in Australia, New Zealand, South Africa and the United Kingdom to demonstrate the efficacy of derquantel-abamectin (DQL-ABA) against a broad spectrum of gastrointestinal and respiratory nematodes of sheep, and to support registration of the combination product. Eleven studies were conducted using natural or experimental parasite infections with unknown or unconfirmed resistance, while eight studies utilised isolates/strains with confirmed or well characterised resistance to one or more currently available anthelmintics, including macrocyclic lactones. All studies included DQL-ABA and negative control groups, and in selected studies one or more reference anthelmintic groups were included. In all studies the commercial formulation of DQL-ABA was administered orally at 2mg/kg DQL and 0.2mg/kg ABA; placebo was administered in the same volume as DQL-ABA; and reference anthelmintics were administered as per label recommendations, except in one instance where levamisole was administered at twice the label dose. Infection, necropsy, worm collection and worm counting procedures were performed using standard techniques. Efficacy was calculated based on the percentage reduction in geometric mean worm count relative to negative control for each nematode species and lifecycle stage targeted. Twenty-two isolates/strains used in the eight studies targeting resistant worms had proven resistance: three to one anthelmintic class, eleven to two classes and eight to three or more classes; of these resistant strains, 16 demonstrated resistance to a macrocyclic lactone anthelmintic. Regardless of resistance status in the 19 studies, DQL-ABA controlled a broad range of economically important gastrointestinal

In vitro anthelmintic effects of Moringa oleifera leaves (FHI 109897 ...

African Journals Online (AJOL)

The need to control helminths resistance to the conventional anthelmintics necessitated the evaluation of phytochemicals in Moringa oleifera leaves for their possible in vitro anthelmintic effects on bovine helminth eggs. The quantitative faecal egg counts (FEC) were determined using the modified McMaster technique.

Anthelmintic-resistant strains of Haemonchus contortus and Trichostrongylus colubriformis isolated from an organic sheep and goat farm in Denmark

DEFF Research Database (Denmark)

Pena-Espinoza, Miguel Angel; Enemark, Heidi L.; Thansborg, Stig M.

2013-01-01

A suspected case of anthelmintic resistance (AR) was investigated in an organic dairy sheep and goat farm. The herd was established in 2007 by purchase of animals from a number of other farms. Selection for the study was based on history of anthelmintic-treatment failure. Forty-eight lambs and 48......) or 10 mg/kg closantel (CLO). Kids were treated with 10 mg/kg FBZ, 0.3 mg/kg MOX, 14 mg/kg LEV, 0.2 mg/kg IVM or 10 mg/kg CLO. FECs were performed at day of treatment and 14 days post treatment. In a subsequent investigation, faeces from adult goats were cultured to obtain 3rd-stage nematode larvae (L3...... %. This is the first isolation of BZ-resistant H. contortus and T. colubriformis in Denmark and highlights the need for continuous surveillance of AR in conventional and organic farms....

Field efficacy of four anthelmintics and confirmation of drug-resistant nematodes by controlled efficacy test and pyrosequencing on a sheep and goat farm in Denmark.

Science.gov (United States)

Peña-Espinoza, Miguel; Thamsborg, Stig M; Demeler, Janina; Enemark, Heidi L

2014-12-15

We describe a case of anthelmintic resistance on one of the largest organic small ruminant farms in Denmark. The flock was established in 2007 by purchase of animals from other Danish farms and had history of clinical parasitism, high mortality of young stock and anthelmintic treatment failure. In October 2011, 40 lambs and 40 kids were selected for a faecal egg count reduction test (FECRT) with fenbendazole (FBZ), ivermectin (IVM), moxidectin (MOX) and levamisole (LEV). Lambs were treated with the recommended sheep dose of each product while kids received the sheep dose of IVM, 1.5× sheep dose of MOX and 2× sheep dose of FBZ and LEV. Untreated lambs and kids were also included and three methods for calculating faecal egg count (FEC) reduction were compared. In a subsequent investigation, a controlled efficacy test (CET) with FBZ and IVM was performed in lambs infected with Haemonchus contortus and Trichostrongylus colubriformis isolated from adult goats on the farm. Recovered specimens of H. contortus were subjected to pyrosequencing for detection of single nucleotide polymorphisms (SNPs) related to benzimidazole (BZ) resistance. During the FECRT, FECs in untreated lambs dropped significantly by 47%. No FEC reduction was detected in untreated kids. After FBZ treatments, FEC reductions in lambs and kids ranged from 15 to 54% and 49-56%, respectively, according to the different calculation methods. Post IVM treatments, FEC reductions in lambs and kids varied between 71-90% and 81-83%, correspondingly. LEV and MOX reduced FECs by 98-100% in both species. In the CET, FBZ reduced H. contortus worm counts by 52-56% and no reduction in T. colubriformis counts were detected after treatment. IVM eliminated 100% of H. contortus and reduced T. colubriformis counts by 84-92%, according to different calculation methods. Pyrosequencing of isolated H. contortus revealed increased frequencies of the BZ resistance-related SNP in codon 200 of the β-tubulin isotype 1 gene

Assessment of gastrointestinal nematode infection, anthelmintic usage and husbandry practices on two small-scale goat farms in Malaysia.

Science.gov (United States)

Wong, Flora; Sargison, Neil

2018-03-01

Haemonchosis is a common problem on goat farms in tropical countries such as Malaysia. Prevention of production losses generally depends on the use of anthelmintic drugs, but is threatened by the emergence of anthelmintic resistance. This study investigates anthelmintic efficacy on small-scale Malaysian goat farms and describes putative risk factors. Adult goats had moderate to high pre-treatment faecal trichostrongyle egg counts, despite being housed on slatted floors and fed on cut-and-carry forage, raising questions about the source of nematode infection. Our results show multiple resistance to benzimidazole and macrocyclic lactone anthelmintic drugs and allow us to discuss the genetic origins of resistance with reference to farm husbandry and management. We conclude that improvement in Malaysian goat production efficiency will require the development of sustainable helminth control strategies, underpinned by a better understanding of the origins and population genetics of anthelmintic resistance.

Resistência anti-helmíntica em rebanhos caprinos no Estado do Ceará Anthelmintic resistance in goat herds in the State of Ceará

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Luiz Silva Vieira

1999-07-01

Full Text Available Um levantamento em nível de campo sobre resistência anti-helmíntica em nematódeos gastrintestinais de caprinos foi realizado em 34 rebanhos no Estado do Ceará. Em cada rebanho foram separados 30 cabritos, de ambos os sexos, com idade variando de 1 a 6 meses, os quais foram individualmente pesados, identificados e distribuídos em três tratamentos: 1 Oxfendazole na dose de 4,75mg/kg; 2 Levamisole na dose de 7,5 mg/kg e 3 Controle (não medicado. Os anti-helmínticos foram administrados de acordo com o peso individual de cada animal e, a dosagem utilizada para cada produto foi a recomendada pelo laboratório fabricante. Foram colhidas fezes dos animais de todos os tratamentos, para OPG e coprocultura, no dia da medicação e 7 dias após. Dos 34 rebanhos avaliados, 7 (20,6% apresentaram resistência aos imidazóis, 6 (17,6% aos benzimidazóis e 12 (35,3% revelaram resistência múltipla. Apenas em 9 rebanhos (26,5%, os nematódeos foram sensíveis aos anti-helmínticos avaliados. Através do questionário aplicado detectou-se que 52,9% dos caprinocultores entrevistados usavam anti-helmínticos de amplo espectro. Os resultados das coproculturas mostraram que os gêneros sobreviventes à medicação com oxfendazole foram principalmente Haemonchus sp, seguido em menor frequência por Oesophagostomum sp, enquanto que ao cloridrato de levamisole sobreviveram Haemonchus sp, Oesophagostomum sp e Trichostrongylus sp.Goats of 45 farms in the State of Ceará, Brazil, were treated with anthelmintics for gastrointestinal nematodes, and their resistance to the anthelmintics was evaluated. On each farm 30 kids were weighed, ear-tagged and divided into three groups of ten. The first group received oxfendazole at 4.75mg/kg, the second levamisole at 7.5mg/kg, and the third group remained untreated as control. All goats were drenched according to their individual body weight. Fecal samples were collected from all animals (treated and control on the day of

Comparative anthelmintic activity investigation of selected ethno-medicinal weeds

Science.gov (United States)

Pueblos, Kirstin Rhys S.; Bajalla, Mark; Pacheco, Dixie; Ganot, Sheila; Paig, Daisy; Tapales, Radyn; Lagare, Jeanne; Quimque, Mark Tristan J.

2017-01-01

Helminth infections are one of the seriously neglected potent diseases in many parts of the world. The problems of parasitic helminthes becoming resistant to currently available anthelmintic drugs pose a challenge for the search - relying on natural products - for new and better anthelmintics. In this paper, four abundant Philippine weeds: Chrysopogon aciculatus Trin. Cyperus brevifolius Rottb., Ruellia tuberosa Linn. and Saccharum spontaneum Linn. were investigated for their anthelmintic activities to establish basis of their folkloric claim. The hexane-soluble and chloroform-soluble extracts were obtained through sequential solvent partitioning of the crude ethanolic extract of the air-dried aerial part of each plant sample. Meanwhile, the decoction was obtained from fresh aerial part of the plant samples. All extracts were then subjected to in vitro anthelmintic screening at different concentration as per method of Ghosh, et al. against African nightcrawler earthworms (Eudrillus euginiae) in which the activity of the extracts was determined by correlation with time. The anthelmintic bioassay results revealed a dose-dependent toxicity relationship. It indicated relatively low anthelmintic activities of the decoction of the four plant samples as compared to their corresponding crude ethanol extracts. Among the crude ethanol extracts, C. brevifolius (CBE) gave fastest time to bring about paralysis and death to the test organisms at all concentrations tested. For the hexane extracts, R. tuberosa (RTH) gave better activity among other plant samples. Lastly, among the chloroform-soluble extracts, both that of C. brevifolius (CBC) and R. tuberosa (RTC) comparably showed strongest anthelmintic activities at all tested concentrations, thus, exhibited best anthelmintic activity that is remarkably comparable to the positive control, Mebendazole at the highest concentration tested. In fact, CBC and RTC showed highest anthelmintic potential compared to all extracts tested in

The extent of anthelmintic Resistance on Nematodes in communally grazed sheep and goats in a Semi-Arid area of North-west Province (RSA) / Tebogo Stanely Ramotshwane

OpenAIRE

Ramotshwane, Tebogo Stanely

2011-01-01

A survey was conducted to investigate the occurrence of anthelmintic resistance of nematodes in communally grazed sheep and goat herds in the Zeerust area of the North-West Province, Republic of South Africa. The fecal egg count reduction test (FECR%) tests were used to assess the sheep and goat small holder farmers. Efficacy of albendazole, ivermectin and closantel was done on both the treatment and control animals. Anthelmintic efficacy of 80% was considered a threshold for ...

Anthelmic Resistance Survey in Commercial Pig Herds in Thika District Kenya

International Nuclear Information System (INIS)

Kagira, J.M.

2002-01-01

The use of anthelmintics is the single most important action taken by the farmers to control worm infections. However due to improper use of these drugs, anthelmintic resistance (AR) has been reported in several countries. Most of the AR reports have been on sheep and goats with very few on pigs. Thus, the occurrence of resistance to three different anthelmintics was studied in four pig herds in Thika District, Kenya by means of faecal egg count reduction test (FECRT) and larval development assay (LDA) test. The FECRT showed that the piperazine and levamisole were less than 95% effective (one farm each) against Oesophagostomum spp., and this was confirmed using the LDA test. A resistant strain of Trichuris suis against levamisole was also detected in one farm. The results show that anthelmintic resistance is present in pig farms in Thika district, and by extension the problem could occur elsewhere in Kenya. Relevant veterinary authorities should advice farmers on strategies to reduce such occurrence

Comparison of biotransformation and efficacy of aminoacetonitrile anthelmintics in vitro.

Science.gov (United States)

Stuchlíková, Lucie; Lecová, Lenka; Jirásko, Robert; Lamka, Jiří; Vokřál, Ivan; Szotáková, Barbora; Holčapek, Michal; Skálová, Lenka

2016-02-01

The present in vitro study was designed to test and compare anthelmintic activity, hepatotoxicity, and biotransformation of four selected aminoacetonitrile derivatives (AADs): monepantel (MOP, anthelmintic approved for the treatment), AAD-970, AAD-1154, and AAD-1336. Micro-agar larval development test, MTT test of cytotoxicity, and biotransformation study coupled with Ultra high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) technique were used for this purpose. Larvae of two Haemonchus contortus strains (drug susceptible and multi-drug resistant) and primary cultures of rat and ovine hepatocytes served as model systems. All AADs (including MOP) exhibited significant larvicidal effect in H. contortus susceptible as well as multi-resistant strains, much higher than those of reference anthelmintics thiabendazole and flubendazole. AAD-1154 provides the best results for most tested parameters among all AADs in this study. The cytotoxicity test showed that all AADs can be considered as nontoxic for hepatocytes. In the biotransformation study, Phase I and Phase II metabolites of AADs were identified and schemes of possible metabolic pathways in ovine hepatocytes were proposed. Biotransformation of MOP was much more extensive than biotransformation of other AADs. Based on obtained results, AAD-1154 and AAD-1336 can be considered as promising candidates for further in vivo testing. Copyright © 2015 John Wiley & Sons, Ltd.

Anthelmintic residues in goat and sheep dairy products

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Jedziniak Piotr

2015-12-01

Full Text Available A multiresidue method (LC-MS/MS for determination of wide range of anthelmintics was developed. The method covered benzimidazoles: albendazole (and metabolites, cambendazole, fenbendazol (and metabolites, flubendazole (and metabolites, mebendazole (and metabolites, oxibendazole, thiabendazole (and metabolites, triclabendazole (and metabolites; macrocyclic lactones: abamectin, doramectin, emamectin, eprinomectin, ivermectin, moxidectin; salicylanilides: closantel, ioxynil, nitroxynil, oxyclosamide, niclosamide, rafoxanid and others: clorsulon, derquantel, imidocarb, monepantel (and metabolites, morantel, praziquantel, and pyrantel. The method was used to examine the potential presence of anthelmintics in goat and sheep milk and dairy products from the Polish market. A total of 120 samples of milk, yoghurt, cottage cheese, cream cheese, and curd were analysed. None of the samples were found positive above CCα (1-10 μg/kg except for one cottage cheese in which traces of albendazole sulfone were detected (5.2 ug/kg and confirmed. The results of the study showed negligible anthelmintic residues in the goat and sheep milk and dairy products and confirm their good quality.

Helminthiasis characterization and anthelmintic efficacy for ewes and lambs raised in tropical semiarid region.

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Bastos, Gabriela Almeida; Fonseca, Leydiana Duarte; de Paiva Ferreira, Adriano Vinícius; Costa, Marco Aurélio Morais Soares; Silva, Maria Luiza França; de Oliveira Vasconcelos, Viviane; de Sousa, Rogério Marcos; Duarte, Eduardo Robson

2017-06-01

In this study the helminthiasis and anthelmintic effectiveness in ewes and lambs were evaluated in a semiarid region of Brazil. Twelve sheep farms were investigated using semi-structured questionnaires and fecal egg count (FEC) reduction test was employed to analyze the profile of anthelmintic resistance. Groups of at least 10 animals with FEC ≥ 300 were selected. After 12 h of fasting, homogeneous groups of lambs or ewes were treated with albendazole, levamisole moxidectin, or oxfendazole and control groups were not treated. Feces were collected before treatments and 14 days after, and larvae genera were identified after cuprocultures in both periods. Extensive grazing was the predominant creation system, using hybrid Santa Ines animals. The separation by age was promoted in 75% of herds; however, maternity pickets there were only in three farms. The strategic treatments were performed only in 8.4% of sheep farms and 16.6% used the anthelmintic efficacy test and alternated anthelmintic classes after 1 year. The initial FEC means for lambs were significantly higher than ewe FEC averages. For lamb tests, moxidectin and levamisole showed higher efficacy (p ≤ 0.05) than benzimidazoles. For ewe tests, moxidectin and levamisole showed efficiencies >75%. Haemonchus spp. and Trichostrongylus spp. were the most frequent nematodes before treatments and the genus Haemonchus was the most prevalent after anthelmintic treatments (p < 0.05). Variations of anthelmintic susceptibility were observed for categories and herds evaluated, which emphasizes the importance of the effectiveness tests for the choice of anthelmintics for ewes and lambs.

Transport of anthelmintic benzimidazole drugs by breast cancer resistance protein (BCRP/ABCG2)

NARCIS (Netherlands)

Merino, G; Jonker, JW; Wagenaar, E; Pulido, MM; Molina, AJ; Alvarez, AI; Schinkel, AH

Methylcarbamate benzimidazoles [albendazole (ABZ), fenbendazole (FBZ), and their respective sulfoxide derivatives, albendazole sulfoxide (ABZSO) and oxfendazole (OXF)] are therapeutically important anthelmintic agents with low bioavailability. We studied their in vitro interaction with the apical

In-vitro anthelmintic activity of Coleus aromaticus root in Indian Adult Earthworm

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Arshad Hussain

2012-05-01

Full Text Available Objective: Anthelmintic resistance creates a major hitch over the decades throughout the world. As per WHO only synthetic drugs are frequently used in the treatment of helminth infestations in human beings but these synthetic drugs are out of reach of millions of people and have a lot of side effects. In view of this, an attempt has been made to study the anthelmintic activity of herbal drug. Methods: All the prototypes and the standard drug solution were freshly prepared before commencement of the experiments. All the earthworms were washed in normal saline solution before they were released into 10 ml of respective formulation as follows, vehicle (2% v/v Tween 80 in normal saline, and Piperazine Citrate (10 mg/ml and prototypes (10, 20 and 50mg/ml. Results: All the investigational extract acquired the anthelmintic activity at minimal dose of 10 mg/ml. its significant activity (P<0.05 at 10 mg/ml for time taken to paralysis and death when compared to the standard drugs Piperzine citrate used at 10 mg/ml respectively. Conclusions: Herbal drugs and synthetic drugs have equally effective in helminth infestations but methanolic extract has the maximum anthelmintic activity potential than other root extract of Coleus aromaticus.

Detection of Strongylus vulgaris in equine faecal samples by real-time PCR and larval culture ? method comparison and occurrence assessment

OpenAIRE

Kaspar, A.; Pfister, K.; Nielsen, M. K.; Silaghi, C.; Fink, H.; Scheuerle, M. C.

2017-01-01

BACKGROUND: Strongylus vulgaris has become a rare parasite in Germany during the past 50 years due to the practice of frequent prophylactic anthelmintic therapy. To date, the emerging development of resistance in Cyathostominae and Parascaris spp. to numerous equine anthelmintics has changed deworming management and the frequency of anthelmintic usage. In this regard, reliable detection of parasitic infections, especially of the highly pathogenic S. vulgaris is essential. In the current study...

Chemical composition, anthelmintic, antibacterial and antioxidant effects of Thymus bovei essential oil.

Science.gov (United States)

Jaradat, Nidal; Adwan, Lina; K'aibni, Shadi; Shraim, Naser; Zaid, Abdel Naser

2016-10-26

It has been recently recognized that oxidative stress, helminth and microbial infections are the cause of much illness found in the underdeveloped, developing and developed countries. The present study was undertaken to identify the chemical composition, and to assess anthelmintic, antimicrobial and antioxidant effects of Thymus bovei essential oil. The chemical composition of the essential oil was analyzed using gas chromatography mass spectrometry (GC-MS). Antimicrobial activity was tested against the selected strains from American Type Culture Collection (ATCC) and clinical isolates such as Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Methicillin Resistant Staphylococcus aureus, Candida albicans using MIC assay. The anthelmintic assay was carried out on adult earthworm (Pheretima posthuma), while antioxidant activity was analyzed using the 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging method. Trans-geraniol (35.38 %), α-citral (20.37 %) and β-citral (14.76 %) were the major compounds comprising 70.51 % of the essential oil. Our results showed that T. bovei essential oil exhibited strong anthelmintic activity, even higher than piperazine citrate, the used reference standard, with potential antioxidant activity almost equal to the Trolox standard. Furthermore, T. bovei essential oil had powerful antibacterial and antifungal activities against the studied pathogens. Essential oil of T. bovei exerted excellent antioxidant, antimicrobial, and anthelmintic activities. Moreover, this study found that T. bovei volatile oil contains active substances that could potentially be used as natural preservatives in food and pharmaceutical industries, these substances could also be employed for developing new anthelmintic, antimicrobial and antioxidant agents.

Metabolic profiling and in vitro assessment of anthelmintic fractions of Picria fel-terrae Lour.

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Rasika Kumarasingha

2016-12-01

Full Text Available Anthelmintic resistance is widespread in gastrointestinal nematode populations, such that there is a consistent need to search for new anthelmintics. However, the cost of screening for new compounds is high and has a very low success rate. Using the knowledge of traditional healers from Borneo Rainforests (Sarawak, Malaysia, we have previously shown that some traditional medicinal plants are a rich source of potential new anthelmintic drug candidates. In this study, Picria fel-terrae Lour. plant extract, which has previously shown promising anthelmintic activities, was fractionated via the use of a solid phase extraction cartridge and each isolated fraction was then tested on free-living nematode Caenorhabditis elegans and the parasitic nematode Haemonchus contortus. We found that a single fraction was enriched for nematocidal activity, killing ≥90% of C. elegans adults and inhibiting the motility of exsheathed L3 of H. contortus, while having minimal cytotoxic activity in mammalian cell culture. Metabolic profiling and chemometric analysis of the effective fraction indicated medium chained fatty acids and phenolic acids were highly represented.

The efficacy of 30% FENBENDAZOLE anthelmintic paste in cyathostominosis in horses

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Corina Badea

2015-06-01

Full Text Available Horse cyathostominosis is a large intestine helminthosis caused by parasites belonging to the family Strongylidae, subfamily Cyathostominae. The cyathostomins (small strongyles represent a challenge for the parasitologists and animal owners due to the different ontogenesis, the high number of parasite species and their ability to develop anthelmintic resistance. The faeces were examined by flotation (Willis method and the infestation level was determined by McMaster method in day 0, 7 and 14 post treatments. The product Vanbendazol (30% fenbendazole had a 97.7% efficacy in the treated horses from Şofronea, Arad County, using the Faecal egg count reduction test (FECRT. Also were performed Presidente and Borgsteede relations and the anthelmintic efficacy was 98.2% for the both relations.

A Novel Application of an Anthelmintic Mixture for Use against Gastrointestinal Parasites of Red Deer (Cervus elaphus

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P. L. Hughes

2018-01-01

Full Text Available A mixture of proprietary anthelmintics delivering 0.5 mg/kg moxidectin, 9.06 mg/kg oxfendazole, 15 mg/kg levamisole, and 0.08 mg/kg selenium on bodyweight basis per os to red deer is investigated. On a deer farm with a history of parasite problems, six weaner red deer were treated orally with a 50/50 mixture of Exodus Pour-On and Oxfen C Plus (Ex/Ox at a dose rate of 1 ml/5 kg bodyweight. Six herd mates were untreated. Eleven days later abomasal worm counts for the untreated deer revealed an arithmetic mean burden of 2,566 Ostertagia-type worms and 300 Trichostrongylus axei. No worms were detected in the abomasa of the treated group. Six yearling red deer were treated with the Ex/Ox combination and sent 39 days later to a slaughter plant where tissue samples were collected for residue analysis. Moxidectin was the only anthelmintic compound to show residues and the concentrations measured were well below maximum residue limits. Laboratory analysis of the Ex/Ox product after six-week storage at ambient temperature indicated good physical and chemical stability. These investigations support the hypothesis that the Ex/Ox combination can be an effective and practical anthelmintic option for use in red deer against a background of widespread gastrointestinal parasite resistance to the registered alternatives.

Voltammetric Determination of a Benzimidazole Anthelmintic Mixture ...

African Journals Online (AJOL)

2002-10-17

Oct 17, 2002 ... Electrochemistry, square-wave voltammetry, benzimidazole anthelmintics, ... potential application, cathodic reduction/anodic oxidation, the type of the electrode ... benzimidazole anthelmintic in 10 cm3 of methanol-formic acid.

Host pharmacokinetics and drug accumulation of anthelmintics within target helminth parasites of ruminants.

Science.gov (United States)

Lifschitz, A; Lanusse, C; Alvarez, L

2017-07-01

pharmacokinetic behaviour and identification of different factors affecting drug activity is important for achieving optimal parasite control and avoiding selection for drug resistance. The search for novel alternatives to deliver enhanced drug concentrations within target helminth parasites may contribute to avoiding misuse, and prolong the lifespan of existing and novel anthelmintic compounds in the veterinary pharmaceutical market.

In vitro screening of six anthelmintic plant products against larval Haemonchus contortus with a modified methyl-thiazolyl-tetrazolium reduction assay.

Science.gov (United States)

Hördegen, P; Cabaret, J; Hertzberg, H; Langhans, W; Maurer, V

2006-11-03

Because of the increasing anthelmintic resistance and the impact of conventional anthelmintics on the environment, it is important to look for alternative strategies against gastrointestinal nematodes. Phytotherapy could be one of the major options to control these pathologies. Extracts or ingredients of six different plant species were tested against exsheathed infective larvae of Haemonchus contortus using a modified methyl-thiazolyl-tetrazolium (MTT) reduction assay. Pyrantel tartrate was used as reference anthelmintic. Bromelain, the enzyme complex of the stem of Ananas comosus (Bromeliaceae), the ethanolic extracts of seeds of Azadirachta indica (Meliaceae), Caesalpinia crista (Caesalpiniaceae) and Vernonia anthelmintica (Asteraceae), and the ethanolic extracts of the whole plant of Fumaria parviflora (Papaveraceae) and of the fruit of Embelia ribes (Myrsinaceae) showed an anthelmintic efficacy of up to 93%, relative to pyrantel tartrate. Based on these results obtained with larval Haemonchus contortus, the modified MTT reduction assay could be a possible method for testing plant products with anthelmintic properties.

Prevalence of anthelmintic resistance on sheep and goat farms in the mid-Atlantic region and comparison of in vivo and in vitro detection methods

Science.gov (United States)

Despite strong economic opportunities and incentives for small ruminant production, their health and productivity are often severely affected by parasitic disease. To combat these effects, most farms administer anthelmintics to their animals at frequent intervals, and without consideration to princ...

Modelling the consequences of targeted selective treatment strategies on performance and emergence of anthelmintic resistance amongst grazing calves

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Zoe Berk

2016-12-01

Full Text Available The development of anthelmintic resistance by helminths can be slowed by maintaining refugia on pasture or in untreated hosts. Targeted selective treatments (TST may achieve this through the treatment only of individuals that would benefit most from anthelmintic, according to certain criteria. However TST consequences on cattle are uncertain, mainly due to difficulties of comparison between alternative strategies. We developed a mathematical model to compare: 1 the most ‘beneficial’ indicator for treatment selection and 2 the method of selection of calves exposed to Ostertagia ostertagi, i.e. treating a fixed percentage of the population with the lowest (or highest indicator values versus treating individuals who exceed (or are below a given indicator threshold. The indicators evaluated were average daily gain (ADG, faecal egg counts (FEC, plasma pepsinogen, combined FEC and plasma pepsinogen, versus random selection of individuals. Treatment success was assessed in terms of benefit per R (BPR, the ratio of average benefit in weight gain to change in frequency of resistance alleles R (relative to an untreated population. The optimal indicator in terms of BPR for fixed percentages of calves treated was plasma pepsinogen and the worst ADG; in the latter case treatment was applied to some individuals who were not in need of treatment. The reverse was found when calves were treated according to threshold criteria, with ADG being the best target indicator for treatment. This was also the most beneficial strategy overall, with a significantly higher BPR value than any other strategy, but its degree of success depended on the chosen threshold of the indicator. The study shows strong support for TST, with all strategies showing improvements on calves treated selectively, compared with whole-herd treatment at 3, 8, 13 weeks post-turnout. The developed model appeared capable of assessing the consequences of other TST strategies on calf populations.

Anthelmintic activity of Pongamia glabra

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R.B. Laware

2007-05-01

Full Text Available Leaves, wood, seed, bark and pericarp of the fruit of Pongamia glabra were separately dried, powdered and extracted with methanol in Soxhlet extractor. Anthelmintic activity of these various extracts was evaluatedon Indian adult earthworms, Pherentima posthuma. Results showed that the seed part of P. glabra took less time to cause paralysis and death of the earthworms; therefore, seeds were extracted successively withpetroleum ether, ethyl acetate and methanol in Soxhlet extractor. Again these extracts were screened for anthelmintic activity. Results showed that the ethyl acetate extract of seeds of P. glabra was most potentfollowed by petroleum ether extract. It can be concluded that anthelmintic activity of the seed of P. glabra is due to the active principles present mostly in the ethyl acetate and petroleum ether extracts.

In vitro screening of six anthelmintic plant products against larval Haemonchus contortus with a modified methyl-thiazolyl-tetrazolium reduction assay

OpenAIRE

Hördegen, P.; Cabaret, J.; Hertzberg, H.; Langhans, W.; Maurer, V.

2006-01-01

Because of the increasing anthelmintic resistance and the impact of conventional anthelmintics on the environment, it is important to look for alternative strategies against gastrointestinal nematodes. Phytotherapy could be one of the major options to control these pathologies. Extracts or ingredients of six different plant species were tested against exsheathed infective larvae of Haemonchus contortus using a modified methyl-thiazolyltetrazolium (MTT) reduction assay. Pyrantel tartrate was u...

Motility in the L3 stage is a poor phenotype for detecting and measuring resistance to avermectin/milbemycin drugs in gastrointestinal nematodes of livestock

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Melissa M. George

2018-04-01

Full Text Available Motility is a commonly used in vitro phenotype for assessing anthelmintic activity of candidate compounds, and for detecting anthelmintic resistance in nematodes. Third-stage larvae (L3 of parasitic nematodes are commonly used in motility-based assays because L3 are simple to obtain and can remain viable in storage for extended periods. To improve the measurement of motility of microscopic stages of nematodes, our laboratory developed the Worminator, which quantitatively measures motility of parasites. Using the Worminator, we compared the dose-response characteristics of several avermectin/milbemycin (AM compounds using L3 from both AM-susceptible and AM-resistant Cooperia spp. (abamectin, doramectin, eprinomectin, ivermectin, moxidectin and Haemonchus contortus (eprinomectin, ivermectin, moxidectin. Concentrations tested with the Worminator ranged from 0.156 to 40 μM. Differences in EC50 between AM-susceptible and AM-resistant isolates of Cooperia spp. and Haemonchus contortus were small, with resistance ratios ranging from 1.00 to 1.34 for Cooperia spp., 0.99 to 1.65 for Haemonchus contortus. Larval migration inhibition assays were conducted using the same isolates and were equally ineffective for detection of resistance with resistance ratios less than 2.0. These results contrast with those of the Larval Development Assay where we obtained a resistance ratio of 16.48 using the same isolates of Haemonchus contortus. Moreover, even at the highest concentration tested (40 μM, 100% inhibition of motility was never achieved and EC50 for Worminator assays were more than 100× higher than peak plasma levels achieved in vivo following treatment. These data demonstrate that dose-response characteristics for inhibition of motility in L3 of gastrointestinal nematodes of livestock do not significantly differ for AM-susceptible and AM-resistant isolates. These data challenge the suitability of motility as a phenotype for detecting and measuring

Developing novel anthelmintics from plant cysteine proteinases

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Stepek Gillian

2008-09-01

Full Text Available Abstract Intestinal helminth infections of livestock and humans are predominantly controlled by treatment with three classes of synthetic drugs, but some livestock nematodes have now developed resistance to all three classes and there are signs that human hookworms are becoming less responsive to the two classes (benzimidazoles and the nicotinic acetylcholine agonists that are licensed for treatment of humans. New anthelmintics are urgently needed, and whilst development of new synthetic drugs is ongoing, it is slow and there are no signs yet that novel compounds operating through different modes of action, will be available on the market in the current decade. The development of naturally-occurring compounds as medicines for human use and for treatment of animals is fraught with problems. In this paper we review the current status of cysteine proteinases from fruits and protective plant latices as novel anthelmintics, we consider some of the problems inherent in taking laboratory findings and those derived from folk-medicine to the market and we suggest that there is a wealth of new compounds still to be discovered that could be harvested to benefit humans and livestock.

Motility in the L3 stage is a poor phenotype for detecting and measuring resistance to avermectin/milbemycin drugs in gastrointestinal nematodes of livestock.

Science.gov (United States)

George, Melissa M; Lopez-Soberal, Lorraine; Storey, Bob E; Howell, Sue B; Kaplan, Ray M

2018-04-01

Motility is a commonly used in vitro phenotype for assessing anthelmintic activity of candidate compounds, and for detecting anthelmintic resistance in nematodes. Third-stage larvae (L3) of parasitic nematodes are commonly used in motility-based assays because L3 are simple to obtain and can remain viable in storage for extended periods. To improve the measurement of motility of microscopic stages of nematodes, our laboratory developed the Worminator, which quantitatively measures motility of parasites. Using the Worminator, we compared the dose-response characteristics of several avermectin/milbemycin (AM) compounds using L3 from both AM-susceptible and AM-resistant Cooperia spp. (abamectin, doramectin, eprinomectin, ivermectin, moxidectin) and Haemonchus contortus (eprinomectin, ivermectin, moxidectin). Concentrations tested with the Worminator ranged from 0.156 to 40 μM. Differences in EC 50 between AM-susceptible and AM-resistant isolates of Cooperia spp. and Haemonchus contortus were small, with resistance ratios ranging from 1.00 to 1.34 for Cooperia spp., 0.99 to 1.65 for Haemonchus contortus. Larval migration inhibition assays were conducted using the same isolates and were equally ineffective for detection of resistance with resistance ratios less than 2.0. These results contrast with those of the Larval Development Assay where we obtained a resistance ratio of 16.48 using the same isolates of Haemonchus contortus. Moreover, even at the highest concentration tested (40 μM), 100% inhibition of motility was never achieved and EC 50 for Worminator assays were more than 100× higher than peak plasma levels achieved in vivo following treatment. These data demonstrate that dose-response characteristics for inhibition of motility in L3 of gastrointestinal nematodes of livestock do not significantly differ for AM-susceptible and AM-resistant isolates. These data challenge the suitability of motility as a phenotype for detecting and measuring resistance to AM

Anthelmintic efficiency of doramectin, fenbendazole, and nitroxynil, in combination or individually, in sheep worm control

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Luciane Holsback

2016-01-01

Full Text Available Abstract The anthelmintic efficiency of doramectin, fenbendazole, and nitroxynil, used individually or in combination, was determined by the Fecal Egg Count Reduction (FECR test and cultivation of larvae of anthelminthic-treated sheep grouped as follows: G1 (doramectin, G2 (fenbendazole, G3 (nitroxynil, G4 (doramectin + fenbendazole, G5 (doramectin + nitroxynil, G6 (fenbendazole + nitroxynil, G7 (doramectin + nitroxynil + fenbendazole, G8 (untreated. In addition to individually used doramectin and fenbendazole, the helminths were also resistant to the combination of doramectin + fenbendazole; nitroxynil + fenbendazole; and doramectin + nitroxynil + fenbendazole, with their FECR rates ranging from 62-83%. The helminths showed possible nitroxynil-resistance, but had low resistance when the drug was administered in combination with doramectin. The evaluation of individual helminth species revealed that fenbendazole was fully effective against Cooperia; doramectin (G1, moderately effective against Haemonchus and insufficiently active againstCooperia; nitroxynil, effective againstHaemonchus and insufficiently active againstCooperia. It was concluded from the results that herd nematodes are resistant to doramectin, fenbendazole, and nitroxynil, and that the combined use of the drugs not only fails to significantly improve the anthelmintic efficiency against Haemonchus andCooperia, but is also cost-ineffective.

Anthelmintic efficiency of doramectin, fenbendazole, and nitroxynil, in combination or individually, in sheep worm control.

Science.gov (United States)

Holsback, Luciane; Luppi, Pedro Alex Ramsey; Silva, Camile Sanches; Negrão, Gustavo Kremer; Conde, Gabriel; Gabriel, Hugo Vinícius; Balestrieri, João Vitor; Tomazella, Lucas

2016-01-01

The anthelmintic efficiency of doramectin, fenbendazole, and nitroxynil, used individually or in combination, was determined by the Fecal Egg Count Reduction (FECR) test and cultivation of larvae of anthelminthic-treated sheep grouped as follows: G1 (doramectin), G2 (fenbendazole), G3 (nitroxynil), G4 (doramectin + fenbendazole), G5 (doramectin + nitroxynil), G6 (fenbendazole + nitroxynil), G7 (doramectin + nitroxynil + fenbendazole), G8 (untreated). In addition to individually used doramectin and fenbendazole, the helminths were also resistant to the combination of doramectin + fenbendazole; nitroxynil + fenbendazole; and doramectin + nitroxynil + fenbendazole, with their FECR rates ranging from 62-83%. The helminths showed possible nitroxynil-resistance, but had low resistance when the drug was administered in combination with doramectin. The evaluation of individual helminth species revealed that fenbendazole was fully effective against Cooperia; doramectin (G1), moderately effective against Haemonchus and insufficiently active against Cooperia; nitroxynil, effective against Haemonchus and insufficiently active against Cooperia. It was concluded from the results that herd nematodes are resistant to doramectin, fenbendazole, and nitroxynil, and that the combined use of the drugs not only fails to significantly improve the anthelmintic efficiency against Haemonchus and Cooperia, but is also cost-ineffective.

In silico exploration of the impact of pasture larvae contamination and anthelmintic treatment on genetic parameter estimates for parasite resistance in grazing sheep.

Science.gov (United States)

Laurenson, Y C S M; Kyriazakis, I; Bishop, S C

2012-07-01

A mathematical model was developed to investigate the impact of level of Teladorsagia circumcincta larval pasture contamination and anthelmintic treatment on genetic parameter estimates for performance and resistance to parasites in sheep. Currently great variability is seen for published correlations between performance and resistance, with estimates appearing to vary with production environment. The model accounted for host genotype and parasitism in a population of lambs, incorporating heritable between-lamb variation in host-parasite interactions, with genetic independence of input growth and immunological variables. An epidemiological module was linked to the host-parasite interaction module via food intake (FI) to create a grazing scenario. The model was run for a population of lambs growing from 2 mo of age, grazing on pasture initially contaminated with 0, 1,000, 3,000, or 5,000 larvae/kg DM, and given either no anthelmintic treatment or drenched at 30-d intervals. The mean population values for FI and empty BW (EBW) decreased with increasing levels of initial larval contamination (IL(0)), with non-drenched lambs having a greater reduction than drenched ones. For non-drenched lambs the maximum mean population values for worm burden (WB) and fecal egg count (FEC) increased and occurred earlier for increasing IL(0), with values being similar for all IL(0) at the end of the simulation. Drenching was predicted to suppress WB and FEC, and cause reduced pasture contamination. The heritability of EBW for non-drenched lambs was predicted to be initially high (0.55) and decreased over time with increasing IL(0), whereas drenched lambs remained high throughout. The heritability of WB and FEC for all lambs was initially low (∼0.05) and increased with time to ∼0.25, with increasing IL(0) leading to this value being reached at faster rates. The genetic correlation between EBW and FEC was initially ∼-0.3. As time progressed the correlation tended towards 0, before

Anthelmintic activity of ethanolic leaf extract of Jasminum mesnyi

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Vibhuti Dullu

2014-02-01

Full Text Available Objective: To evaluate the anthelmintic activity of ethanolic extract of leaves of Jasminum mesnyi. Methods: Anthelmintic activity was carried out on adult Indian earthworm Eisenia fetida. In this study, 20 mg/mL and 40 mg/mL concentrations of plant extract were tested which involved the time of paralysis and time of death of worm. Albendazole (10 mg/mL was taken as reference standard drug whereas distilled water was used as control. Results: The extract exhibited significant anthelmintic acitivity which was more in higher concentration extract. Conclusions: It was concluded from the present study that the plant exhibited significant anthelmintic activity.

In vitro anthelmintic activity of active compounds of the fringed rue Ruta chalepensis against dairy ewe gastrointestinal nematodes.

Science.gov (United States)

Ortu, E; Sanna, G; Scala, A; Pulina, G; Caboni, P; Battacone, G

2017-07-01

Infections by gastrointestinal nematodes negatively affect small ruminant health and at the same time cause substantial economic losses worldwide. Because resistance to conventional anthelmintic compounds is growing, target studies evaluating the effectiveness of alternative ingredients of botanical origin on gastrointestinal nematodes are needed. In this study, we evaluated the in vitro anthelmintic activity of Ruta chalepensis L. extracts on the third-stage larvae of sheep gastrointestinal nematodes. A methanol extract showed the highest anthelmintic activity, with an EC50 = 0.10 ± 0.06 mg/ml after 96 h, while the essential oil had an EC50 = 1.45 ± 1.22 mg/ml after 48 h. Moreover, three secondary metabolites of the essential oil, i.e. 2-decanone, 2-nonanone and 2-undecanone, showed EC50 values of 0.07 ± 0.06, 0.25 ± 0.29 and 0.88 ± 0.73 mg/ml at 24 h, respectively. The present study indicated that the R. chalepensis methanol extract, the essential oil and its metabolites 2-decanone, 2-nonanone and 2-undecanone showed promising anthelmintic activity on gastrointestinal nematodes.

Anthelmintic resistance impact on tropical beef cattle productivity: effect on weight gain of weaned calves.

Science.gov (United States)

Borges, Fernando A; Almeida, Gabriel D; Heckler, Rafael P; Lemes, Raul T; Onizuka, Marcel K V; Borges, Dyego G L

2013-03-01

The performance of grazing cattle in tropical areas is deeply influenced by parasitism, and the increasing reports of resistance are a threat to effective nematode control. The present study aimed to evaluate the effect of avermectins on the performance of weaned calves naturally infected by ivermectin-resistant gastrointestinal nematodes. The effect of four commercial endectocides (ivermectin 2.25 % + abamectin 1.25 %, ivermectin 3.15 %, doramectin 3.15 %, and doramectin 1 %) on parasitism and performance of a hundred weaned Nellore calves were evaluated during 112 days. The most effective anthelmintic showed efficacy of 84 % and resulted in an increase (P < 0.05) of live weight gain of 11.85 kg, compared to untreated group, 9.05 and 9.41 kg compared to those treated with more ineffective avermectins which showed efficacy of 0 and 48.2 %, respectively. A significant (P < 0.05) and weak negative correlation (r = -0.22) between the eggs per gram (EPG) and body weight was observed, indicating that even the low mean EPG (175 ± 150) observed at day 0 in the control group, with predominance of Haemonchus sp., was responsible for production losses. These results indicate that control of nematode parasites in beef cattle in the weaning phase may not result in increased productivity when carried out without technical criteria.

An ayurvedic formulation Sankat Mochan: A potent anthelmintic medicine

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Khomendra Kumar Sarwa

2017-01-01

Full Text Available Aim and Object: Sankat Mochan is an ayurvedic formulation used in the urban and rural area of India. This polyherbal formulation is used for general stomach problems including abdominal cramping and diarrhea. The present investigation evaluated the anthelmintic activity of an aqueous solution of an ayurvedic medicine Sankat Mochan. Materials and Method: Various concentrations (1%, 5%, and 10% of medicine were used for anthelmintic activity on Pheretima posthuma. Piperazine citrate (10 mg/ml was used as a reference standard and distilled water as a control. Result and Conclusion: The result showed that the Sankat Mochan possess anthelmintic activity more potent than that of piperazine citrate. Thus, Sankat Mochan may be used as a potent anthelmintic agent against helminthiasis.

Ethyl 3-oxo-2-(2,5-dioxopyrrolidin-3-yl)butanoate derivatives: Anthelmintic and cytotoxic potentials, antimicrobial and docking studies

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Mahmood, Fawad; Jan, Muhammad S.; Ahmad, Sajjad; Rashid, Umer; Ayaz, Muhammad; Ullah, Farhat; Hussain, Fida; Ahmad, Ashfaq; Khan, Arif-ullah; Aasim, Muhammad; Sadiq, Abdul

2017-12-01

The development of novel and more effective drugs is slow asthe resistance produced by pathogens.From the current scenario it can be imagine that this field of research will enter into the pre-antibiotic era. This work aims to study and evaluate the preliminary antibacterial, anthelmintic and cytotoxic potentials of ethyl 3-oxo-2-(2,5-dioxopyrrolidin-3-yl)butanoates.Among all of the four compounds, compound 2 has displayed remarkable potency with MIC values of 0.125, 0.083, 0.073 and 0.109 mg/ml against E. sakazakii, E. coli. S. aureus and K. pneumonia respectively. Compared to etoposide (LC50 9.8 µg/ml), the compounds demonstrated LC50 values from 280 to 765 µg/ml. For anthelmintic assay, three concentrations of each compound and standard drug were studied in determination of time of death of the two species. Excellent anthelmintic activity was observed by all four compounds against P. posthuma and A. gallibetter than standard albendazole. High GOLD fitness score data from docking analysis towards the targets represent better protein–ligand binding affinity and thus indicate a high propensity for all the active compounds to bind to the active site.Thepromisingin-vitro antimicrobial, anthelmintic activity and cytotoxicity data conclusively revealed that these compounds may serve as viable lead compounds for the treatment of bacterial and parasitic infections, and therefore, could help the medicinal chemists to design future chemotherapeutic agents to avoid rapid drug resistance.

Anthelmintics efficacy against intestinal strongyles in horses of Sardinia, Italy

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G. Sanna

2016-06-01

Full Text Available Intestinal strongyles (IS are the most important parasites of equids, due to their high prevalence worldwide, pathogenicity and the spread of drug-resistant populations. Despite the large number of horses bred in Sardinia Island, Italy, no data are available on the efficacy of anthelmintic compounds in the control of horse strongylosis. Therefore the aim of the present study was to evaluate the efficacy of five commercial anthelmintic formulations containing fenbendazole (FBZ, pyrantel (PYR, moxidectin (MOX and two ivermectin formulations (IVM1 and IVM2 against IS in Sardinia by performing a fecal egg count reduction test (FECRT and investigating the egg reappearance period (ERP after treatment. In total, 74 horses from 7 farms were examined. Coprocultures performed for individual fecal samples collected at the day of the treatment revealed that cyathostomins were the predominant parasitic species (98.6%. The FECR for all horses belonging to the treatment groups after two weeks was ≥95% with a 95% C.I. >90%. The expected ERP did not decrease in any of the treatment group as FECR values 90% for the entire duration of the trial until D150. The results of the present survey indicate that drug-resistant cyathostomin populations are not present in the examined horse population, contrariwise to what observed in other Italian and European regions. The reasons and implications of these results are discussed.

Phytochemical screening and anthelmintic activities of andrachne cordifolia

International Nuclear Information System (INIS)

Ajaib, M.; Wahla, S.Q.; Wahla, U.G.

2017-01-01

The present work was done to assess the phytochemical screening and anthelmintic potential of leaves and bark of Andrachne cordifolia (Wall. ex Decne.) Muell. The phytochemical screening for reducing sugars, terpenoids, cardiac glycosides, flavonoids, saponins, anthraquinones and alkaloids was performed. Saponins, terpenoids and tannins were reported in all the extracts. Anthelmintic activity of the extracts was carried out at four concentrations 20, 50, 80, 100 mg/mL. The time taken for death and paralysis of Haemonchus contortous were determined. Significance anthelmintic potential was shown by all the macerates which was dose dependent and compared to standard piperazine citrate. Chloroform macerate of leaf and petroleum ether extract of bark showed good activity. This may be because of the vicinity of phytochemical constituents like terpenoids, saponins and tannins in the plants. The results indicated that plant has secondary metabolites that have broad anthelmintic properties and plant might be a novel source of pharmaceutical drugs against helminthes. (author)

ANTHELMINTIC RESISTANCE TO BENZIMIDAZOLE IN GASTROINTESTINAL NEMATODES FROM SMALL RUMINANTS OF SEMI-ARID BRAZILIAN NORTHEAST RESISTÊNCIA AOS ANTI-HELMÍNTICOS BENZIMIDAZÓIS EM NEMATÓIDES GASTRINTESTINAIS DE PEQUENOS RUMINANTES DO SEMIÁRIDO NORDESTINO BRASILEIRO

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Ana Carolina Fonseca Lindoso Melo

2009-04-01

Full Text Available Resistance to benzimidazole anthelmintics is reported as an old and persistent problem in many parts of the world. Resistance development depends on the presence of resistance promoters and there are operational, genetic and bioecological factors. The objective of this work was to determine the prevalence of benzimidazole resistance and to study some variables associated with resistance development in small ruminant farms in the Brazilian northeastern semi-arid area. The work was accomplished in 25 sheep and goat farms in Limoeiro do Norte, Palhano, Jaguaruana, Itaiçaba, Aracati, Alto Santo, Morada Nova and Jaguaribe municipalities, in the state of Ceará, Brazil. The procedure used to detect anthelmintic resistant nematodes was the fecal egg count reduction test. In addition, a questionnaire about management practices, infrastructure, anthelmintic usage, flocks sanitary state and veterinary assistance was applied. Data were analyzed using RESO statistical program. The questionnaires were analyzed using Spearman correlation and the simple GLM. In sheep farms, the prevalence of benzimidazole resistance was 88% and in goat farms, it was 87.5%. In sheep and goats farms, Haemonchus spp was the most prevalent genus, followed by Trichostrongylus spp and Oesophagostomum spp. Among variables studied, treatment in the dry season was statistically significant (P = 0.03, pasture rotation was not significant (P = 0.17 but has a predictable value in resistance development.

KEY WORDS: Associated factors, benzimidazole, Ceará, resistance development.
A resistência a anti-helmínticos benzimidazóis é relatada como um antigo e persistente problema em diversas partes do mundo. O desenvolvimento da resistência depende da presença de promotores, os quais podem ser fatores operacionais, genéticos e bioecológicos. O objetivo do presente estudo foi determinar a prevalência da resistência a anti-helmínticos benzimidazóis e estudar algumas

Synthesis and evaluation of antimicrobial and anthelmintic activity of ...

Indian Academy of Sciences (India)

compounds were screened for antimicrobial activity and anthelmintic activity. The structural assignments of compounds were made on the basis of spectroscopic data and elemental analysis. Keywords. 10H-phenothiazines; Smiles rearrangement; sulphones; ribofuranosides; antimicrobial activity; anthelmintic activity. 1.

Single vs. double dose of copper oxide wire particles (COWP) for treatment of anthelmintic resistant Haemonchus contortus in weanling lambs.

Science.gov (United States)

Schweizer, Nikki M; Foster, Derek M; Knox, William B; Sylvester, Hannah J; Anderson, Kevin L

2016-10-15

Haemonchus contortus parasitism is a major disease of sheep, with these parasites frequently demonstrating multi-drug class anthelmintic resistance. Copper oxide wire particles (COWP) have shown potential as adjuncts or alternatives to anthelmintics in resistant flocks. The purpose of this study was to compare the efficacy of two different COWP treatment regimens or placebo in the control of H. contortus in weaned lambs within a flock historically shown to have multi-drug resistant H. contortus using the DrenchRite ® assay. Data from 43 lambs within 3 treatment groups in a double blind study were included in the experiment. Treatments were administered as a total of 2 boluses, each given on separate occasions (day 0 and day 42), so that each lamb received either 2 placebos, a single dose of 2g COWP followed by placebo, or two doses of 1g COWP. Strongyle-type fecal egg counts (FEC) were performed at initial treatment (day 0), on day 10, at second treatment (day 42), on day 52, and at study end (day 84). At the start of the trial, mean±standard deviation FEC were 1634.4±825.2, 2241.7±1496.8, and 2013.3±1194.2epg for the 2g, 1g×2, and control groups, respectively. At the end of the trial, FEC were 757.1±825.3, 483.4±557.2, and 1660.0±1345.3epg for the 2g, 1g×2, and control groups, respectively. Lambs given a 2g single dose of COWP or a 1g dose of COWP twice had reductions in strongyle-type FEC (p≤0.01) from trial start to trial end, whereas lambs given placebo did not. Average daily gains did not differ significantly among groups. Although copper is potentially toxic to sheep, no signs of toxicity were observed during this trial, which was consistent with similar studies at this treatment dose. The study indicated that administering COWP to lambs at weaning reduced FEC. Copyright © 2016 Elsevier B.V. All rights reserved.

Anthelmintic activity of acetone extracts from South African plants used on egg hatching of Haemonchus contortus

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Gerda Fouche

2016-07-01

Full Text Available The nematode, Haemonchus contortus, is responsible for major economic losses in the livestock industry. The management of parasites such as H. contortus has been through the use of synthetic parasiticides. This has resulted in the presence of residues in meat and milk, which affects food safety. The development of resistance to available anthelmintics coupled with their high cost has further complicated matters. This has led to the investigation of alternative methods to manage nematodes, including the use of plants and plant extracts as a potential source of novel anthelmintics. Acetone extracts were prepared from 15 South African plant species and their anthelmintic activity determined using the egg hatch assay (EHA. The leaf extract of Cleome gynandra had the best inhibitory activity (68% ± 3% at a concentration of 2.5 mg/mL, followed by the stem extract of Maerua angolensis (65% ± 5%. The extracts had a relatively low toxicity on Vero cells determined by the MTT (3-(4,5-dimethylthiazol-2-yl-2,5- diphenyltetrazolium bromide cellular assay.

Transcriptome analysis reveals molecular anthelmintic effects of procyanidins in C. elegans.

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Verena Spiegler

Full Text Available Worldwide, more than 1 billion people are affected by infestations with soil-transmitted helminths and also in veterinary medicine helminthiases are a severe threat to livestock due to emerging resistances against the common anthelmintics. Proanthocyanidins have been increasingly investigated for their anthelmintic properties, however, except for an interaction with certain proteins of the nematodes, not much is known about their mode of action. To investigate the anthelmintic activity on a molecular level, a transcriptome analysis was performed in Caenorhabditis elegans after treatment with purified and fully characterized oligomeric procyanidins (OPC. The OPCs had previously been obtained from a hydro-ethanolic (1:1 extract from the leaves of Combretum mucronatum, a plant which is traditionally used in West Africa for the treatment of helminthiasis, therefore, also the crude extract was included in the study. Significant changes in differential gene expression were observed mainly for proteins related to the intestine, many of which were located extracellularly or within cellular membranes. Among the up-regulated genes, several hitherto undescribed orthologues of structural proteins in humans were identified, but also genes that are potentially involved in the worms' defense against tannins. For example, T22D1.2, an orthologue of human basic salivary proline-rich protein (PRB 2, and numr-1 (nuclear localized metal responsive were found to be strongly up-regulated. Down-regulated genes were mainly associated with lysosomal activity, glycoside hydrolysis or the worms' innate immune response. No major differences were found between the groups treated with purified OPCs versus the crude extract. Investigations using GFP reporter gene constructs of T22D1.2 and numr-1 corroborated the intestine as the predominant site of the anthelmintic activity. The current findings support previous hypotheses of OPCs interacting with intestinal surface proteins

Anthelmintic potential of Calotropis procera, Azadirachta indica and Punica granatum against Gastrothylax indicus.

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Aggarwal, Rama; Kaur, Kiranjeet; Suri, Mansi; Bagai, Upma

2016-12-01

Anthelmintic activity of both ethanolic and aqueous extracts of Calotropis procera flowers, Azadirachta indica leaves and Punica granatum fruit peel in comparison with albendazole was evaluated through in vitro studies by the worm motility inhibition assay. Significant anthelmintic effects (p granatum ethanolic and aqueous extracts respectively, whereas it was 29.23 μg/ml ± 4.51 for albendazole. The mean mortality index (MI) was 1.0 and 0.90 for C. procera , 0.90 for A. indica and 0.73 and 0.80 for P. granatum ethanolic and aqueous extracts respectively whereas for albendazole it was 1.0. Percent mean worm motility inhibition (%WMI) was observed to be between 70 and 100 % for different extracts.Various concentrations (5-5000 μg/ml) of all the plant extracts and albendazole were used to detect their cytotoxic effects against HeLa cell line to determine CC-50 by MTT assay. CC-50 values, of all the plant extracts were determined to be >1000 μg/ml and for albendazole it was found to be >10 μM. All the three plants can be potential sources for novel anthelmintics.

Anthelmintic effects of forage chicory against parasitic nematodes in cattle

DEFF Research Database (Denmark)

Pena-Espinoza, Miguel Angel; Williams, Andrew; Thamsborg, Stig Milan

BACKGROUND: Chicory (Cichorium intybus) has potential as a natural anthelmintic in livestock, however evidence of efficacy against cattle nematodes is lacking. Here, we investigated anthelmintic effects of chicory in stabled calves. METHODS: Jersey male calves (2-4 months) were stratified by live...

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

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Silvestre A.

2004-06-01

Full Text Available 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.

Anthelmintic Potential of Thymoquinone and Curcumin on Fasciola gigantica.

Science.gov (United States)

Ullah, Rizwan; Rehman, Abdur; Zafeer, Mohd Faraz; Rehman, Lubna; Khan, Yasir A; Khan, M A Hannan; Khan, Shahper N; Khan, Asad U; Abidi, S M A

2017-01-01

Fasciolosis an economically important global disease of ruminants in the temperate and tropical regions, caused by Fasciola hepatica and F. gigantica, respectively, also poses a potential zoonotic threat. In India alone it causes huge losses to stakeholders. Anthelmintics including triclabendazole have been used to control this menace but the emerging resistance against the available compounds necessitates identification of novel and alternative therapeutic measures involving plant derived natural compounds for their anthelmintic potential. Thymoquinone (T) and curcumin (C), the active ingredients of Nigella sativa and Curcuma longa respectively have been used as antiparasitic agents but the information on their flukicidal effect is very limited. Adult flukes of F. gigantica were in vitro exposed to different concentrations of thymoquinone and curcumin separately for 3h at 37+ 1°C. A significant (p<0.05) reduction in the worm motility at 60 μM concentration of both T and C was observed though all the worms remained alive after 3h exposure, whereas the effect on egg shedding was statistically insignificant. Pronounced tegumental disruptions and erosion of spines in the posterior region and around the acetabulum was evident. A significant (p<0.05) decrease in glutathione-S-transferase and superoxide dismutase activity and reduced glutathione (GSH) level was observed, while protein carbonylation increased differentially. A significant inhibition of CathepsinL (CatL) gene expression in thymoquinone treated worms was also evident. Further, in silico molecular docking of T and C with CatL revealed a stronger interaction of curcumin with the involvement of higher number of amino acids as compared to thymoquinone that could be more effective in inhibiting the antioxidant enzymes of F. gigantica. It is concluded that both the compounds understudy will decrease the detoxification ability of F. gigantica, while inhibition of CatL will significantly affect their virulence

Anthelmintic Potential of Thymoquinone and Curcumin on Fasciola gigantica.

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Rizwan Ullah

Full Text Available Fasciolosis an economically important global disease of ruminants in the temperate and tropical regions, caused by Fasciola hepatica and F. gigantica, respectively, also poses a potential zoonotic threat. In India alone it causes huge losses to stakeholders. Anthelmintics including triclabendazole have been used to control this menace but the emerging resistance against the available compounds necessitates identification of novel and alternative therapeutic measures involving plant derived natural compounds for their anthelmintic potential. Thymoquinone (T and curcumin (C, the active ingredients of Nigella sativa and Curcuma longa respectively have been used as antiparasitic agents but the information on their flukicidal effect is very limited. Adult flukes of F. gigantica were in vitro exposed to different concentrations of thymoquinone and curcumin separately for 3h at 37+ 1°C. A significant (p<0.05 reduction in the worm motility at 60 μM concentration of both T and C was observed though all the worms remained alive after 3h exposure, whereas the effect on egg shedding was statistically insignificant. Pronounced tegumental disruptions and erosion of spines in the posterior region and around the acetabulum was evident. A significant (p<0.05 decrease in glutathione-S-transferase and superoxide dismutase activity and reduced glutathione (GSH level was observed, while protein carbonylation increased differentially. A significant inhibition of CathepsinL (CatL gene expression in thymoquinone treated worms was also evident. Further, in silico molecular docking of T and C with CatL revealed a stronger interaction of curcumin with the involvement of higher number of amino acids as compared to thymoquinone that could be more effective in inhibiting the antioxidant enzymes of F. gigantica. It is concluded that both the compounds understudy will decrease the detoxification ability of F. gigantica, while inhibition of CatL will significantly affect

RESISTÊNCIA ANTI-HELMÍNTICA EM REBANHOS OVINOS DA REGIÃO DA ASSOCIAÇÃO DOS MUNICÍPIOS DO ALTO IRANI (AMAI, OESTE DE SANTA CATARINA ANTHELMINTIC RESISTANCE ON SHEEP FLOCKS FROM ASSOCIATION OF THE MUNICIPALITIES OF THE ALTO IRANI REGION (AMAI, WEST OF SANTA CATARINA STATE, BRAZIL

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Ingrid Kelly Zanchet

2007-09-01

reduce FEC up to 95% were considered effective. The mo-lecules used were: levamisole (7.5 mg/kg, closantel (7.5 mg/Kg, albendazole (10 and 5 mg/Kg, ivermectin and moxidectin (0.2 mg/Kg. It was detected resistance of the gastrointestinal nematodes to all the anthelmintics tested. One hundred percent of the farms showed resistance to the ivermectin, 66.7% to the moxidectin, 44.4% to levamisole e 75% to albendazole. To the macrocyclic lactones and al-bendazol, both Haemonchus sp. and Trichostrongylus sp. were resistant. To levamisole, resistance is restricted to the genus Trichostrongylus. It was also detected one population of Haemonchus sp. resistant to Closantel and one of Nematodirus sp. resistant to albendazole. These data show the urgency to spread means of integrated control of para-sites, with the objective to enlarge the life of the few drugs that remain effective.

KEY-WORDS: Anthelmintic resistance, Santa Catarina, SHEEP.

Anthelmintic properties of traditional African and Caribbean medicinal plants

DEFF Research Database (Denmark)

Williams, Andrew R.; Soelberg, Jens; Jäger, Anna

2016-01-01

Ascariasis affects more than 1 billion people worldwide, mainly in developing countries, causing substantial morbidity. Current treatments for Ascaris infection are based on mass drug administration (MDA) with synthetic anthelmintic drugs such as albendazole, however continual re-infection and th......Ascariasis affects more than 1 billion people worldwide, mainly in developing countries, causing substantial morbidity. Current treatments for Ascaris infection are based on mass drug administration (MDA) with synthetic anthelmintic drugs such as albendazole, however continual re...

Anthelmintic efficacy on Parascaris equorum in foals on Swedish studs

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Christensson Dan

2009-11-01

Full Text Available Abstract Background In the last few years stud farms have experienced increasing problems with Parascaris equorum infections in foals despite intensive deworming programs. This has led to the question as to whether the anthelmintic drugs used against this parasite are failing. This study aimed to investigate the efficacy of ivermectin, fenbendazole and pyrantel on the faecal output of ascarid eggs of foals. Methods A Faecal Egg Count Reduction Test (FECRT was performed on nine large studs in Sweden. Anthelmintic drugs were given orally and faecal samples were examined for ascarid eggs on the day of deworming and 14 days later. Faecal Egg Count Reductions (FECRs were calculated on arithmetic means of transformed individual FECRs and on arithmetic means of individual FECRs. Results Seventy-nine (48% out of a total of 165 foals sampled were positive for P. equorum eggs before deworming and 66 of these met the criteria for being used in the efficacy assessment. It was shown that there was no, or very low activity of ivermectin on the output of ascarid eggs in the majority of the foals, whereas for fenbendazole and pyrantel it was >90%. Conclusion Ivermectin resistance was shown in 5 out of 6 farms. Therefore, ivermectin should not be the drug of choice in the control of P. equorum infections in foals. According to the results of this study, fenbendazole or pyrantel are still effective and should be used against this parasite.

Drug resistance makes new control measures of stomach parasites in small ruminants necessary = Resistentie maakt nieuwe aanpak van de bestrijding van maagdarmwormen bij kleine herkauwers noodzakelijk

NARCIS (Netherlands)

Eysker, M.; Borgsteede, F.H.M.; Ploeger, H.W.; Vellema, P.

2005-01-01

Control of Parasitic gastroenteritis in small ruminants is threatened by the worldwide growing problem of anthelmintic resistance. Therfore, alternativeapproaches for worm control are imperative. Of utmost importance is to slow down selection pressure for anthelmintic resistance by using alternative

Anthelmintic potential of various extracts of the rhizomes of Curcuma amada Roxb.

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Maharudra S. Rakh

2014-02-01

Full Text Available Objective: To investigate the anthelmintic potential of various extracts of the rhizomes of Curcuma amada Roxb. Methods: Cyclohexane, ethyl acetate, methanol and aqueous extracts of Curcuma amada Roxb. against Pheretima posthuma, an Indian species of earthworm. Various concentrations (25, 50 and 100 mg/mL of these extract were evaluated for anthelmintic activity by recording the time required for paralysis and death of worms. Albendazole was used as a reference standard drug. Results: Methanol and aqueous extracts shows most significant anthelmintic activity compared with standard drug. Conclusions: Since methanol and aqueous extracts were tested by different chemical tests. It shows presence of carbohydrates, steroid, alkaloids, glycosides and flavanoids. These phytoconstituents may be responsible for the said activities.

Selection and characterisation of monepantel resistance in Teladorsagia circumcincta isolates.

Science.gov (United States)

Bartley, D J; Devin, L; Nath, M; Morrison, A A

2015-08-01

Monepantel (MPTL) is one of two new anthelmintic compounds introduced onto the sheep market to control gastro-intestinal nematodes. Resistance to this compound is rare but has been reported. In order to preserve the efficacy of this and other anthelmintics, it is essential to understand both (a) the mechanisms involved in the selection of resistance and (b) how the parasites evolve to deal with these compounds. To address these questions three MPTL-resistant Teladorsagia circumcincta isolates (MTci2-11, MTci5-13 and MTci7-12) have been artificially selected in vivo from phenotypically characterised parent isolates (MTci2, MTci5, MTci7 respectively). The selection process involved collecting and culturing eggs from surviving worms from sheep administered sub-optimal dosages of MPTL (Zolvix®) to provide infective larvae to infect further sheep until resistant isolates were generated (between 9 and 13 rounds of selection). A controlled efficacy test was conducted using the original parental isolates and the newly generated MPTL resistant isolates (n = 5 per group). Selected isolates were assessed both under anthelmintic stress (Zolvix®, 2.5 mg/kg bodyweight; MTci-MPTL) and at rest (untreated, MTci-CON). A number of life-history traits were assessed, namely, worm establishment rates, time to patency, faecal egg output, body length of adults and eggs in utero. The estimated resistance status of the selected isolates was confirmed with 48%, 28% and 9% reductions in worm burden at 7-days post Zolvix® administration for MTci2-11-MPTL, MTci5-13-MPTL and MTci7-12-MPTL, respectively, compared with untreated controls. One of the selected isolates MTci7-12-CON showed significantly greater total worm burden (p = 0.025), greater establishment rate (p = 0.033), decreased time to patency (p = 0.048), higher cumulative egg outputs (p = 0.002) compared with its parental derivative MTci7. The trial results suggest that anthelmintic selection in T

Introgression of ivermectin resistance genes into a susceptible Haemonchus contortus strain by multiple backcrossing.

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Elizabeth Redman

2012-02-01

Full Text Available Anthelmintic drug resistance in livestock parasites is already widespread and in recent years there has been an increasing level of anthelmintic drug selection pressure applied to parasitic nematode populations in humans leading to concerns regarding the emergence of resistance. However, most parasitic nematodes, particularly those of humans, are difficult experimental subjects making mechanistic studies of drug resistance extremely difficult. The small ruminant parasitic nematode Haemonchus contortus is a more amenable model system to study many aspects of parasite biology and investigate the basic mechanisms and genetics of anthelmintic drug resistance. Here we report the successful introgression of ivermectin resistance genes from two independent ivermectin resistant strains, MHco4(WRS and MHco10(CAVR, into the susceptible genome reference strain MHco3(ISE using a backcrossing approach. A panel of microsatellite markers were used to monitor the procedure. We demonstrated that after four rounds of backcrossing, worms that were phenotypically resistant to ivermectin had a similar genetic background to the susceptible reference strain based on the bulk genotyping with 18 microsatellite loci and individual genotyping with a sub-panel of 9 microsatellite loci. In addition, a single marker, Hcms8a20, showed evidence of genetic linkage to an ivermectin resistance-conferring locus providing a starting point for more detailed studies of this genomic region to identify the causal mutation(s. This work presents a novel genetic approach to study anthelmintic resistance and provides a "proof-of-concept" of the use of forward genetics in an important model strongylid parasite of relevance to human hookworms. The resulting strains provide valuable resources for candidate gene studies, whole genome approaches and for further genetic analysis to identify ivermectin resistance loci.

Synthesis and anthelmintic activity of some hybrid Benzimidazolyl ...

African Journals Online (AJOL)

Erah

parasites a major economic and food security issue. In this context ... a new hybrid chemical profile of chalcone and benzimidazole .... standard anthelmintic drugs (fenbendazole ... laboratory according to the classical Claisen-. Schmidt using.

Direct anthelmintic effects of condensed tannins from diverse plant sources against Ascaris suum.

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Andrew R Williams

Full Text Available Ascaris suum is one of the most prevalent nematode parasites in pigs and causes significant economic losses, and also serves as a good model for A. lumbricoides, the large roundworm of humans that is ubiquitous in developing countries and causes malnutrition, stunted growth and compromises immunity to other pathogens. New treatment options for Ascaris infections are urgently needed, to reduce reliance on the limited number of synthetic anthelmintic drugs. In areas where Ascaris infections are common, ethno-pharmacological practices such as treatment with natural plant extracts are still widely employed. However, scientific validation of these practices and identification of the active compounds are lacking, although observed effects are often ascribed to plant secondary metabolites such as tannins. Here, we extracted, purified and characterised a wide range of condensed tannins from diverse plant sources and investigated anthelmintic effects against A. suum in vitro. We show that condensed tannins can have potent, direct anthelmintic effects against A. suum, as evidenced by reduced migratory ability of newly hatched third-stage larvae and reduced motility and survival of fourth-stage larvae recovered from pigs. Transmission electron microscopy showed that CT caused significant damage to the cuticle and digestive tissues of the larvae. Furthermore, we provide evidence that the strength of the anthelmintic effect is related to the polymer size of the tannin molecule. Moreover, the identity of the monomeric structural units of tannin polymers may also have an influence as gallocatechin and epigallocatechin monomers exerted significant anthelmintic activity whereas catechin and epicatechin monomers did not. Therefore, our results clearly document direct anthelmintic effects of condensed tannins against Ascaris and encourage further in vivo investigation to determine optimal strategies for the use of these plant compounds for the prevention and

Molecular Detection of Antimicrobial Resistance

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Fluit, Ad C.; Visser, Maarten R.; Schmitz, Franz-Josef

2001-01-01

The determination of antimicrobial susceptibility of a clinical isolate, especially with increasing resistance, is often crucial for the optimal antimicrobial therapy of infected patients. Nucleic acid-based assays for the detection of resistance may offer advantages over phenotypic assays. Examples are the detection of the methicillin resistance-encoding mecA gene in staphylococci, rifampin resistance in Mycobacterium tuberculosis, and the spread of resistance determinants across the globe. However, molecular assays for the detection of resistance have a number of limitations. New resistance mechanisms may be missed, and in some cases the number of different genes makes generating an assay too costly to compete with phenotypic assays. In addition, proper quality control for molecular assays poses a problem for many laboratories, and this results in questionable results at best. The development of new molecular techniques, e.g., PCR using molecular beacons and DNA chips, expands the possibilities for monitoring resistance. Although molecular techniques for the detection of antimicrobial resistance clearly are winning a place in routine diagnostics, phenotypic assays are still the method of choice for most resistance determinations. In this review, we describe the applications of molecular techniques for the detection of antimicrobial resistance and the current state of the art. PMID:11585788

Anthelmintic activity of Cocos nucifera L. against sheep gastrointestinal nematodes.

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Oliveira, L M B; Bevilaqua, C M L; Costa, C T C; Macedo, I T F; Barros, R S; Rodrigues, A C M; Camurça-Vasconcelos, A L F; Morais, S M; Lima, Y C; Vieira, L S; Navarro, A M C

2009-01-22

The development of anthelmintic resistance has made the search for alternatives to control gastrointestinal nematodes of small ruminants imperative. Among these alternatives are several medicinal plants traditionally used as anthelmintics. This work evaluated the efficacy of Cocos nucifera fruit on sheep gastrointestinal parasites. The ethyl acetate extract obtained from the liquid of green coconut husk fiber (LGCHF) was submitted to in vitro and in vivo tests. The in vitro assay was based on egg hatching (EHT) and larval development tests (LDT) with Haemonchus contortus. The concentrations tested in the EHT were 0.31, 0.62, 1.25, 2.5 and 5 mg ml(-1), while in the LDT they were 5, 10, 20, 40 and 80 mg ml(-1). The in vivo assay was a controlled test. In this experiment, 18 sheep infected with gastrointestinal nematodes were divided into three groups (n=6), with the following doses administered: G1-400 mg kg(-1) LGCHF ethyl acetate extract, G2-0.2 mg kg(-1) moxidectin (Cydectin) and G3-3% DMSO. The worm burden was analyzed. The results of the in vitro and in vivo tests were submitted to ANOVA and analyzed by the Tukey and Kruskal-Wallis tests, respectively. The extract efficacy in the EHT and LDT, at the highest concentrations tested, was 100% on egg hatching and 99.77% on larval development. The parameters evaluated in the controlled test were not statistically different, showing that despite the significant results of the in vitro tests, the LGCHF ethyl acetate extract showed no activity against sheep gastrointestinal nematodes.

Ethnobotanical survey of medicinal plants used as anthelmintic remedies in Gabon.

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Bajin Ba Ndob, Idensi; Mengome, Line Edwige; Bourobou Bourobou, Henri-Paul; Lossangoye Banfora, Yvon; Bivigou, Francis

2016-09-15

In this article, we report on an ethnobotanical survey realized at the Peyrie market in Libreville on Gabonese medicinal plants used to treat helminthiasis. While several alerts about cases of resistance to conventional anthelmintic treatments are causing to fear a public and animal health issue, the search for new sources of active compounds becomes an urgent issue. In Gabon like in many developing countries, people regularly turn to traditional medicine in case of physical ailments and/or spiritual healing therapies. To determine which medicinal plants are traditionally used by the populations of Libreville to fight against nematodes, medicinal plant traders were interviewed with standardized questionnaires. The surveys were conducted in the main market of Libreville. Ethnobotanical data such as frequency and percentage of families, species, administrations pathways, modes of preparations and parts of plants used were analyzed and summarized. Thirty-four (34) traders were interviewed belonging to five (5) different ethnic groups. Twenty-four 24 plants used to treat intestinal, cutaneous and ocular helminthiasis were listed. The healers mainly turned towards to ligneous species. The parts of the plant used are mostly leaves and trunk bark. Most of the traditional remedies are prepared directly in water and four (4) principal routes were used for administration namely, oral, rectal, ocular and dermal. This study allowed us to list anthelmintic species which will be subjected to a series of chemical and pharmacological assays. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

HIV resistance testing and detected drug resistance in Europe

DEFF Research Database (Denmark)

Schultze, Anna; Phillips, Andrew N; Paredes, Roger

2015-01-01

to Southern Europe. CONCLUSIONS: Despite a concurrent decline in virological failure and testing, drug resistance was commonly detected. This suggests a selective approach to resistance testing. The regional differences identified indicate that policy aiming to minimize the emergence of resistance......OBJECTIVES: To describe regional differences and trends in resistance testing among individuals experiencing virological failure and the prevalence of detected resistance among those individuals who had a genotypic resistance test done following virological failure. DESIGN: Multinational cohort...... study. METHODS: Individuals in EuroSIDA with virological failure (>1 RNA measurement >500 on ART after >6 months on ART) after 1997 were included. Adjusted odds ratios (aORs) for resistance testing following virological failure and aORs for the detection of resistance among those who had a test were...

Nippostronglylus brasiliensis infection in the rat: effect of iron and protein deficiency and dexamethasone on the efficacy of benzimidazole anthelmintics.

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Duncombe, V M; Bolin, T D; Davis, A E; Kelly, J D

1977-01-01

Malnutrition, anaemia, and gut parasites are commonly interrelated. Using the Nippostrongylus brasiliensis-rat model, the effect of iron and protein deficiency on the efficacy of benzimidazole anthelmintics was studied. It was demonstrated that the anthelmintics mebendazole and fenbendazole were significantly less effective in eradicating parasites when animals were deficient in iron and protein. This decreased efficacy of anthelmintics in iron and protein deficiency could not be overcome by intraperitoneal administration of the drug. Since nutritional deficiencies may act via impairment of the immune response, anthelmintic efficacy was determined in adequately nourished rats treated with the immunosuppressive drug dexamethasone. A similar decrease in efficacy of mebendazole was shown when these animals were treated with dexamethasone. Thus it is possible that lowered anthelmintic efficacy in iron and protein deficient animals is mediated by immune deficiency. These findings may be relevant to anthelmintic programmes in malnourished communities. PMID:590849

In vitro anthelmintic efficacy of Borassus flabellifer Linn. (Palmae against Pheretima posthuma

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Prasad Govindrao Jamkhande

2014-02-01

Full Text Available Objective: To investigate the anthelmintic activity of Borassus flabellifer (B. flabellifer Linn. leaves against Indian adult earth worms (Pheretima posthuma. Methods: The B. flabellifer leaves and Indian adult earth worms (Pheretima posthuma were collected and authenticated. Earth worms were grouped and treated with extract at 10, 20 and 50 mg/mL concentration, albendazole (10 mg/mL as a standard and normal saline. The paralysis time and mortality time was considered as indicator of anthelmintic efficacy. Results: All the extracts showed concentration dependent activity but significant activity was observed at 50 mg/mL. At concentration 50 mg/mL extract pertained better activity with paralysis time (13.3 min and death times (17.92 min when compared to standard albendazole. Conclusions: The study findings reveal that the methanolic extract of B. flabellifer leaves has effective anthelmintic activity against Indian adult earth worms.

Anthelmintic activity of Trianthema portulacastrum L. and Musa paradisiaca L. against gastrointestinal nematodes of sheep.

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Hussain, Altaf; Khan, Muhammad Nisar; Iqbal, Zafar; Sajid, Muhammad Sohail; Khan, Muhammad Kasib

2011-06-30

Evaluation of anthelmintic effects of Trianthema (T.) portulacastrum L. (Aizoaceae) whole plant and Musa (M.) paradisiaca L. (Musaceae) leaves against prevalent gastrointestinal worms of sheep was done that may justify their traditional use in veterinary clinical medicine. In vitro anthelmintic activity of the crude aqueous methanolic extract (CAME) of both the plants was determined using mature female Haemonchus (H.) contortus and their eggs in adult motility assay (AMA) and egg hatch test (EHT), respectively. In vivo anthelmintic activity of crude powder (CP) and CAME in increasing doses (1.0-8.0 g kg(-1)) was determined in sheep naturally infected with mixed species of nematodes using fecal egg count reduction test (FECRT) and larval counts. The study design also included untreated as well as treated controls. Fecal egg count reduction and larval counts from coprocultures were performed pre- and post-treatments to assess the anthelmintic activity of the plants. CAME of T. portulacastrum and M. paradisiaca showed a strong in vitro anthelmintic activity and pronounced inhibitory effects on H. contortus egg hatching as observed through AMA and EHT, respectively. Both plants exhibited dose and time dependent anthelmintic effects on live worms as well as egg hatching. M. paradisiaca (LC(50)=2.13 μg mL(-1)) was found to be more potent than T. portulacastrum (LC(50)=2.41 μg mL(-1)) in EHT. However, in vivo, maximum reduction in eggs per gram (EPG) of faeces was recorded as 85.6% and 80.7% with CAME of T. portulacastrum and M. paradisiaca at 8.0 g kg(-1) on 15th day post-treatment, respectively as compared to that of Levamisole (7.5 mg kg(-1)) that caused 97.0% reduction in EPG. All the species of gastrointestinal nematodes (GINs), i.e. Haemonchus contortus, Trichostronglyus spp., Oesophagostomum columbianum and Trichuris ovis which were prevalent, found susceptible (Pparadisiaca possess strong anthelmintic activity in vitro and in vivo, thus, justifying their use in

Anthelmintic properties of traditional African and Caribbean medicinal plants: identification of extracts with potent activity against Ascaris suum in vitro

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Williams Andrew R.

2016-01-01

Full Text Available Ascariasis affects more than 1 billion people worldwide, mainly in developing countries, causing substantial morbidity. Current treatments for Ascaris infection are based on mass drug administration (MDA with synthetic anthelmintic drugs such as albendazole, however continual re-infection and the threat of drug resistance mean that complementary treatment options would be highly valuable. Here, we screened ethanolic extracts from 29 medicinal plants used in Africa (Ghana and the Caribbean (US Virgin Islands for in vitro anthelmintic properties against Ascaris suum, a swine parasite that is very closely related to the human A. lumbricoides. A wide variety of activities were seen in the extracts, from negligible to potent. Extracts from Clausena anisata, Zanthoxylum zanthoxyloides and Punica granatum were identified as the most potent with EC50 values of 74, 97 and 164 μg/mL, respectively. Our results encourage further investigation of their use as complementary treatment options for ascariasis, alongside MDA.

In vitro anthelmintic activity of Barleria buxifolia on Indian adult earthworms and estimation of total flavonoid content

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Purna A. Chander

2014-02-01

Full Text Available Objective: To study the anthelmintic activity of Barleria buxifolia leaf and to estimate the total flavonoid content. Methods: The aqueous and ethanolic leaf extracts were prepared and these were analyzed for total flavonoid content by aluminium chloride colorimetric method and Pheretima posthuma was used for anthelmintic activity by using the different concentrations (10, 20, 40, 80 and 100 mg/mL. Results: All the investigational extracts showed an anthelmintic activity at concentration of 10 mg/mL. The ethanolic extract of 100 mg/mL has produced an significant effect (P<0.001 when compared to aqueous extract. The total flavonoid content was found to be 5.67 mg QE/100 g. Conclusions: From the above study, the leaf extract has shown a good anthelmintic activity.

Resistência anti-helmíntica de nematóides gastrintestinais em ovinos, Mato Grosso do Sul Anthelmintic resistance of gastrointestinal nematodes in sheep, Mato Grosso do Sul, Brazil

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Eurico A. Sczesny-Moraes

2010-03-01

, the use of chemicals is the most common. However, the continued, and indiscriminate, use of these products has selected populations of resistant helminths to anthelmintics, a phenomenon reported in the whole world. This study aimed to identify the species of gastrointestinal parasites and diagnose the status of anthelmintic resistance in sheep in the State of Mato Grosso do Sul Brazil. Feacal egg count reduction tests (FECRT were performed in flocks of sixteen farms, and the seven formulations used contained the following pharmacological bases: Albendazole, Ivermectin, Levamizol, Trichlorfon, Moxidectin, Closantel and one containing the first three in association. The species identified at necropsy, in adult sheep, were: Haemonchus contortus, Trichostrongylus colubriformis, Cooperia curticei, C. punctata, C. pectinata and Oesophagostomum columbianum, in order of prevalence. The formulations containing Albendazole and Ivermectin did not show efficacy in reducing the EPG in the flocks tested, with average reductions of 0.7 and -19.6%, respectively. Closantel presented an average efficacy of 6.7%; Levamisolee, Moxidectin and Trichlorfon, 28.7, 26.8 and 65% respectively, the combination of three bases (Albendazole, Ivermectin and Levamizol, an average efficacy of 55.8 %. The average percentages of infective larvae recovered in the faecal cultures, pre and post treatment were similar, indicating that resistance to the bases tested is present in all species cited, to a greater or lesser degree. The two genera predominantly resistant are Haemonchus sp., with 86.9%, followed by Trichostrongylus sp., with an average of 47.5%, Strongyloides sp. 33.6%, Oesophagostomum sp. 21.4% and Cooperia sp. 19.7%.

Cytotoxic and anthelmintic potential of crude saponins isolated from Achillea Wilhelmsii C. Koch and Teucrium Stocksianum boiss

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Ali Niaz

2011-11-01

Full Text Available Abstract Background Saponins isolated from plant sources have a number of traditional and industrial applications. Saponins have pharmacological effects like anti-inflammatory, molluscicidal, antimicrobial, antispasmodic, antidiabetic, anticancer, anticonvulsant, anthelmintic, antitussive and cytotoxic activities. The current work describes the anthelmintic and cytotoxic activities of crude saponins of Achillea Wilhelmsii and Teucrium Stocksianum as these plants are rich with saponins. Methods Brine shrimp cytotoxic activity of crude saponins was determined by Meyer et al. (1982 at test concentrations of 1000 μg/ml, 100 μg/ml, 10 μg/ml, 7.5 μg/ml, 5.0 μg/ml, 2.5 μg/ml and 1.25 μg/ml. Percentage mortality of test concentrations was determined. Similarly, in vitro anthelmintic activity was determined against roundworms, tapeworms and earthworms. Albendazole and piperazine citrate at concentration 10 mg/ml were used as standard anthelmintic drugs. Results Crude saponins of Achillea wilhelmsii (CSA and Teucrium stocksianum (CST had, respectively, cytotoxic activity with LC50 values 2.3 ± 0.16 and 5.23 ± 0. 34 μg/ml. For in vitro anthelmintic activity, time for paralysis and death of parasites (parasiticidal activity was noted. At concentration 40 mg/ml, crude saponins of Achillea wilhelmsii are 1.96 and 2.12 times more potent than albendazole against Pheretima posthuma and Raillietina spiralis, respectively. Similarly, at concentration 40 mg/ml, crude saponins of Teucrium stocksianum (CST has 1.89, 1.96 and 1.37 times more parasiticidal activity than albendazole against Pheretima posthuma, Raillietina spiralis and Ascardia galli, respectively. Conclusion Crude saponins of Achillea wilhelmsii and Teucrium stocksianum have cytotoxic and anthelmintic activity. The crude saponins may be excellent sources of cytotoxic and anthelmintic constituents that warrant its isolation and purification for new drug development.

In vitro and in vivo anthelmintic activity of pumpkin seeds and pomegranate peels extracts against Ascaridia galli

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Amer R. Abdel Aziz

2018-06-01

Full Text Available Pumpkin seeds (Cucurbita pepo and Pomegranate peel (Punica granatum have anthelmintic properties. The aim of this study was to compare the anthelmintic efficacy of pumpkin seeds ethanolic extract and pomegranate peel aqueous extract against Ascaridia galli in vitro and in vivo in Baladi chicks. On adult worms, the extracts of the two herbs were compared in vitro at concentrations of 25, 50, and 75 mg/ml with fenbendazole at a concentration of 5 mg/ml. Chicks were infected with Ascaridia galli eggs containing second stage larva and treated with 2000 mg/kg of each of the extracts compared with 100 mg/kg fenbendazole. In vitro, all concentrations of pumpkin seed extract and the concentration of 75 mg/ml pomegranate peel extract exhibited a nearly similar effect to fenbendazole. In vivo, the mortality rate of the worms extracted from the 2000 mg/kg pumpkin seeds extract-treated chicken was non-significantly different from that of fenbendazole for 48 h. While pomegranate peels extract exhibited a lower lethal effect than fenbendazole. The anthelmintic efficacy was dependent on time and concentration. The study presented the anthelmintic efficacy of the pumpkin seeds and pomegranate peel extracts on Ascaridia galli. Pumpkin seed extract was more effective than pomegranate peel extract. Future studies to determine the optimal dose to maximize their effectiveness especially for pumpkin seeds as anthelmintic therapeutic are required. Keywords: Pomegranate peel, Pumpkin seeds, Anthelmintic, Ascaridia galli, In vitro, In vivo

Evaluation of anthelmintic activity of Iris hookeriana against gastrointestinal nematodes of sheep.

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Tariq, K A; Chishti, M Z; Ahmad, F; Shawl, A S; Tantray, M A

2008-06-01

The objective of this study was to evaluate the anthelmintic efficacy of Iris hookeriana Linn. rhizome against gastrointestinal nematodes of sheep. A worm motility inhibition assay was used for in vitro study and a faecal egg count reduction assay was used for an in vivo study. The in vitro study revealed anthelmintic effects of crude aqueous extracts and crude ethanolic extracts on live Trichuris ovis worms (P < or = 0.05) as evident from their paralysis and/or death at 8 h after exposure. The aqueous extracts of I. hookeriana resulted in a mean worm motility inhibition of 54.0%, while ethanolic extracts resulted in a mean worm motility inhibition of 84.6%. The mean mortality index of aqueous extracts was 0.55, while for ethanolic extracts it was 0.85. The lethal concentration 50 for aqueous extracts was 0.45 mg ml- 1 and for ethanolic extracts it was 0.15 mg ml- 1. The in vivo anthelmintic activity of aqueous and ethanolic extracts of I. hookeriana in sheep naturally infected with mixed species of gastrointestinal nematodes demonstrated a maximum (45.62%) egg count reduction in sheep treated with ethanolic extracts at 2 g kg- 1 body weight on day 10 after treatment, closely followed by ethanolic extracts at 1 g kg- 1 body weight on day 10 after treatment (43.54% egg count reduction). The aqueous extracts resulted in a maximum of 31.53% reduction in faecal egg counts on day 10 after treatment with 1 g kg- 1 body weight. Thus ethanolic extracts exhibited greater anthelmintic activity under both in vitro and in vivo conditions; this could be due to the presence of alcohol-soluble active ingredients in I. hookeriana. From the present study it can be suggested that I. hookeriana rhizome exhibited significant anthelmintic activity against gastrointestinal nematodes of sheep and has the potential to contribute to the control of gastrointestinal nematode parasites of small ruminants.

Anthelmintic resistance in a dairy cattle farm in the State of Minas Gerais Resistência anti-helmíntica em uma propriedade de bovinos leiteiros em Minas Gerais

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Maria do Socorro Veloso Leite Ferraz da Costa

2011-06-01

Full Text Available Eighty-four half-blood Gir × Holstein (F1 calves aged six months who were naturally infected by gastrointestinal helminths and maintained in rotational grazing received different anthelmintic treatments. Group A received anthelmintics according to the usual management in the property (eight treatments, seven including a macrocyclic lactone agent. Group B received strategic treatment (ivermectin 3.15% at the beginning and at the end of the rainy period. Eggs per gram of feces (EPG counts and genus of larvae from fecal cultures were determined on a monthly basis from April 2002 to December 2003. There was no significant reduction (p > 0.05 in EPG counts in any group after anthelminthic treatment, and the larvae in fecal cultures observed were Cooperia, Haemonchus, Oesophagostomum and a few Trichostrongylus. Cooperia was the most prevalent genus in the first four months of the experiment and Haemonchus in the following months. In 2003, tracer calves were introduced onto the pastures monthly and they showed high nematode burden many times throughout the year, and Cooperia punctata and Haemonchus contortus were the main species identified. The results suggest that there is anthelminthic resistance in this farm, mainly to macrocyclic lactones, and the development of immunity by crossbred animals was vital to reduce nematode burden.Oitenta e quatro bezerras meio sangue Gir × holandês (F1 com seis meses de idade, naturalmente infectadas por helmintos gastrintestinais e mantidas em pastejo rotacionado receberam diferentes tratamentos anti-helmínticos. O grupo A recebeu anti-helmínticos segundo manejo empregado na propriedade (oito tratamentos, sete com produtos à base de lactonas macrocíclicas. O grupo B recebeu tratamento estratégico (ivermectina 3,15% no inicio e final de período chuvoso. Mensalmente, no período de abril de 2002 a dezembro de 2003, foram realizadas contagens de ovos por grama de fezes (OPG e coproculturas. Não houve redu

Hepatic biotransformation pathways and ruminal metabolic stability of the novel anthelmintic monepantel in sheep and cattle.

Science.gov (United States)

Ballent, M; Virkel, G; Maté, L; Viviani, P; Lanusse, C; Lifschitz, A

2016-10-01

Monepantel (MNP) is a new amino-acetonitrile derivative anthelmintic drug used for the treatment of gastrointestinal (GI) nematodes in sheep. The present work investigated the main enzymatic pathways involved in the hepatic biotransformation of MNP in sheep and cattle. The metabolic stability in ruminal fluid of both the parent drug and its main metabolite (monepantel sulphone, MNPSO2 ) was characterized as well. Additionally, the relative distribution of both anthelmintic molecules between the fluid and particulate phases of the ruminal content was studied. Liver microsomal fractions from six (6) rams and five (5) steers were incubated with a 40 μm of MNP. Heat pretreatment (50 °C for 2 min) of liver microsomes was performed for inactivation of the flavin-monooxygenase (FMO) system. Additionally, MNP was incubated in the presence of 4, 40, and 80 μm of methimazole (MTZ), a FMO inhibitor, or equimolar concentrations of piperonyl butoxide (PBx), a well-known general cytochrome P450 (CYP) inhibitor. In both ruminant species, MNPSO2 was the main metabolite detected after MNP incubation with liver microsomes. The conversion rate of MNP into MNPSO2 was fivefold higher (P ruminal contents of both species showed a high chemical stability without evident metabolism and/or degradation as well as an extensive degree of adsorption (83% to 90%) to the solid phase of the ruminal content. Overall, these results are a further contribution to the understanding of the metabolic fate of this anthelmintic drug in ruminants. © 2016 John Wiley & Sons Ltd.

A cost-benefit analysis of pre- and post-lambing anthelmintic treatments to twin-bearing ewes on commercial farms in the southern North Island of New Zealand.

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Garland, C B; Leathwick, D M

2015-07-01

lower net benefit than a CRC containing albendazole administered pre-lambing (p<0.05). A positive financial return resulting from the anthelmintic treatment of adult ewes around lambing is neither consistent nor predictable, and is often not achieved. Given that the additional costs of accelerating the development of anthelmintic resistance were not included in these calculations, farmers need to consider carefully the merits of administering anthelmintics to ewes around lambing.

Prevalence of drug-resistant gastrointestinal nematodes in an organized sheep farm

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Ambalathaduvar Meenakshisundaram

2014-12-01

Full Text Available Aim: The present study was aimed to determine the resistance against albendazole, fenbendazole, levamisole and closantel in gastrointestinal (GI nematodes of sheep. Introduction: Anthelmintics are used traditionally as an integral part of helminthic control strategies for grazing livestock to prevent production losses from parasitic infections. The continuous and indiscriminate use of the same anthelmintics over years together as the sole means of control are now failing due to the emergence of resistance strains of helminths. Resistance to the commonly used anthelmintics in GI nematodes of sheep has become an increasingly widespread problem throughout the world. Materials and Methods: Fifty-five naturally infected Madras Red lambs of 6-12 months of age were selected and distributed randomly into five treatment groups of 11 animals each. Four groups were treated orally with albendazole (5 mg/kg, fenbendazole (7 mg/kg, levamisole (7.5 mg/kg and closantel (10 mg/kg respectively, whereas the fifth group served as untreated control. Fecal samples were collected per rectum of each lamb just prior to treatment (pre-treatment and on 7, 14, 21 and 28 days post-treatment. The anthelmintic resistance was evaluated by in vivo fecal egg count reduction test (FECRT, post-treatment larval culture and in vitro egg hatch assay. Results: In the FECRT, albendazole reduced the faecal egg count by 86.50%, 84.81%, 85.28% and 84.47% respectively for 4 weeks after treatment. Fecal egg count reduction using fenbendazole was 92.64, 93.04, 90.80 and 90.06% respectively for 4 weeks after treatment. The percent efficacy for levamisole and closantel was more than 95%. The post-treatment larval culture contained only Haemonchus contortus. In the in vitro egg hatch assay, the ED50 value for benzimidazole was 0.299 μg albendazole/ml and levamisole showed an ED50 value of 0.283 μg/ml. Conclusion: Our study confirmed the resistance of H. contortus to benzimidazole in sheep. .

ANTHELMINTIC AND ANTIOXIDANT ACTIVITY OF ALCOHOLIC EXTRACTS OF DIFFERENT PARTS OF COLEUS AMBOINICUS LOUR

OpenAIRE

Bhattacharjee Prasenjit; Hullatti K.K.; Vijay Kumar M.L.

2011-01-01

The present study reports anthelmintic activity of alcoholic extracts of leaf, stem and root of Coleus amboinicus against Indian earthworms Pheritima posthuma. The results revealed that all the tested extracts of Coleus amboinicus possessed significant anthelmintic activity in a dose-dependent manner. The activities were comparable with the reference drug Piperazine citrate and Albendazole. Among the tested extracts, the leaf extract was found to be more promising in comparison to stem and ro...

Anthelmintic Effect of Moringa oleifera Lam. in Wild- caught Achatina ...

African Journals Online (AJOL)

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This study assessed the anthelmintic effect of Moringa oleifera in A. achatina. Using dissecting and microscopic ..... Ecosystems and Sustainable Tourism. Available: ... commercially in Australia: a report for the Rural. Industries Research and ...

Metabolism of albendazole, ricobendazole and flubendazole in Haemonchus contortus adults: Sex differences, resistance-related differences and the identification of new metabolites

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Lucie Raisová Stuchlíková

2018-04-01

Full Text Available Haemonchus contortus (family Trichostrongylidae, Nematoda, a hematophagous gastrointestinal parasite found in small ruminants, has a great ability to develop resistance to anthelmintic drugs. We studied the biotransformation of the three benzimidazole anthelmintics: albendazole (ABZ, ricobendazole (albendazole S-oxide; RCB and flubendazole (FLU in females and males of H. contortus in both a susceptible ISE strain and resistant IRE strain. The ex vivo cultivation of living nematodes in culture medium with or without the anthelmintics was used. Ultrasensitive UHPLC/MS/MS analysis revealed 9, 7 and 12 metabolites of ABZ, RCB and FLU, respectively, with most of these metabolites now described in the present study for the first time in H. contortus. The structure of certain metabolites shows the presence of biotransformation reactions not previously reported in nematodes. There were significant qualitative and semi-quantitative differences in the metabolites formed by male and female worms. In most cases, females metabolized drugs more extensively than males. Adults of the IRE strain were able to form many more metabolites of all the drugs than adults of the ISE strain. Some metabolites were even found only in adults of the IRE strain. These findings suggest that increased drug metabolism may play a role in resistance to benzimidazole drugs in H. contortus. Keywords: Drug resistance, Drug metabolism, Anthelmintics, Benzimidazole, Nematode

Effectiveness testing of some vegetal extracts comparing with clasical anthelmintics

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Ilie M.S.,

2007-12-01

Full Text Available We have tested the efficacy of some vegetal extracts (Parazitol –Medica Laboratories, a natural product with an anthelmintic effect and a Cucurbita sp. oil extract compared to the classic anthelmintics (Rombendazol – Romvac and Dehelman – KRKA Slovenia at domestic poultry, whose parasitical status had been previously established through animal killing and necropsies. Parazitol and the pumpkin oil have had a lower efficacy than the levamisole and albendazole upon the species Ascaridia galli. Heterakis gallinarum was not affected by the pumpkin oil. Parazitol have a moderate efficacy (36%, while levamisole and albendazole were very efficient (100%. The treatments with albendazole upon the cestods belonging to the genus Raillietina have had a 100% efficacy. In cestods, Parazitol had a better efficacy (57% than the pumpkin oil (14%.

Anthelmintic activity of chicory (Cichorium intybus): in vitro effects on swine nematodes and relationship to sesquiterpene lactone composition

DEFF Research Database (Denmark)

Williams, Andrew; Pena-Espinoza, Miguel Angel; Boas, Ulrik

2016-01-01

of a specific anthelmintic effect rather than generalized toxicity. Ultra-high liquid chromatography-mass spectrometry analysis revealed that the purified extracts were rich in sesquiterpene lactones (SL), and that the SL profile differed significantly between cultivars. This is the first report of anthelmintic...

In vitro anthelmintic activity of Heliotropium indicum, Senna fistula and Spigelia anthelmia used as worm expeller in South West Nigeria

OpenAIRE

Oluwakemi K. Sobiyi; A.O. Tom Ashafa

2015-01-01

The anthelmintic potential of Heliotropium indicum, Senna fistula and Spigelia anthelmia were investigated using nematodes larvae from sheep and adult earthworms in order to justify the folkloric claim of the plants as worm expeller in south west of Nigeria. The acetone, ethanol, hydro-alcohol and distilled water extracts showed dose-dependent anthelmintic activities at the different concentrations (0.25, 0.50, 1.0 mg/ml) when tested against nematodes larvae. The order of anthelmintic effect ...

Detection and semi-quantification of Strongylus vulgaris DNA in equine faeces by real-time quantitative PCR.

Science.gov (United States)

Nielsen, Martin K; Peterson, David S; Monrad, Jesper; Thamsborg, Stig M; Olsen, Susanne N; Kaplan, Ray M

2008-03-01

Strongylus vulgaris is an important strongyle nematode with high pathogenic potential infecting horses world-wide. Several decades of intensive anthelmintic use has virtually eliminated clinical disease caused by S. vulgaris, but has also caused high levels of anthelmintic resistance in equine small strongyle (cyathostomin) nematodes. Recommendations aimed at limiting the development of anthelmintic resistance by reducing treatment intensity raises a simultaneous demand for reliable and accurate diagnostic tools for detecting important parasitic pathogens. Presently, the only means available to differentiate among strongyle species in a faecal sample is by identifying individual L3 larvae following a two week coproculture procedure. The aim of the present study is to overcome this diagnostic obstacle by developing a fluorescence-based quantitative PCR assay capable of identifying S. vulgaris eggs in faecal samples from horses. Species-specific primers and a TaqMan probe were designed by alignment of published ribosomal DNA sequences of the second internal transcribed spacer of cyathostomin and Strongylus spp. nematodes. The assay was tested for specificity and optimized using genomic DNA extracted from identified male worms of Strongylus and cyathostomin species. In addition, eggs were collected from adult female worms and used to evaluate the quantitative potential of the assay. Statistically significant linear relationships were found between egg numbers and cycle of threshold (Ct) values. PCR results were unaffected by the presence of cyathostomin DNA in the sample and there was no indication of PCR inhibition by faecal sources. A field evaluation on faecal samples obtained from four Danish horse farms revealed a good agreement with the traditional larval culture (kappa-value=0.78), but with a significantly higher performance of the PCR assay. An association between Ct values and S. vulgaris larval counts was statistically significant. The present assay can

Anthelmintic efficacy of pawpaw ( Carica papaya ) seeds in ...

African Journals Online (AJOL)

The anthelmintic efficacy of the aqueous and crude extract of Carica papaya seeds was studied in 40 Isa Brown commercial layers infected naturally with nematodes. They were randomly divided into 4 groups: A, B, C and D with 10 birds per group. Group A birds were untreated, while groups B, C and D were treated orally ...

Anthelmintic drug dispensing in South Africa: An analysis of ...

African Journals Online (AJOL)

More than two billion people worldwide are infected with helminths. The primary aim of the study was to evaluate the dispensing patterns of anthelmintic products using dispensing data of South African community pharmacies. A retrospective, cross-sectional drug utilisation study was conducted on a 2013 community ...

Evaluation of the anthelmintic activity and toxicity of an aqueous extract of Chenopodium ambrosioides in goats

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Gisele Dias da Silva

2016-10-01

Full Text Available ABSTRACT. da Silva G.D., Botura M.B., de Lima H.G., de Oliveira J.V.A., Moreira E.L.T., Santos F.O., de Souza T.S., de Almeida M.A.O. & Batatinha M.J.M. Evaluation of the anthelmintic activity and toxicity of an aqueous extract of Chenopodium ambrosioides in goats. [Avaliação da atividade anti-helmíntica e toxicidade do extrato aquoso de Chenopodium ambrosioides em caprinos.] Revista Brasileira de Medicina Veterinária, 38(Supl.1:156-162, 2016. Programa de Pós-Graduação em Ci- ência Animal nos Trópicos, Universidade Federal da Bahia, Av. Ademar de Barros, 500, Ondina, Salvador, BA 40170-110, Brasil. E-mail: mjmb@ufba.br The objective of this study was to evaluate the anthelmintic activity of an aqueous extract (AE from Chenopodium ambrosioides on goat gastrointestinal nematodes (GINs and its toxic effects. The anthelmintic activity in vitro was investigated using the inhibition of egg hatching assay (EHA, while cytotoxicity on Vero cells was evaluated using the MTT test. In vivo, thirty goats that were naturally infected with GINs were divided into three groups: group I, treated with a daily dose of AE C. ambrosioides (700mg/kg for eight days; group II (positive control, treated with a single dose of levamisole phosphate (6.3mg/kg; and Group III, untreated (negative control. Treatment efficacy was assessed on the basis of egg counts (FEC, faecal cultures and post-mortem worm burden counts. Clinical and laboratory evaluations were performed to detect toxic effects associated with treatment. In the EHA, the EC50 and EC90 corresponded to 1.6 and 1.9mg/mL, respectively. The AE promoted a slight reduction in cell viability in the cytotoxicity test. The AE reduced (p <0.05 the number of infective larvae of the genera Haemonchus and Oesophagostomum. The anthelmintic treatment of goats with AE C.ambrosioides resulted in moderate efficacy against infective larvae, but revealed neither ovicidal nor toxic activity towards adult nematodes. No toxic

Phytochemical screening and in vitro anthelmintic activity of methanol extract of Terminalia citrina leaves

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Narhari Das

2015-06-01

Full Text Available Objective: To evaluate anthelmintic activity of methanolic extract of leaves of Terminalia citrina (T. citrina plant belonging to the Combretaceae family. Methods: The tests of phytochemical screening included alkaloids, flavonoids, tannins, saponins, quinines, anthocyanins, glycosides, carbohydrates and reducing sugars. The anthelmintic activity of methanolic extract of leaves of T. citrina was evaluated against Pheretima posthuma at three different concentrations (25 mg/mL, 50 mg/mL and 100 mg/mL of extracts which involved determination of time of paralysis and time of death of worms. Results: The phytochemical screening of T. citrina leaves revealed the presence of flavonoids, tannins, alkaloids, carbohydrates and reducing sugars. The present study indicated that methanolic extract significantly exhibited paralysis and also caused death of worms especially at highest concentration of 100 mg/mL, as compared to standard reference Albendazole (10 mg/mL. Conclusions: This study suggests that the leaves of T. citrina possess potent anthelmintic activity.

In vitro anthelmintic effect of two medicinal plants (Anogeissus ...

African Journals Online (AJOL)

STORAGESEVER

2009-09-15

Sep 15, 2009 ... A study was conducted to evaluate Anogeissus leiocarpus leaf and Daniellia oliveri stem barks as effective remedy for gastrointestinal parasites. The anthelmintic activity of these extracts on eggs, first stage larvae and adults of Haemonchus contortus was examined by in vitro tests. The extracts were.

[Recent findings on the genetics of gastro-intestinal nematode resistance in ruminants].

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Carta, A; Scala, A

2004-06-01

The control of helminthiases in ruminants raised in open pasture has been mainly undertaken by using prophylactic measures in the environment, but these are often inadequate due to incorrect application. With the appearance of anthelmintics, the strategy for controlling these parasitoses, passed to pharmacological treatments which became effective in reducing their impact. However, the frequent and incorrect utilisation of these molecules resulted in resistance to anthelmintics and the presence of chemical residues in animal products for human consumption. Anthelmintic resistance is widespread throughout the world, heterogeneous and probably underestimated. This has encouraged the introduction of homeopathic agents and products derived from plants whose effectiveness has not been scientifically assessed. It is well known that it is possible to detect differences in resistance to the most important parasites between breeds. In Europe, it has been reported that some ovine autochthonous breeds, Scottish Blackface and Lacaune, showed higher resistance. The implementation of breeding strategies aimed at obtaining animals with naturally low susceptibility to nematode infestations could therefore play an increasingly important role. Standard animal breeding techniques have been largely successful in improving the performance of domestic animals in the last century. Standard quantitative selection requires field data on: i) individual phenotype performance; ii) expected covariance among animals due to blood relationship between them. The whole process of predicting the breeding value of animals in order to select subsequently the genetically superior parents of the next generation is entirely based on sophisticated computations (BLUP-animal model). In sheep, the main objective is always selecting for milk yield and sometimes, in addition, milk composition. However, due to the evolution of the EU agricultural policy and consumer demand in terms of healthy and organic food

Plasma disposition, concentration in the hair, and anthelmintic efficacy of eprinomectin after topical administration in donkeys.

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Gokbulut, Cengiz; Di Loria, Antonio; Gunay, Necati; Masucci, Roberto; Veneziano, Vincenzo

2011-12-01

To investigate plasma disposition, concentration in the hair, and anthelmintic efficacy of eprinomectin after topical administration in donkeys. 12 donkeys naturally infected with strongyle nematodes. The pour-on formulation of eprinomectin approved for use in cattle was administered topically to donkeys at a dosage of 0.5 mg/kg. Heparinized blood samples and hair samples were collected at various times between 1 hour and 40 days after administration. Samples were analyzed via high-performance liquid chromatography with fluorescence detection. Fecal strongyle egg counts were performed by use of a modified McMaster technique before and at weekly intervals for 8 weeks after treatment. Plasma concentration and systemic availability of eprinomectin were relatively higher in donkeys, compared with values reported for other animal species. Concerning the anthelmintic efficacy against strongyle nematodes, eprinomectin was completely effective (100%) on days 7 and 14 and highly effective (> 99%) until the end of the study at 56 days after treatment. No abnormal clinical signs or adverse reactions were observed for any donkeys after treatment. Eprinomectin had excellent safety. The relatively high plasma concentration after topical administration could result in use of eprinomectin for the control and treatment of parasitic diseases in donkeys.

Anthelmintic and Other Pharmacological Activities of the Root Bark ...

African Journals Online (AJOL)

The anthelmintic activity of water, methanol and chloroform extracts of the root bark of Albizia anthelmintica on strongyle-type sheep nematode eggs and larvae were examined in vitro. In addition, pharmacological tests were carried out on the water extract to confirm other ethnomedical uses of the plant. The water extract ...

Anthelmintic treatment in horses : the extra-label use of products and the danger of under-dosing

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S. Matthee

2003-06-01

Full Text Available Anthelmintic products form the basis of helminth control practices on horse stud farms at present. Regular evaluation of the efficacy of these products is advisable, as it will provide information on the worm egg reappearance period and the resistance status in the worm population. The aim of this study was to evaluate the efficacy of doramectin, pyrantel pamoate, ivermectin and moxidectin on a Thoroughbred stud farm in the Western Cape Province, South Africa. The study also compared the anthelmintic efficacy of two moxidectin formulations administered at their recommended dosages (an injectable, at 0.2 mg / kg, not registered for horses, and an oral gel at 0.4 mg / kg, registered for horses. Two mixed-sex groups of 30 yearlings and 40 weaners were tested in 2001 and 2002, respectively, divided into 3 and 4 groups of equal size. In 2001, moxidectin was one of 3 drugs administered orally and at a dose rate of 0.4 mg / kg. In 2002, pyrantel pamoate and ivermectin were orally administered at 19 and 0.2 mg / kg. Moxidectin and doramectin (the latter not registered for horses were administered by intramuscular injection at a dose of 0.2 mg / kg, the dosage registered for other host species. The faecal egg count reduction test was used to determine the anthelmintic efficacies in both years. Each animal acted as its own control and the arithmetic mean faecal egg count and lower 95 % confidence limit was calculated for each of the groups. A 100 % reduction in the faecal egg counts and a 100 % lower 95 % confidence limit was recorded for moxidectin (0.4 mg / kg in 2001. In 2002, a 99 % and 96% reduction was recorded for pyrantel pamoate and ivermectin, respectively. In the same year doramectin and moxidectin (both injectable and given at 0.2 mg / kg did not have any effect on worm egg counts. Of the 4 drugs tested in 2002, only pyrantel pamoate recorded lower 95 % confidence limits above 90 %.

Efficacy of some anthelmintics used in porcine practice in Ibadan ...

African Journals Online (AJOL)

Ninety three (93) pigs (crosses of Large white, Landrace, Hampshire and Duroc) were screened for gastrointestinal worms before and after treatment with the following drugs: Levamisole, Albendazole, Morantel citrate, Piperazine., thiabendazole and Ivermectin. The anthelmintic efficacy (measure as reduction egg per gram ...

Evaluation of antibacterial and anthelmintic activities with total phenolic contents of Piper betel leaves

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Kazi Akter

2014-09-01

Full Text Available Objective: The study was conducted to investigate the antibacterial and anthelmintic activities and to determine total phenolic contents of methanolic extract of Piper betel leaves. Materials and Methods: The extract was subjected to assay for antibacterial activity using both gram positive and gram negative bacterial strains through disc diffusion method; anthelmintic activity with the determination of paralysis and death time using earthworm (Pheritima posthuma at five different concentrations and the determination of total phenolic contents using the Folin-ciocalteau method. Results: The extract showed significant (p

Dry selection and wet evaluation for the rational discovery of new anthelmintics

Science.gov (United States)

Marrero-Ponce, Yovani; Castañeda, Yeniel González; Vivas-Reyes, Ricardo; Vergara, Fredy Máximo; Arán, Vicente J.; Castillo-Garit, Juan A.; Pérez-Giménez, Facundo; Torrens, Francisco; Le-Thi-Thu, Huong; Pham-The, Hai; Montenegro, Yolanda Vera; Ibarra-Velarde, Froylán

2017-09-01

Helminths infections remain a major problem in medical and public health. In this report, atom-based 2D bilinear indices, a TOMOCOMD-CARDD (QuBiLs-MAS module) molecular descriptor family and linear discriminant analysis (LDA) were used to find models that differentiate among anthelmintic and non-anthelmintic compounds. Two classification models obtained by using non-stochastic and stochastic 2D bilinear indices, classified correctly 86.64% and 84.66%, respectively, in the training set. Equation 1(2) correctly classified 141(135) out of 165 [85.45%(81.82%)] compounds in external validation set. Another LDA models were performed in order to get the most likely mechanism of action of anthelmintics. The model shows an accuracy of 86.84% in the training set and 94.44% in the external prediction set. Finally, we carry out an experiment to predict the biological profile of our 'in-house' collections of indole, indazole, quinoxaline and cinnoline derivatives (∼200 compounds). Subsequently, we selected a group of nine of the theoretically most active structures. Then, these chemicals were tested in an in vitro assay and one good candidate (VA5-5c) as fasciolicide compound (100% of reduction at concentrations of 50 and 10 mg/L) was discovered.

An Update Review on the Anthelmintic Activity of Bitter Gourd, Momordica charantia.

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Poolperm, Sutthaya; Jiraungkoorskul, Wannee

2017-01-01

Momordica charantia (Family: Cucurbitales ), as known as bitter melon or gourd, is a daily consumption as food and traditional medicinal plant in Southeast Asia and Indo-China. It has been shown to possess anticancer, antidepressant, antidiabetic, anti-inflammatory, antimicrobial, antiobesity, antioxidant, and antiulcer properties. Its common phytochemical components include alkaloids, charantin, flavonoids, glycosides, phenolics, tannins, and terpenoids. This plant is rich in various saponins including momordicin, momordin, momordicoside, karavilagenin, karaviloside, and kuguacin, all of which have been reported to contribute to its remedial properties including antibacterial, antifungal, antiviral, and antiparasitic infections. Based on established literature on the anthelmintic activity of M. charantia and possible mode of action, this review article has attempted to compile M. charantia could be further explored for the development of potential anthelmintic drug.

Evaluation of antibacterial and anthelmintic activities with total phenolic contents of Piper betel leaves

Science.gov (United States)

Akter, Kazi Nahid; Karmakar, Palash; Das, Abhijit; Anonna, Shamima Nasrin; Shoma, Sharmin Akter; Sattar, Mohammad Mafruhi

2014-01-01

Objective: The study was conducted to investigate the antibacterial and anthelmintic activities and to determine total phenolic contents of methanolic extract of Piper betel leaves. Materials and Methods: The extract was subjected to assay for antibacterial activity using both gram positive and gram negative bacterial strains through disc diffusion method; anthelmintic activity with the determination of paralysis and death time using earthworm (Pheritima posthuma) at five different concentrations and the determination of total phenolic contents using the Folin-ciocalteau method. Results: The extract showed significant (pbetel leaves extract, therefore it may be processed for further drug research. PMID:25386394

Resistance to benzimidazoles and levamisole in nematode parasites of sheep in Nyandarua district of Kenya

DEFF Research Database (Denmark)

Maingi, N.; Bjørn, H.; Gichohi, V.M.

1998-01-01

The occurrence of anthelmintic resistance on 25 sheep farms in the Nyandarua District of Kenya was investigated, using the faecal egg count reduction test (FECRT), the egg hatch assay (EHA) and a larval development assay (LDA). In the FECRT, resistance to both benzimidazoles (BZs) and levamisole...

Phenotypic- and Genotypic-Resistance Detection for Adaptive Resistance Management in Tetranychus urticae Koch.

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Deok Ho Kwon

Full Text Available Rapid resistance detection is necessary for the adaptive management of acaricide-resistant populations of Tetranychus urticae. Detection of phenotypic and genotypic resistance was conducted by employing residual contact vial bioassay (RCV and quantitative sequencing (QS methods, respectively. RCV was useful for detecting the acaricide resistance levels of T. urticae, particularly for on-site resistance detection; however, it was only applicable for rapid-acting acaricides (12 out of 19 tested acaricides. QS was effective for determining the frequencies of resistance alleles on a population basis, which corresponded to 12 nonsynonymous point mutations associated with target-site resistance to five types of acaricides [organophosphates (monocrotophos, pirimiphos-methyl, dimethoate and chlorpyrifos, pyrethroids (fenpropathrin and bifenthrin, abamectin, bifenazate and etoxazole]. Most field-collected mites exhibited high levels of multiple resistance, as determined by RCV and QS data, suggesting the seriousness of their current acaricide resistance status in rose cultivation areas in Korea. The correlation analyses revealed moderate to high levels of positive relationships between the resistance allele frequencies and the actual resistance levels in only five of the acaricides evaluated, which limits the general application of allele frequency as a direct indicator for estimating actual resistance levels. Nevertheless, the resistance allele frequency data alone allowed for the evaluation of the genetic resistance potential and background of test mite populations. The combined use of RCV and QS provides basic information on resistance levels, which is essential for choosing appropriate acaricides for the management of resistant T. urticae.

Evaluation of anthelmintic activity and in silico PASS assisted prediction of Cordia dichotoma (Forst.) root extract.

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Jamkhande, Prasad G; Barde, Sonal R

2014-01-01

Worm infection and associated complications are severe problems that afflict a large population worldwide. Failure of synthetic drugs in worm infections because of drug resistance has made alternative drug therapy desirable. Cordia dichotoma (Forst.) is an ethnomedicinal plant which is rich in several secondary metabolites. Traditionally, the plant has been claimed to have high medicinal properties including antimicrobial activity and cytotoxicity. The study begun with an aim to explore plant-based natural anthelmintic agents against Pheretima posthuma, an Indian earthworm. Methanol extract of the drug was obtained by successive soxhlet extraction. The extract was tested for different phytochemicals. Worms were exposed to 10 mg/ml, 25 mg/ml, 50 mg/ml, and 75 mg/ml concentrations of extract and standard drug, albendazole. A software-based tool, prediction of activity spectra for substances was used to estimate anthelmintic efficacy of plant metabolites. The phytochemical analysis revealed presence of alkaloids, tannins, glycosides, saponins, flavonoids, and phenols. The extract showed dose-dependent effects, affecting worm motility, viability, and mortality. It was also found that the biological activity spectrum of the plant phytoconstituents such as octacosanol, lupeol, caffeic acid, and hentricontanol were >0.5 (probable activity > 0.5). The findings of the present work suggest that the extract of C. dichotoma significantly interferes with motility pattern of P. posthuma. The paralysis and mortality of P. posthuma might be due to the combined effects different phytoconstituents. The extract of C. dichotoma promises natural sources to control worm infection.

Effects of feeding on the plasma disposition kinetics of the anthelmintic albendazole in laying hens.

Science.gov (United States)

Bistoletti, M; Alvarez, L; Lanusse, C; Moreno, L

2014-01-01

1. To optimise the use of albendazole (ABZ) as an anthelmintic in hens, the effects of fasting and type of diet on the plasma kinetics of ABZ and its metabolites were evaluated. 2. Twenty-four hens were distributed into 4 groups: In experiment I the Fed group were fed ad libitum, while the Fasted group was fasted over a 12-h period. In experiment II the Pelleted group was fed with pelleted commercial food, while the Grain group was fed with cereal grains. All the groups were treated with ABZ by oral route. Blood samples were taken and plasma analysed by HPLC. 3. ABZ and its metabolites albendazole-sulphoxide (ABZSO) and albendazole-sulphone (ABZSO2) were recovered in plasma in all the groups. The 12-h fasting period did not modify the disposition kinetics of ABZ in hens. The type of feed affected ABZ kinetics. ABZSO concentration profile was higher and detected for longer in the Grain group compared to the Pelleted group. Statistical differences were not found for AUC0-∞ values, whereas the T1/2for and T1/2el were different between groups. 4. Factors affecting ABZ kinetic behaviour should be taken into account to optimise its use to ensure the sustainability of the limited available anthelmintic therapeutic tools in avian parasite control.

Selection and characterisation of monepantel resistance in Teladorsagia circumcincta isolates

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D.J. Bartley

2015-08-01

To address these questions three MPTL-resistant Teladorsagia circumcincta isolates (MTci2-11, MTci5-13 and MTci7-12 have been artificially selected in vivo from phenotypically characterised parent isolates (MTci2, MTci5, MTci7 respectively. The selection process involved collecting and culturing eggs from surviving worms from sheep administered sub-optimal dosages of MPTL (Zolvix® to provide infective larvae to infect further sheep until resistant isolates were generated (between 9 and 13 rounds of selection. A controlled efficacy test was conducted using the original parental isolates and the newly generated MPTL resistant isolates (n = 5 per group. Selected isolates were assessed both under anthelmintic stress (Zolvix®, 2.5 mg/kg bodyweight; MTci-MPTL and at rest (untreated, MTci-CON. A number of life-history traits were assessed, namely, worm establishment rates, time to patency, faecal egg output, body length of adults and eggs in utero. The estimated resistance status of the selected isolates was confirmed with 48%, 28% and 9% reductions in worm burden at 7-days post Zolvix® administration for MTci2-11-MPTL, MTci5-13-MPTL and MTci7-12-MPTL, respectively, compared with untreated controls. One of the selected isolates MTci7-12-CON showed significantly greater total worm burden (p = 0.025, greater establishment rate (p = 0.033, decreased time to patency (p = 0.048, higher cumulative egg outputs (p = 0.002 compared with its parental derivative MTci7. The trial results suggest that anthelmintic selection in T. circumcincta, albeit under experimental conditions, can select for more prolific/fecund and quicker maturing populations. These data provide an insight into how parasites evolve in response to anthelmintic pressure.

Phenolics and Terpenoids; the Promising New Search for Anthelmintics: A Critical Review.

Science.gov (United States)

Mukherjee, Niladri; Mukherjee, Suprabhat; Saini, Prasanta; Roy, Priya; Babu, Santi P Sinha

2016-01-01

Ailments caused by helminth parasites are global causing different types of clinical complications with permanent and long term morbidity in humans. Although huge advances have been made in medical sciences the effectiveness of available anthelmintics are still quite limited. Starting from the 50's, most importance was given to synthetic compounds for developing remedies from them, however, the traditional knowledge of medicine of different countries continued to provide us clues against this widespread health problem. Natural products or structural analogs with diverse structures are always been the major sources for discovering new therapeutics and in recent past different active compounds have also been identified form these plant sources having anthelmintic properties. Although compounds of diverse chemical nature and classes were identified, most active ones belong to either phenol or terpene in broad chemical nature. The mechanism of action of these phytotherapeutics is usually multi-targeted and can act against the helminth parasites through diverse spectrum of activities. In this review we summarized the effective anthelmintics belong to either phenolics or terpenoids and highlighted the major way of their effectiveness. This also highlights the recent development of new therapeutic strategies against helminth parasites in the light of recent advances of knowledge. In addition, developing efficient strategies to promote apoptosis and disturbing redox status in them by natural products can provide us a clue in antifilarial drug developmental research and crucial unmet medical need.

Study on the interaction between three benzimidazole anthelmintics and eosin Y by high performance liquid chromatography associating with resonance light scattering and its application.

Science.gov (United States)

Pan, Ziyu; Peng, Jingdong; Zang, Xu; Lei, Gang; He, Yan; Liu, Di

2016-07-01

A novel, highly selective, and sensitive resonance light scattering (RLS) detection approach coupled with high performance liquid chromatography (HPLC) was researched and developed for the synchronous analysis of three kinds of benzimidazole anthelmintics, including mebendazole (MBZ), albendazole (ABZ), and fenbendazole (FBZ) for the first time. In the pH range of 3.5-3.7 Britton-Robinson buffer medium, three kinds of anthelmintics, which were separated by HPLC, reacted with eosin Y (EY) to form 1:1 ion-association complexes, resulting in significantly enhanced RLS signals and the maximum peak located at 335 nm. The enhanced RLS intensity was in proportion to the MBZ, ABZ, and FBZ concentration in the range 0.2-25, 0.2-23, and 0.15-20 μg/mL, respectively. The limit of detection was in the range of 0.064-0.16 μg/mL. In addition, human urine was determined to validate the proposed method by spiked samples and real urine samples. Satisfactory results were obtained by HPLC-RLS method. Graphical Abstract The diagram mechanism of generating resonance between emitted light and scattered light.

Detection of Strongylus vulgaris in equine faecal samples by real-time PCR and larval culture - method comparison and occurrence assessment.

Science.gov (United States)

Kaspar, A; Pfister, K; Nielsen, M K; Silaghi, C; Fink, H; Scheuerle, M C

2017-01-11

Strongylus vulgaris has become a rare parasite in Germany during the past 50 years due to the practice of frequent prophylactic anthelmintic therapy. To date, the emerging development of resistance in Cyathostominae and Parascaris spp. to numerous equine anthelmintics has changed deworming management and the frequency of anthelmintic usage. In this regard, reliable detection of parasitic infections, especially of the highly pathogenic S. vulgaris is essential. In the current study, two diagnostic methods for the detection of infections with S. vulgaris were compared and information on the occurrence of this parasite in German horses was gained. For this purpose, faecal samples of 501 horses were screened for S. vulgaris with real-time PCR and an additional larval culture was performed in samples of 278 horses. A subset of 26 horses underwent multiple follow-up examinations with both methods in order to evaluate both the persistence of S. vulgaris infections and the reproducibility of each diagnostic method. The real-time PCR revealed S. vulgaris-DNA in ten of 501 investigated equine samples (1.9%). The larval culture demonstrated larvae of S. vulgaris in three of the 278 samples (1.1%). A direct comparison of the two methods was possible in 321 samples including 43 follow-up examinations with the result of 11 S. vulgaris-positive samples by real-time PCR and 4 S. vulgaris-positive samples by larval culture. The McNemar's test (p-value = 0.016) revealed a significant difference and the kappa values (0.525) showed a moderate agreement between real-time PCR and larval culture. The real-time PCR detected a significantly higher proportion of positives of S. vulgaris compared to larval culture and should thus be considered as a routine diagnostic method for the detection of S. vulgaris in equine samples.

New solid state forms of antineoplastic 5-fluorouracil with anthelmintic piperazine

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Moisescu-Goia, C.; Muresan-Pop, M.; Simon, V.

2017-12-01

The aim of the present study was to asses the formation of solid forms between the 5-fluorouracil chemotherapy drug and the anthelmintic piperazine. Two new solid forms of antineoplastic agent 5-fluorouracil with anthelmintic piperazine were obtained by liquid assisted ball milling and slurry crystallization methods. The Nsbnd H hydrogen bonding donors and C = O hydrogen bonding acceptors of 5-fluorouracil allow to form co-crystals with other drugs delivering improved properties for medical applications, as proved for other compounds of pharmaceutical interest. Both new solid forms were investigated using X-ray powder diffraction (XRD), differential thermal analysis (DTA) and Fourier transform infrared (FTIR) spectroscopy. The XRD results show that by both methods were successfully synthesized new solid forms of 5-fluorouracil with piperazine. According to FTIR results the form prepared by lichid assisted grinding process was obtained as co-crystal and the other one, prepared by slurry method, resulted as a salt.

In vitro anthelmintic activity of Heliotropium indicum, Senna fistula and Spigelia anthelmia used as worm expeller in South West Nigeria

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Oluwakemi K. Sobiyi

2015-06-01

Full Text Available The anthelmintic potential of Heliotropium indicum, Senna fistula and Spigelia anthelmia were investigated using nematodes larvae from sheep and adult earthworms in order to justify the folkloric claim of the plants as worm expeller in south west of Nigeria. The acetone, ethanol, hydro-alcohol and distilled water extracts showed dose-dependent anthelmintic activities at the different concentrations (0.25, 0.50, 1.0 mg/ml when tested against nematodes larvae. The order of anthelmintic effect for the plants was H. indicum> S. fistula> S. anthelmia. Acetone and ethanol extracts of the three plants showed the most effective activity (100% mortality against adult earth worm (Pheretima posthuma after 30 min of exposure The reference standard drug (Vermox® showed less effectiveness compared to the medicinal plants used in the study. Overall, the study indicates that Heliotropium indicum, Senna fistula and Spigelia anthelmia are potential anthelmintic herbal drugs, which in turn validates the use of the three species as worm expellers by the Yorubas tribe of South West Nigeria.

In vivo and in vitro studies of Cry5B and nicotinic acetylcholine receptor agonist anthelmintics reveal a powerful and unique combination therapy against intestinal nematode parasites.

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Yan Hu

2018-05-01

Full Text Available The soil-transmitted nematodes (STNs or helminths (hookworms, whipworms, large roundworms infect the intestines of ~1.5 billion of the poorest peoples and are leading causes of morbidity worldwide. Only one class of anthelmintic or anti-nematode drugs, the benzimidazoles, is currently used in mass drug administrations, which is a dangerous situation. New anti-nematode drugs are urgently needed. Bacillus thuringiensis crystal protein Cry5B is a powerful, promising new candidate. Drug combinations, when properly made, are ideal for treating infectious diseases. Although there are some clinical trials using drug combinations against STNs, little quantitative and systemic work has been performed to define the characteristics of these combinations in vivo.Working with the hookworm Ancylostoma ceylanicum-hamster infection system, we establish a laboratory paradigm for studying anti-nematode combinations in vivo using Cry5B and the nicotinic acetylcholine receptor (nAChR agonists tribendimidine and pyrantel pamoate. We demonstrate that Cry5B strongly synergizes in vivo with both tribendimidine and pyrantel at specific dose ratios against hookworm infections. For example, whereas 1 mg/kg Cry5B and 1 mg/kg tribendimidine individually resulted in only a 0%-6% reduction in hookworm burdens, the combination of the two resulted in a 41% reduction (P = 0.020. Furthermore, when mixed at synergistic ratios, these combinations eradicate hookworm infections at doses where the individual doses do not. Using cyathostomin nematode parasites of horses, we find based on inhibitory concentration 50% values that a strongylid parasite population doubly resistant to nAChR agonists and benzimidazoles is more susceptible or "hypersusceptible" to Cry5B than a cyathostomin population not resistant to nAChR agonists, consistent with previous Caenhorhabditis elegans results.Our study provides a powerful means by which anthelmintic combination therapies can be examined in vivo

Evaluation of pharmaceuticals and personal care products with emphasis on anthelmintics in human sanitary waste, sewage, hospital wastewater, livestock wastewater and receiving water.

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Sim, Won-Jin; Kim, Hee-Young; Choi, Sung-Deuk; Kwon, Jung-Hwan; Oh, Jeong-Eun

2013-03-15

We investigated 33 pharmaceuticals and personal care products (PPCPs) with emphasis on anthelmintics and their metabolites in human sanitary waste treatment plants (HTPs), sewage treatment plants (STPs), hospital wastewater treatment plants (HWTPs), livestock wastewater treatment plants (LWTPs), river water and seawater. PPCPs showed the characteristic specific occurrence patterns according to wastewater sources. The LWTPs and HTPs showed higher levels (maximum 3000 times in influents) of anthelmintics than other wastewater treatment plants, indicating that livestock wastewater and human sanitary waste are one of principal sources of anthelmintics. Among anthelmintics, fenbendazole and its metabolites are relatively high in the LWTPs, while human anthelmintics such as albendazole and flubendazole are most dominant in the HTPs, STPs and HWTPs. The occurrence pattern of fenbendazole's metabolites in water was different from pharmacokinetics studies, showing the possibility of transformation mechanism other than the metabolism in animal bodies by some processes unknown to us. The river water and seawater are generally affected by the point sources, but the distribution patterns in some receiving water are slightly different from the effluent, indicating the influence of non-point sources. Copyright © 2013 Elsevier B.V. All rights reserved.

Evaluation of anthelmintic activity and in silico PASS assisted prediction of Cordia dichotoma (Forst.) root extract

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Jamkhande, Prasad G.; Barde, Sonal R.

2014-01-01

Background: Worm infection and associated complications are severe problems that afflict a large population worldwide. Failure of synthetic drugs in worm infections because of drug resistance has made alternative drug therapy desirable. Cordia dichotoma (Forst.) is an ethnomedicinal plant which is rich in several secondary metabolites. Traditionally, the plant has been claimed to have high medicinal properties including antimicrobial activity and cytotoxicity. Materials and Methods: The study begun with an aim to explore plant-based natural anthelmintic agents against Pheretima posthuma, an Indian earthworm. Methanol extract of the drug was obtained by successive soxhlet extraction. The extract was tested for different phytochemicals. Worms were exposed to 10 mg/ml, 25 mg/ml, 50 mg/ml, and 75 mg/ml concentrations of extract and standard drug, albendazole. A software-based tool, prediction of activity spectra for substances was used to estimate anthelmintic efficacy of plant metabolites. Result: The phytochemical analysis revealed presence of alkaloids, tannins, glycosides, saponins, flavonoids, and phenols. The extract showed dose-dependent effects, affecting worm motility, viability, and mortality. It was also found that the biological activity spectrum of the plant phytoconstituents such as octacosanol, lupeol, caffeic acid, and hentricontanol were >0.5 (probable activity > 0.5). Conclusion: The findings of the present work suggest that the extract of C. dichotoma significantly interferes with motility pattern of P. posthuma. The paralysis and mortality of P. posthuma might be due to the combined effects different phytoconstituents. The extract of C. dichotoma promises natural sources to control worm infection. PMID:25737609

In vitro and in vivo anthelmintic effects of Caesalpinia bonducella (L. Roxb. leaf extract on Hymenolepis diminuta (Cestoda and Syphacia obvelata (Nematoda

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Shyamalima Gogoi

2016-12-01

Full Text Available Background: Leaves of Caesalpinia bonducella (L. Roxb., have been traditionally used as herbal remedy against intestinal helminths helminth infections in traditional medicine of India. Aim: This study was undertaken to evaluate the potential in vitro and in vivo anthelmintic effects of C. bonducella leaf extract against Syphacia obvelata (Nematoda and Hymenolepis diminuta (Cestoda. Materials and Methods: The in vitro anthelmintic activity of the extract was investigated on adult Syphacia obvelata (Nematoda and Hymenolepis diminuta (Cestoda worms in terms of physical motility and mortality of parasites. The in vivo study was performed in H. diminuta-rat model and S. obvelata-mice model, by monitoring the egg per gram (EPG of faeces count and worm count of animals following the treatment with different doses of plant extract. Results: The study recorded significant and dose-dependent anthelmintic effects of extract on both the parasites. In the in vitro study, 30 mg/ml concentration of extract caused mortality of H. diminuta in 2.5±0.2 h and S. obvelata in 3.57± 0.16 h. In the in vivo study, the extract showed a comparatively better efficacy on S. obvelata, where its 800 mg/kg dose revealed 93% reduction of worm load in mice, as compared to 85% reduction of H. diminuta worms in load. Conclusions: The findings suggest that leaf extract of C. bonducella possesses significant anthelmintic effects, and supports its use as an anthelmintic in traditional medicine. This appears to be the first report of in vivo anthelmintic activity of Caesalpinia bonducella against these parasites [J Complement Med Res 2016; 5(4.000: 427-433

Atividade anti-helmíntica de plantas nativas do continente americano: uma revisão Anthelmintic activity of native American plants: a review

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R.G. Sousa

2013-01-01

Full Text Available O presente trabalho faz uma revisão de plantas nativas do continente americano com atividade anti-helmíntica com a finalidade de subsidiar pesquisas e o desenvolvimento de novos medicamentos na área de produtos naturais. Na literatura, foram citadas 22 famílias de plantas, tais como: Moraceae, Asteraceae e Cucurbitaceae, e estudadas cerca de 35 espécies, como a Ficus sp, Cucurbita máxima, e Ruta graveolens, usadas por sua alegada atividade anti-helmíntica. Destas espécies, 26 foram ativas contra alguns helmintos: Strongyloides stercolaris, Ancylostomidae, Trichostrongylus sp, Capillaria sp, Ascaris lumbricoides, Taenia saginata, Haemonchus contortus, e Hymenolepsis diminuta. Foi verificado maior número de estudos com parasitas de animais, possivelmente devido ao desenvolvimento de resistência dos helmintos aos tratamentos convencionais disponíveis.This paper is a review of native American plants showing anthelmintic activity in order to give support to research and development of new drugs in the field of natural products. In the literature, 22 plant families were cited, such as: Moraceae, Asteraceae and Cucurbitaceae, and around 35 species were studied, including Ficus sp, Cucurbita maxima and Ruta graveolens, used for their putative anthelmintic activity. From these species, 26 were active against some helminthes: Strongyloides stercolaris, Ancylostomidae, Trichostrongylus sp, Capillaria sp, Ascaris lumbricoides, Taenia saginata, Haemonchus contortus and Hymenolepsis diminuta. Data indicated a larger number of studies with animal parasites, probably due to the resistance of these helminthes to the conventional treatments available.

Anthelmintic Activity of a Herbal Formulation Against Gastrointestinal Nematodes of Sheep

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Muhammad Arfan Zaman*§, Zafar Iqbal, Muhammad Nisar Khan and Ghulam Muhammad1

2012-01-01

Full Text Available This study was carried out to evaluate the anthelmintic activity of a herbal formulation (HF based on aqueous extracts of leaves of Azadirachta indica and Nicotiana tabacum, flowers of Calotropis procera and seeds of Trachyspermum ammi. In vitro, eggs and adult Haemonchus contortus were exposed to different concentrations of HF following the standard procedures of egg hatch test (EHT; 50 to 0.024414 mg ml-1 and adult motility assay (AMA; 200-0.1953125mg ml-1, respectively. The reference drugs used in the study were oxfendazole (0.0056704 to 0.0000027 mg ml-1 and levamisole (1.50 mg ml-1 for EHT and AMA, respectively. In vivo, pre and post-treatment (4 mg, 2 mg and 500 µg kg-1 body weight fecal egg counts were determined following standard fecal egg count reduction test in sheep naturally parasitized with mixed species of gastrointestinal nematodes. In EHT, LC50 values of HF and oxfendazole (reference drug were 275.1 and 0.016 µg ml-1, respectively. In AMA, 100% mortality of H. contortus was observed 6 hr post-exposure to 3.125-200 mg ml-1 concentrations of HF and 2 hr post-exposure to levamisole. In vivo, maximum (96.2% fecal egg count (EPG reduction was recorded in sheep treated with HF @ 4 mg kg-1 body weight; whereas, 89.3% reduction in EPG was recorded in sheep treated with levamisole @ 7.5 mg kg-1 body weight. A graded dose response was noted in all the tests used in the present study to evaluate the anthelmintic activity of HF. Therefore, HF seems to be promising as an anthelmintic for animals. Large scale trials on efficacy and safety, however, are recommended before the HF is considered for commercialization in crude form.

Modulation of porcine biotransformation enzymes by anthelmintic therapy with fenbendazole and flubendazole.

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Savlík, M; Fimanová, K; Szotáková, B; Lamka, J; Skálová, L

2006-06-01

Fenbendazole (FEN) and flubendazole (FLU) are benzimidazole anthelmintics often used in pig management for the control of nematodoses. The in vivo study presented here was designed to test the influence of FLU and FEN on cytochrome P4501A and other cytochrome P450 (CYP) isoforms, UDP-glucuronosyl transferase and several carbonyl reducing enzymes. The results indicated that FEN (in a single therapeutic dose as well as in repeated therapeutic doses) caused significant induction of pig CYP1A, while FLU did not show an inductive effect towards this isoform. Some of the other hepatic and intestinal biotransformation enzymes that were assayed were moderately influenced by FEN or FLU. Strong CYP1A induction following FEN therapy in pigs may negatively affect the efficacy and pharmacokinetics of FEN itself or other simultaneously or consecutively administered drugs. From the perspective of biotransformation enzyme modulation, FLU would appear to be a more convenient anthelmintic therapy of pigs than FEN.

Assessment of the anthelmintic activity of medicinal plant extracts and purified condensed tannins against free-living and parasitic stages of Oesophagostomum dentatum

DEFF Research Database (Denmark)

Williams, Andrew Richard; Ropiak, Honorata M.; Fryganas, Christos

2014-01-01

BackgroundPlant-derived condensed tannins (CT) show promise as a complementary option to treat gastrointestinal helminth infections, thus reducing reliance on synthetic anthelmintic drugs. Most studies on the anthelmintic effects of CT have been conducted on parasites of ruminant livestock. Oesop...

Diagnosis, Treatment and Management of Haemonchus contortus in Small Ruminants.

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Besier, R B; Kahn, L P; Sargison, N D; Van Wyk, J A

2016-01-01

Haemonchus contortus is a highly pathogenic, blood-feeding nematode of small ruminants, and a significant cause of mortalities worldwide. Haemonchosis is a particularly significant threat in tropical, subtropical and warm temperate regions, where warm and moist conditions favour the free-living stages, but periodic outbreaks occur more widely during periods of transient environmental favourability. The clinical diagnosis of haemonchosis is based mostly on the detection of anaemia in association with a characteristic epidemiological picture, and confirmed at postmortem by the finding of large numbers of H. contortus in the abomasum. The detection of impending haemonchosis relies chiefly on periodic monitoring for anaemia, including through the 'FAMACHA' conjunctival-colour index, or through faecal worm egg counts and other laboratory procedures. A range of anthelmintics for use against H. contortus is available, but in most endemic situations anthelmintic resistance significantly limits the available treatment options. Effective preventative programmes vary depending on environments and enterprise types, and according to the scale of the haemonchosis risk and the local epidemiology of infections, but should aim to prevent disease outbreaks while maintaining anthelmintic efficacy. Appropriate strategies include animal management programmes to avoid excessive H. contortus challenge, genetic and nutritional approaches to enhance resistance and resilience to infection, and the monitoring of H. contortus infection on an individual animal or flock basis. Specific strategies to manage anthelmintic resistance centre on the appropriate use of effective anthelmintics, and refugia-based treatment schedules. Alternative approaches, such as biological control, may also prove useful, and vaccination against H. contortus appears to have significant potential in control programmes. Copyright © 2016 Elsevier Ltd. All rights reserved.

In vitro anthelmintic activity and chemical composition of methanol extracts and fractions of Croton paraguayensis and Vernonia brasiliana against Eisenia fetida

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Andrea Leticia Cáceres

2017-02-01

Full Text Available Objective: To evaluate the chemical composition and the anthelmintic activity of the methanol extracts and the acid and basic fractions of Croton paraguayensis (C. paraguayensis and Vernonia brasiliana (V. brasiliana against Eisenia fetida. Methods: A preliminary phytochemical analysis was performed to assess the presence of groups of secondary metabolites. The plants were extracted with methanol to obtain the crude extracts. A differential pH extraction was performed to isolate basic compounds like alkaloids. The methanolic extracts and the fractions obtained were tested for anthelmintic activity against Eisenia fetida, using albendazole as positive control. Results: The phytochemical test demonstrated the presence of alkaloids in the crude extracts and alkaline fractions, along with flavonoids, coumarins, steroids/triterpenes and tannins. The anthelmintic activity of the extracts and fractions of C. paraguayensis and V. brasiliana showed a statistically significant decrease of the times for paralysis and death compared to albendazole. Conclusions: The methanolic extracts and fractions of C. paraguayensis and V. brasiliana contain compounds that possess anthelmintic activity. The isolation of the substances responsible for the biological effect described could result in the development of new drugs to treat helminth diseases.

Characterisation of an acetylcholine receptor gene of Haemonchus contortus in relation to levamisole resistance

NARCIS (Netherlands)

Hoekstra, R.; Visser, A.; Wiley, L.; Weiss, A.S.; Sangster, N.C.; Roos, M.H.

1997-01-01

The anthelmintic drug levamisole is thought to bind to nicotinic acetylcholine receptors of nematodes. It is possible that resistance to this drug is associated with either a change in binding characteristics or a reduction in the number of nicotinic acetylcholine receptors. Therefore, the molecular

Molecular detection methods of resistance to antituberculosis drugs in Mycobacterium tuberculosis.

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Brossier, F; Sougakoff, W

2017-09-01

Molecular methods predict drug resistance several weeks before phenotypic methods and enable rapid implementation of appropriate therapeutic treatment. We aimed to detail the most representative molecular tools used in routine practice for the rapid detection of resistance to antituberculosis drugs among Mycobacterium tuberculosis strains. The molecular diagnosis of resistance to antituberculosis drugs in clinical samples or from in vitro cultures is based on the detection of the most common mutations in the genes involved in the development of resistance in M. tuberculosis strains (encoding either protein targets of antibiotics, or antibiotic activating enzymes) by commercial molecular kits or by sequencing. Three hypotheses could explain the discrepancies between the genotypic results and the phenotypic drug susceptibility testing results: a low percentage of resistant mutants precluding the detection by genotypic methods on the primary culture; a low level of resistance not detected by phenotypic testing; and other resistance mechanisms not yet characterized. Molecular methods have varying sensitivity with regards to detecting antituberculosis drug resistance; that is why phenotypic susceptibility testing methods are mandatory for detecting antituberculosis drug-resistant isolates that have not been detected by molecular methods. The questionable ability of existing phenotypic and genotypic drug susceptibility testing to properly classify strains as susceptible or resistant, and at what level of resistance, was raised for several antituberculosis agents. Copyright © 2017 Elsevier Masson SAS. All rights reserved.

Determination of anthelmintic efficacy against Toxocara canis in dogs by use of capsule endoscopy.

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Lee, Alice C Y; Epe, Christian; Bowman, Dwight D

2015-09-15

Industry guidelines for anthelmintic testing call for postmortem inspection of animals to verify treatment efficacy. A previous study showed that capsule endoscopy (CE) can be performed on dogs in vivo to quantify hookworms in the small intestine. Adoption of a minimally invasive procedure such as this could reduce the need for necropsy in efficacy trials. The present study employed CE to enumerate Toxocara canis in dogs, with two main goals: to determine if multiple capsule examinations improves the accuracy of worm counts compared to a single examination, and to establish if the efficacy of an anthelmintic compound is the same whether calculated using CE or necropsy data. To avoid needless animal sacrifice, the study was carried out on beagle dogs already in a product development trial with a planned terminal endpoint. Dogs were infected by oral inoculation with T. canis eggs. Untreated control dogs (n=8) were evaluated by CE three times while dogs treated with test compounds (3 groups of 4) were examined only once. Utilizing either the average count or just the last complete capsule examination, a robust correlation was found between CE and postmortem numbers (r=0.94, p<0.001). Calculated anthelmintic efficacy was essentially identical for the two enumeration methods, ranging from 94% to 100% for the three research compounds. CE may therefore be a viable alternative to necropsy for T. canis parasiticide trials. Copyright © 2015 Elsevier B.V. All rights reserved.

In vitro anthelmintic effects of Spigelia anthelmia protein fractions against Haemonchus contortus.

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Sandra Alves Araújo

Full Text Available Gastrointestinal nematodes are a significant concern for animal health and well-being, and anthelmintic treatment is mainly performed through the use of chemical products. However, bioactive compounds produced by plants have shown promise for development as novel anthelmintics. The aim of this study is to assess the anthelmintic activity of protein fractions from Spigelia anthelmia on the gastrointestinal nematode Haemonchus contortus. Plant parts were separated into leaves, stems and roots, washed with distilled water, freeze-dried and ground into a fine powder. Protein extraction was performed with sodium phosphate buffer (75 mM, pH 7.0. The extract was fractionated using ammonium sulfate (0-90% and extensively dialyzed. The resulting fractions were named LPF (leaf protein fraction, SPF (stem protein fraction and RPF (root protein fraction, and the protein contents and activities of the fractions were analyzed. H. contortus egg hatching (EHA, larval exsheathment inhibition (LEIA and larval migration inhibition (LMIA assays were performed. Proteomic analysis was conducted, and high-performance liquid chromatography (HPLC chromatographic profiles of the fractions were established to identify proteins and possible secondary metabolites. S. anthelmia fractions inhibited H. contortus egg hatching, with LPF having the most potent effects (EC50 0.17 mg mL-1. During LEIA, SPF presented greater efficiency than the other fractions (EC50 0.25 mg mL-1. According to LMIA, the fractions from roots, stems and leaves also reduced the number of larvae, with EC50 values of 0.11, 0.14 and 0.21 mg mL-1, respectively. Protein analysis indicated the presence of plant defense proteins in the S. anthelmia fractions, including protease, protease inhibitor, chitinase and others. Conversely, secondary metabolites were absent in the S. anthemia fractions. These results suggest that S. anthelmia proteins are promising for the control of the gastrointestinal nematode H

Direct detection of Mycobacterium tuberculosis and drug resistance in respiratory specimen using Abbott Realtime MTB detection and RIF/INH resistance assay.

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Tam, Kingsley King-Gee; Leung, Kenneth Siu-Sing; To, Sabrina Wai-Chi; Siu, Gilman Kit-Hang; Lau, Terrence Chi-Kong; Shek, Victor Chi-Man; Tse, Cindy Wing-Sze; Wong, Samson Sai-Yin; Ho, Pak-Leung; Yam, Wing-Cheong

2017-10-01

Abbott RealTime MTB (Abbott-RT) in conjunction with Abbott RealTime MTB RIF/INH Resistance (Abbott-RIF/INH) is a new, high-throughput automated nucleic acid amplification platform (Abbott-MDR) for detection of Mycobacterium tuberculosis complex (MTBC) and the genotypic markers for rifampicin (RIF) and isoniazid (INH) resistance directly from respiratory specimens. This prospective study evaluated the diagnostic performance of this new platform for MTBC and multidrug-resistant tuberculosis (MDR-TB) using 610 sputum specimens in a tuberculosis high-burden setting. Using conventional culture results and clinical background as reference standards, Abbott-RT exhibited an overall sensitivity and specificity of 95.2% and 99.8%, respectively. Genotypic RIF/INH resistance of 178 "MTB detected" specimens was subsequently analyzed by Abbott-RIF/INH. Compared to phenotypic drug susceptibility test results, Abbott-RIF/INH detected resistance genotypic markers in 84.6% MDR-TB, 80% mono-RIF-resistant and 66.7% mono-INH-resistant specimens. Two of the RIF-resistant specimens carried a novel single, nonsense mutation at rpoB Q513 and in silico simulation demonstrated that the truncated RpoB protein failed to bind with other subunits for transcription. Overall, Abbott-MDR platform provided high throughput and reliable diagnosis of MDR-TB within a TB high-burden region. Copyright © 2017 Elsevier Inc. All rights reserved.

The efficacy of a slow-release albendazole capsule against Haemonchus contortus with known resistance to albendazole

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Alan D. Fisher

2013-12-01

Full Text Available Controlled-release albendazole capsules (CRCs are currently registered for use in Australia and New Zealand as anthelmintic treatment in sheep. However, reports on the efficacy of such products on resistant parasite populations are sometimes controversial. This is the first study to report on the efficacy of such products under South African field conditions in sheep harbouring a population of Haemonchus contortus with known multiple anthelmintic resistance, including to albendazole. Treatment groups were comprised of CRC-treated and single dose albendazole-treated sheep, as well as negative controls. Groups were compared by using faecal egg count reduction tests, FAMACHA© anaemia scoring, conception rates and comparative weight gains over three and a half months. Based on a comparison of faecal egg counts, no advantage could be found using CRCs. Moreover, the use of the product actually decreased weight gain when compared with the control group animals.

Mixed methods evaluation of targeted selective anthelmintic treatment by resource-poor smallholder goat farmers in Botswana

Science.gov (United States)

Walker, Josephine G.; Ofithile, Mphoeng; Tavolaro, F. Marina; van Wyk, Jan A.; Evans, Kate; Morgan, Eric R.

2015-01-01

Due to the threat of anthelmintic resistance, livestock farmers worldwide are encouraged to selectively apply treatments against gastrointestinal nematodes (GINs). Targeted selective treatment (TST) of individual animals would be especially useful for smallholder farmers in low-income economies, where cost-effective and sustainable intervention strategies will improve livestock productivity and food security. Supporting research has focused mainly on refining technical indicators for treatment, and much less on factors influencing uptake and effectiveness. We used a mixed method approach, whereby qualitative and quantitative approaches are combined, to develop, implement and validate a TST system for GINs in small ruminants, most commonly goats, among smallholder farmers in the Makgadikgadi Pans region of Botswana, and to seek better understanding of system performance within a cultural context. After the first six months of the study, 42 out of 47 enrolled farmers were followed up; 52% had monitored their animals using the taught inspection criteria and 26% applied TST during this phase. Uptake level showed little correlation with farmer characteristics, such as literacy and size of farm. Herd health significantly improved in those herds where anthelmintic treatment was applied: anaemia, as assessed using the five-point FAMACHA© scale, was 0.44–0.69 points better (95% confidence interval) and body condition score was 0.18–0.36 points better (95% C.I., five-point scale) in treated compared with untreated herds. Only targeting individuals in greatest need led to similar health improvements compared to treating the entire herd, leading to dose savings ranging from 36% to 97%. This study demonstrates that TST against nematodes can be implemented effectively by resource-poor farmers using a community-led approach. The use of mixed methods provides a promising system to integrate technical and social aspects of TST programmes for maximum uptake and effect. PMID

Potentilla fulgens (Family Rosaceae), a medicinal plant of north-east India: a natural anthelmintic?

Science.gov (United States)

Roy, Bishnupada; Swargiary, Ananta; Syiem, D; Tandon, V

2010-10-01

The cestode parasite, Raillietina echinobothrida and the trematode, Gastrothylax crumenifer were exposed to the ethanolic root peel extract of Potentilla fulgens, an antiparasitic local medicinal plant of Meghalaya, India, to evaluate the anthelmintic efficacy of the plant. The parasites were incubated in 1, 5, 10, 20, 50 and 100 mg crude alcoholic extract per ml of phosphate buffered saline (PBS) at a temperature of 37 ± 1°C. Paralysis and death were observed at 2.00 ± 0.05 and 2.80 ± 0.06 h for the cestode and 1.21 ± 0.06 and 2.18 ± 0.04 h for the trematode parasites at the highest test concentration of the plant extract. The commercial anthelmintic, Praziquantel (PZQ) showed higher activity at the tested concentration (0.02 mg/ml). To further investigate the efficacy of the plant extract, vital tegumental enzymes of the parasite viz. Acid phosphatase (AcPase), Alkaline phosphatase (AlkPase) and Adenosine triphosphatase (ATPase) were studied. Quantitatively, the total enzyme activity of AcPase, AlkPase and ATPase was found to be reduced significantly by 69.20, 66.43 and 29.63% for R. echinobothrida and 47.96, 51.79 and 42.63% for G. crumenifer, respectively compared to the respective controls; histochemical study also showed reduction in the visible staining of the enzymes. The reference drug, PZQ also showed more or less similar effect like that of the plant extract. The result suggests that phytochemicals of P. fulgens have anthelmintic potential.

Molecular Methods for Detection of Antimicrobial Resistance

DEFF Research Database (Denmark)

Anjum, Muna F.; Zankari, Ea; Hasman, Henrik

2017-01-01

The increase in bacteria harboring antimicrobial resistance (AMR) is a global problem because there is a paucity of antibiotics available to treat multidrug-resistant bacterial infections in humans and animals. Detection of AMR present in bacteria that may pose a threat to veterinary and public...

ANTHELMINTIC EFFECTS OF DRIED GROUND BANANA PLANT LEAVES (MUSA SPP.) FED TO SHEEP ARTIFICIALLY INFECTED WITH HAEMONCHUS CONTORTUS AND TRICHOSTRONGYLUS COLUBRIFORMIS.

Science.gov (United States)

Gregory, Lilian; Yoshihara, Eidi; Silva, Leandro Kataoaka Fernandes; Marques, Eduardo Carvalho; Ribeiro, Bruno Leonardo Mendonça; de Souza Meira, Enoch Brandão; Rossi, Rodolfo Santos; do Amarante, Alessandro Francisco Talamini; Hasegawa, Marjorie Yumi

2017-01-01

Helminths is a endoparasites that cause the major losses for profitable sheep production in Brazil. The increased development of resistant strains of endoparasites have enforced the search for sustainable alternatives. The aim of this paper was to provide information about endoparasites control with banana leaves in infected sheep as alternative control strategies and see its viability. In this study, we performed two trials to investigate the anthelmintic properties of banana leaves on endoparasites in sheep. In Trial 1, twelve sheep were artificially infected with Trichostrongylus colubriformis ; in Trial 2, eleven sheep were artificially infected with Haemonchus contortus . Clinical examinations, packed cell volume, total protein, faecal egg counts (FECs) and egg hatchability tests (EHTs) were performed. At the end of the trials, the sheep were humanely slaughtered, and total worm counts were performed. In Trial 1 and 2, no significant FEC decreases were note but significant diference in EHTs were observed. Total worm counts, clinical and haematological parameters did not reveal significant changes between the treatment and control groups. These results suggest that feeding dried ground banana plant leaves to sheep may reduce the viability of Trichostrongylus colubriformis eggs, and this anthelmintic activity is potentially exploitable as part of an integrated parasite management programme. However, further investigation is needed to establish the optimal dosage, develop a convenient delivery form and confirm the economic feasibility of using banana plantation byproducts as feed for ruminant species. Abbreviations: Coproculture test (CT)., Faecal egg count (FEC)., Egg hatchability test (EHT).

Anastomotic leak detection by electrolyte electrical resistance.

Science.gov (United States)

DeArmond, Daniel T; Cline, Adam M; Johnson, Scott B

2010-08-01

To characterize a new method of postoperative gastrointestinal leak detection based on electrical resistance changes due to extravasated electrolyte contrast. Postoperative gastrointestinal leak results in increased patient morbidity, mortality, and hospital costs that can be mitigated by early diagnosis. A sensitive and specific diagnostic test that could be performed at the bedside has the potential to shorten the time to diagnosis and thereby improve the quality of treatment. Anaesthetized rats underwent celiotomy and creation of a 5-mm gastrotomy. In experimental animals, electrical resistance changes were measured with a direct current ohmmeter after the introduction of 5 cc of 23.4% NaCl electrolyte solution via gavage and measured with a more sensitive alternating current ohmmeter after the gavage of 1-5 cc of 0.9% NaCl. Comparison was made to negative controls and statistical analysis was performed. Leakage from the gastrotomy induced by as little as 1 cc of gavage-delivered 0.9% NaCl contrast solution was detectable as a statistically significant drop in electrical resistance when compared to results from negative controls. Electrical resistance change associated with electrolyte-gated leak detection is highly sensitive and specific and has the potential to be rapidly translated into clinical settings.

Haemonchotolerance in West African Dwarf goats: contribution to sustainable, anthelmintics-free helminth control in traditionally managed Nigerian dwarf goats

Science.gov (United States)

Chiejina, Samuel N.; Behnke, Jerzy M.; Fakae, Barineme B.

2015-01-01

West African Dwarf (WAD) goats are extremely important in the rural village economy of West Africa, but still little is known about their biology, ecology and capacity to cope with gastrointestinal nematode (GIN) infections. Here, we summarise the history of this breed and explain its economic importance in rural West Africa. We review recent work showing that Nigerian WAD goats are highly trypanotolerant and resist infections with Haemonchus contortus more effectively than other breeds of domestic goat (haemonchotolerance). We believe that haemonchotolerance is largely responsible for the generally low level GIN infections and absence of clinical haemonchosis in WADs under field conditions, and has contributed to the relatively successful and sustainable, anthelmintics-free, small-scale system of goat husbandry in Nigeria’s humid zone, and is immunologically based and genetically controlled. If haemonchotolerance can be shown to be genetically controlled, it should be possible to exploit the underlying genes to improve GIN resistance among productive fibre and milk producing breeds of goats, most of which are highly susceptible to nematode infections. Genetic resistance to GIN and trypanosome infections would obviate the need for expensive chemotherapy, mostly unaffordable to small-holder farmers in Africa, and a significant cost of goat husbandry in more developed countries. Either introgression of resistance alleles into susceptible breeds by conventional breeding, or transgenesis could be used to develop novel parasite-resistant, but highly productive breeds, or to improve the resistance of existing breeds, benefitting the local West African rural economy as well as global caprine livestock agriculture. PMID:25744655

Haemonchotolerance in West African Dwarf goats: contribution to sustainable, anthelmintics-free helminth control in traditionally managed Nigerian dwarf goats.

Science.gov (United States)

Chiejina, Samuel N; Behnke, Jerzy M; Fakae, Barineme B

2015-01-01

West African Dwarf (WAD) goats are extremely important in the rural village economy of West Africa, but still little is known about their biology, ecology and capacity to cope with gastrointestinal nematode (GIN) infections. Here, we summarise the history of this breed and explain its economic importance in rural West Africa. We review recent work showing that Nigerian WAD goats are highly trypanotolerant and resist infections with Haemonchus contortus more effectively than other breeds of domestic goat (haemonchotolerance). We believe that haemonchotolerance is largely responsible for the generally low level GIN infections and absence of clinical haemonchosis in WADs under field conditions, and has contributed to the relatively successful and sustainable, anthelmintics-free, small-scale system of goat husbandry in Nigeria's humid zone, and is immunologically based and genetically controlled. If haemonchotolerance can be shown to be genetically controlled, it should be possible to exploit the underlying genes to improve GIN resistance among productive fibre and milk producing breeds of goats, most of which are highly susceptible to nematode infections. Genetic resistance to GIN and trypanosome infections would obviate the need for expensive chemotherapy, mostly unaffordable to small-holder farmers in Africa, and a significant cost of goat husbandry in more developed countries. Either introgression of resistance alleles into susceptible breeds by conventional breeding, or transgenesis could be used to develop novel parasite-resistant, but highly productive breeds, or to improve the resistance of existing breeds, benefitting the local West African rural economy as well as global caprine livestock agriculture. © S.N. Chiejina et al., published by EDP Sciences, 2015.

Detection and semi-quantification of Strongylus vulgaris DNA in equine faeces by real-time quantitative PCR

DEFF Research Database (Denmark)

Nielsen, Martin Krarup; Peterson, David S.; Monrad, Jesper

2008-01-01

Strongylus vulgaris is an important strongyle nematode with high pathogenic potential infecting horses world-wide. Several decades of intensive anthelminitic use has virtually eliminated clinical disease caused by S. vulgaris, but has also causes high levels of anthelmintic resistance in equine...

The anthelmintic levamisole is an allosteric modulator of human neuronal nicotinic acetylcholine receptors.

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Levandoski, Mark M; Piket, Barbara; Chang, Jane

2003-06-13

L-[-]-2,3,5,6-Tetrahydro-6-phenylimidazo[2,1b]-thiazole hydrochloride (levamisole) is an anthelmintic that targets the nicotinic acetylcholine receptors of parasitic nematodes. We report here the effects of levamisole on human neuronal alpha 3 beta 2 and alpha 3 beta 4 nicotinic receptors, heterologously expressed in Xenopus oocytes and studied with the voltage clamp method. Applied alone, levamisole was a very weak partial agonist for the two subunit combinations. When co-applied with acetylcholine, micromolar concentrations of levamisole potentiated responses, while millimolar concentrations inhibited them; these effects were complex functions of both acetylcholine and levamisole concentrations. The differences in the levamisole effects on the two receptor combinations suggest that the effects are mediated by the beta subunit. Several combinations of agonist and anthelmintic gave the dual potentiation/inhibition behavior, suggesting that the modulatory effects are general. Levamisole inhibition showed macroscopic characteristics of open channel block. Several results led us to conclude that levamisole potentiation occurs through noncompetitive binding to the receptor. We propose pseudo-site binding for noncompetitive potentiation by levamisole.

Detection of antibiotic resistance and tetracycline resistance genes in Enterobacteriaceae isolated from the Pearl rivers in South China

International Nuclear Information System (INIS)

Tao Ran; Ying Guangguo; Su Haochang; Zhou Hongwei; Sidhu, Jatinder P.S.

2010-01-01

This study investigated antibiotic resistance profiles and tetracycline resistance genes in Enterobacteriaceae family isolates from the Pearl rivers. The Enterobacteriaceae isolates were tested for susceptibility to seven antibiotics ampicillin, chloramphenicol, ciprofloxacin, levofloxacin, sulphamethoxazole/trimethoprim, tetracycline and trimethoprim. In Liuxi reservoir, with an exception to ampicillin resistant strains (11%) no other antibiotic resistance bacterial strains were detected. However, multiple drug resistance in bacterial isolates from the other sites of Pearl rivers was observed which is possibly due to sewage discharge and input from other anthropogenic sources along the rivers. Four tetracycline resistance genes tet A, tet B, tet C and tet D were detected in the isolates from the rivers. The genes tet A and tet B were widely detected with the detection frequencies of 43% and 40% respectively. Ciprofloxacin and levofloxacin resistant enteric bacteria were also isolated from the pig and duck manures which suggest a wider distribution of human specific drugs in the environment. This investigation provided a baseline data on antibiotic resistance profiles and tetracycline resistance genes in the Pearl rivers delta. - High rates of antibiotic resistance in Enterobacteriaceae from river water are attributed to wastewater contamination.

Detection of antibiotic resistance and tetracycline resistance genes in Enterobacteriaceae isolated from the Pearl rivers in South China

Energy Technology Data Exchange (ETDEWEB)

Tao Ran [State Key Laboratory of Organic Geochemistry, Guangzhou Institute of Geochemistry, Chinese Academy of Sciences, 511 Kehua Street, Tianhe District, Guangzhou 510640 (China); Ying Guangguo, E-mail: guangguo.ying@gmail.co [State Key Laboratory of Organic Geochemistry, Guangzhou Institute of Geochemistry, Chinese Academy of Sciences, 511 Kehua Street, Tianhe District, Guangzhou 510640 (China); Su Haochang [State Key Laboratory of Organic Geochemistry, Guangzhou Institute of Geochemistry, Chinese Academy of Sciences, 511 Kehua Street, Tianhe District, Guangzhou 510640 (China); Zhou Hongwei [Department of Environmental Health, School of Public Health and Tropical Medicine, Southern Medical University, 1838 North Guangzhou Street, Baiyun District, Guangzhou 510515 (China); Sidhu, Jatinder P.S. [CSIRO Land and Water, Queensland Bioscience Precinct, 306 Carmody Road, St Lucia QLD 4067 (Australia)

2010-06-15

This study investigated antibiotic resistance profiles and tetracycline resistance genes in Enterobacteriaceae family isolates from the Pearl rivers. The Enterobacteriaceae isolates were tested for susceptibility to seven antibiotics ampicillin, chloramphenicol, ciprofloxacin, levofloxacin, sulphamethoxazole/trimethoprim, tetracycline and trimethoprim. In Liuxi reservoir, with an exception to ampicillin resistant strains (11%) no other antibiotic resistance bacterial strains were detected. However, multiple drug resistance in bacterial isolates from the other sites of Pearl rivers was observed which is possibly due to sewage discharge and input from other anthropogenic sources along the rivers. Four tetracycline resistance genes tet A, tet B, tet C and tet D were detected in the isolates from the rivers. The genes tet A and tet B were widely detected with the detection frequencies of 43% and 40% respectively. Ciprofloxacin and levofloxacin resistant enteric bacteria were also isolated from the pig and duck manures which suggest a wider distribution of human specific drugs in the environment. This investigation provided a baseline data on antibiotic resistance profiles and tetracycline resistance genes in the Pearl rivers delta. - High rates of antibiotic resistance in Enterobacteriaceae from river water are attributed to wastewater contamination.

P-glycoprotein-9 and macrocyclic lactone resistance status in selected strains of the ovine gastrointestinal nematode, Teladorsagia circumcincta.

Science.gov (United States)

Turnbull, Frank; Jonsson, Nicholas N; Kenyon, Fiona; Skuce, Philip J; Bisset, Stewart A

2018-04-01

The Teladorsagia circumcincta P-glycoprotein-9 (Tci-pgp-9) gene has previously been implicated in multiple-anthelmintic resistance in this parasite. Here we further characterise genetic diversity in Tci-pgp-9 and its possible role in ivermectin (IVM) and multi-drug resistance using two UK field isolates of T. circumcincta, one susceptible to anthelmintics (MTci2) and the other resistant to most available anthelmintics including IVM (MTci5). A comparison of full-length Tci-pgp-9 cDNA transcripts from the MTci2 and MTci5 isolates (∼3.8 kb in both cases) indicated that they shared 95.6% and 99.5% identity at the nucleotide and amino acid levels, respectively. Nine non-synonymous SNPs were found in the MTci5 sequences relative to their MTci2 counterparts. Twelve genomic sequence variants of the first internucleotide binding domain of Tci-pgp-9 were identified and up to 10 of these were present in some individual worms, strongly supporting previous evidence that amplification of this gene has occurred in T. circumcincta. On average, fewer distinct sequence variants of Tci-pgp-9 were present in individual worms of the MTci5 isolate than in those of the MTci2 isolate. A further reduction in the number of sequence variants was observed in individuals derived from an IVM-treated sub-population of MTci5. These findings suggest that Tci-pgp-9 was under purifying selection in the face of IVM treatment in T. circumcincta, with some sequence variants being selected against. Copyright © 2018. Published by Elsevier Ltd.

Preparation of stir cake sorptive extraction based on polymeric ionic liquid for the enrichment of benzimidazole anthelmintics in water, honey and milk samples

Energy Technology Data Exchange (ETDEWEB)

Wang, Yulei [State Key Laboratory of Marine Environmental Science, Key Laboratory of the Ministry of Education for Coastal and Wetland Ecosystem, College of the Environment and Ecology, Xiamen University, Siming Road, P.O. Box 1009, Xiamen 361005 (China); Zhang, Jie [Key Lab of Urban Environment and Health, Institute of Urban Environment, Chinese Academy of Sciences (China); Huang, Xiaojia, E-mail: hxj@xmu.edu.cn [State Key Laboratory of Marine Environmental Science, Key Laboratory of the Ministry of Education for Coastal and Wetland Ecosystem, College of the Environment and Ecology, Xiamen University, Siming Road, P.O. Box 1009, Xiamen 361005 (China); Yuan, Dongxing [State Key Laboratory of Marine Environmental Science, Key Laboratory of the Ministry of Education for Coastal and Wetland Ecosystem, College of the Environment and Ecology, Xiamen University, Siming Road, P.O. Box 1009, Xiamen 361005 (China)

2014-08-20

Highlights: • A new polymeric ionic liquid-based monolith was prepared. • The monolith was used as the extractive medium of stir cake sorptive extraction. • The SCSE–AMIIDB can extract benzimidazole anthelmintics (BAs) effectively. • A combination of SCSE–AMIIDB–LD–HPLC/DAD was developed. • The combination was applied to monitor trace BAs in water, milk and honey samples. - Abstract: In this work, a new stir cake sorptive extraction (SCSE) using polymeric ionic liquid monolith as sorbent was prepared. The sorbent was obtained by in situ copolymerization of an ionic liquid, 1-allyl-3-methylimidazolium bis[(trifluoro methyl)sulfonyl]imide (AMII) and divinylbenzene (DB) in the presence of N,N-dimethylformamide. The influence of the content of ionic liquid and the porogen in the polymerization mixture on extraction performance was studied thoroughly. The physicochemical properties of the polymeric ionic liquid were characterized by infrared spectroscopy, elemental analysis, scanning electron microscopy and mercury intrusion porosimetry. The usefulness of SCSE–AMIIDB was demonstrated by the enrichment of trace benzimidazole anthelmintics. Several parameters affecting the extraction efficiency were investigated, and under the optimized conditions, a simple and effective method for the determination of trace benzimidazoles residues in water, milk and honey samples was established by coupling SCSE–AMIIDB with high performance liquid chromatography/diode array detection (SCSE–AMIIDB–HPLC/DAD). Results indicated that the limits of detection (S/N = 3) for target compounds were 0.020–0.072 μg L{sup −1}, 0.035–0.10 μg L{sup −1} and 0.026–0.076 μg L{sup −1} in water, milk and honey samples, respectively. In addition, an acceptable reproducibility was achieved by evaluating the repeatability and intermediate precision with relative standard deviations (RSD) of less than 9% and 11%, respectively. Finally, the established AMII�

Phytochemicals, antioxidant, and anthelmintic activity of selected traditional wild edible plants of lower Assam.

Science.gov (United States)

Swargiary, Ananta; Daimari, Abhijita; Daimari, Manita; Basumatary, Noymi; Narzary, Ezekiel

2016-01-01

Clerodendrum viscosum , Eryngium foetidum , Lippia javanica , and Murraya koenigii are one among the common wild edible plants in Northeast India which are also used as antidiabetic, stomach-ache relieving drugs, etc., The present study was aimed to reveal the phytochemical, antioxidant, and anthelmintic activity of the plants. The antioxidant capacity of methanolic extract of plants was studied by 1,1-diphenyl-2-picrylhydrazyl (DPPH), ferric reducing antioxidant power, TBARS, and total antioxidant activity (TAA). Total phenolics, flavonoids, Vitamin C, carbohydrate, and protein are also estimated following standard protocols. Anthelmintic activity of the extracts has also been studied in vitro against trematode parasites. The result showed that the methanolic extracts of plants possess a substantial quantity of alkaloids, phenolics, flavonoids, proteins, carbohydrates, and Vitamin C. Phenolics, flavonoids, and Vitamin C contents were found higher in C. viscosum followed by M. koenigii , L. javanica , and E. foetidum . The in vitro antioxidant assays revealed substantial free radical scavenging property in all the plants. TAA increased in the order C. viscosum > M. koenigii > L. javanica > E. foetidum . Similarly, C. viscosum displayed a better antioxidant capacity with IC 50 values 29.74 ± 3.63 μg and 148.77 ± 18.38 μg for DPPH and thiobarbituric acid reactive species, respectively. In addition, the plant extracts also showed good anthelmintic activity against Paramphistomum sp. Time taken for paralysis and death were 0:56 ± 0:09 h and 1:35 ± 0:07 h for L. javanica at 50 mg/mL concentration. The study therefore suggests the importance of tested plants as a natural source of free radical scavenger and plausible veterinary uses.

The anthelmintic effect of aqueous methanol extract of Combretum molle (R. Br. x. G. Don) (Combretaceae) in lambs experimentally infected with Haemonchus contortus.

Science.gov (United States)

Simon, M K; Ajanusi, O J; Abubakar, M S; Idris, A L; Suleiman, M M

2012-06-08

The aqueous methanol extract from the stem-bark of Combretum molle was evaluated for anthelmintic activity in lambs infected with Haemonchus contortus using faecal egg count (FEC) reduction assay. The extract showed a dose-dependent reduction in FEC in infected animals. At doses of 500, 1000 and 2000 mg kg(-1), the extract caused FEC reduction of 63%, 69.25% and 96.23%, respectively. Similarly, the standard anthelmintic (albendazole) at a dose of 200 mg kg(-1) produced FEC reduction of 99.24%. FEC reduction produced by the extract at doses of 500 and 1000 mg kg(-1) is below the minimum standard of 90% FEC recommended by the World Association for the Advancement of Veterinary Parasitology (WAAVP). However, there was no significant (P>0.05) difference between the means of groups treated with 1000 mg kg(-1) and 2000 mg kg(-1) compared to that of albendazole. In this study, C. molle has shown a promising anthelmintic activity against experimental haemonchosis. Nonetheless, further studies to evaluate its detailed toxicity are required for the plant extract to be developed into a useful anthelmintic drug. There is also the need to evaluate other parts of the plant (root, leaves, fruits, etc.) for the same effect. Copyright © 2012 Elsevier B.V. All rights reserved.

Determination of anthelmintic drug residues in milk using ultra high performance liquid chromatography-tandem mass spectrometry with rapid polarity switching.

Science.gov (United States)

Whelan, Michelle; Kinsella, Brian; Furey, Ambrose; Moloney, Mary; Cantwell, Helen; Lehotay, Steven J; Danaher, Martin

2010-07-02

A new UHPLC-MS/MS (ultra high performance liquid chromatography coupled to tandem mass spectrometry) method was developed and validated to detect 38 anthelmintic drug residues, consisting of benzimidazoles, avermectins and flukicides. A modified QuEChERS-type extraction method was developed with an added concentration step to detect most of the analytes at keeper to ensure analytes remain in solution. Using rapid polarity switching in electrospray ionisation, a single injection was capable of detecting both positively and negatively charged ions in a 13 min run time. The method was validated at two levels: the unapproved use level and at the maximum residue level (MRL) according to Commission Decision (CD) 2002/657/EC criteria. The decision limit (CCalpha) of the method was in the range of 0.14-1.9 and 11-123 microg kg(-1) for drugs validated at unapproved and MRL levels, respectively. The performance of the method was successfully verified for benzimidazoles and levamisole by participating in a proficiency study.

Preliminary analysis of the relationship between structure and anthelmintic activity of condensed tannins in cattle nemaotdes

DEFF Research Database (Denmark)

Desrues, Olivier; Larsen Enemark, Heidi; Mueller-Harvey, Irene

2013-01-01

Some plant secondary metabolites as tannins have direct anthelminthic properties and may play a role in the control of nematodes in livestock. However, their great diversity in structural characteristics and different levels of content in plants are responsible for a highly variable response...... in anthelmintic activity, as measured in vitro. The aim of the present study was to assess the relationship between structure and anthelmintic activity using an in vitro assay. We used a series of purified tannins (from 65% to 100% of purity) characterized for their degree of polymerization (mDP), prodelphinidin....../procyanidin ratio and cis/trans ratio by thiolytic degradation. Tannins diluted in two concentrations in water, epigallocatechin gallate, positive (ivermectin) and negative (water) controls were examined by the Larval Feeding Inhibition Assay (LFIA) with first stage larvae (L1) of the cattle nematode Cooperia...

Preliminary analysis of the relationship between structure and anthelmintic activity of condensed tannins in cattle nematodes

DEFF Research Database (Denmark)

Desrues, Oliver; Enemark, Heidi L.; Mueller-Harvey, I.

2013-01-01

Some plant secondary metabolites as tannins have direct anthelminthic properties and may play a role in the control of nematodes in livestock. However, their great diversity in structural characteristics and different levels of content in plants are responsible for a highly variable response...... in anthelmintic activity, as measured in vitro. The aim of the present study was to assess the relationship between tannin structure and anthelmintic activity using an in vitro assay. We used a series of purified tannins (from 65% to 100% of purity) characterized for their degree of polymerization (m......DP), prodelphinidin/procyanidin (PC/PD) ratio and cis/trans ratio by thiolytic degradation. Tannins diluted in two concentrations in water, epigallocatechin gallate (EGCG), positive (ivermectin) and negative (water) controls were examined by the Larval Feeding Inhibition Assay (LFIA) with first stage larvae (L1...

Oxfendazole Resistance in Gastrointestinal Nematodes of Beetal Goats at Livestock Farms of Punjab (Pakistan

Directory of Open Access Journals (Sweden)

M. Saeed

2007-01-01

Full Text Available This study was carried out to screen goat farms for anthelmintic resistance (AR against oxfendazole (OXF and to determine contributory factors for its development. For this purpose, Beetal goat farms (n = 18 were randomly selected, with natural mixed gastrointestinal nematodosis infection. In vivo (faecal egg count reduction test and in vitro (egg hatch assay tests were used to ascertain the presence of AR while a scorecard was used to determine the role of possible contributory factors for oxfendazole resistance. For in vivo test, the experimental animals were divided into two groups of 10 animals each; one group received OXF treatment, while the other served as control. Pre- and post-treatment coproculture was performed to identify the species and genera of nematodes. Egg hatch assay (EHA was used to confirm the results of FECRT. Fecal egg count reduction test (FECRT revealed the development of resistance on six farms and post-treatment larval cultures indicated Haemonchus contortus, Trichostrongylus colubriformis, Cooperia curticei, Teladorsagia circumcincta and Oesophagostomum spp. as dominant species with resistance. Furthermore, EHA confirmed the results of FECRT. Among the presumptive factors for AR, the highest composite score was for rotation of anthelmintics followed by treatment frequency, dose rate and nature of medication. The scorecard for the development of AR, used in this study, may be helpful for the assessment of contributory factors of AR.

Resistance delaying strategies on UK sheep farms: A cost benefit analysis.

Science.gov (United States)

Learmount, Jane; Glover, Mike J; Taylor, Mike A

2018-04-30

UK guidelines for the sustainable control of parasites in sheep (SCOPS) were formulated with the primary aim of delaying development of anthelmintic resistance (AR) on UK sheep farms. Promoting their use requires the engagement and commitment of stakeholders. An important driver for behavioural change in sheep farmers is evidence of economic benefits. A recent evaluation of SCOPS guidance in practice demonstrated a significant reduction in anthelmintic use, suggesting economic benefits through a direct reduction in product and labour costs. However, in order to maintain production, a range of alternative control strategies are advised, resulting in additional costs to farmers and so a full cost benefit analysis of best practice management was undertaken. We allocated financial values to the management recommendations described in the SCOPS technical manual. Benefits were calculated using data for production variables and anthelmintic use measured during studies to evaluate the effect of SCOPS recommendations on 16 UK sheep farms and from other published work. As SCOPS control is not prescriptive and a range of different diagnostics are available, best and worst case scenarios were presented, comparing the cheapest methods (e.g. egg counts without larval culture) and management situations (e.g closed flocks not requiring quarantine treatments) with the most laborious and expensive. Simulations were run for farms with a small, medium or large flock (300; 1000; 1900 ewes) as well as comparing scenarios with and without potential production benefits from using effective wormers. Analysis demonstrated a moderate cost for all farms under both scenarios when production benefits were not included. A cost benefit was demonstrated for medium and large farms when production benefits were included and the benefit could be perceived as significant in the case of the large farms for the best case scenario (>£5000 per annum). Despite a significant potential reduction in

Practical aspects of equine parasite control: a review based upon a workshop discussion consensus.

Science.gov (United States)

Nielsen, M K; Fritzen, B; Duncan, J L; Guillot, J; Eysker, M; Dorchies, P; Laugier, C; Beugnet, F; Meana, A; Lussot-Kervern, I; von Samson-Himmelstjerna, G

2010-07-01

Development of resistance of several important equine parasites to most of the available anthelmintic drug classes has led to a reconsideration of parasite control strategies in many equine establishments. Routine prophylactic treatments based on simple calendar-based schemes are no longer reliable and veterinary equine clinicians are increasingly seeking advice and guidance on more sustainable approaches to equine parasite control. Most techniques for the detection of equine helminth parasites are based on faecal analysis and very few tests have been developed as diagnostic tests for resistance. Recently, some molecular and in vitro based diagnostic assays have been developed and have shown promise, but none of these are currently available for veterinary practice. Presently, the only reliable method for the detection of anthelmintic resistance is a simple faecal egg count reduction test, and clinicians are urged to perform such tests on a regular basis. The key to managing anthelmintic resistance is maintaining parasite refugia and this concept is discussed in relation to treatment strategies, drug rotations and pasture management. It is concluded that treatment strategies need to change and more reliance should now be placed on surveillance of parasite burdens and regular drug efficacy tests are also recommended to ensure continuing drug efficacy. The present review is based upon discussions held at an equine parasite workshop arranged by the French Equine Veterinary Association (Association Vétérinaire Equine Française, AVEF) in Reims, France, in October 2008.

Anthelmintics, used for the treatment of fascioliasis as uncouplers of oxidative phosphorylation in warm blooded animals

NARCIS (Netherlands)

Miert, A.S.J.P.A.M. van; Groeneveld, H.W.

1969-01-01

Dinitrophenol and liverfluke anthelmintics such as niclofolan, nitroxynil, bromophenophos and hexachlorophene increased ATP-ase activity of rat-liver mitochondria and caused a contracture of striated muscle in vitro. After lethal doses, rigidity occured soon after deatth. It is postulated that these

Synthesis and anthelmintic activity of arctigenin derivatives against Dactylogyrus intermedius in goldfish.

Science.gov (United States)

Hu, Yang; Liu, Lei; Liu, Guang-Lu; Tu, Xiao; Wang, Gao-Xue; Ling, Fei

2017-08-01

To control the parasitic disease of Dactylogyrus intermedius, a series of new arctigenin derivatives were designed, synthesized and tested in our study. The anthelmintic activity of most of the derivatives ranged from 1 to 10mg/L. Compared to traditional drug praziquantel (EC 50 =2.69mg/L), ether derivatives 2g and 2h exhibited slightly higher anti-parasitic activity, with the EC 50 values of 2.48 and 1.52mg/L, respectively. Furthermore, the arctigenin-imidazole hybrids 4a and 4b also removed D. intermedius effectively, with the EC 50 values of 2.13 and 2.07mg/L, respectively. The structure-activity relationship analysis indicated that four carbon atoms length of linker and imidazole substitute group could significantly increase the anthelmintic activity, and reduced the toxicity. Through the scanning electron microscope observation, compounds 4a and 4b caused the D. intermedius tegumental damage such as intensive wrinkles, holes and nodular structures. Overall, the structural optimization analysis of arctigenin suggested that 4a and 4b can be used for preventing and controlling Dactylogyrus infections and considered as promising lead compounds for the development of commercial drugs. Copyright © 2017 Elsevier Ltd. All rights reserved.

Anthelmintic efficacies of a tablet formula of ivermectin-praziquantel on horses experimentally infected with three Strongylus species.

Science.gov (United States)

Bonneau, Stephane; Maynard, Laurence; Tomczuk, Krzysztof; Kok, Dawid; Eun, Hyone-Myong

2009-09-01

In this blinded randomized and controlled study, the anthelmintic efficacy of a tablet formula of ivermectin-praziquantel was evaluated in horses experimentally infected with three species of Strongylus larvae. Eighteen previously dewormed horses were inoculated on study day 0 with third-stage larvae of Strongylus vulgaris, Strongylus equinus, and Strongylus edentatus. The horses were randomly allocated to three groups (n = 6): test-drug (tablet formula), positive-control (reference gel), and negative-control (placebo tablet). On day 56, the horses were treated once with the respective drugs. On day 95, the horses were sacrificed, and necropsy examinations were performed to assess the status of the parasite burden (L4 and immature L5) and pathological lesions on selected organs and tissues. By the criteria of worm counts, the test-drug and positive-control showed, respectively, 100% and 97.3% anthelmintic efficacies on S. vulgaris, 100% and 81.4% on S. equinus, and equally 100% on S. edentatus. However, the efficacies on S. equinus and S. edentatus should be taken only as face values considering their respective low worm counts in the placebo group. The S. vulgaris-induced arterial lesions were also reduced in the test-drug and positive-control groups with efficacies of 73.9% and 62.9%, respectively. No adverse reactions were observed with either of the drugs. Our data demonstrate that the Equimax tablet formula was as safe and efficacious as the gel formula anthelmintic on large strongyles in horses.

Detection of multidrug-resistant tuberculosis from stored DNA Samples: A multicenter study

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Marie Sylvianne Rabodoarivelo

2018-01-01

Full Text Available Background: In low-income countries, rapid detection of tuberculosis (TB drug resistance is often restricted by the difficulties of transporting and storing sputum samples from remote health centers to the reference laboratories where molecular tests are available. The aim of this study was to evaluate the performance of four transport and storage systems for molecular detection of rifampicin (RIF and isoniazid (INH resistance. Methods: This was a multicenter study. Molecular detection of RIF and INH resistance was performed directly from smear-positive TB sputa spotted on a slide, FTA card, GenoCard, and ethanol using the Genotype MTBDRplus assay. The performance of the DNA extraction method from each storage support to detect drug resistance was assessed by calculating their sensitivity and specificity compared to the phenotypic method. Results: From all sites, the overall sensitivity and specificity for RIF-resistance detection was 88% and 85%, respectively, for slides, 86% and 92%, respectively, for GenoCard, 87% and 89%, respectively, for FTA card, and 88% and 92%, respectively, for ethanol. For INH-resistance detection, the overall sensitivity and specificity was 82% and 90%, respectively, for slides, 85% and 96%, respectively, for GenoCard, 86% and 92%, respectively, for FTA card, and 86% and 94%, respectively, for ethanol. Conclusion: Smear slides and filter cards showed to be very useful tools to facilitate DNA extraction from sputum samples with the potential to accelerate the detection of drug resistance in remote areas.

Detection of multidrug-resistant tuberculosis from stored DNA Samples: A multicenter study.

Science.gov (United States)

Rabodoarivelo, Marie Sylvianne; Brandao, A; Cergole Novella, M C; C Bombonatte, A G; Imperiale, B; Rakotosamimanana, N; Morcillo, N; Rasolofo, V; Palomino, J C; Martin, A

2018-01-01

In low-income countries, rapid detection of tuberculosis (TB) drug resistance is often restricted by the difficulties of transporting and storing sputum samples from remote health centers to the reference laboratories where molecular tests are available. The aim of this study was to evaluate the performance of four transport and storage systems for molecular detection of rifampicin (RIF) and isoniazid (INH) resistance. This was a multicenter study. Molecular detection of RIF and INH resistance was performed directly from smear-positive TB sputa spotted on a slide, FTA card, GenoCard, and ethanol using the Genotype MTBDRplus assay. The performance of the DNA extraction method from each storage support to detect drug resistance was assessed by calculating their sensitivity and specificity compared to the phenotypic method. From all sites, the overall sensitivity and specificity for RIF-resistance detection was 88% and 85%, respectively, for slides, 86% and 92%, respectively, for GenoCard, 87% and 89%, respectively, for FTA card, and 88% and 92%, respectively, for ethanol. For INH-resistance detection, the overall sensitivity and specificity was 82% and 90%, respectively, for slides, 85% and 96%, respectively, for GenoCard, 86% and 92%, respectively, for FTA card, and 86% and 94%, respectively, for ethanol. Smear slides and filter cards showed to be very useful tools to facilitate DNA extraction from sputum samples with the potential to accelerate the detection of drug resistance in remote areas.

Evaluation of the larval migration inhibition assay for detecting macrocyclic lactone resistance in Dirofilaria immitis.

Science.gov (United States)

Evans, Christopher C; Moorhead, Andrew R; Storey, Bobby E; Blagburn, Byron L; Wolstenholme, Adrian J; Kaplan, Ray M

2017-11-15

Anthelmintics of the macrocyclic lactone (ML) drug class are widely used as preventives against the canine heartworm (Dirofilaria immitis). Over the past several years, however, reports of ML lack of efficacy (LOE) have emerged, in which dogs develop mature heartworm infection despite the administration of monthly prophylactics. More recently, isolates from LOE cases have been used to infect laboratory dogs and the resistant phenotype has been confirmed by the establishment of adult worms in the face of ML treatment at normally preventive dosages. Testing for and monitoring resistance in D. immitis requires a validated biological or molecular diagnostic assay. In this study, we assessed a larval migration inhibition assay (LMIA) that we previously optimized for use with D. immitis third-stage larvae (L 3 ). We used this assay to measure the in vitro ML susceptibilities of a known-susceptible laboratory strain of D. immitis and three highly suspected ML-resistant isolates originating from three separate LOE cases; progeny from two of these isolates have been confirmed ML-resistant by treatment of an infected dog in a controlled setting. A nonlinear regression model was fit to the dose-response data, from which IC 50 values were calculated. The D. immitis LMIA yielded consistent and reproducible dose-response data; however, no statistically significant differences in drug susceptibility were observed between control and LOE parasites. Additionally, the drug concentrations needed to paralyze the L 3 were much higher than those third- and fourth-stage larvae would experience in vivo. IC 50 values ranged from 1.57 to 5.56μM (p≥0.19). These data could suggest that ML resistance in this parasite is not mediated through a reduced susceptibility of L 3 to the paralytic effects of ML drugs, and therefore motility-based assays are likely not appropriate for measuring the effects of MLs against D. immitis in this target stage. Published by Elsevier B.V.

Evaluation of resistance in a selected field strain of Haemonchus contortus to ivermectin and moxidectin using the Larval Migration on Agar Test

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Fernanda S. Fortes

2013-02-01

Full Text Available Haemonchus contortus is one of the most common and economically significant causes of disease in small ruminants worldwide, and the control programs of parasitic nematodes - including H. contortus - rely mostly on the use of anthelmintic drugs. The consequence of the use of this, as the sole sanitary strategy to avoid parasite infections, was the reduction of the efficacy of all chemotherapeutic products with a heavy selection for resistance. The widespread of anthelmintic resistance and the difficulty of its early diagnosis has been a major concern for the sustainable parasite management on farms. The objective of this research was to determine and compare the ivermectin (IVM and moxidectin (MOX effect in a selected field strain of H. contortus with a known resistance status, using the in vitro larval migration on agar test (LMAT. Third stage larvae of the selected isolate were obtained from faecal cultures of experimentally infected sheep and incubated in eleven increasing diluted concentrations of IVM and MOX (6, 12, 24, 48, 96, 192, 384, 768, 1536, 3072 and 6144µg/mL. The dose-response sigmoidal curves were obtained using the R² value of >0.90 and the lethal concentration (LC50 dose for the tested anthelmintic drugs using a four-parameter logistic model. The LC50 value for MOX was significantly lower than IVM (1.253µg/mL and 91.06µg/mL, identifying the H. contortus isolate as considerably less susceptible to IVM compared to MOX. Furthermore, the LMAT showed a high consistency (p<0.0001 and provided to be a useful diagnostic tool for monitoring the resistance status of IVM and MOX in H. contortus field isolate, as well as it may be used for official routine drug monitoring programs under the Ministry of Agriculture (MAPA guidance.

Evaluation of anthelmintic resistance in livestock parasites using observational data and hierarchical models

DEFF Research Database (Denmark)

Nielsen, Martin Krarup; Vidyashankar, Anand N.; Hanlon, Bret

= 200 eggs per gram (EPG) and were treated. Post treatment samples and information on age, gender and farm zip code were collected for each horse. In addition, individual coprocultureswere performed on all pretreatment fecal samples to determine the presence of Strongylus vulgaris, with 31farms (48.......4 %) testing positive. The efficacy of pyrantel was unaffected by the presence of S. vulgaris on the farm. The cutoff LCL values used for classifying farms as pyrantel resistant were: >92%: no resistance, 88-92%: suspect resistance, and

Modified DNA extraction for rapid PCR detection of methicillin-resistant staphylococci

International Nuclear Information System (INIS)

Japoni, A.; Alborzi, A.; Rasouli, M.; Pourabbas, B.

2004-01-01

Nosocomial infection caused by methicillin-resistant staphylococci poses a serious problem in many countries. The aim of this study was to rapidly and reliably detect methicillin-resistant-staphylococci in order to suggest appropriate therapy. The presence or absence of the methicillin-resistance gene in 115 clinical isolates of staphylococcus aureus and 50 isolates of coagulase negative staphylococci was examined by normal PCR. DNA extraction for PCR performance was then modified by omission of achromopeptadiase and proteinase K digestion, phenol/chloroform extraction and ethanol precipitation. All isolates with Mic>8 μ g/ml showed positive PCR. No differences in PCR detection have been observed when normal and modified DNA extractions have been performed. Our modified DNA extraction can quickly detect methicillin-resistant staphylococci by PCR. The advantage of rapid DNA extraction extends to both reduction of time and cost of PCR performance. This modified DNA extraction is suitable for different PCR detection, when staphylococci are the subject of DNA analysis

Detection of low frequency multi-drug resistance and novel putative maribavir resistance in immunocompromised paediatric patients with cytomegalovirus

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Charlotte Jane Houldcroft

2016-09-01

Full Text Available Human cytomegalovirus (HCMV is a significant pathogen in immunocompromised individuals, with the potential to cause fatal pneumonitis and colitis, as well as increasing the risk of organ rejection in transplant patients. With the advent of new anti-HCMV drugs there is therefore considerable interest in using virus sequence data to monitor emerging resistance to antiviral drugs in HCMV viraemia and disease, including the identification of putative new mutations. We used target-enrichment to deep sequence HCMV DNA from 11 immunosuppressed paediatric patients receiving single or combination anti-HCMV treatment, serially sampled over 1-27 weeks. Changes in consensus sequence and resistance mutations were analysed for three ORFs targeted by anti-HCMV drugs and the frequencies of drug resistance mutations monitored. Targeted-enriched sequencing of clinical material detected mutations occurring at frequencies of 2%. Seven patients showed no evidence of drug resistance mutations. Four patients developed drug resistance mutations a mean of 16 weeks after starting treatment. In two patients, multiple resistance mutations accumulated at frequencies of 20% or less, including putative maribavir and ganciclovir resistance mutations P522Q (UL54 and C480F (UL97. In one patient, resistance was detected 14 days earlier than by PCR. Phylogenetic analysis suggested recombination or superinfection in one patient. Deep sequencing of HCMV enriched from clinical samples excluded resistance in 7 of eleven subjects and identified resistance mutations earlier than conventional PCR-based resistance testing in 2 patients. Detection of multiple low level resistance mutations was associated with poor outcome.

Evaluation of anthelmintic potential of the Ethiopian medicinal plant Embelia schimperi Vatke in vivo and in vitro against some intestinal parasites.

Science.gov (United States)

Debebe, Yared; Tefera, Mesfin; Mekonnen, Walelign; Abebe, Dawit; Woldekidan, Samuel; Abebe, Abiy; Belete, Yehualashet; Menberu, Temesgen; Belayneh, Bethelhem; Tesfaye, Berhanu; Nasir, Ibrahim; Yirsaw, Kidist; Basha, Hirut; Dawit, Asrat; Debella, Asfaw

2015-06-18

Embelia schimperi has been used for the treatment of intestinal parasites especially tapeworm infestations for centuries in Ethiopia. However, there is lack of scientific based evidences regarding the efficacy, safety and phytochemical analysis of this plant despite its frequent use as an anthelmintic. This study has therefore evaluated the efficacy and acute toxicity of E. schimperi thereby generating relevant preclinical information. The anthelmintic activities of the crude hydroalcoholic extract of E. schimperi and the isolated compound, embelin, were conducted using in vivo and in vitro models against the dwarf tapeworm, Hymenolepis nana, and the hookworm, Necator americanus, respectively. LD50 of the crude hydroalcoholic extract was determined using Swiss albino mice following the OECD guidelines. Chemical characterization of the isolated embelin was conducted using UV-spectroscopy, HPLC and NMR. In the acute toxicity study no prominent signs of toxicity and mortality were recorded among the experimental animals at the highest administered dose. Hence the LD50 of the plant was found to be higher than 5000 mg/kg. In vivo cestocidal activity of the crude hydroalcoholic extract of E. schimperi showed 100% parasite clearance at 1000 mg/kg, while the diammonium salt of embelin showed 85.3% parasite clearance at 750 mg/kg. The in vitro anthelminthic activity study revealed that the LC50 value of the crude extract and albendazole were 228.7 and 51.33 μg/mL, respectively. The results clearly indicated that the hydroalcoholic extract of E. schimperi and the diammonium salt of the isolated compound embelin had anthelmintic activity against hookworm larva in vitro and H. nana in vivo. Hence the findings of this study showed Embelia schimperi appears to possess some anthelmintic activity that may support the usage of these plants by local traditional healers to treat helminthic infestations.

Monitoring of anthelmintic resistance in small strongyles in the Czech Republic in the years 2006–2009

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Štěpán Bodeček

2013-01-01

Full Text Available The aim of the field study performed in 2006 was to investigate the occurrence and distribution of intestinal helminths in horses based on pre-treatment faecal egg counts. In total, 948 horses bred on 37 farms were tested. Thirty six (97.2% farms tested were positive for cyathostomins; horses in 9 (24.3%, 6 (16.2% and 1 (2.7% different herds tested were positive for Parascaris equorum, Anoplocephala perfoliata and Strongyloides westeri, respectively. In 21 herds, 344 horses with values exceeding 100 eggs per gram were included in the trial for the presence of drug resistant cyathostomins by a faecal egg count reduction test. Horses were treated orally with recommended doses of fenbendazole and ivermectin. Resistance to fenbendazole was detected on 20 farms (95.24% with values of faecal egg count reduction test ranging from 0 to 90%. Ivermectin remained effective in all tested herds with the value of faecal egg count reduction test 96–100%. In autumn 2008, 178 horses on 10 farms were examined. Of these, only seven horses tested were negative for cyathostomins. One farm was tested positively for Anoplocephala perfoliata, and one for Parascaris equorum. In spring 2009, six farms were examined, four of which were the same farms as in 2006. We found a decreased number of eggs per gram in all horses, but an increase in benzimidazole resistance, which was found in 5 farms out of 6 (faecal egg count reduction test 15.2–84.6%. This is the first wide survey in horses from the Czech Republic. Based on this study, we can conclude that benzimidazole resistant cyathostomins in horses are widespread but ivermectin is still fully effective.

PCR detection of oxytetracycline resistance genes otr(A) and otr(B) in tetracycline-resistant streptomycete isolates from diverse habitats

NARCIS (Netherlands)

Nikolakopoulou, T; Egan, S; van Overbeek, L; Guillaume, G; Heuer, H; Wellington, EMH; van Elsas, JD; Collard, JM; Smalla, K; Karagouni, A

2005-01-01

A range of European habitats was screened by PCR for detection of the oxytetracycline resistance genes otr(A) and otr(B), found in the oxytetracycline-producing strain Streptomyces rimosus. Primers were developed to detect these otr genes in tetracycline-resistant (Tc-R) streptomycete isolates from

In vitro anthelmintic and cytotoxicity activities the Digitaria insularis (Poaceae).

Science.gov (United States)

Santos, Francianne Oliveira; de Lima, Hélimar Gonçalves; de Souza Santos, Nathália Silva; Serra, Taiane Menezes; Uzeda, Rosângela Soares; Reis, Isabella Mary Alves; Botura, Mariana Borges; Branco, Alexsandro; Batatinha, Maria José Moreira

2017-10-15

This study aimed to evaluate the in vitro activity of D. insularis extracts and fractions against gastrointestinal nematodes of goats and its cytotoxicity on Vero cells. The egg hatch (EHT) and larval motility (LMT) tests were conducted to investigate the anthelmintic effects of the crude hydroethanolic (CH), ethyl acetate (EA), butanolic (BT) and residual hydroethanolic (RH) extracts. The elution of the active extract (EA) on column chromatography (SiO 2 ) using organic solvents furnished six fractions (FR1 to FR6), which were also tested. Cytotoxicity was determined using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and Trypan Blue exclusion assays. All extracts, FR2 and FR3, inhibited egg hatching in a concentration-dependent manner. The EHT led to EC 50 values (effective concentration 50%) of 0.64; 0.69; 0.77; 0.96; 0.27 and 0.65mg/mL for CH, EA, BT, RH, FR2 and FR3, respectively. However, the extracts exhibited low effect on the motility of L 3. In the cytotoxicity evaluation (MTT assay), the IC 50 (inhibitory concentration 50%) was 1.18 (EA), 1.65 (FR2) and 1.59mg/mL (FR3), which was relatively high (low toxicity) in comparison to the EC 50 values in EHT, mainly for FR2. The chemical analyses of most active fractions (FR2) by Liquid Chromatography coupled to Mass Spectrometry (LC-MS) led the characterization of the flavones tricin and diosmetin. These results showed the high anthelmintic effect and low cytotoxicity of D. insularis and also that the flavones can be probably responsible for the nematocidal activity of this plant. Copyright © 2017 Elsevier B.V. All rights reserved.

Molecular detection of disease resistance genes to powdery mildew ...

African Journals Online (AJOL)

A study was conducted to detect the presence of disease resistance genes to infection of wheat powdery mildew (Blumeria graminis f. sp. tritici) in selected wheat cultivars from China using molecular markers. Genomic DNA of sixty cultivars was extracted and tested for the presence of selected prominent resistance genes to ...

Impact of chemical structure of flavanol monomers and condensed tannins on in vitro anthelmintic activity against bovine nematodes

DEFF Research Database (Denmark)

Desrues, Olivier; Fryganas, Christos; Ropiak, Honorata M.

2016-01-01

Plants containing condensed tannins (CT) may have potential to control gastrointestinal nematodes (GIN) of cattle. The aim was to investigate the anthelmintic activities of four flavan-3-ols, two galloyl derivatives and 14 purified CT fractions, and to define which structural features of CT deter...

Study on drug resistance of mycobacterium tuberculosis in patients with pulmonary tuberculosis by drug resistance gene detecting

International Nuclear Information System (INIS)

Wang Wei; Li Hongmin; Wu Xueqiong; Wang Ansheng; Ye Yixiu; Wang Zhongyuan; Liu Jinwei; Chen Hongbing; Lin Minggui; Wang Jinhe; Li Sumei; Jiang Ping; Feng Bai; Chen Dongjing

2004-01-01

To investigate drug resistance of mycobacterium tuberculosis in different age group, compare detecting effect of two methods and evaluate their the clinical application value, all of the strains of mycobacterium tuberculosis were tested for resistance to RFP, INH SM PZA and EMB by the absolute concentration method on Lowenstein-Jensen medium and the mutation of the rpoB, katG, rpsL, pncA and embB resistance genes in M. tuberculosis was tested by PCR-SSCP. In youth, middle and old age group, the rate of acquired drug resistance was 89.2%, 85.3% and 67.6% respectively, the gene mutation rate was 76.2%, 81.3% and 63.2% respectively. The rate of acquired drug resistance and multiple drug resistance in youth group was much higher than those in other groups. The gene mutation was correlated with drug resistance level of mycobacterium tuberculosis. The gene mutation rate was higher in strains isolated from high concentration resistance than those in strains isolated from low concentration resistance. The more irregular treatment was longer, the rate of drug resistance was higher. Acquired drug resistance varies in different age group. It suggested that surveillance of drug resistence in different age group should be taken seriously, especially in youth group. PCR - SSCP is a sensitive and specific method for rapid detecting rpoB, katG, rpsL, pncA and embB genes mutations of MTB. (authors)

Elisa development for detection of glyphosat resistant gm soybean

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Владислав Геннадійович Спиридонов

2015-11-01

Full Text Available During research we have utilized recombinant enzyme 5-enolpyruvylshikimate-3-phosphate synthase (CP4 EPSPS, conferring resistance to glyphosate for GM soybean, for the hen immunization and obtaining specific yolk antibodies IgY. Stages of ELISA development that can detect at least 0,1 % of GM-soybean resistant to glyphosate were present

Effectiveness of Ivermectin and Albendazole against Haemonchus contortus in Sheep in West Java, Indonesia.

Science.gov (United States)

Puspitasari, Silvia; Farajallah, Achmad; Sulistiawati, Erni; Muladno

2016-02-01

Administering a half dose of an anthelmintic is a simple method for detecting resistance in parasites infesting small ruminants. When a single anthelmintic fails in native sheep from Indonesia, a combination of anthelmintics from different chemical classes with different modes of action are administered as an alternative parasite-control strategy. This study compared the anthelmintic efficacy of ivermectin (IVM) and albendazole (ABZ) given either separately as a single dose or half dose or co-administered to sheep naturally infected with Haemonchus contortus. Twelve sheep from Bogor, West Java, Indonesia were divided into the following six treatment groups: half-dose IVM, full-dose IVM, half-dose ABZ, full-dose ABZ, combined IVM + ABZ, and control. The treatment efficacy was determined using the faecal egg count reduction test (FECRT) at day 0 (pre-treatment) and post-treatment at days 7, 14, 21, 28, 35, and 42. The efficacies of half-dose IVM, full-dose IVM, half-dose ABZ, full-dose ABZ, and the combination treatment ranged from -1900% to 100%, 99% to 100%, -167% to 100%, -467% to 89%, and -200% to 100%, respectively. The FECRT for the half-dose IVM, half-dose ABZ, full-dose ABZ showed that H. contortus is resistant to half-dose IVM and ABZ. Full-dose IVM was effective against H. contortus. The combined treatment was more effective against H. contortus than ABZ alone.

Search Engine for Antimicrobial Resistance: A Cloud Compatible Pipeline and Web Interface for Rapidly Detecting Antimicrobial Resistance Genes Directly from Sequence Data.

Science.gov (United States)

Rowe, Will; Baker, Kate S; Verner-Jeffreys, David; Baker-Austin, Craig; Ryan, Jim J; Maskell, Duncan; Pearce, Gareth

2015-01-01

Antimicrobial resistance remains a growing and significant concern in human and veterinary medicine. Current laboratory methods for the detection and surveillance of antimicrobial resistant bacteria are limited in their effectiveness and scope. With the rapidly developing field of whole genome sequencing beginning to be utilised in clinical practice, the ability to interrogate sequencing data quickly and easily for the presence of antimicrobial resistance genes will become increasingly important and useful for informing clinical decisions. Additionally, use of such tools will provide insight into the dynamics of antimicrobial resistance genes in metagenomic samples such as those used in environmental monitoring. Here we present the Search Engine for Antimicrobial Resistance (SEAR), a pipeline and web interface for detection of horizontally acquired antimicrobial resistance genes in raw sequencing data. The pipeline provides gene information, abundance estimation and the reconstructed sequence of antimicrobial resistance genes; it also provides web links to additional information on each gene. The pipeline utilises clustering and read mapping to annotate full-length genes relative to a user-defined database. It also uses local alignment of annotated genes to a range of online databases to provide additional information. We demonstrate SEAR's application in the detection and abundance estimation of antimicrobial resistance genes in two novel environmental metagenomes, 32 human faecal microbiome datasets and 126 clinical isolates of Shigella sonnei. We have developed a pipeline that contributes to the improved capacity for antimicrobial resistance detection afforded by next generation sequencing technologies, allowing for rapid detection of antimicrobial resistance genes directly from sequencing data. SEAR uses raw sequencing data via an intuitive interface so can be run rapidly without requiring advanced bioinformatic skills or resources. Finally, we show that SEAR

The diterpenoid 7-keto-sempervirol, derived from Lycium chinense, displays anthelmintic activity against both Schistosoma mansoni and Fasciola hepatica.

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Jennifer Edwards

2015-03-01

Full Text Available BACKGROUND: Two platyhelminths of biomedical and commercial significance are Schistosoma mansoni (blood fluke and Fasciola hepatica (liver fluke. These related trematodes are responsible for the chronic neglected tropical diseases schistosomiasis and fascioliasis, respectively. As no vaccine is currently available for anti-flukicidal immunoprophylaxis, current treatment is mediated by mono-chemical chemotherapy in the form of mass drug administration (MDA (praziquantel for schistosomiasis or drenching (triclabendazole for fascioliasis programmes. This overreliance on single chemotherapeutic classes has dramatically limited the number of novel chemical entities entering anthelmintic drug discovery pipelines, raising significant concerns for the future of sustainable blood and liver fluke control. METHODOLOGY/ PRINCIPLE FINDINGS: Here we demonstrate that 7-keto-sempervirol, a diterpenoid isolated from Lycium chinense, has dual anthelmintic activity against related S. mansoni and F. hepatica trematodes. Using a microtiter plate-based helminth fluorescent bioassay (HFB, this activity is specific (Therapeutic index = 4.2, when compared to HepG2 cell lines and moderately potent (LD50 = 19.1 μM against S. mansoni schistosomula cultured in vitro. This anti-schistosomula effect translates into activity against both adult male and female schistosomes cultured in vitro where 7-keto-sempervirol negatively affects motility/behaviour, surface architecture (inducing tegumental holes, tubercle swelling and spine loss/shortening, oviposition rates and egg morphology. As assessed by the HFB and microscopic phenotypic scoring matrices, 7-keto-sempervirol also effectively kills in vitro cultured F. hepatica newly excysted juveniles (NEJs, LD50 = 17.7 μM. Scanning electron microscopy (SEM evaluation of adult F. hepatica liver flukes co-cultured in vitro with 7-keto-sempervirol additionally demonstrates phenotypic abnormalities including breaches in tegumental

Molecular approaches for detection of the multi-drug resistant tuberculosis (MDR-TB in Bangladesh.

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Tafsina Haque Aurin

Full Text Available The principal obstacles in the treatment of tuberculosis (TB are delayed and inaccurate diagnosis which often leads to the onset of the drug resistant TB cases. To avail the appropriate treatment of the patients and to hinder the transmission of drug-resistant TB, accurate and rapid detection of resistant isolates is critical. Present study was designed to demonstrate the efficacy of molecular techniques inclusive of line probe assay (LPA and GeneXpert MTB/RIF methods for the detection of multi-drug resistant (MDR TB. Sputum samples from 300 different categories of treated and new TB cases were tested for the detection of possible mutation in the resistance specific genes (rpoB, inhA and katG through Genotype MTBDRplus assay or LPA and GeneXpert MTB/RIF tests. Culture based conventional drug susceptibility test (DST was also carried out to measure the efficacy of the molecular methods employed. Among 300 samples, 191 (63.7% and 193 (64.3% cases were found to be resistant against rifampicin in LPA and GeneXpert methods, respectively; while 189 (63% cases of rifampicin resistance were detected by conventional DST methods. On the other hand, 196 (65.3% and 191 (63.7% isolates showed isoniazid resistance as detected by LPA and conventional drug susceptibility test (DST, respectively. Among the drug resistant isolates (collectively 198 in LPA and 193 in conventional DST, 189 (95.6% and 187 (96.9% were considered to be MDR as examined by LPA and conventional DST, respectively. Category-II and -IV patients encountered higher frequency of drug resistance compared to those from category-I and new cases. Considering the higher sensitivity, specificity and accuracy along with the required time to results significantly shorter, our study supports the adoption of LPA and GeneXpert assay as efficient tools in detecting drug resistant TB in Bangladesh.

EVALUATION OF ANTHELMINTIC ACTIVITY OF PINEAPPLE FRUIT EXTRACT USING INDIAN EARTHWORM (PHERITIMA POSTHUMA)

OpenAIRE

Dey P; Debnath P; Bhakta T

2013-01-01

Pineapple is a favorite for the lovers of fruit in its fresh forms as well as in preserves like jams, jellies and squashes. Two varieties of Pineapples (Queen and Kew) are available in Tripura during mid-May to mid-September. The plant is well known for its different folk medicines like the root and fruit are either eaten or applied topically as an anti-inflammatory, digestive and anthelmintic. It was observed that the people of Tripura especially the Tribes are use the juice of matured root ...

Detection of multidrug-resistant tuberculosis from stored DNA Samples: A multicenter study

OpenAIRE

Marie Sylvianne Rabodoarivelo; A Brandao; M C Cergole Novella; A G C. Bombonatte; B Imperiale; N Rakotosamimanana; N Morcillo; V Rasolofo; J C Palomino; A Martin

2018-01-01

Background: In low-income countries, rapid detection of tuberculosis (TB) drug resistance is often restricted by the difficulties of transporting and storing sputum samples from remote health centers to the reference laboratories where molecular tests are available. The aim of this study was to evaluate the performance of four transport and storage systems for molecular detection of rifampicin (RIF) and isoniazid (INH) resistance. Methods: This was a multicenter study. Molecular detection of ...

Anthelmintic efficiency of doramectin, fenbendazole, and nitroxynil, in combination or individually, in sheep worm control

OpenAIRE

Holsback, Luciane; Luppi, Pedro Alex Ramsey; Silva, Camile Sanches; Negrão, Gustavo Kremer; Conde, Gabriel; Gabriel, Hugo Vinícius; Balestrieri, João Vitor; Tomazella, Lucas

2016-01-01

Abstract The anthelmintic efficiency of doramectin, fenbendazole, and nitroxynil, used individually or in combination, was determined by the Fecal Egg Count Reduction (FECR) test and cultivation of larvae of anthelminthic-treated sheep grouped as follows: G1 (doramectin), G2 (fenbendazole), G3 (nitroxynil), G4 (doramectin + fenbendazole), G5 (doramectin + nitroxynil), G6 (fenbendazole + nitroxynil), G7 (doramectin + nitroxynil + fenbendazole), G8 (untreated). In addition to individually used ...

Detecting and treating breast cancer resistance to EGFR inhibitors

Science.gov (United States)

Moonlee, Sun-Young; Bissell, Mina J.; Furuta, Saori; Meier, Roland; Kenny, Paraic A.

2016-04-05

The application describes therapeutic compositions and methods for treating cancer. For example, therapeutic compositions and methods related to inhibition of FAM83A (family with sequence similarity 83) are provided. The application also describes methods for diagnosing cancer resistance to EGFR inhibitors. For example, a method of diagnosing cancer resistance to EGFR inhibitors by detecting increased FAM83A levels is described.

Evaluation of Anthelmintic Activity and Composition of Pumpkin (Cucurbita pepo L. Seed Extracts—In Vitro and in Vivo Studies

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Maciej Grzybek

2016-09-01

Full Text Available A significant number of studies report growing resistance in nematodes thriving in both humans and livestock. This study was conducted to evaluate the in vitro and in vivo anthelmintic efficiency of Curcubita pepo (C. pepo L. hot water extract (HWE, cold water extract (CWE or ethanol extract (ETE on two model nematodes: Caenorhabditis elegans (C. elegans and Heligmosoides bakeri (H. bakeri. Methods: Raman, IR and LC-MS spectroscopy analyses were performed on the studied plant material to deliver qualitative and quantitative data on the composition of the obtained extracts: ETE, HWE and CWE. The in vitro activity evaluation showed an impact of C. pepo extracts on C. elegans and different developmental stages of H. bakeri. The following in vivo experiments on mice infected with H. bakeri confirmed inhibitory properties of the most active pumpkin extract selected by the in vitro study. All of the extracts were found to contain cucurbitine, aminoacids, fatty acids, and-for the first time-berberine and palmatine were identified. All C. pepo seed extracts exhibited a nematidicidal potential in vitro, affecting the survival of L1 and L2 H. bakeri larvae. The ETE was the strongest and demonstrated a positive effect on H. bakeri eggs hatching and marked inhibitory properties against worm motility, compared to a PBS control. No significant effects of pumpkin seed extracts on C. elegans integrity or motility were found. The EtOH extract in the in vivo studies showed anthelmintic properties against both H. bakeri fecal egg counts and adult worm burdens. The highest egg counts reduction was observed for the 8 g/kg dose (IC50 against H. bakeri = 2.43; 95% Cl = 2.01–2.94. A decrease in faecal egg counts (FEC was accompanied by a significant reduction in worm burden of the treated mice compared to the control group. Conclusions: Pumpkin seed extracts may be used to control of Gastrointestinal (G.I. nematode infections. This relatively inexpensive alternative

Assessment of the anthelmintic efficacy of albendazole in school children in seven countries where soil-transmitted helminths are endemic.

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Vercruysse, Jozef; Behnke, Jerzy M; Albonico, Marco; Ame, Shaali Makame; Angebault, Cécile; Bethony, Jeffrey M; Engels, Dirk; Guillard, Bertrand; Nguyen, Thi Viet Hoa; Kang, Gagandeep; Kattula, Deepthi; Kotze, Andrew C; McCarthy, James S; Mekonnen, Zeleke; Montresor, Antonio; Periago, Maria Victoria; Sumo, Laurentine; Tchuenté, Louis-Albert Tchuem; Dang, Thi Cam Thach; Zeynudin, Ahmed; Levecke, Bruno

2011-03-29

The three major soil-transmitted helminths (STH) Ascaris lumbricoides, Trichuris trichiura and Necator americanus/Ancylostoma duodenale are among the most widespread parasites worldwide. Despite the global expansion of preventive anthelmintic treatment, standard operating procedures to monitor anthelmintic drug efficacy are lacking. The objective of this study, therefore, was to define the efficacy of a single 400 milligram dose of albendazole (ALB) against these three STH using a standardized protocol. Seven trials were undertaken among school children in Brazil, Cameroon, Cambodia, Ethiopia, India, Tanzania and Vietnam. Efficacy was assessed by the Cure Rate (CR) and the Fecal Egg Count Reduction (FECR) using the McMaster egg counting technique to determine fecal egg counts (FEC). Overall, the highest CRs were observed for A. lumbricoides (98.2%) followed by hookworms (87.8%) and T. trichiura (46.6%). There was considerable variation in the CR for the three parasites across trials (country), by age or the pre-intervention FEC (pre-treatment). The latter is probably the most important as it had a considerable effect on the CR of all three STH. Therapeutic efficacies, as reflected by the FECRs, were very high for A. lumbricoides (99.5%) and hookworms (94.8%) but significantly lower for T. trichiura (50.8%), and were affected to different extents among the 3 species by the pre-intervention FEC counts and trial (country), but not by sex or age. Our findings suggest that a FECR (based on arithmetic means) of >95% for A. lumbricoides and >90% for hookworms should be the expected minimum in all future surveys, and that therapeutic efficacy below this level following a single dose of ALB should be viewed with concern in light of potential drug resistance. A standard threshold for efficacy against T. trichiura has yet to be established, as a single-dose of ALB is unlikely to be satisfactory for this parasite. ClinicalTrials.gov NCT01087099.

Evaluation of Anthelmintic Activity and Composition of Pumpkin (Cucurbita pepo L.) Seed Extracts-In Vitro and in Vivo Studies.

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Grzybek, Maciej; Kukula-Koch, Wirginia; Strachecka, Aneta; Jaworska, Aleksandra; Phiri, Andrew M; Paleolog, Jerzy; Tomczuk, Krzysztof

2016-09-01

A significant number of studies report growing resistance in nematodes thriving in both humans and livestock. This study was conducted to evaluate the in vitro and in vivo anthelmintic efficiency of Curcubita pepo (C. pepo) L. hot water extract (HWE), cold water extract (CWE) or ethanol extract (ETE) on two model nematodes: Caenorhabditis elegans (C. elegans) and Heligmosoides bakeri (H. bakeri). Raman, IR and LC-MS spectroscopy analyses were performed on the studied plant material to deliver qualitative and quantitative data on the composition of the obtained extracts: ETE, HWE and CWE. The in vitro activity evaluation showed an impact of C. pepo extracts on C. elegans and different developmental stages of H. bakeri. The following in vivo experiments on mice infected with H. bakeri confirmed inhibitory properties of the most active pumpkin extract selected by the in vitro study. All of the extracts were found to contain cucurbitine, aminoacids, fatty acids, and-for the first time-berberine and palmatine were identified. All C. pepo seed extracts exhibited a nematidicidal potential in vitro, affecting the survival of L1 and L2 H. bakeri larvae. The ETE was the strongest and demonstrated a positive effect on H. bakeri eggs hatching and marked inhibitory properties against worm motility, compared to a PBS control. No significant effects of pumpkin seed extracts on C. elegans integrity or motility were found. The EtOH extract in the in vivo studies showed anthelmintic properties against both H. bakeri fecal egg counts and adult worm burdens. The highest egg counts reduction was observed for the 8 g/kg dose (IC50 against H. bakeri = 2.43; 95% Cl = 2.01-2.94). A decrease in faecal egg counts (FEC) was accompanied by a significant reduction in worm burden of the treated mice compared to the control group. Pumpkin seed extracts may be used to control of Gastrointestinal (G.I.) nematode infections. This relatively inexpensive alternative to the currently available

Evaluation of Anthelmintic Activity and Composition of Pumpkin (Cucurbita pepo L.) Seed Extracts—In Vitro and in Vivo Studies

Science.gov (United States)

Grzybek, Maciej; Kukula-Koch, Wirginia; Strachecka, Aneta; Jaworska, Aleksandra; Phiri, Andrew M.; Paleolog, Jerzy; Tomczuk, Krzysztof

2016-01-01

A significant number of studies report growing resistance in nematodes thriving in both humans and livestock. This study was conducted to evaluate the in vitro and in vivo anthelmintic efficiency of Curcubita pepo (C. pepo) L. hot water extract (HWE), cold water extract (CWE) or ethanol extract (ETE) on two model nematodes: Caenorhabditis elegans (C. elegans) and Heligmosoides bakeri (H. bakeri). Methods: Raman, IR and LC-MS spectroscopy analyses were performed on the studied plant material to deliver qualitative and quantitative data on the composition of the obtained extracts: ETE, HWE and CWE. The in vitro activity evaluation showed an impact of C. pepo extracts on C. elegans and different developmental stages of H. bakeri. The following in vivo experiments on mice infected with H. bakeri confirmed inhibitory properties of the most active pumpkin extract selected by the in vitro study. All of the extracts were found to contain cucurbitine, aminoacids, fatty acids, and-for the first time-berberine and palmatine were identified. All C. pepo seed extracts exhibited a nematidicidal potential in vitro, affecting the survival of L1 and L2 H. bakeri larvae. The ETE was the strongest and demonstrated a positive effect on H. bakeri eggs hatching and marked inhibitory properties against worm motility, compared to a PBS control. No significant effects of pumpkin seed extracts on C. elegans integrity or motility were found. The EtOH extract in the in vivo studies showed anthelmintic properties against both H. bakeri fecal egg counts and adult worm burdens. The highest egg counts reduction was observed for the 8 g/kg dose (IC50 against H. bakeri = 2.43; 95% Cl = 2.01–2.94). A decrease in faecal egg counts (FEC) was accompanied by a significant reduction in worm burden of the treated mice compared to the control group. Conclusions: Pumpkin seed extracts may be used to control of Gastrointestinal (G.I.) nematode infections. This relatively inexpensive alternative to the

Clinical laboratory detection of carbapenem-resistant and carbapenemase-producing Enterobacteriaceae.

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Miller, Shelley; Humphries, Romney M

2016-08-01

Carbapenemases, enzymes that hydrolyze carbapenem-class antimicrobials, pose serious clinical and diagnostic challenges, including their recent rapid spread among members of the Enterobacteriaceae, a family with no inherent carbapenem resistance. Currently there is no one-size-fits-all method for detecting carbapenem-resistant Enterobacteriaceae (CRE) in the laboratory, nor how to differentiate carbapenemase-producers (CP) from isolates that are carbapenem-resistant via other or combined mechanisms. This article reviews definitions for CRE and CP-CRE, and discusses current phenotypic and molecular methods available to the clinical laboratory for the detection of both CP and non-CP CRE. Expert commentary: Routine evaluation of carbapenem resistance mechanism by the routine clinical laboratory are not necessary for patient care, as clinical breakpoints best predict response. However, evaluation for carbapenemase is integral to infection control efforts, and laboratories should have the capacity to do such testing, either in house or by submitting isolates to a reference laboratory.

IN VITRO ANTHELMINTIC EFFECT OF METHANOLIC LEAF EXTRACT OF Gliricidia sepium AGAINST GASTROINTESTINALE NEMATODES OF SHEEP

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Manuel Mateo Hernandez-Villegas

2014-04-01

Full Text Available The use of local resources for food and health care of animals is a highly profitable and sustainable strategy. Among these resources are native trees and shrubs which in addition to providing good quality nutrients, produce secondary metabolites with anthelmintic (AH effect. Therefore the aim of this study was to evaluate the in vitro AH effect of Gliricidia sepium leaves methanol extract (GSME, through the egg hatch inhibition assay (EHA. Three concentrations of the extracts were tested: 125, 250 and 500 μg/mL.  Also a negative control (distilled water and a positive control (levamisole 2 mg/mL were included. The GSME showed significant differences P<0.05 when compared with the positive control. The GSME also showed a dose-dependent response in inhibition of eggs hatching. Effectiveness percentages found were: 27.7%, 46.2%, 49.7% of inhibition at 125, 250, and 500 μg/mL respectively. The average dose (ED50 obtained through probit analysis was 394.96 μg/mL. These results suggest that the ME of leaves of G. sepium has anthelmintic activity against eggs of gastrointestinal nematodes.

Serum Strongylus vulgaris-specific antibody responses to anthelmintic treatment in naturally infected horses.

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Nielsen, Martin K; Vidyashankar, Anand N; Bellaw, Jennifer; Gravatte, Holli S; Cao, Xin; Rubinson, Emily F; Reinemeyer, Craig R

2015-02-01

Strongylus vulgaris is the most pathogenic helminth parasite of horses, causing verminous endarteritis with thromboembolism and infarction. A serum enzyme-linked immunosorbent assay (ELISA) has been validated for detection of antibodies to an antigen produced by migrating larvae of this parasite. The aim was to evaluate ELISA responses to anthelmintic treatment in cohorts of naturally infected horses. Fifteen healthy horses harboring patent S. vulgaris infections were turned out for communal grazing in May 2013 (day 0). On day 55, horses were ranked according to ELISA titers and randomly allocated to the following three groups: no treatment followed by placebo pellets daily; ivermectin on day 60 followed by placebo pellets daily; or ivermectin on day 60 followed by daily pyrantel tartrate. Fecal and serum samples were collected at ∼28-day intervals until study termination on day 231. Increased ELISA values were observed for the first 53 days following ivermectin treatment. Titers were significantly reduced 80 days after ivermectin treatment. Horses receiving daily pyrantel tartrate maintained lower ELISA values from 137 days post ivermectin treatment until trial termination. These results illustrate that a positive ELISA result is indicative of either current or prior exposure to larval S. vulgaris infection within the previous 5 months.

Mechanistic and single-dose in vivo therapeutic studies of Cry5B anthelmintic action against hookworms.

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Yan Hu

Full Text Available Hookworm infections are one of the most important parasitic infections of humans worldwide, considered by some second only to malaria in associated disease burden. Single-dose mass drug administration for soil-transmitted helminths, including hookworms, relies primarily on albendazole, which has variable efficacy. New and better hookworm therapies are urgently needed. Bacillus thuringiensis crystal protein Cry5B has potential as a novel anthelmintic and has been extensively studied in the roundworm Caenorhabditis elegans. Here, we ask whether single-dose Cry5B can provide therapy against a hookworm infection and whether C. elegans mechanism-of-action studies are relevant to hookworms.To test whether the C. elegans invertebrate-specific glycolipid receptor for Cry5B is relevant in hookworms, we fed Ancylostoma ceylanicum hookworm adults Cry5B with and without galactose, an inhibitor of Cry5B-C. elegans glycolipid interactions. As with C. elegans, galactose inhibits Cry5B toxicity in A. ceylanicum. Furthermore, p38 mitogen-activated protein kinase (MAPK, which controls one of the most important Cry5B signal transduction responses in C. elegans, is functionally operational in hookworms. A. ceylanicum hookworms treated with Cry5B up-regulate p38 MAPK and knock down of p38 MAPK activity in hookworms results in hypersensitivity of A. ceylanicum adults to Cry5B attack. Single-dose Cry5B is able to reduce by >90% A. ceylanicum hookworm burdens from infected hamsters, in the process eliminating hookworm egg shedding in feces and protecting infected hamsters from blood loss. Anthelmintic activity is increased about 3-fold, eliminating >97% of the parasites with a single 3 mg dose (∼30 mg/kg, by incorporating a simple formulation to help prevent digestion in the acidic stomach of the host mammal.These studies advance the development of Cry5B protein as a potent, safe single-dose anthelmintic for hookworm therapy and make available the information of how

In vitro thrombolytic, anthelmintic, anti-oxidant and cytotoxic activity with phytochemical screening of methanolic extract of Xanthium indicum leaves

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Antara Ghosh

2015-12-01

Full Text Available Xanthium indicum is an important medicinal plant traditionally used in Bangladesh as a folkloric treatment. The current study was undertaken to evaluate thrombolytic, anthelmintic, anti-oxidant, cytotoxic properties with phytochemical screening of methanolic extract of X. indicum leaves. The analysis of phytochemical screening confirmed the existence of phytosetrols and diterpenes. In thrombolytic assay, a significant clot lysis was observed at four concentrations of plant extract compare to the positive control streptokinase (30,000 IU, 15,000 IU and negative control normal saline. The extract revealed potent anthelmintic activity at different concentrations. In anti-oxidant activity evaluation by two potential experiments namely total phenolic content determination and free radical scavenging assay by 2, 2-diphenylpicrylhydrazyl (DPPH, the leaves extract possess good anti-oxidant property. In the brine shrimp lethality bioassay, the crude extract showed potent (LC50 1.3 μg/mL cytotoxic activity compare to the vincristine sulfate as a positive control (LC50 0.8 μg/mL.

Synthesis, Spectral and Anthelmintic Activity Studies on Some Novel Imidazole Derivatives

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Rajiv Dahiya

2008-01-01

Full Text Available Present study describes the synthesis of a novel series of 3,5-diiodo-4-(5-nitro-1H-2-imidazolylbenzoyl amino acids and di/tri/tetrapeptides using diisopropylcarbodiimide/dicyclohexylcarbodiimide (DIPC/DCC as coupling agents and N-methylmorpholine/triethylamine (NMM/TEA as bases. Structure elucidation of all the newly synthesized compounds was done by elemental analysis and IR, 1H NMR, 13C NMR and mass spectral data. Synthesized imidazolopeptides were screened for their anthelmintic activity and found to possess moderate to good bioactivity against earthworms Megascoplex konkanensis, Pontoscotex corethruses and Eudrilus eugeniea when compared to reference drugs - albendazole and mebendazole at dose level of 2 mg mL−1.

Molecular Detection of Helicobacter pylori and its Antimicrobial Resistance in Brazzaville, Congo.

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Ontsira Ngoyi, Esther Nina; Atipo Ibara, Blaise Irénée; Moyen, Rachelle; Ahoui Apendi, Philestine Clausina; Ibara, Jean Rosaire; Obengui, O; Ossibi Ibara, Roland Bienvenu; Nguimbi, Etienne; Niama, Rock Fabien; Ouamba, Jean Maurille; Yala, Fidèle; Abena, Ange Antoine; Vadivelu, Jamuna; Goh, Khean Lee; Menard, Armelle; Benejat, Lucie; Sifre, Elodie; Lehours, Philippe; Megraud, Francis

2015-08-01

Helicobacter pylori infection is involved in several gastroduodenal diseases which can be cured by antimicrobial treatment. The aim of this study was to determine the prevalence of H. pylori infection and its bacterial resistance to clarithromycin, fluoroquinolones, and tetracycline in Brazzaville, Congo, by using molecular methods. A cross- sectional study was carried out between September 2013 and April 2014. Biopsy specimens were obtained from patients scheduled for an upper gastrointestinal endoscopy and were sent to the French National Reference Center for Campylobacters and Helicobacters where they were tested by molecular methods for detection of H. pylori and clarithromycin resistance by real-time PCR using a fluorescence resonance energy transfer-melting curve analysis (FRET-MCA) protocol, for detection of tetracycline resistance by real-time PCR on 16S rRNA genes (rrnA and rrnB), for detection of point mutations in the quinolone resistance-determining regions (QRDR) of H. pylori gyrA gene, associated with resistance to quinolones, by PCR and sequencing. This study showed a high H. pylori prevalence (89%), low rates of clarithromycin and tetracycline resistance (1.7% and 2.5%, respectively), and a high rate of quinolone resistance (50%). Therefore, the use of standard clarithromycin-based triple therapy is still possible as an empiric first-line treatment as well as prescription of bismuth-based quadruple therapy, which includes tetracycline, but not a levofloxacin-based triple therapy because of the high rate of resistance to fluoroquinolones. © 2015 John Wiley & Sons Ltd.

Molecular detection of drug resistance in microbes by isotopic techniques: The IAEA experience

International Nuclear Information System (INIS)

Dar, L.; Boussaha, A.; Padhy, A.K.; Khan, B.

2003-01-01

The International Atomic Energy Agency (IAEA) supports various programmes on the uses of radionuclide techniques in the management of human communicable diseases. An important issue, being addressed through several technology transfer projects, is the detection of drug resistance in microbes by radioisotope based molecular-biology diagnostic procedures. The techniques employed include dot blot hybridisation with P-32 labelled oligonucleotide probes to detect point mutations, associated with drug resistance, in microbial genes amplified by the polymerase chain reaction (PCR). Molecular methods have been used for the detection of drug resistance in the malarial parasite, Plasmodium falciparum, and in Mycobacterium tuberculosis. Radioisotope based molecular-biology methods have been demonstrated to have comparative advantages in being sensitive, specific, cost-effective, and suitable for application to large-scale molecular surveillance for drug resistance. (author)

A molecular diagnostic tool to replace larval culture in conventional faecal egg count reduction testing in sheep.

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Florian Roeber

Full Text Available The accurate diagnosis of parasitic nematode infections in livestock (including sheep and goats is central to their effective control and the detection of the anthelmintic resistance. Traditionally, the faecal egg count reduction test (FECRT, combined with the technique of larval culture (LC, has been used widely to assess drug-susceptibility/resistance in strongylid nematodes. However, this approach suffers from a lack of specificity, sensitivity and reliability, and is time-consuming and costly to conduct. Here, we critically assessed a specific PCR assay to support FECRT, in a well-controlled experiment on sheep with naturally acquired strongylid infections known to be resistant to benzimidazoles. We showed that the PCR results were in close agreement with those of total worm count (TWC, but not of LC. Importantly, albendazole resistance detected by PCR-coupled FECRT was unequivocally linked to Teladorsagia circumcincta and, to lesser extent, Trichostrongylus colubriformis, a result that was not achievable by LC. The key findings from this study demonstrate that our PCR-coupled FECRT approach has major merit for supporting anthelmintic resistance in nematode populations. The findings also show clearly that our PCR assay can be used as an alternative to LC, and is more time-efficient and less laborious, which has important practical implications for the effective management and control strongylid nematodes of sheep.

Effectiveness of Ivermectin and Albendazole against Haemonchus contortus in Sheep in West Java, Indonesia

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Puspitasari, Silvia; Farajallah, Achmad; Sulistiawati, Erni; Muladno

2016-01-01

Administering a half dose of an anthelmintic is a simple method for detecting resistance in parasites infesting small ruminants. When a single anthelmintic fails in native sheep from Indonesia, a combination of anthelmintics from different chemical classes with different modes of action are administered as an alternative parasite-control strategy. This study compared the anthelmintic efficacy of ivermectin (IVM) and albendazole (ABZ) given either separately as a single dose or half dose or co-administered to sheep naturally infected with Haemonchus contortus. Twelve sheep from Bogor, West Java, Indonesia were divided into the following six treatment groups: half-dose IVM, full-dose IVM, half-dose ABZ, full-dose ABZ, combined IVM + ABZ, and control. The treatment efficacy was determined using the faecal egg count reduction test (FECRT) at day 0 (pre-treatment) and post-treatment at days 7, 14, 21, 28, 35, and 42. The efficacies of half-dose IVM, full-dose IVM, half-dose ABZ, full-dose ABZ, and the combination treatment ranged from −1900% to 100%, 99% to 100%, −167% to 100%, −467% to 89%, and −200% to 100%, respectively. The FECRT for the half-dose IVM, half-dose ABZ, full-dose ABZ showed that H. contortus is resistant to half-dose IVM and ABZ. Full-dose IVM was effective against H. contortus. The combined treatment was more effective against H. contortus than ABZ alone. PMID:27019686

Identification and detection of methicillin resistance in Non-Epidermidis coagulase-negative staphylococci

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Carina Secchi

Full Text Available The NCCLS (2004 presented a new methodology to detect, by disk-diffusion agar, oxacillin-resistance using a cefoxitin disk. We identified coagulase-negative staphylococci (SCoN to the species level and compared the use of cefoxitin disks (30 µg with oxacillin disks (1 µg, agar dilution (minimum inhibitory concentration of oxacillin and mecA gene detection in isolates of coagulase-negative bacteria other than Staphylococcus epidermidis (SCoNne. A total of 238 SCoNne was evaluated; oxacillin-resistance (the mecA gene was detected in 71% of the isolates. All methods gave 100% sensitivity, based on presence of the mecA gene. The specificity of the cefoxitin disk was 100%, while the oxacillin disk gave a specificity of 91% and agar dilution oxacillin gave a specificity of 88%. We conclude that the cefoxitin disk is an efficient test, and it is an easy method for use in clinical laboratories to detect oxacillin-resistance in staphylococci.

An in vitro larval migration assay for assessing anthelmintic activity of different drug classes against Ascaris suum

DEFF Research Database (Denmark)

Zhao, Jianguo; Williams, Andrew R; Hansen, Tina Vicky Alstrup

2017-01-01

and agar gel larval migration assay to test the effect of benzimidazole and tetrahydropyrimidin/imidazothiazole anthelmintics against nine isolates of A. suum collected from locations in China and Denmark. Drugs tested were thiabendazole, fenbendazole, mebendazole, levamisole, and pyrantel. The percentages...

Germination test as a fast method to detect glyphosate-resistant sourgrass

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Marcos Altomani Neves Dias

2015-01-01

Full Text Available The occurrence of weed species with different levels of resistance to glyphosate has increasingly spread in agricultural areas. In Brazil, sourgrass is among the main species presenting issues in this regard. Thus, fast and reliable methods to detect glyphosate resistance are of special interest for this specie, either for research or rational management purposes. This study was carried out to verify the feasibility of using the germination test to detect glyphosate resistance in sourgrass. The experiment was conducted with two sourgrass biotypes, with different levels of susceptibility to glyphosate. The seeds were previously imbibed in solutions composed of 0, 0.1875%, 0.25%, 0.75%, 1.5%, 3% and 6% of glyphosate during two periods, five and ten minutes, and submitted to germination tests. The results indicate the germination test as a feasible and time-saving approach to evaluate glyphosate-resistant sourgrass, with results available in seven days.

Germination test as a fast method to detect glyphosate-resistant sourgrass

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Marcos Altomani Neves Dias

2015-09-01

Full Text Available The occurrence of weed species with different levels of resistance to glyphosate has increasingly spread in agricultural areas. In Brazil, sourgrass is among the main species presenting issues in this regard. Thus, fast and reliable methods to detect glyphosate resistance are of special interest for this specie, either for research or rational management purposes. This study was carried out to verify the feasibility of using the germination test to detect glyphosate resistance in sourgrass. The experiment was conducted with two sourgrass biotypes, with different levels of susceptibility to glyphosate. The seeds were previously imbibed in solutions composed of 0, 0.1875%, 0.25%, 0.75%, 1.5%, 3% and 6% of glyphosate during two periods, five and ten minutes, and submitted to germination tests. The results indicate the germination test as a feasible and time-saving approach to evaluate glyphosate-resistant sourgrass, with results available in seven days.

In vitro and in vivo anthelmintic activity of extracts from Artemisia parviflora and A. sieversiana

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Irum S.

2017-09-01

Full Text Available In the northern areas of Pakistan, the use of Artemisia based therapeutics is a common practice. Plants of genus Artemisia are known to possess anthelmintic and therapeutic effect. Infections caused by gastrointestinal nematodes are major threat to livestock industry across the world resulting in loss of production and indirect economic losses due to high cost of anthelmintic drugs. Present study was carried out to evaluate in vitro and in vivo effect of Artemisia sieversiana and Artemisia parviflora on Haemonchus contortus, a parasitic nematode of small ruminants. Methanolic plant extract was tested against three different developmental stages using an egg hatch assay, infective larvae and adult worm motility assay. Different concentrations were used for the bioassays and post exposure mortality was recorded after 8 hr for adult worms and infective larvae, while egg inhibition percentage was observed after 27 hr. A highly significant ability to inhibit the egg hatching (100 % was recorded for both plant extracts while, the highest activity for adult worm assay and larvicidal assay was 90 % for A. sieversiana. The highest activity for adult motility and larvicidal assay for A. parviflora was 89 % and 86.6 % respectively. For in vivo trials maximum parentage reduction was 77.0 % for A. sieversiana and 73.6 % for A. parviflora. It is concluded that selected plant extracts were effective in reducing worm burden in animals.

Direct anthelmintic effects of Cereus jamacaru (Cactaceae) on trichostrongylid nematodes of sheep: in vivo studies.

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Vatta, A F; Kandu-Lelo, C; Ademola, I O; Eloff, J N

2011-08-25

Following claims of anthelmintic activity of Cereus jamacaru DC (Cactaceae) by a commercial farmer, in vivo studies were conducted to determine the possible direct anthelmintic effects of the plant on ovine gastrointestinal nematodes. Eighteen sheep were infected with 4000 Haemonchus contortus and 6000 Trichostrongylus colubriformis larvae given in three divided doses over a period of three days. Once the infections were patent, the sheep were allocated to three groups and were drenched once a week for six weeks with fresh blended C. jamacaru plant material at a single (32.3g/sheep) or double dose (64.6g/sheep) or they remained as undrenched controls. Faeces were collected from individual animals on the day of treatment and three days thereafter on a weekly basis for seven weeks for faecal egg count. While there were no statistically significant differences in the egg counts between the groups, a double dose of C. jamacaru was effective in reducing the egg counts in the sheep by 18-65% over the 49 days of the experiment. Given that all animals remained in good health throughout the course of the experiment, with no adverse events occurring during the study, further experiments using higher doses or administering the plant material for a longer period of time than in the present study would be warranted. Copyright © 2011 Elsevier B.V. All rights reserved.

In Vitro Anthelmintic Activity of Crude Extracts of Aerial Parts of Cissus quadrangularis L. and Leaves of Schinus molle L. against Haemonchus contortus.

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Zenebe, Selamawit; Feyera, Teka; Assefa, Solomon

2017-01-01

Haemonchus contortus, the causative agent of Haemonchosis, is the most economically important parasite in small ruminant production. Control with chemotherapy has not been successful due to rapid emergence of drug-resistant strains. There is a continuous search for alternative leads particularly from plants. The study aimed to evaluate the anthelmintic activity of crude methanolic extracts of leaves of Schinus molle and aerial parts of Cissus quadrangularis against H. contortus. Methods . Adult motility test and egg hatching inhibition assay were employed to investigate the in vitro adulticidal and egg hatching inhibitory effects of the extracts. Higher concentrations of the extracts (10 and 5 mg/ml) had a significantly superior adulticidal activity ( p molle and C. quadrangularis extracts indicated a maximum of 96% and 88% egg hatch inhibition, respectively, within the 48 hrs of exposure at 1 mg/ml. The current study evidenced that the crude methanolic extracts of the plants have promising adulticidal and egg hatching inhibitory effects against H. contortus .

Comparative evaluation of anthelmintic and antibacterial activities in leaves and fruits of Garcinia cambogia (Gaertn. desr. and Garcinia indica (Dupetit-Thouars choisy

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C. Tharachand

2015-06-01

Full Text Available This study aimed to evaluate the in vitro anthelmintic activity and antibacterial activity of the extracts from the leaves and fruits of Garcinia indica (Dupetit-Thouars Choisy and Garcinia cambogia(Gaertn. Desr. using the Indian earthworm Pheretima posthuma. Two concentrations (25 and 50 mg/mL of various extracts such as petroleum ether, chloroform, ethyl acetate, methanol and water were tested. Albendazole at the concentrations of 25 and 50 mg/mL was used as the standard reference. Significant anthelmintic effects of the fruits and leaves of G. cambogia and G. indica (P<0.05 were observed and the results were expressed in terms of paralysis and death time. All the extracts showed the dose dependent paralysis and death of earthworms. Among all the extracts used, methanol extract exhibited the highest activity. G. cambogia leaf extract (50 mg/mL had 30% faster paralysis effect on earthworms than the standard reference. Furthermore, the antimicrobial activity of the methanol extracts of the fruits and leaves showed significant (P<0.05 activity against Gram-positive and Gram-negative bacteria. At a concentration of 500 µg/mL, G. indica fruit extract presented higher zones of inhibition against Pseudomonas aeruginosa and Staphylococcus aureus. Hence, it could be concluded that the leaves and fruits of G. indica and G. cambogia contained active anthelmintic and antibacterial phytochemicals, which could find their applications in pharmaceuticals.

The anthelmintic efficacy of fenbendazole in the control of Moniezia expansa and Trichuris ovis in sheep.

Science.gov (United States)

Townsend, R B; Kelly, J D; James, R; Weston, I

1977-11-01

The anthelmintic efficacy of fenbendazole (methyl 5-(phenyl-thio)-2-benzimidazole-carbamate) against Moniezia expansa and Trichuris ovis was tested. At dose rates of 5 mg per kg and above, efficacies were found to be greater than 91 percent against M expansa and greater than 92 per cent against T ovis. At these dose rates efficacy on egg suppression was 100 per cent for Moniezia and greater than 97 per cent for Trichuris.

[Detection of CRISPR and its relationship to drug resistance in Shigella].

Science.gov (United States)

Wang, Linlin; Wang, Yingfang; Duan, Guangcai; Xue, Zerun; Guo, Xiangjiao; Wang, Pengfei; Xi, Yuanlin; Yang, Haiyan

2015-04-04

To detect clustered regularly interspaced short palindromic repeats (CRISPR) in Shigella, and to analyze its relationship to drug resistance. Four pairs of primers were used for the detection of convincing CRISPR structures CRISPR-S2 and CRISPR-S4, questionable CRISPR structures CRISPR-S1 and CRISPR-S3 in 60 Shigella strains. All primers were designed using sequences in CRISPR database. CRISPR Finder was used to analyze CRISPR and susceptibilities of Shigella strains were tested by agar diffusion method. Furthermore, we analyzed the relationship between drug resistance and CRISPR-S4. The positive rate of convincing CRISPR structures was 95%. The four CRISPR loci formed 12 spectral patterns (A-L), all of which contained convincing CRISPR structures except type K. We found one new repeat and 12 new spacers. The multi-drug resistance rate was 53. 33% . We found no significant difference between CRISPR-S4 and drug resistant. However, the repeat sequence of CRISPR-S4 in multi- or TE-resistance strains was mainly R4.1 with AC deletions in the 3' end, and the spacer sequences of CRISPR-S4 in multi-drug resistance strains were mainly Sp5.1, Sp6.1 and Sp7. CRISPR was common in Shigella. Variations df repeat sequences and diversities of spacer sequences might be related to drug resistance in Shigella.

Performance of Four Transport and Storage Systems for Molecular Detection of Multidrug-Resistant Tuberculosis

Science.gov (United States)

Rabodoarivelo, Marie Sylvianne; Imperiale, Bélen; andrianiavomikotroka, Rina; Brandao, Angela; Kumar, Parveen; Singh, Sarman; Ferrazoli, Lucilaine; Morcillo, Nora; Rasolofo, Voahangy; Palomino, Juan Carlos; Vandamme, Peter; Martin, Anandi

2015-01-01

Background Detection of drug-resistant tuberculosis is essential for the control of the disease but it is often hampered by the limitation of transport and storage of samples from remote locations to the reference laboratory. We performed a retrospective field study to evaluate the performance of four supports enabling the transport and storage of samples to be used for molecular detection of drug resistance using the GenoType MTBDRplus. Methods Two hundred Mycobacterium tuberculosis strains were selected and spotted on slides, FTA cards, GenoCards, and in ethanol. GenoType MTBDRplus was subsequently performed with the DNA extracted from these supports. Sensitivity and specificity were calculated and compared to the results obtained by drug susceptibility testing. Results For all supports, the overall sensitivity and specificity for detection of resistance to RIF was between 95% and 100%, and for INH between 95% and 98%. Conclusion The four transport and storage supports showed a good sensitivity and specificity for the detection of resistance to RIF and INH in M. tuberculosis strains using the GenoType MTBDRplus. These supports can be maintained at room temperature and could represent an important alternative cost-effective method useful for rapid molecular detection of drug-resistant TB in low-resource settings. PMID:26431352

In vitro anthelmintic activity of Combretum molle (R. Br. ex G. Don) (Combretaceae) against Haemonchus contortus ova and larvae.

Science.gov (United States)

Ademola, I O; Eloff, J N

2010-04-19

Parasitic nematodes, especially Haemonchus contortus (Rudolphi), are among the most common and economically important causes of disease in sheep and goats owned by pastoralists and small holder farmers in Africa. The control of these infections relies mainly on the use of anthelmintic drugs. However, herbal preparations are widely used by pastoralists and small holder farmers for the treatment of their livestock against helminth parasites. The anthelmintic effect of acetone leaf extract and fractions of Combretum molle was investigated to determine the relative efficacy of the components against gastrointestinal sheep nematodes. The fractions were obtained by solvent:solvent extraction from the acetone extract. These were evaluated for nematocidal activity by means of an egg hatch (EHA) and larval a development and viability assay (LDVA) in vitro. The effect of the test extracts on the hatchability of eggs and development of first to third stage larvae and the survival rate of the third stage larvae. H. contortus, were used to determine the relative bioactivities. Best-fit LC(50) values were computed using global model of nonlinear regression curve-fitting. The extracts inhibited egg hatching and development of the larvae of H. contortus in a concentration-dependent manner. Best-fit LC(50) values for the egg hatch test were 0.866, 0.333, 0.833, 0.747, and 0.065mg/mL for acetone extract, n-butanol, hexane, chloroform, and 35% water in methanol fractions, respectively. The best-fit LC(50) values for the LDVA were 0.604, 0.362, 1.077, 0.131 and 0.318mg/mL for the acetone extract, butanol, hexane, chloroform, and 35% water in methanol fractions, respectively. In the EHA the 35% water in methanol fraction was significantly more active than all the other fractions (pmolle leaf could find application in anthelmintic therapy in veterinary practice.

World association for the advancement of veterinary parasitology (WAAVP): Second edition of guidelines for evaluating the efficacy of anthelmintics in swine

DEFF Research Database (Denmark)

Hennessy, D.R.; Bauer, C.; Boray, J.C.

2006-01-01

Guidelines are provided for evaluating the efficacy of anthelmintics in swine which, in conjunction with other sets of guidance such as those of the International Cooperation on Harmonization of Technical Requirements for Registration of Veterinary Medicinal Products (VICH GL7 and VICH GL16), sho...

Rapid detection of multidrug-resistant Mycobacterium tuberculosis using the malachite green decolourisation assay

Science.gov (United States)

Coban, Ahmet Yilmaz; Uzun, Meltem

2013-01-01

Early detection of drug resistance in Mycobacterium tuberculosis isolates allows for earlier and more effective treatment of patients. The aim of this study was to investigate the performance of the malachite green decolourisation assay (MGDA) in detecting isoniazid (INH) and rifampicin (RIF) resistance in M. tuberculosis clinical isolates. Fifty M. tuberculosis isolates, including 19 multidrug-resistant, eight INH-resistant and 23 INH and RIF-susceptible samples, were tested. The sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV) and agreement of the assay for INH were 92.5%, 91.3%, 92.5%, 91.3% and 92%, respectively. Similarly, the sensitivity, specificity, PPV, NPV and agreement of the assay for RIF were 94.7%, 100%, 100%, 96.8% and 98%, respectively. There was a major discrepancy in the tests of two isolates, as they were sensitive to INH by the MGDA test, but resistant by the reference method. There was a minor discrepancy in the tests of two additional isolates, as they were sensitive to INH by the reference method, but resistant by the MGDA test. The drug susceptibility test results were obtained within eight-nine days. In conclusion, the MGDA test is a reliable and accurate method for the rapid detection of INH and RIF resistance compared with the reference method and the MGDA test additionally requires less time to obtain results. PMID:24402143

Rapid detection of multidrug-resistant Mycobacterium tuberculosis using the malachite green decolourisation assay

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Ahmet Yilmaz Coban

2013-12-01

Full Text Available Early detection of drug resistance in Mycobacterium tuberculosis isolates allows for earlier and more effective treatment of patients. The aim of this study was to investigate the performance of the malachite green decolourisation assay (MGDA in detecting isoniazid (INH and rifampicin (RIF resistance in M. tuberculosis clinical isolates. Fifty M. tuberculosis isolates, including 19 multidrug-resistant, eight INH-resistant and 23 INH and RIF-susceptible samples, were tested. The sensitivity, specificity, positive predictive value (PPV, negative predictive value (NPV and agreement of the assay for INH were 92.5%, 91.3%, 92.5%, 91.3% and 92%, respectively. Similarly, the sensitivity, specificity, PPV, NPV and agreement of the assay for RIF were 94.7%, 100%, 100%, 96.8% and 98%, respectively. There was a major discrepancy in the tests of two isolates, as they were sensitive to INH by the MGDA test, but resistant by the reference method. There was a minor discrepancy in the tests of two additional isolates, as they were sensitive to INH by the reference method, but resistant by the MGDA test. The drug susceptibility test results were obtained within eight-nine days. In conclusion, the MGDA test is a reliable and accurate method for the rapid detection of INH and RIF resistance compared with the reference method and the MGDA test additionally requires less time to obtain results.

Usefulness of detection of clarithromycin-resistant Helicobacter pylori from fecal specimens for young adults treated with eradication therapy.

Science.gov (United States)

Osaki, Takako; Mabe, Katsuhiro; Zaman, Cynthia; Yonezawa, Hideo; Okuda, Masumi; Amagai, Kenji; Fujieda, Shinji; Goto, Mitsuhide; Shibata, Wataru; Kato, Mototsugu; Kamiya, Shigeru

2017-10-01

To prevent Helicobacter pylori infection in the younger generation, it is necessary to investigate the prevalence of antibiotic-resistant H. pylori. The aim of this study was to evaluate the method of PCR-based sequencing to detect clarithromycin (CAM) resistance-associated mutations using fecal samples as a noninvasive method. DNA extracted from fecal specimens and isolates from gastric biopsy specimens were collected from patients with H. pylori infection. Antibiotic resistance to CAM was analyzed by molecular and culture methods. The detection rates of CAM resistance-associated mutations (A2142C or A2143G) were compared before and after eradication therapy. With CAM resistance of H. pylori evaluated by antibiotic susceptibility test as a gold standard, the sensitivity and the specificity of gene mutation detection from fecal DNA were 80% and 84.8%, respectively. In contrast, using DNA of isolated strains, the sensitivity and the specificity were 80% and 100%. Of the seven cases in which eradication was unsuccessful by triple therapy including CAM, CAM-resistant H. pylori, and resistance-associated mutations were detected in three cases, CAM-resistant H. pylori without the mutation was detected in two patients, and resistance-associated mutation was only detected in one patient. PCR-based sequencing to detect CAM resistance-associated mutations using isolates or fecal samples was useful for finding antibiotic-resistant H. pylori infection. Although the specificity of the detection from fecal samples compared with antibiotic susceptibility testing was lower than that from isolates, this fecal detection method is suitable especially for asymptomatic subjects including children. Further improvement is needed before clinical application. © 2017 John Wiley & Sons Ltd.

Rapid detection of drug resistance and mutational patterns of extensively drug-resistant strains by a novel GenoType® MTBDRsl assay

Directory of Open Access Journals (Sweden)

A K Singh

2013-01-01

Full Text Available Background: The emergence of extensively drug-resistant tuberculosis (XDR-TB is a major concern in the India. The burden of XDR-TB is increasing due to inadequate monitoring, lack of proper diagnosis, and treatment. The GenoType ® Mycobacterium tuberculosis drug resistance second line (MTBDRsl assay is a novel line probe assay used for the rapid detection of mutational patterns conferring resistance to XDR-TB. Aim: The aim of this study was to study the rapid detection of drug resistance and mutational patterns of the XDR-TB by a novel GenoType ® MTBDRsl assay. Materials and Methods: We evaluated 98 multidrug-resistant (MDR M. tuberculosis isolates for second line drugs susceptibility testing by 1% proportion method (BacT/ALERT 3D system and GenoType ® MTBDRsl assay for rapid detection of conferring drug resistance to XDR-TB. Results: A total of seven (17.4% were identified as XDR-TB by using standard phenotypic method. The concordance between phenotypic and GenoType ® MTBDRsl assay was 91.7-100% for different antibiotics. The sensitivity and specificity of the MTBDRsl assay were 100% and 100% for aminoglycosides; 100% and 100% for fluoroquinolones; 91.7% and 100% for ethambutol. The most frequent mutations and patterns were gyrA MUT1 (A90V in seven (41.2% and gyrA + WT1-3 + MUT1 in four (23.5%; rrs MUT1 (A1401G in 11 (64.7%, and rrs WT1-2 + MUT1 in eight (47.1%; and embB MUT1B (M306V in 11 (64.7% strains. Conclusions: These data suggest that the GenoType ® MTBDRsl assay is rapid, novel test for detection of resistance to second line anti-tubercular drugs. This assay provides additional information about the frequency and mutational patterns responsible for XDR-TB resistance.

Efficacy of pyrantel pamoate and ivermectin for the treatment of canine nematodes

Directory of Open Access Journals (Sweden)

Ana Paula Jesus

2015-12-01

Full Text Available This study evaluated the efficacy of pyrantel pamoate and ivermectin on gastrointestinal nematodes in dogs. Fecal egg counts per gram (EPG were measured by the fecal egg count reduction test (FECRT in order to evaluate the anthelmintic efficiency and fecal float exams were also performed to assess the concordance between coproparasitological techniques. A total of 45 naturally infected dogs in the city of Bandeirantes, Paraná State, were selected and divided into three groups: Group 1, 15 animals that received pyrantel pamoate (145 mg in a single dose; Group 2, 15 animals that received ivermectin (3 mg; and Group 3, 15 animals that comprised an untreated control group. Fecal testing was performed two and 10 days after treatment. Toxocara was the most prevalent genus, followed by Ancylostoma and Trichuris. Ancylostoma had low resistance to ivermectin and pyrantel pamoate treatment, while Toxocara were resistant to both treatments. Statistical correlation testing to compare coproparasitogical techniques revealed moderate concordance, substantial and almost perfect concordance for detection of Ancylostoma, Trichuris, and Toxocara, respectively. The results of this study suggest that the gender Ancylostoma had low resistance and Toxocara is resistant to both drugs and because of their high prevalence in young animals means that others anthelmintic drugs may be recommended to combat infections. Additionally, the Gordon and Whitlock modified and Willis-Mollay techniques are effective for detection particularly of Toxocara in dogs.

Resistance pattern and detection of metallo‑beta‑lactamase genes ...

African Journals Online (AJOL)

Materials and Methods: Two hundred nonduplicate, consecutive isolates of P. aeruginosa from clinical samples submitted to the Medical Microbiology Laboratory of National Hospital, Abuja were screened for carbapenem resistance using imipenem and meropenem. Phenotypic detection of MBL‑producing strains was ...

Detection of antibiotic resistance in clinical bacterial strains from pets

OpenAIRE

Poeta, P.; Rodrigues, J.

2008-01-01

The identification of different bacterial strains and the occurrence of antibiotic resistance were investigated in several infection processes of pets as skin abscess with purulent discharge, bronco alveolar fluid, earwax, urine, mammary, and eye fluid. Streptococcus spp. and Staphylococcus spp. were the most detected in the different samples. A high frequency of antimicrobial resistance has been observed and this could reflect the wide use of antimicrobials in pets, making the effectiveness ...

Effects of anthelmintic treatment and feed supplementation on grazing Tuli weaner steers naturally infected with gastrointestinal nematodes

Directory of Open Access Journals (Sweden)

A. Magaya

2000-07-01

Full Text Available A study was carried out to determine the epidemiology of gastrointestinal nematodes in indigenous Tuli cattle and the effect of dietary protein supplementation and anthelmintic treatment on productivity in young growing cattle. Forty steers with an average age of 18 months were divided into 4 groups; 1 fenbendazole (slow release bolus and cottonseed meal (FCSM group, 2 fenbendazole (FBZ group, 3 cottonseed meal (CSM group and 4 control (no cottonseed meal and no fenbendazole (control group. Performance parameters measured included wormeggs per gram of faeces (EPG, packed cell volume (PCV, albumin and live-weight gain. Results showed that faecal worm egg counts were lower and PCV was higher in the FCSM and FBZ groups than in the CSM and control groups (P < 0.01. Weight gains were higher in the CSMand FCSM groups than in the FBZ and control groups (P < 0.05. The cost benefits of anthelmintic treatment and dietary supplementation were apparent in this study. The improved growth performance of the FCSM, FBZ and CSM groups reflected a financial gain over the controls on termination of the study. The dominant genera of gastrointestinal nematodes on faecal culture, pasture larval counts and necropsy were Cooperia and Haemonchus. The incidences of Trichostrongylus, Oesophagostomum and Bunostomum were low.

PCR-based Approaches for the Detection of Clinical Methicillin-resistant Staphylococcus aureus

Science.gov (United States)

Liu, Ying; Zhang, Jiang; Ji, Yinduo

2016-01-01

Staphylococcus aureus is an important pathogen that can cause a variety of infections, including superficial and systematic infections, in humans and animals. The persistent emergence of multidrug resistant S. aureus, particularly methicillin-resistant S. aureus, has caused dramatically economic burden and concerns in the public health due to limited options of treatment of MRSA infections. In order to make a correct choice of treatment for physicians and understand the prevalence of MRSA, it is extremely critical to precisely and timely diagnose the pathogen that induces a specific infection of patients and to reveal the antibiotic resistant profile of the pathogen. In this review, we outlined different PCR-based approaches that have been successfully utilized for the rapid detection of S. aureus, including MRSA and MSSA, directly from various clinical specimens. The sensitivity and specificity of detections were pointed out. Both advantages and disadvantages of listed approaches were discussed. Importantly, an alternative approach is necessary to further confirm the detection results from the molecular diagnostic assays. PMID:27335617

Effect of combinations of marketed human anthelmintic drugs against Trichuris muris in vitro and in vivo

Directory of Open Access Journals (Sweden)

Keiser Jennifer

2012-12-01

Full Text Available Abstract Background Soil-transmitted helminth (STH infections are responsible for a huge public health burden, however treatment options are limited. The discovery and development of novel efficacious drugs or drug combinations for the treatment of STH infections therefore has a high research priority. Methods We studied drug combination effects using the main standard anthelmintics, albendazole, mebendazole, levamisole, pyrantel pamoate and ivermectin in the Trichuris muris model. Drug combinations were first tested in vitro and additive and synergistic combinations investigated further in vivo in female mice using ratios based on the ED50 of the respective drugs. Results In vitro all 10 combinations of the standard anthelmintics tested against T. muris revealed synergistic behavior. We identified three drug combinations in vivo as strongly synergistic, namely mebendazole-ivermectin (Combination index (CI=0.16, mebendazole-levamisole (CI=0.17 and albendazole-mebendazole (CI=0.23. For albendazole-ivermectin, moderate synergism was observed (CI=0.81 and for albendazole-levamisole a nearly additive effect was documented (CI=0.93 in vivo. Five combinations (albendazole-pyrantel pamoate, mebendazole-pyrantel pamoate, levamisole-pyrantel pamoate, levamisole-ivermectin and pyrantel pamoate-ivermectin were antagonistic in vivo. Conclusion Our results strengthen the evidence that combination chemotherapy might play a role in the treatment of Trichuris infections. Albendazole-mebendazole should be studied in greater detail in preclinical studies.

Laboratory methods for diagnosis and detection of drug resistant ...

African Journals Online (AJOL)

Data source: Published series of peer reviewed journals and manuals written on laboratory methods that are currently used for diagnosis and detection of drug resistance of Mycobacterium tuberculosis complex were reviewed using the index medicus, pubmed and medline search. Conventional bacteriological microscopy ...

Detection of underground mined voids using line electrode resistivity technique - case study

Energy Technology Data Exchange (ETDEWEB)

Peng, S.S.; Ziaie, F. (West Virginia University, Morgantown, WV (USA))

1991-06-01

A new resistivity method was developed and tested in three phases; simulated model, similitude model, and field survey. This resistivity method was a combination of the Bristow arrangement and line electrode method. Three line electrodes were chosen so that the sinkhole electrode was emplaced at a far distance from the other two electrodes. Any of the two electrodes and the sinkhole electrode were activated and several resistivity profiles perpendicular to the line electrode prepared for different electrodes activation. Subsurface cavities caused resistivity anomalies which were interpreted to locate their sources (cavities) and estimate the depths and dimension of the cavities. A coal mine site employing the room and pillar mining system was selected to confirm the results of the laboratory. The results of the interpretation indicated that the entry with a dimension of 135 cm high and 5.40 m wide at a depth of 25.50 m can be detected by this method. The resolution of the detectability of this method proved a great success when compared to other resistivity techniques. 6 refs., 6 figs.

PCR-based detection of resistance genes in anaerobic bacteria isolated from intra-abdominal infections.

Science.gov (United States)

Tran, Chau Minh; Tanaka, Kaori; Watanabe, Kunitomo

2013-04-01

Little information is available on the distribution of antimicrobial resistance genes in anaerobes in Japan. To understand the background of antimicrobial resistance in anaerobes involved in intra-abdominal infections, we investigated the distribution of eight antimicrobial resistance genes (cepA, cfiA, cfxA, ermF, ermB, mefA, tetQ, and nim) and a mutation in the gyrA gene in a total of 152 organisms (Bacteroides spp., Prevotella spp., Fusobacterium spp., Porphyromonas spp., Bilophila wadsworthia, Desulfovibrio desulfuricans, Veillonella spp., gram-positive cocci, and non-spore-forming gram-positive bacilli) isolated between 2003 and 2004 in Japan. The cepA gene was distributed primarily in Bacteroides fragilis. Gene cfxA was detected in about 9 % of the Bacteroides isolates and 75 % of the Prevotella spp. isolates and did not appear to contribute to cephamycin resistance. Two strains of B. fragilis contained the metallo-β-lactamase gene cfiA, but they did not produce the protein product. Gene tetQ was detected in about 81, 44, and 63 % of B. fragilis isolates, other Bacteroides spp., and Prevotella spp. isolates, respectively. The ermF gene was detected in 25, 13, 56, 64, and 16 % of Bacteroides spp., Prevotella spp., Fusobacterium spp., B. wadsworthia, and anaerobic cocci, respectively. Gene mefA was found in only 10 % of the B. fragilis strains and 3 % of the non-B. fragilis strains. Genes nim and ermB were not detected in any isolate. Substitution at position 82 (Ser to Phe) in gyrA was detected in B. fragilis isolates that were less susceptible or resistant to moxifloxacin. This study is the first report on the distribution of resistance genes in anaerobes isolated from intra-abdominal infections in Japan. We expect that the results might help in understanding the resistance mechanisms of specific anaerobes.

Detection of antibiotic resistance in probiotics of dietary supplements

KAUST Repository

Wong, Aloysius Tze

2015-09-14

Background Probiotics are live microorganisms that confer nutrition- and health-promoting benefits if consumed in adequate amounts. Concomitant with the demand for natural approaches to maintaining health is an increase in inclusion of probiotics in food and health products. Since probiotic bacteria act as reservoir for antibiotic resistant determinants, the transfer of these genes to pathogens sharing the same intestinal habitat is thus conceivable considering the fact that dietary supplements contain high amounts of often heterogeneous populations of probiotics. Such events can confer pathogens protection against commonly-used drugs. Despite numerous reports of antibiotic resistant probiotics in food and biological sources, the antibiogram of probiotics from dietary supplements remained elusive. Findings Here, we screened five commercially available dietary supplements for resistance towards antibiotics of different classes. Probiotics of all batches of products were resistant towards vancomycin while batch-dependent resistance towards streptomycin, aztreonam, gentamycin and/or ciprofloxacin antibiotics was detected for probiotics of brands Bi and Bn, Bg, and L. Isolates of brand Cn was also resistant towards gentamycin, streptomycin and ciprofloxacin antibiotics. Additionally, we also report a discrepancy between the enumerated viable bacteria amounts and the claims of the manufacturers. Conclusions This short report has highlighted the present of antibiotic resistance in probiotic bacteria from dietary supplements and therefore serves as a platform for further screenings and for in-depth characterization of the resistant determinants and the molecular machinery that confers the resistance.

Effective anthelmintic therapy of residents living in endemic area of high prevalence for Hookworm and Schistosoma mansoni infections enhances the levels of allergy risk factor anti-Der p1 IgE

Directory of Open Access Journals (Sweden)

Sabrina S. Campolina

2015-01-01

Full Text Available In this work were investigated the relationship between Hookworm/Schistosoma mansoni infections and allergy related risk factors in two endemic areas with distinct prevalence of infections and co-infection. The intensity of infections, eosinophilia, allergy risk factors, infections status and anti-Der p1 IgE levels before and 2 years (population 1 and 3 years (population 2 after anthelmintic treatment, were evaluated. It was observed that the population with lower prevalence and intensity of infection (population 2 had lower eosinophils counts (>600/mm3 and higher animal contact than the population with higher parasites intensity (population 1. After anthelmintic treatment the intensity of S. mansoni single infection decreased, but no changes were observed in Hookworm and co-infected individuals. The anthelmintic treatment also enhanced anti-Der p1 IgE optical density in ELISA on the subgroups that became negative for helminth infection regardless of their previous infection condition in population 1. Facing that, we evaluated the anti-Der p1 IgE reactivity index, and the ratio (after/before treatment was significantly higher in patients co-infected before treatment. On the other hand, no association between anti-Der p1 IgE reactivity index and the intensity of infections were observed. In conclusion, effective anthelmintic therapy of subjects from endemic areas with high prevalence of Hookworm and S. mansoni infections enhances anti-Der p1 IgE levels.

Anti-parasitic activity of pelleted sainfoin (Onobrychis viciifolia) against Ostertagia ostertagi and Cooperia oncophora in calves

DEFF Research Database (Denmark)

Desrues, O.; Pena-Espinoza, Miguel Angel; Hansen, T. V.

2016-01-01

BACKGROUND: Increasing anthelmintic-resistance in nematodes of ruminants emphasises the need for sustainable parasite control. Condensed tannin-containing legume forages such as sainfoin (Onobrychis viciifolia) have shown promising anthelmintic properties in small ruminants but this has never been...

Ice detection in heat pumps and coolers. [By thermal resistance and capacitance detection

Energy Technology Data Exchange (ETDEWEB)

Buick, T R; McMullan, J T; Morgan, R; Murray, R B

1978-01-01

Some methods are discussed for detecting the formation of ice on the evaporators of air-source heat pumps and air coolers by electronic means. The sensing of thermal resistance caused by ice build-up can be done by measuring temperature differences between the evaporator and the air, and analyses are presented of the effect of using both linear and non-linear temperature sensors for this purpose. The direct detection of the presence of ice can be done by measuring the capacitance of a suitably-placed pair of plates, and the performance of such a system is analyzed. Preliminary reports are presented of the use of both of these methods of ice detection in the defrosting of an experimental heat pump.

Field efficacy of four anthelmintics and confirmation of drug-resistant nematodes by controlled efficacy test and pyrosequencing on a sheep and goat farm in Denmark

DEFF Research Database (Denmark)

Pena-Espinoza, Miguel Angel; Thamsborg, Stig M.; Demeler, Janina

2014-01-01

and Trichostrongylus colubriformis isolated from adult goats on the farm. Recovered specimens of H. contortus were subjected to pyrosequencing for detection of single nucleotide polymorphisms (SNPs) related to benzimidazole (BZ) resistance. During the FECRT, FECs in untreated lambs dropped significantly by 47%. No FEC...

Evaluation of different methods to detect methicillin resistance in Staphylococcus aureus (MRSA).

Science.gov (United States)

Alipour, Farzad; Ahmadi, Malahat; Javadi, Shahram

2014-01-01

The studies suggest that dogs living with human are potential risk of becoming MRSA carrier and increased risk of infections caused by MRSA. Phenotypic methods to detect methicillin resistance in Staphylococcus aureus (MRSA) are inadequate. The objective of the present study was to determine methicillin resistance in S. aureus by phenotypic susceptibility test (oxacillin disk diffusion, cefoxitin disk diffusion, oxacillin screen agar) and molecular methods (PCR as a gold standard) and the latex agglutination test for the detection of PBP2a and to evaluate the results of these tests for its sensitivity and specificity. A total of 100 swab samples were taken from muzzle site, in more contact with human, of dogs and MRSA were isolated. Oxacillin (1 μg), cefoxitin (30 μg) disk diffusion and oxacillin screen agar method were used. The isolates were also subjected to latex agglutination test for detection of PBP2a and PCR to detect mecA gene. By PCR 37% of isolates show the presence of mecA. Latex agglutination was found to be the most sensitive (97.29%) and cefoxitin disk diffusion to be the most specific (96.82%) tests for detection of MRSA. Our finding showed that combining oxacillin screen agar or cefoxitin disk diffusion with latex agglutination improves sensitivity and specificity to detect methicillin resistance S. aureus (MRSA) isolates. Copyright © 2014 King Saud Bin Abdulaziz University for Health Sciences. Published by Elsevier Ltd. All rights reserved.

Development of leak detection system using high temperature-resistant microphones

International Nuclear Information System (INIS)

Morishita, Yoshitsugu; Mochizuki, Hiroyasu; Watanabe, Kenshiu; Nakamura, Takahisa; Nakazima, Yoshiaki; Yamauchi, Tatsuya

1995-01-01

This report describes the development and testing of a coolant leak detection system for an inlet feeder pipe of an advanced thermal reactor (ATR) using high temperature-resistant microphones. Such microphones must be resistant to both high temperatures and high radiation doses. Leakage sound characteristics, attenuation of the sound level in a heat insulating box for the inlet feeder pipes, and background noise were investigated using the experimental facility and the prototype ATR 'FUGEN'. The optimum frequency ranges for the microphone were then determined based on the observed leakage sound and background noise. The ability of the microphone to discriminate between leaks and other burst-type noises was also investigated by statistical analyses. Finally, it was confirmed that the present method could detect a leak within a couple of seconds. (author)

Discovery of genomic intervals that underlie nematode responses to benzimidazoles.

Science.gov (United States)

Zamanian, Mostafa; Cook, Daniel E; Zdraljevic, Stefan; Brady, Shannon C; Lee, Daehan; Lee, Junho; Andersen, Erik C

2018-03-01

Parasitic nematodes impose a debilitating health and economic burden across much of the world. Nematode resistance to anthelmintic drugs threatens parasite control efforts in both human and veterinary medicine. Despite this threat, the genetic landscape of potential resistance mechanisms to these critical drugs remains largely unexplored. Here, we exploit natural variation in the model nematodes Caenorhabditis elegans and Caenorhabditis briggsae to discover quantitative trait loci (QTL) that control sensitivity to benzimidazoles widely used in human and animal medicine. High-throughput phenotyping of albendazole, fenbendazole, mebendazole, and thiabendazole responses in panels of recombinant lines led to the discovery of over 15 QTL in C. elegans and four QTL in C. briggsae associated with divergent responses to these anthelmintics. Many of these QTL are conserved across benzimidazole derivatives, but others show drug and dose specificity. We used near-isogenic lines to recapitulate and narrow the C. elegans albendazole QTL of largest effect and identified candidate variants correlated with the resistance phenotype. These QTL do not overlap with known benzimidazole target resistance genes from parasitic nematodes and present specific new leads for the discovery of novel mechanisms of nematode benzimidazole resistance. Analyses of orthologous genes reveal conservation of candidate benzimidazole resistance genes in medically important parasitic nematodes. These data provide a basis for extending these approaches to other anthelmintic drug classes and a pathway towards validating new markers for anthelmintic resistance that can be deployed to improve parasite disease control.

Efficacy of closantel against ivermectin- and fenbendazole-resistant Haemonchus sp. in sheep in Ontario, Canada.

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Westers, T; Jones-Bitton, A; Menzies, P; Van Leeuwen, J; Poljak, Z; Peregrine, A S

2016-09-15

In Ontario, Canada, widespread resistance to ivermectin and fenbendazole, the only readily available ovine anthelmintics, has been documented, primarily in Haemonchus sp. In other parts of the world, closantel has been used to control such infections; however, the drug was not currently licensed for use in Canada and the USA. A randomized controlled trial was conducted on six client-owned farms in Ontario in 2013 and 2014 to determine the efficacy of closantel (Flukiver ® 5% Oral Suspension, Elanco Animal Health, 10mg/kg bodyweight) against ivermectin- and fenbendazole-resistant Haemonchus sp. infections in periparturient ewes and grazing lambs. Three farms were randomly assigned to treat all ewes, and three farms were randomly assigned to selectively treat individual ewes at lambing, using predetermined criteria. Fecal samples were collected from a minimum of 15 randomly selected ewes and 13 lambs per group on each farm at the time of treatment and approximately 14days later. Trichostrongyle-type fecal egg counts (FEC) were performed using a modified McMaster technique with a lower detection limit of 8.3 eggs per gram of feces (epg). Haemonchus-specific FECs were determined by multiplying FECs by the proportion of Haemonchus sp. identified from coproculture for each farm; Haemonchus-specific FEC reductions were calculated for each farm. Twenty grazing lambs had FECs conducted monthly, and when mean monthly FECs surpassed 200 epg, all lambs were randomly allocated to either closantel, positive control (ivermectin, fenbendazole, or levamisole) or negative control groups. Pre-treatment Haemonchus-specific mean FECs ranged from 27 to 3359 epg in ewes and 0-5698 epg in lambs. Efficacy of closantel against Haemonchus sp. ranged from 99% (95% CI: 97%-99%) to 100% in recently lambed ewes on all farms in both years (total n=274 ewes), and from 99% (95% CI: 98%-99%) to 100% in grazing lambs in both years on all but one farm (total n=171 lambs). On the latter farm, a whole

Development of a leak detection system using high temperature-resistant microphones

International Nuclear Information System (INIS)

Morishita, Yoshitsugu; Mochizuki, Hiroyasu; Watanabe, Kenshiu; Nakamura, Takahisa; Nakajima, Yoshiaki; Yamauchi, Tatsuya

1991-01-01

This report describes the development of a detection system of coolant leak from an inlet feeder pipe of an Advanced Thermal Reactor (ATR) with high temperature-resistant microphones. A microphone having resistance to both high temperature and high radiation dose has been developed at first. The characteristics with regard to leakage sound, attenuation of sound level in a heat insulating box for the inlet feeder pipes and background noise were clarified by laboratory experiments and measurements in the prototype ATR 'Fugen'. On the basis of these experimental findings, appropriate frequency ranges were surveyed to detect the leakage sound with a high S/N ratio under the background noise. Reliability of the system to a malfunction caused by burst-type noises observed in the plant was also investigated by statistical analyses. Finally, it was confirmed that the present method could detect a leak within a couple of seconds. (author)

Factors affecting fecal egg counts in peri-parturient Katahdin ewes and their lambs

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Selection for low fecal egg counts (FEC) can be used to genetically enhance resistance to gastrointestinal nematode parasites in growing lambs, thereby reducing the frequency of use of anthelmintics, facilitating marketing of organic lamb, and reducing the risk of development of anthelmintic resista...

Examination of commercially available copper oxide wire particles in combination with albendazole for control of gastrointestinal nematodes in lambs

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Alternatives to synthetic anthelmintics remain critical due to the prevalence of anthelmintic resistance. The objective of the experiment was to determine the efficacy of copper oxide wire particles (COWP) from three commercial sources to control Haemonchus contortus in lambs. Naturally infected Ka...

Microfluidic platform for electrophysiological recordings from host-stage hookworm and Ascaris suum larvae: A new tool for anthelmintic research

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Janis C. Weeks

2016-12-01

Full Text Available The screening of candidate compounds and natural products for anthelmintic activity is important for discovering new drugs against human and animal parasites. We previously validated in Caenorhabditis elegans a microfluidic device (‘chip’ that records non-invasively the tiny electrophysiological signals generated by rhythmic contraction (pumping of the worm's pharynx. These electropharyngeograms (EPGs are recorded simultaneously from multiple worms per chip, providing a medium-throughput readout of muscular and neural activity that is especially useful for compounds targeting neurotransmitter receptors and ion channels. Microfluidic technologies have transformed C. elegans research and the goal of the current study was to validate hookworm and Ascaris suum host-stage larvae in the microfluidic EPG platform. Ancylostoma ceylanicum and A. caninum infective L3s (iL3s that had been activated in vitro generally produced erratic EPG activity under the conditions tested. In contrast, A. ceylanicum L4s recovered from hamsters exhibited robust, sustained EPG activity, consisting of three waveforms: (1 conventional pumps as seen in other nematodes; (2 rapid voltage deflections, associated with irregular contractions of the esophagus and openings of the esophogeal-intestinal valve (termed a ‘flutter’; and (3 hybrid waveforms, which we classified as pumps. For data analysis, pumps and flutters were combined and termed EPG ‘events.’ EPG waveform identification and analysis were performed semi-automatically using custom-designed software. The neuromodulator serotonin (5-hydroxytryptamine; 5HT increased EPG event frequency in A. ceylanicum L4s at an optimal concentration of 0.5 mM. The anthelmintic drug ivermectin (IVM inhibited EPG activity in a concentration-dependent manner. EPGs from A. suum L3s recovered from pig lungs exhibited robust pharyngeal pumping in 1 mM 5HT, which was inhibited by IVM. These experiments validate the use of A

Drug resistance detection and mutation patterns of multidrug resistant tuberculosis strains from children in Delhi

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Jyoti Arora

2017-06-01

Full Text Available A total of 312 sputum samples from pediatric patients presumptive of multidrug resistant tuberculosis were tested for the detection of drug resistance using the GenoTypeMTBDRplus assay. A total of 193 (61.8% patients were smear positive and 119 (38.1% were smear negative by Ziehl–Neelsen staining. Line probe assay (LPA was performed for 208 samples/cultures (193 smear positive samples and 15 cultures from smear negative samples. Valid results were obtained from 198 tests. Of these, 125/198 (63.1% were sensitive to both rifampicin (RIF and isoniazid (INH. 73/198 (36.9% were resistant to at least INH/RIF, out of which 49 (24.7% were resistant to both INH and RIF (multidrug resistant. Children with tuberculosis are often infected by someone close to them, so strengthening of contact tracing in the program may help in early diagnosis to identify additional cases within the household. There is a need to evaluate newer diagnostic assays which have a high sensitivity in the case of smear negative samples, additional samples other than sputum among young children not able to expectorate, and also to fill the gap between estimated and reported cases under the program.

Development of Radiometric and Allied Analytical Methods and Strategies to Strengthen National Residue Control Programmes for Antibiotic and Anthelmintic Veterinary Drug Residues. Final Report of a Coordinated Research Project

International Nuclear Information System (INIS)

2016-08-01

Awareness of food safety is rising among consumers, and many importing countries implement food control regulations to guarantee the quality and safety of imported foods for their consumers. Many developing countries have also taken steps to put in place control systems that encourage responsible use of veterinary medicines to combat possible drug resistance, control drug residues and ensure compliance with international and national standards. However, these countries still require the necessary know-how and skills to protect local consumers and to access international markets. One significant constraint is the capacity of laboratory services to generate surveillance data using reliable and cost effective analytical methods validated to national and international standards. The IAEA coordinated research project (CRP) on the Development of Radiometric and Allied Analytical Methods to Strengthen National Residue Control Programmes for Antibiotic and Anthelmintic Veterinary Drug Residues was initiated in 2009 to conduct work on robust nuclear and related technologies suitable for the screening and confirmatory analysis of residues of veterinary medicines, including antimicrobials and anthelmintics commonly used in animal production, with public health and trade significance. The CRP also explored mechanisms to enhance networking among research institutions involved in research on pharmacologically active veterinary drug residues in food (primarily) and environmental samples. The project was implemented by the Joint FAO/IAEA Division of Nuclear Techniques in Food and Agriculture between 2009 and 2014 and involved eleven research contracts and one technical contract, five research agreements and one institution. The CRP was a continuation of the CRP on the Development of Strategies for the Effective Monitoring of Veterinary Drug Residues in Livestock and Livestock Products in Developing Countries and the key findings are also summarized in this publication.

An analysis of genetic diversity and inbreeding in Wuchereria bancrofti: implications for the spread and detection of drug resistance.

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Thomas S Churcher

2008-04-01

Full Text Available Estimates of genetic diversity in helminth infections of humans often have to rely on genotyping (immature parasite transmission stages instead of adult worms. Here we analyse the results of one such study investigating a single polymorphic locus (a change at position 200 of the beta-tubulin gene in microfilariae of the lymphatic filarial parasite Wuchereria bancrofti. The presence of this genetic change has been implicated in benzimidazole resistance in parasitic nematodes of farmed ruminants. Microfilariae were obtained from patients of three West African villages, two of which were sampled prior to the introduction of mass drug administration. An individual-based stochastic model was developed showing that a wide range of allele frequencies in the adult worm populations could have generated the observed microfilarial genetic diversity. This suggests that appropriate theoretical null models are required in order to interpret studies that genotype transmission stages. Wright's hierarchical F-statistic was used to investigate the population structure in W. bancrofti microfilariae and showed significant deficiency of heterozygotes compared to the Hardy-Weinberg equilibrium; this may be partially caused by a high degree of parasite genetic differentiation between hosts. Studies seeking to quantify accurately the genetic diversity of helminth populations by analysing transmission stages should increase their sample size to account for the variability in allele frequency between different parasite life-stages. Helminth genetic differentiation between hosts and non-random mating will also increase the number of hosts (and the number of samples per host that need to be genotyped, and could enhance the rate of spread of anthelmintic resistance.

Evaluation of in-vitro anthelmintic activity of Heliotropium indicum Linn. leaves in Indian adult earthworm

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Kabita Mahato

2014-02-01

Full Text Available Objective: To carry out an exhaustive study with a view to substantiate the therapeutic potential of the plant in terms of its anthelmintic activity against Pheretima posthuma using mebendazole as a reference standard. Methods: Forty five worms were collected and were divided into nine groups and were washed in normal saline before they were released into 10 mL of respective drug solutions using distilled water as vehicle. Different concentrations (25 mg/mL, 50 mg/mL and 100 mg/mL of the test (MEHI and AEHI and standard solution were prepared before the commencement of the experiment. Time for paralysis and time for death were recorded for each group. Results: Crude methanolic extract with concentrations of 25 mg/mL, 50 mg/mL, 100 mg/mL produced dose-dependent paralysis. Time of paralysis and death increased with concentration of 25 mg/mL, 50 mg/mL as compared with that of standard drug mebendazole. Methanolic extract of Heliotropium indicum Linn. (H. indicum gave shorter paralysis and death time at 100 mg/mL as compared to aqueous extract H. indicum Linn. Results are expressed as mean依SEM (P<0.05 of 5 worms in each group. Conclusions: From the investigation, conclusion can be drawn that the methanolic extract of H. indicum (Boraginaceae showed better activity than aqueous extract of the same to treat intestinal worm infections. In comparison with the standard drug mebendazole, methanolic extract of H. indicum showed significant anthelmintic efficacy.

Transmission and selection of macrolide resistant Mycoplasma genitalium infections detected by rapid high resolution melt analysis.

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Jimmy Twin

Full Text Available BACKGROUND: Mycoplasma genitalium (MG causes urethritis, cervicitis and pelvic inflammatory disease. The MG treatment failure rate using 1 g azithromycin at an Australian Sexual Health clinic in 2007-9 was 31% (95%CI 23-40%. We developed a rapid high resolution melt analysis (HRMA assay targeting resistance mutations in the MG 23S rRNA gene, and validated it against DNA sequencing by examining pre- and post-treatment archived samples from MG-infected patients. METHODOLOGY/PRINCIPAL FINDINGS: Available MG-positive pre-treatment (n = 82 and post-treatment samples from individuals with clinical treatment failure (n = 20 were screened for 23S rRNA gene mutations. Sixteen (20% pre-treatment samples possessed resistance mutations (A2058G, A2059G, A2059C, which were significantly more common in patients with symptomatic azithromycin-treatment failure (12/26; 44% than in those clinically cured (4/56; 7%, p<0.001. All 20 patients experiencing azithromycin-failure had detectable mutations in their post-treatment samples. In 9 of these cases, the same mutational types were present in both pre- and post-treatment samples indicating transmitted resistance, whilst in 11 of these cases (55%, mutations were absent in pre-treatment samples indicating likely selection of resistant isolates have occurred. HRMA was able to detect all mutational changes determined in this study by DNA sequencing. An additional HRMA assay incorporating an unlabelled probe was also developed to detect type 4 single-nucleotide polymorphisms found in other populations, with a slightly lower sensitivity of 90%. CONCLUSIONS/SIGNIFICANCE: Treatment failure is associated with the detection of macrolide resistance mutations, which appear to be almost equally due to selection of resistant isolates following exposure to 1 g azithromycin and pre-existing transmitted resistance. The application of a rapid molecular assay to detect resistance at the time of initial detection of infection allows

Synthesis and antimicrobial, antifungal and anthelmintic activities of 3H-1,5-benzodiazepine derivatives

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RAJESH KUMAR

2008-10-01

Full Text Available The diazonium salt of 4-amino-1-methyl-3-propyl-1H-pyrazole-5-carboxamide in the presence of sodium hydroxide was condensed with different β-diketones/β-ketoesters, 3a–e, to obtain new β-diketones/β-ketoesters, 4a–e. The β-diketones/β-ketoesters 4a–e were condensed with o-phenylenediamine (o-PDA in presence of p-toluenesulfonic acid/SiO2 to give biologically active 3H-1,5-benzodiazepines, 5a–e. All the newly synthesized compounds were characterized by elemental analysis and spectral studies. The compounds 5a–e was screened for their antimicrobial, antifungal and anthelmintic activities.

Production benefits from pre- and post-lambing anthelmintic treatment of ewes on commercial farms in the southern North Island of New Zealand.

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Miller, C M; Ganesh, S; Garland, C B; Leathwick, D M

2015-07-01

To measure the magnitude and variability in production responses to anthelmintic treatments administered to adult ewes around lambing. Ewes carrying twin lambs, from sheep and beef farms (eight in Year 1 and six in Year 2) in the Wairarapa region of New Zealand, were enrolled in 14 trials (part of an experiment carried out on one farm in one year). Experiment 1 compared ewes treated 2-4 weeks pre-lambing with a controlled release capsule (CRC) containing abamectin, albendazole, Se and Co, to ewes injected pre-lambing with a long-acting Se plus vitamin B12 product, and to untreated ewes. Experiment 2 included these treatments, plus a CRC administered at pregnancy scanning. Experiment 3 included the same treatments as Experiment 1, plus administration of a CRC containing albendazole, Se and Co, injectable moxidectin or oral derquantel plus abamectin, all administered pre-lambing, or oral derquantel plus abamectin administered 4-6 weeks after lambing. Variables compared were ewe liveweight at weaning and pre-mating, lamb liveweight at weaning, total weight of lamb weaned per ewe and ewe dag score at weaning. Ewes treated with a CRC pre-lambing were heavier than untreated ewes (mean 3.2 kg) at weaning in 12/14 trials, and pre-mating (mean 2.8 kg) in 9/14 trials (pewes the mean difference was 2.8 kg pre-lambing (9/14 trials) and 1.7 kg pre-weaning (6/14 trials). Lambs reared by treated ewes were heavier (mean 1.55 kg) at weaning in 6/14 trials (pewe (p=0.507). Variation in weight of lamb weaned per ewe was largely explained by differences in lamb survival from birth to weaning (p0.65). Treatment of ewes with a CRC at pregnancy scanning was neither better nor worse than a pre-lambing treatment (p=0.065). There was no difference in the response from treatment with either of the two CRC or moxidectin. Treatment with short-acting oral anthelmintics resulted in no consistent benefit. Anthelmintic treatments administered to ewes around lambing resulted in variable responses

Nonradioactive heteroduplex tracking assay for the detection of minority-variant chloroquine-resistant Plasmodium falciparum in Madagascar

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Juliano, Jonathan J; Randrianarivelojosia, Milijaona; Ramarosandratana, Benjamin; Ariey, Frédéric; Mwapasa, Victor; Meshnick, Steven R

2009-01-01

Background Strains of Plasmodium falciparum genetically resistant to chloroquine (CQ) due to the presence of pfcrt 76T appear to have been recently introduced to the island of Madagascar. The prevalence of such resistant genotypes is reported to be low (chloroquine resistant parasites were described in Malawian patients using an isotopic heteroduplex tracking assay (HTA), which can detect pfcrt 76T-bearing P. falciparum minority variants in individual patients that were undetectable by conventional PCR. However, as this assay required a radiolabeled probe, it could not be used in many resource-limited settings. Methods This study describes a digoxigenin (DIG)-labeled chemiluminescent heteroduplex tracking assay (DIG-HTA) to detect pfcrt 76T-bearing minority variant P. falciparum. This assay was compared to restriction fragment length polymorphism (RFLP) analysis and to the isotopic HTA for detection of genetically CQ-resistant parasites in clinical samples. Results Thirty one clinical P. falciparum isolates (15 primary isolates and 16 recurrent isolates) from 17 Malagasy children treated with CQ for uncomplicated malaria were genotyped for the pfcrt K76T mutation. Two (11.7%) of 17 patients harboured genetically CQ-resistant P. falciparum strains after therapy as detected by HTA. RFLP analysis failed to detect any pfcrt K76T-bearing isolates. Conclusion These findings indicate that genetically CQ-resistant P. falciparum are more common than previously thought in Madagascar even though the fitness of the minority variant pfcrt 76T parasites remains unclear. In addition, HTAs for malaria drug resistance alleles are promising tools for the surveillance of anti-malarial resistance. The use of a non-radioactive label allows for the use of HTAs in malaria endemic countries. PMID:19291288

Detection of Macrolide, Lincosamide and Streptogramin Resistance among Methicillin Resistant Staphylococcus aureus (MRSA in Mumbai

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Arunagiri Subramanian

2015-01-01

Full Text Available Background: The increase in incidence of Methicillin Resistant Staphyloccocus aureus (MRSA and its extraordinary potential to develop antimicrobial resistance has highlighted the need for better agents to treat such infections. This has led to a renewed interest in use of new drugs for treatment with clindamycin and quinuprsitin-dalfopristin being the preferred choice for treatment. Aim & Objectives: This study was undertaken to detect the prevalence of MacrolideLincosamide-Streptogramin (MLS resistance among clinical isolates of MRSA.Material and Methods:Two hundred and thirty clinical isolates of S. aureus were subjected to routine antibiotic susceptibility testing including cefoxitin, erythromycin and quinupristindalfopristin. Inducible resistance to clindamycin was tested by 'D' test as per Clinical and Laboratory Standards Institute (CLSI guidelines. Results: Out of all S. aureus isolates, 93.91% were identified as MRSA. In the disc diffusion testing, 81.5% of isolates showed erythromycin resistance. Among these, the prevalence of constitutive (cMLS , inducible (iMLS b b and MS-phenotype were 35.80%, 31.82% and 32.39% respectively by the D-test method. 77.8% of isolates were resistant to quinupristin-dalfopristin and the Minimum Inhibitory Concentration (MIC ranged from 4–32 µg/ml. 89.20% of isolates were resistant to both quinupristin-dalfopristin and erythromycin of which 35.03%, 35.67% and 29.30% belonged to iMLS , cMLS and MS phenotype respectively. Conclusion: The emergence of quinupristindalfopristin resistance and MLS phenotypes brings b about the need for the simple and reliable D-test in routine diagnosis and further susceptibility testing for proper antimicrobial therapy.

Mechanisms of antibiotic resistance in Mycobacterium tuberculosis, validation of methods BACTECTM MGIT 960 and AnyplexM TII MTB / MDR / XDR Detection for detection of antibiotic resistance to first and second line in Mycobacterium tuberculosis strains

International Nuclear Information System (INIS)

Centeno Urena, Yadel

2014-01-01

A literature review is developed of drug-resistant TB in the world and in Costa Rica. The mechanisms of resistance to antibiotics are studied of the bacterium that causes tuberculosis; drug resistance to first-line and second-line, treatment regimen according to the World Health Organization and edge detection methods available in the market. The agreement between the results is studied by the phenotypic detection system of resistance of M. tuberculosis BACTEC MGIT960 and PCR, in real-time of commercial kit Anyplex II MTB/MDR/XDR, for genotypic identification of M. tuberculosis and related mutations to resistance with the referring results to thirty strains provided by the Pan American Health Organization, allowing a significant shortening in the time of obtaining reliable results. The results obtained have allowed to suggest a possible implementation at the Centro Nacional de Referencia en Micobacteriologia (CNRM), to perform antibiotic susceptibility testing and genotypic testing of multidrug cases respectively. The study results have allowed the implementation of the technology of genotypic detection of M. tuberculosis in the CNRM, obtaining for the first time in Costa Rica, information about genes of M. tuberculosis related to the generation of resistance to the major drugs of Primary treatment scheme as well as testing of resistance to second-line drug for resistant strains referred to the Centro Nacional de Referencia en Micobacteriologia in 2013. (author) [es

Performance of Molecular Approaches for Aspergillus Detection and Azole Resistance Surveillance in Cystic Fibrosis

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Hélène Guegan

2018-03-01

Full Text Available Aspergillus fumigatus triazole resistance is an emerging concern for treating chronically infected/colonized patients. This study sought to evaluate the performance of PCR assays to detect Aspergillus fungi together with azole resistance in sputum samples from cystic fibrosis (CF patients. In total, 119 sputum samples from 87 CF patients were prospectively processed for Aspergillus detection by means of mycological culture and four qPCR assays, 2 in-house methods and two commercial multiplex real-time PCR assays simultaneously detecting Aspergillus and the most relevant cyp51A gene mutations (MycoGENIE® and AsperGenius®. Azole susceptibility of A. fumigatus isolates was assessed using Etest® method and cyp51A gene mutation were characterized by sequencing. The overall rate of Aspergillus detection with the four qPCR assays ranged from 47.9 to 57.1%, contrasting with 42/119 (35.3% positive cultures with A. fumigatus. The high sensitivity of PCR on sputum could then contribute to more effective grading of Aspergillus disease in CF patients. Five out of 41 isolated strains (12.2% exhibited azole-resistant MIC patterns, three of which harbored cyp51A mutations and only 1/3 with the sequence TR34/L98H. Combined with culture, PCR assay achieved high sensitivity Aspergillus screening in CF samples. However, cyp51A targeting was only moderately effective for azole resistance monitoring, while Aspergillus resistance remains of great concern.

Performance of Molecular Approaches for Aspergillus Detection and Azole Resistance Surveillance in Cystic Fibrosis

OpenAIRE

Hélène Guegan; Sylviane Chevrier; Chantal Belleguic; Eric Deneuville; Florence Robert-Gangneux; Florence Robert-Gangneux; Jean-Pierre Gangneux; Jean-Pierre Gangneux

2018-01-01

Aspergillus fumigatus triazole resistance is an emerging concern for treating chronically infected/colonized patients. This study sought to evaluate the performance of PCR assays to detect Aspergillus fungi together with azole resistance in sputum samples from cystic fibrosis (CF) patients. In total, 119 sputum samples from 87 CF patients were prospectively processed for Aspergillus detection by means of mycological culture and four qPCR assays, 2 in-house methods and two commercial multiplex...

Monitoring of Non-Ferrous Wear Debris in Hydraulic Oil by Detecting the Equivalent Resistance of Inductive Sensors

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Lin Zeng

2018-03-01

Full Text Available Wear debris in hydraulic oil contains important information on the operation of equipment, which is important for condition monitoring and fault diagnosis in mechanical equipment. A micro inductive sensor based on the inductive coulter principle is presented in this work. It consists of a straight micro-channel and a 3-D solenoid coil wound on the micro-channel. Instead of detecting the inductance change of the inductive sensor, the equivalent resistance change of the inductive sensor is detected for non-ferrous particle (copper particle monitoring. The simulation results show that the resistance change rate caused by the presence of copper particles is greater than the inductance change rate. Copper particles with sizes ranging from 48 μm to 150 μm were used in the experiment, and the experimental results are in good agreement with the simulation results. By detecting the inductive change of the micro inductive sensor, the detection limit of the copper particles only reaches 70 μm. However, the detection limit can be improved to 48 μm by detecting the equivalent resistance of the inductive sensor. The equivalent resistance method was demonstrated to have a higher detection accuracy than conventional inductive detection methods for non-ferrous particle detection in hydraulic oil.

The in vitro anthelmintic effects of plumbagin on newly excysted and 4-weeks-old juvenile parasites of Fasciola gigantica.

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Lorsuwannarat, Natcha; Piedrafita, David; Chantree, Pathanin; Sansri, Veerawat; Songkoomkrong, Sineenart; Bantuchai, Sirasate; Sangpairot, Kant; Kueakhai, Pornanan; Changklungmoa, Narin; Chaichanasak, Pannigan; Chansela, Piyachat; Sobhon, Prasert

2014-01-01

The effect of plumbagin (PB, 5-hydroxy-2-methyl-1,4-naphthoquinone) against newly excysted juveniles (NEJs) and 4-weeks-old immature parasites of Fasciola gigantica were compared with triclabendazole (TCZ). The anthelmintic efficacy of 1, 10 and 100μg/ml of PB or TCZ following incubation in vitro for 1-24h was compared using a combination of relative motility (RM), survival index (SI) and larval migration inhibition (LMI) assays for parasite viability. The RM and SI values of the PB-treated group decreased at a more rapid rate than the TCZ-treated group. For NEJs, the decreased RM values were first observed at 1h incubation with 1μg/ml PB, and 90% of flukes were killed at 24h. In contrast, in TCZ-treated groups a 10-fold higher concentration of TCZ (10μg/ml) resulted in only 9% dead parasites after 24h incubation. In 4-weeks-old juvenile parasites, PB reduced the RM value at 10μg/ml with 100% of flukes dead after 3h, while TCZ decreased RM values at the concentration of 100μg/ml but with only 5% of flukes killed at 24h. NEJs treated with PB exhibited 88%, 99% and 100% of LMIs at the concentrations of 1, 10 and 100μg/ml, respectively. NEJs incubated with TCZ have an LMI of only 32% at the highest concentration of 100μg/ml. Similarly PB had a significantly greater killing of immature 4weeks juvenile stages than TCZ at all concentrations; however, 4-weeks-old juvenile parasites were more resistant to killing by PB or TCZ at all concentrations when compared to NEJs. Further studies were carried out to investigate the alterations of the parasite tegument by scanning electron microscope (SEM). PB caused similar tegumental alterations in 4-weeks-old juveniles as those observed in TCZ treatment but with greater damage at comparative time points, comprising of swelling, blebbing and rupture of the tegument, loss of spines, and eventual erosion, lesion and desquamation of the total tegument. These data indicate that PB had a greater fasciolicidal effect against

Use of medicinal plants to control Haemonchus contortus infection in small ruminants

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Sawleha Qadir1

Full Text Available Haemonchus contortus is singly the most important of all the gastrointestinal nematodes that constrain the survival and productivity of sheep and goats owned by rural poor farmers in the developing world. This haematophagus parasite is infamous throughout the humid tropics/subtropics, being responsible for acute disease outbreaks with high levels of mortalities, particularly in young animals. Costs associated with control of this parasite in India, have been estimated to be US$ 103 million. H. contortus is also prominent amongst the reports of anthelmintic resistance that has emerged in all countries of the world that produce small ruminants. This emergence of multiple anthelmintic resistances has provided a spur for research on alternative forms of control. Recent surveys in developing countries have identified many plants that are intended and have the potential to be used as anthelmintics. This paper reviews the use of some medicinal plants as anthelmintics against H. contortus infection in small ruminants. [Veterinary World 2010; 3(11.000: 515-518

Suspension Array for Multiplex Detection of Eight Fungicide-Resistance Related Alleles in Botrytis cinerea

OpenAIRE

Zhang, Xin; Xie, Fei; Lv, Baobei; Zhao, Pengxiang; Ma, Xuemei

2016-01-01

A simple and high-throughput assay to detect fungicide resistance is required for large-scale monitoring of the emergence of resistant strains of Botrytis cinerea. Using suspension array technology performed on a Bio-Plex 200 System, we developed a single-tube allele-specific primer extension (ASPE) assay that can simultaneously detect eight alleles in one reaction. These eight alleles include E198 and 198A of the β-Tubulin gene (BenA), H272 and 272Y of the Succinate dehydrogenase iron–sulfur...

Novel Aminoglycoside Resistance Transposons and Transposon-Derived Circular Forms Detected in Carbapenem-Resistant Acinetobacter baumannii Clinical Isolates

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Dwibedi, Chinmay Kumar; Sjöström, Karin; Edquist, Petra; Wai, Sun Nyunt; Uhlin, Bernt Eric

2016-01-01

Acinetobacter baumannii has emerged as an important opportunistic pathogen equipped with a growing number of antibiotic resistance genes. Our study investigated the molecular epidemiology and antibiotic resistance features of 28 consecutive carbapenem-resistant clinical isolates of A. baumannii collected throughout Sweden in 2012 and 2013. The isolates mainly belonged to clonal complexes (CCs) with an extensive international distribution, such as CC2 (n = 16) and CC25 (n = 7). Resistance to carbapenems was related to blaOXA-23 (20 isolates), blaOXA-24/40-like (6 isolates), blaOXA-467 (1 isolate), and ISAba1-blaOXA-69 (1 isolate). Ceftazidime resistance was associated with blaPER-7 in the CC25 isolates. Two classical point mutations were responsible for resistance to quinolones in all the isolates. Isolates with high levels of resistance to aminoglycosides carried the 16S rRNA methylase armA gene. The isolates also carried a variety of genes encoding aminoglycoside-modifying enzymes. Several novel structures involved in aminoglycoside resistance were identified, including Tn6279, ΔTn6279, Ab-ST3-aadB, and different assemblies of Tn6020 and TnaphA6. Importantly, a number of circular forms related to the IS26 or ISAba125 composite transposons were detected. The frequent occurrence of these circular forms in the populations of several isolates indicates a potential role of these circular forms in the dissemination of antibiotic resistance genes. PMID:26824943

PCR detection of oxytetracycline resistance genes from diverse habitats in total community DNA and in streptomycete isolates.

NARCIS (Netherlands)

Nikolakopoulou, T.L.; Egan, S.; Overbeek, van L.S.; Guillaume, G.; Heuer, H.; Wellington, E.M.H.; Elsas, van J.D.; Collard, J.M.; Smalla, K.; Karagouni, A.D.

2005-01-01

A range of European habitats was screened by PCR for detection of the oxytetracycline resistance genes otr(A) and otr(B), found in the oxytetracycline-producing strain Streptomyces rimosus. Primers were developed to detect these otr genes in tetracycline-resistant (TcR) streptomycete isolates from

Detection of Chloramphenicol Resistance Genes (cat in Clinical Isolates of Pseudomonas aeruginosa with Polymerase Chain Reaction Method

Directory of Open Access Journals (Sweden)

Tiana Milanda

2014-12-01

Full Text Available Pseudomonas aeruginosa is an opportunistic Gram negative bacteria, which may cause infection in eyes, ears, skin, bones, central nervous system, gastrointestinal tract, circulatory system, heart, respiratory system, and urinary tract. Recently, chloramphenicol is no longer used as the main option of the therapy due of its resistance case. The aim of this research was to detect the presence of gene which is responsible to chloramphenicol resistance in clinical isolates of P.aeruginosa. These bacteria isolated from pus of external otitis patients in Hasan Sadikin Hospital in Bandung City. Polymerase Chain Reaction (PCR method (colony-PCR and DNA-PCR were performed to detect this resistance gene. Electropherogram from PCR products showed that the chloramphenicol resistance in clinical isolates of P. aeruginosa was caused by cat gene (317 bp. Based on this research, cat gene may be used to detect the chloramphenicol resistance in patients with external ostitis.

HIGH RESOLUTION RESISTIVITY LEAK DETECTION DATA PROCESSING and EVALUATION MEHTODS and REQUIREMENTS

International Nuclear Information System (INIS)

SCHOFIELD JS

2007-01-01

This document has two purposes: (sm b ullet) Describe how data generated by High Resolution REsistivity (HRR) leak detection (LD) systems deployed during single-shell tank (SST) waste retrieval operations are processed and evaluated. (sm b ullet) Provide the basic review requirements for HRR data when Hrr is deployed as a leak detection method during SST waste retrievals

Autoradiographic detection of mutation to exotoxin-A resistance in mouse fibroblasts treated with ethyl methanesulfonate, X-rays and ultraviolet light

International Nuclear Information System (INIS)

Tiah, M.; Ronen, A.

1989-01-01

P. aeruginosa exotoxin-A (PE) blocks protein synthesis in mammalian cells by inactivating elongation factor 2 (EF-2). Toxin-resistant mutant cells can be detected autoradioraphically, in cultures grown on microscope coverslips in the presence of PE, and exposed to [ 3 H]leucine. The frequency of PE-resistant cells detected by the autoradiographic assay in non-mutagenized cells of the established mouse cell line LTKA is 9.7 j 0.6 x 10 -5 . Upon treatment with ethyl methanesulfonate (EMS), X-rays of ultraviolet (UV) light it increases in a dose-dependent fashion. The mutational nature of the resistance detected by the assay is indicated by its clonal inheritance, and by the dose-dependent increase in the frequency of resistant cells after utagenesis. On the basis of the high frequency of PE-resistant cells detected by the autoradiographic assay, and their cross-resistance to diphteria toxin (DT), the authors suggest that the PE-resistant mutants detected by the autoradiographic assay are of class II, i.e., they are altered in the structural gene for EF-2. (author). 27 refs.; 5 figs.; 3 tabs

An electrical resistivity monitor for the detection of composition changes in Pb-17Li

International Nuclear Information System (INIS)

Hubberstey, P.; Barker, M.G.; Sample, T.

1991-01-01

An electrical resistivity monitor for the detection of composition changes in the lithium-lead eutectic alloy, Pb-17Li, has been developed. A miniature electromagnetic pump is used to sample alloy continuously from a pool or loop system and force it through a capillary section, within which the necessary resistance measurements are made, prior to its return to the bulk source. To calibrate the monitor, detailed resistivity-temperature and resistivity-composition data have been determined for Pb-Li alloys at temperatures from 600 to 800K and compositions from 0 to 20.5 at% Li. The resistivity increases with both temperature and composition; for Pb-17li at 723 K, dρ/dT=0.054x10 -8 ΩmK -1 , and dρ/d[Li]=1.27x10 -8 Ωm(at% Li) -1 . The sensitivity of the monitor is such that changes in composition of as little as ±0.05 at% Li can be detected and its response time is limited soley by the rate of sampling. (orig.)

Detecting and monitoring of water inrush in tunnels and coal mines using direct current resistivity method: A review

Directory of Open Access Journals (Sweden)

Shucai Li

2015-08-01

Full Text Available Detecting, real-time monitoring and early warning of underground water-bearing structures are critically important issues in prevention and mitigation of water inrush hazards in underground engineering. Direct current (DC resistivity method is a widely used method for routine detection, advanced detection and real-time monitoring of water-bearing structures, due to its high sensitivity to groundwater. In this study, the DC resistivity method applied to underground engineering is reviewed and discussed, including the observation mode, multiple inversions, and real-time monitoring. It is shown that a priori information constrained inversion is desirable to reduce the non-uniqueness of inversion, with which the accuracy of detection can be significantly improved. The focused resistivity method is prospective for advanced detection; with this method, the flanking interference can be reduced and the detection distance is increased subsequently. The time-lapse resistivity inversion method is suitable for the regions with continuous conductivity changes, and it can be used to monitor water inrush in those regions. Based on above-mentioned features of various methods in terms of benefits and limitations, we propose a three-dimensional (3D induced polarization method characterized with multi-electrode array, and introduce it into tunnels and mines combining with real-time monitoring with time-lapse inversion and cross-hole resistivity method. At last, the prospective applications of DC resistivity method are discussed as follows: (1 available advanced detection technology and instrument in tunnel excavated by tunnel boring machine (TBM, (2 high-resolution detection method in holes, (3 four-dimensional (4D monitoring technology for water inrush sources, and (4 estimation of water volume in water-bearing structures.

Analytical and clinical performance characteristics of the Abbott RealTime MTB RIF/INH Resistance, an assay for the detection of rifampicin and isoniazid resistant Mycobacterium tuberculosis in pulmonary specimens.

Science.gov (United States)

Kostera, Joshua; Leckie, Gregor; Tang, Ning; Lampinen, John; Szostak, Magdalena; Abravaya, Klara; Wang, Hong

2016-12-01

Clinical management of drug-resistant tuberculosis patients continues to present significant challenges to global health. To tackle these challenges, the Abbott RealTime MTB RIF/INH Resistance assay was developed to accelerate the diagnosis of rifampicin and/or isoniazid resistant tuberculosis to within a day. This article summarizes the performance of the Abbott RealTime MTB RIF/INH Resistance assay; including reliability, analytical sensitivity, and clinical sensitivity/specificity as compared to Cepheid GeneXpert MTB/RIF version 1.0 and Hain MTBDRplus version 2.0. The limit of detection (LOD) of the Abbott RealTime MTB RIF/INH Resistance assay was determined to be 32 colony forming units/milliliter (cfu/mL) using the Mycobacterium tuberculosis (MTB) strain H37Rv cell line. For rifampicin resistance detection, the Abbott RealTime MTB RIF/INH Resistance assay demonstrated statistically equivalent clinical sensitivity and specificity as compared to Cepheid GeneXpert MTB/RIF. For isoniazid resistance detection, the assay demonstrated statistically equivalent clinical sensitivity and specificity as compared to Hain MTBDRplus. The performance data presented herein demonstrate that the Abbott RealTime MTB RIF/INH Resistance assay is a sensitive, robust, and reliable test for realtime simultaneous detection of first line anti-tuberculosis antibiotics rifampicin and isoniazid in patient specimens. Copyright © 2016 The Author. Published by Elsevier Ltd.. All rights reserved.

In Vitro Anthelmintic Activity of Crude Extracts of Aerial Parts of Cissus quadrangularis L. and Leaves of Schinus molle L. against Haemonchus contortus

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Selamawit Zenebe

2017-01-01

Full Text Available Background. Haemonchus contortus, the causative agent of Haemonchosis, is the most economically important parasite in small ruminant production. Control with chemotherapy has not been successful due to rapid emergence of drug-resistant strains. There is a continuous search for alternative leads particularly from plants. The study aimed to evaluate the anthelmintic activity of crude methanolic extracts of leaves of Schinus molle and aerial parts of Cissus quadrangularis against H. contortus. Methods. Adult motility test and egg hatching inhibition assay were employed to investigate the in vitro adulticidal and egg hatching inhibitory effects of the extracts. Results. Higher concentrations of the extracts (10 and 5 mg/ml had a significantly superior adulticidal activity (p<0.05 compared to the negative control and lower concentration levels, which was comparable to albendazole. Similarly, the relative egg hatch inhibition efficacy of S. molle and C. quadrangularis extracts indicated a maximum of 96% and 88% egg hatch inhibition, respectively, within the 48 hrs of exposure at 1 mg/ml. Conclusion. The current study evidenced that the crude methanolic extracts of the plants have promising adulticidal and egg hatching inhibitory effects against H. contortus.

In situ detection of tree root distribution and biomass by multi-electrode resistivity imaging.

Science.gov (United States)

Amato, Mariana; Basso, Bruno; Celano, Giuseppe; Bitella, Giovanni; Morelli, Gianfranco; Rossi, Roberta

2008-10-01

Traditional methods for studying tree roots are destructive and labor intensive, but available nondestructive techniques are applicable only to small scale studies or are strongly limited by soil conditions and root size. Soil electrical resistivity measured by geoelectrical methods has the potential to detect belowground plant structures, but quantitative relationships of these measurements with root traits have not been assessed. We tested the ability of two-dimensional (2-D) DC resistivity tomography to detect the spatial variability of roots and to quantify their biomass in a tree stand. A high-resolution resistivity tomogram was generated along a 11.75 m transect under an Alnus glutinosa (L.) Gaertn. stand based on an alpha-Wenner configuration with 48 electrodes spaced 0.25 m apart. Data were processed by a 2-D finite-element inversion algorithm, and corrected for soil temperature. Data acquisition, inversion and imaging were completed in the field within 60 min. Root dry mass per unit soil volume (root mass density, RMD) was measured destructively on soil samples collected to a depth of 1.05 m. Soil sand, silt, clay and organic matter contents, electrical conductivity, water content and pH were measured on a subset of samples. The spatial pattern of soil resistivity closely matched the spatial distribution of RMD. Multiple linear regression showed that only RMD and soil water content were related to soil resistivity along the transect. Regression analysis of RMD against soil resistivity revealed a highly significant logistic relationship (n = 97), which was confirmed on a separate dataset (n = 67), showing that soil resistivity was quantitatively related to belowground tree root biomass. This relationship provides a basis for developing quick nondestructive methods for detecting root distribution and quantifying root biomass, as well as for optimizing sampling strategies for studying root-driven phenomena.

Anthelmintic efficacy of gold nanoparticles derived from a phytopathogenic fungus, Nigrospora oryzae.

Directory of Open Access Journals (Sweden)

Pradip Kumar Kar

Full Text Available Exploring a green chemistry approach, this study brings to the fore, the anthelmintic efficacy of gold nanoparticles, highlighting the plausible usage of myconanotechnology. Gold nanoparticles of ∼6 to ∼18 nm diameter were synthesized by treating the mycelia-free culture filtrate of the phytopathogenic fungus with gold chloride. Their size and morphology were confirmed by UV-Vis spectroscopy, DLS data, AFM and TEM images. The XRD studies reveal a crystalline nature of the nanoparticles, which are in cubic phase. The FTIR spectroscopic studies before and after the formation of nanoparticles show the presence of possible functional groups responsible for the bio-reduction and capping of the synthesized gold nanoparticles. The latter were tested as vermifugal agents against a model cestode Raillietina sp., an intestinal parasite of domestic fowl. Further, ultrastructural and biochemical parameters were used to corroborate the efficacy study.

Standard method for detecting Bombyx mori nucleopolyhedrovirus disease-resistant silkworm varieties

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Yang Qiong

Full Text Available ABSTRACT Bombyx mori nucleopolyhedrovirus (BmNPV disease is one of the most serious silkworm diseases, and it has caused great economic losses to the sericulture industry. So far, the disease has not been controlled effectively by therapeutic agents. Breeding resistant silkworm varieties breeding may be an effective way to improve resistance to BmNPV and reduce economic losses. A precise resistance-detection method will help to accelerate the breeding process. For this purpose, here we described the individual inoculation method (IIM. Details of the IIM include pathogen BmNPV preparation, mulberry leaf size, pathogen volume, rearing conditions, course of infection, and breeding conditions. Finally, a resistance comparison experiment was performed using the IIM and the traditional group inoculation method (GIM. The incidence of BmNPV infection and the within-group variance results showed that the IIM was more precise and reliable than the GIM.

Performance of conventional pcr for screening for strongylus vulgaris on horse farms

OpenAIRE

Nielsen, Marianne K.; Wøhlk, Chamilla B.M.; Petersen, Stig L.; Nielsen, Martin Krarup

2010-01-01

  Strongyle parasites are ubiquitous in grazing horses. Of these, the bloodworm Strongylus vulgaris is regarded most pathogenic. Increasing levels of anthelmintic resistance in strongyle parasites has lead to recommendations of decreased treatment intensities, and there is now a pronounced need for reliable tools for detection of parasite burdens in general and S. vulgaris in particular. The only method currently available is the larval culture, which is laborious and time-consuming, so veter...

Nematóides resistentes a alguns anti-helmínticos em rebanhos caprinos no Cariri Paraibano Nematode resistant to some anthelmintics in dairy goats in Cariri Paraibano, Brazil

Directory of Open Access Journals (Sweden)

Wagner C. Lima

2010-12-01

Full Text Available Objetivou-se avaliar a resistência de nematódeos a alguns anti-helmínticos em rebanhos caprinos no Cariri Paraibano. Foram utilizados 144 animais no período seco e 120 animais no período chuvoso, fêmeas com idade acima de oito meses de vida e lactantes, distribuídos em 4 grupos: Grupo I não tratado, Grupo II tratado com albendazole a 10%, Grupo III tratado com ivermectina a 1% e Grupo IV tratado com fosfato de levamisole a 18,8%, nas doses recomendadas pelos fabricantes. Para avaliar a resistência, aplicou-se o teste de redução na contagem de ovos por grama de fezes (RCOF e o cultivo de larvas de helmintos. As amostras fecais foram coletadas no dia do tratamento (dia base e 7, 14 e 21 dias após o tratamento. No grupo tratado com Albendazole, observaram-se eficácias de 61%, 11% e 24% no período seco e de 55%, 14% e 12% no período chuvoso, aos 7, 14, e 21 dias, respectivamente. No grupo tratado com Ivermectina, a eficácia foi de 14%, 70% e 66% para o período seco, e de 76%, 34% e 71% para o período chuvoso, aos 7, 14 e 21 dias, respectivamente. O grupo tratado com fosfato de Levamisole apresentou percentuais de eficácia de 89%, 79% e 73% no período seco e de 76%, 69% e 67% no período chuvoso, aos 7, 14 e 21 dias, respectivamente. Os resultados obtidos indicam que os nematódeos gastrintestinais de alguns rebanhos caprinos no Cariri Paraibano não são sensíveis aos princípios ativos Albendazole, Levamisole e Ivermectina. Durante o período de estudo foi identificada a presença de parasitas dos gêneros Haemonchus, Trichostrongylus, Oesophagostomum e Strongyloides.The aim of this paper was to evaluate the resistance of nematodes to some anthelmintics in dairy goats in Cariri Paraibano, Brazil. A total of 144 female goats, over 8 months of age, were used in the dry season and 120 ones during the rainy season, divided into four groups: Group I untreated, Group II treated with albendazole 10%, Group III treated with ivermectin

Comparison of two long acting pre-lambing anthelmintic treatments on the productivity of ewes in low body condition.

Science.gov (United States)

Bingham, C; Hodge, A; Mariadass, B

2017-05-01

To determine if there was a benefit from treating ewes with a low body condition score (BCS) with long acting anthelmintic products pre-lambing and to compare the effects of two commonly used treatment options. The study was conducted on a single commercial hill country sheep and beef property in the central North Island of New Zealand. Mixed age twin-bearing ewes were preselected by the farmer as being in poor condition 4 weeks before the planned start of lambing, and were sequentially drafted into three equal groups identified with coloured ear tags. The negative control group (n=199) received no anthelmintic treatment; the other two groups received either a controlled release capsule (CRC) containing abamectin, albendazole, Se and Co (n=200) or a long-acting injection of moxidectin (n=200). All ewes were body condition scored (1-5 scale) and weighed at pre-lambing, docking (65 days after treatment) and at weaning (127 days after treatment). Faecal nematode egg counts (FEC) were carried out on 10 ewes from each group at these three times. Most lambs were matched to the ewe treatment groups at weaning, and weighed. At weaning the mean body weight of ewes treated with moxidectin was 3.2 (95% CI=2.3-4.3) kg heavier than controls, and of ewes treated with CRC was 3.6 (95% CI=2.5-4.5) kg heavier than control ewes (pewes had a BCS≥3. At weaning, more ewes treated with CRC (140/194; 72%) or moxidectin (122/187; 65%) had a BCS≥3 than control ewes (55/179 (31%); pewes from the treatment groups, the mean weight at weaning of lambs from ewes treated with moxidectin was 2.6 (95% CI=1.9-3.3) kg heavier, and from ewes treated with CRC was 2.6 (95% CI=1.9-3.4) kg heavier than lambs from control ewes (pewes with low BCS pre-lambing with long acting anthelmintic treatments (moxidectin long acting injection or CRC) resulted in an increase in mean body weight of the ewes and lambs at weaning. There were no significant differences between the two pre-lambing treatments used

Pyrosequencing for Rapid Detection of Mycobacterium tuberculosis Resistance to Rifampin, Isoniazid, and Fluoroquinolones ▿

Science.gov (United States)

Bravo, Lulette Tricia C.; Tuohy, Marion J.; Ang, Concepcion; Destura, Raul V.; Mendoza, Myrna; Procop, Gary W.; Gordon, Steven M.; Hall, Geraldine S.; Shrestha, Nabin K.

2009-01-01

After isoniazid and rifampin (rifampicin), the next pivotal drug class in Mycobacterium tuberculosis treatment is the fluoroquinolone class. Mutations in resistance-determining regions (RDR) of the rpoB, katG, and gyrA genes occur with frequencies of 97%, 50%, and 85% among M. tuberculosis isolates resistant to rifampin, isoniazid, and fluoroquinolones, respectively. Sequences are highly conserved, and certain mutations correlate well with phenotypic resistance. We developed a pyrosequencing assay to determine M. tuberculosis genotypic resistance to rifampin, isoniazid, and fluoroquinolones. We characterized 102 M. tuberculosis clinical isolates from the Philippines for susceptibility to rifampin, isoniazid, and ofloxacin by using the conventional submerged-disk proportion method and validated our pyrosequencing assay using these isolates. DNA was extracted and amplified by using PCR primers directed toward the RDR of the rpoB, katG, and gyrA genes, and pyrosequencing was performed on the extracts. The M. tuberculosis H37Rv strain (ATCC 25618) was used as the reference strain. The sensitivities and specificities of pyrosequencing were 96.7% and 97.3%, 63.8% and 100%, and 70.0% and 100% for the detection of resistance to rifampin, isoniazid, and ofloxacin, respectively. Pyrosequencing is thus a rapid and accurate method for detecting M. tuberculosis resistance to these three drugs. PMID:19846642

Evaluation of Eight Different Cephalosporins for Detection of Cephalosporin Resistance in Salmonella enterica and Escherichia coli

DEFF Research Database (Denmark)

Aarestrup, Frank Møller; Hasman, Henrik; Veldman, K

2010-01-01

This study evaluates the efficacy of eight different cephalosporins for detection of cephalosporin resistance mediated by extended spectrum beta-lactamases (ESBL) and plasmidic AmpC beta-lactamases in Salmonella and Escherichia coli. A total of 138 E. coli and 86 Salmonella isolates with known beta......-resistant but cephalosporin-susceptible, 56 ESBL isolates and 19 isolates with plasmidic AmpC, as well as 10 ampC hyper-producing E. coli. The minimum inhibitory concentration distributions and zone inhibitions varied with the tested compound. Ampicillin-resistant isolates showed reduced susceptibility to the cephalosporins...... compared to ampicillin-susceptible isolates. Cefoperazone, cefquinome, and cefuroxime were not useful in detecting isolates with ESBL or plasmidic AmpC. The best substances for detection were cefotaxime, cefpodoxime, and ceftriaxone, whereas ceftazidime and ceftiofur were not as efficient. Ceftriaxone may...

Performance of PCR-reverse blot hybridization assay for detection of rifampicin-resistant Mycobacterium leprae.

Science.gov (United States)

Wang, Hye-young; Kim, Hyunjung; Kim, Yeun; Bang, Hyeeun; Kim, Jong-Pill; Hwang, Joo Hwan; Cho, Sang-Nae; Kim, Tae Ue; Lee, Hyeyoung

2015-10-01

Drug resistance in Mycobacterium leprae is a significant problem in countries where leprosy is endemic. A sensitive, specific, and high-throughput reverse blot hybridization assay (REBA) for the detection of genotypic resistance to rifampicin (RIF) was designed and evaluated. It has been shown that resistance to RIF in M. leprae involves mutations in the rpoB gene encoding the -subunit of the RNA polymerase. The PCR-REBA simultaneously detects both 6 wild-type regions and 5 different mutations (507 AGC, 513 GTG, 516 TAT, 531 ATG, and 531 TTC) including the most prevalent mutations at positions 507 and 531. Thirty-one clinical isolates provided by Korea Institute of Hansen-s Disease were analyzed by PCR-REBA with RIF resistance of rpoB gene. As a result, missense mutations at codons 507 AGC and 531 ATG with 2-nucleotide substitutions were found in one sample, and a missense mutation at codon 516 TAT and ΔWT6 (deletion of 530-534) was found in another sample. These cases were confirmed by DNA sequence analysis. This rapid, simple, and highly sensitive assay provides a practical alternative to sequencing for genotypic evaluation of RIF resistance in M. leprae.

Impact of manure fertilization on the abundance of antibiotic-resistant bacteria and frequency of detection of antibiotic resistance genes in soil and on vegetables at harvest.

Science.gov (United States)

Marti, Romain; Scott, Andrew; Tien, Yuan-Ching; Murray, Roger; Sabourin, Lyne; Zhang, Yun; Topp, Edward

2013-09-01

Consumption of vegetables represents a route of direct human exposure to bacteria found in soil. The present study evaluated the complement of bacteria resistant to various antibiotics on vegetables often eaten raw (tomato, cucumber, pepper, carrot, radish, lettuce) and how this might vary with growth in soil fertilized inorganically or with dairy or swine manure. Vegetables were sown into field plots immediately following fertilization and harvested when of marketable quality. Vegetable and soil samples were evaluated for viable antibiotic-resistant bacteria by plate count on Chromocult medium supplemented with antibiotics at clinical breakpoint concentrations. DNA was extracted from soil and vegetables and evaluated by PCR for the presence of 46 gene targets associated with plasmid incompatibility groups, integrons, or antibiotic resistance genes. Soil receiving manure was enriched in antibiotic-resistant bacteria and various antibiotic resistance determinants. There was no coherent corresponding increase in the abundance of antibiotic-resistant bacteria enumerated from any vegetable grown in manure-fertilized soil. Numerous antibiotic resistance determinants were detected in DNA extracted from vegetables grown in unmanured soil. A smaller number of determinants were additionally detected on vegetables grown only in manured and not in unmanured soil. Overall, consumption of raw vegetables represents a route of human exposure to antibiotic-resistant bacteria and resistance determinants naturally present in soil. However, the detection of some determinants on vegetables grown only in freshly manured soil reinforces the advisability of pretreating manure through composting or other stabilization processes or mandating offset times between manuring and harvesting vegetables for human consumption.

Utility of capsule endoscopy for evaluating anthelmintic efficacy in fully conscious dogs.

Science.gov (United States)

Lee, Alice C Y; Epe, Christian; Simpson, Kenneth W; Bowman, Dwight D

2011-11-01

The current accepted standard for evaluating the efficacy of gastrointestinal anthelmintic drugs is necropsy of infected animals followed by a comparison of worm counts between treated and non-treated groups. In this study capsule endoscopy, a minimally invasive method of imaging the small intestine of humans, is evaluated as a possible alternative to necropsy for the purposes of worm quantification in dogs. Eighteen Beagle dogs were included in this study. These dogs were part of a separate trial intended to determine the efficacy of various candidate parasiticides against Ancylostoma caninum via the necropsy standard. Dogs were inoculated with A. caninum L3s 4 weeks prior to treatment with one of the candidate compounds; a control group (n=8) received no treatment. Capsule endoscopy was performed 6-14 days post-treatment, followed by necropsy the following day. Seventeen dogs had complete examinations, i.e. the capsule traversed the small intestine and reached the colon within the battery life of the capsule. A strong correlation (r(s)=0.87, Pcapsule endoscopy and necropsy. There was no clear relationship between the ability of the capsule endoscope to detect hookworms and either visibility of the intestinal lumen or small intestinal transit time. Generation of a virtual spatial record of hookworm location from the capsule endoscopy data revealed a temporal trend, with the majority of worms present in the proximal small intestine in the morning versus the central to distal small intestine in the afternoon. Worm distribution as determined by capsule endoscopy closely resembled post-mortem findings. In conclusion, capsule endoscopy shows promise as an alternative to necropsy for the enumeration of A. caninum in the canine small intestine, although further work is required to improve completion rates and optimise intestinal examination. Copyright © 2011 Australian Society for Parasitology Inc. Published by Elsevier Ltd. All rights reserved.

A novel automatic molecular test for detection of multidrug resistance tuberculosis in sputum specimen: A case control study.

Science.gov (United States)

Li, Qiang; Ou, Xi C; Pang, Yu; Xia, Hui; Huang, Hai R; Zhao, Bing; Wang, Sheng F; Zhao, Yan L

2017-07-01

MiniLab tuberculosis (ML TB) assay is a new automatic diagnostic tool for diagnosis of multidrug resistance tuberculosis (MDR-TB). This study was conducted with aims to know the performance of this assay. Sputum sample from 224 TB suspects was collected from tuberculosis suspects seeking medical care at Beijing Chest hospital. The sputum samples were directly used for smear and ML TB test. The left sputum sample was used to conduct Xpert MTB/RIF, Bactec MGIT culture and drug susceptibility test (DST). All discrepancies between the results from DST, molecular and phenotypic methods were confirmed by DNA Sequencing. The sensitivity and specificity of ML TB test for detecting MTBC from TB suspects were 95.1% and 88.9%, respectively. The sensitivity for smear negative TB suspects was 64.3%. For detection of RIF resistance, the sensitivity and specificity of ML TB test were 89.2% and 95.7%, respectively. For detection of INH resistance, the sensitivity and specificity of ML TB test were 78.3% and 98.1%, respectively. ML TB test showed similar performance to Xpert MTB/RIF for detection of MTBC and RIF resistance. In addition, ML TB also had good performance for INH resistance detection. Copyright © 2017. Published by Elsevier Ltd.

Simplified Paper Format for Detecting HIV Drug Resistance in Clinical Specimens by Oligonucleotide Ligation

Science.gov (United States)

Panpradist, Nuttada; Beck, Ingrid A.; Chung, Michael H.; Kiarie, James N.; Frenkel, Lisa M.; Lutz, Barry R.

2016-01-01

Human immunodeficiency virus (HIV) is a chronic infection that can be managed by antiretroviral treatment (ART). However, periods of suboptimal viral suppression during lifelong ART can select for HIV drug resistant (DR) variants. Transmission of drug resistant virus can lessen or abrogate ART efficacy. Therefore, testing of individuals for drug resistance prior to initiation of treatment is recommended to ensure effective ART. Sensitive and inexpensive HIV genotyping methods are needed in low-resource settings where most HIV infections occur. The oligonucleotide ligation assay (OLA) is a sensitive point mutation assay for detection of drug resistance mutations in HIV pol. The current OLA involves four main steps from sample to analysis: (1) lysis and/or nucleic acid extraction, (2) amplification of HIV RNA or DNA, (3) ligation of oligonucleotide probes designed to detect single nucleotide mutations that confer HIV drug resistance, and (4) analysis via oligonucleotide surface capture, denaturation, and detection (CDD). The relative complexity of these steps has limited its adoption in resource-limited laboratories. Here we describe a simplification of the 2.5-hour plate-format CDD to a 45-minute paper-format CDD that eliminates the need for a plate reader. Analysis of mutations at four HIV-1 DR codons (K103N, Y181C, M184V, and G190A) in 26 blood specimens showed a strong correlation of the ratios of mutant signal to total signal between the paper CDD and the plate CDD. The assay described makes the OLA easier to perform in low resource laboratories. PMID:26751207

Analysis of Constraints of Chemoprophylactic Control of Livestock Helminthosis in Kenya

International Nuclear Information System (INIS)

Wanyangu, S.W; Rugut, M.K; Nginyi, J.M; Onyango-Abuye, J.A; Mugambi, J.M; Bain, A.M; Monteiro, F; Jackson, Mackellar F

1999-01-01

Three studies were carried out to find out how livestock anthelmintics are used in Kenya. Using questioners in the first study, 342 farms were surveyed. In the second study a survey on anthelmintic resistance was carried out in 42 farms comprising of small and large scale concerns. The third study was carried out on 9 commonly used anthelmintic brands sold in Kenya. These were brought from agrochemical shops and pharmacies with the aim of determining their pharmaceutical quality. The results indicated that, farmers were ignorant of good anthelmintic practice. They either under or overdose their livestock. Few farmers recognized the value of drenching suckling or weaned animals. A prevalence of 75% of anthelmintic resistance was recorded. This was mainly against levamisoles and benzimidazoles in sheep and goats. Resistance was associated with increased dosing rates on both smallholder and large-scale farms. An analysis of the pharmaceutical quality of the drugs showed that there were some substandard drugs available on the market. Four out of seven brand names claiming to contain levamisole had the concentration at a much lower level than claimed on the label. Indeed two products did not contain any trace of levamisole. One product supposed to contain mebendazole had a drug at a substandard level. Two products supposed to contain oxyclosanide had the drug concentration at a satisfactory level although one had the concentration ata much higher level. The concentration of levamisole in two substandard drugs varied significantly between different batch products. These varied from 0% to 73.6%-85.4% of the concentration shown on the labels

Targeted next generation sequencing for the detection of ciprofloxacin resistance markers using molecular inversion probes

Science.gov (United States)

2016-07-06

ecological studies have shown development of antibiotic resistance in bacterial pathogens caused by increased antibiotic usage in animals , food, and...et al. Danish Integrated Antimicrobial Resistance Monitoring and Research Program. Emerging Infectious Diseases 13, 1633-1639, doi:10.3201...F. J. Molecular detection of antimicrobial resistance . Clin. Microbiol. Rev. 14, 836-871, table of contents, doi:10.1128/CMR.14.4.836-871.2001

[Application value of Xpert MTB/RIF in diagnosis of spinal tuberculosis and detection of rifampin resistance].

Science.gov (United States)

Jin, Yang-Hui; Shi, Shi-Yuan; Zheng, Qi; Shen, Jian; Ying, Xiao-Zhang; Wang, Yi-Fan

2017-09-25

To investigate the application value of Xpert MTB/RIF in diagnosis of spinal tuberculosis and detection of rifampin resistance. The 109 pus specimens were obtained from patients who were primaryly diagnosed as spinal tuberculosis. All of the pus specimens were detected by acid-fast stain, liquid fast culturing by BACTEC MGIT 960 and Xpert MTB/RIF assay to definite the differences in sensitivity and specificity of mycobacterium tuberculosis among detecting methods. Pus specimens obtained by different methods were deteceded by MTB/RIF test to analyze the self-influence on Xpert MTB/RIF test. The result of liquid fast culturing by BACTEC MGIT 960 was used as the gold standard; and the value of Xpert MTB/RIF assay in detecting rifampin resistance was analyzed. The sensitivity of acid-fast stain, liquid fast culturing by BACTEC MGIT 960 and Xpert MTB/RIF assay were 25.92%, 48.15%, 77.78%, respectively. The sensitivity of pus specimens obtained from open surgery, ultrasound positioning puncture and biopsy the sensitivity were 83.78%, 76.47%, 44.68% respectively deteceded by MTB/RIF test. According to the gold standard of the results of liquid fast culturing by BACTEC MGIT 960 assay, the sensitivity and specificity of Xpert MTB/RIF assay in detecting rifampin resistance were 80%(4/5) and 90.70%(39/43), respectively. Xpert MTB/RIF assay has higher value in diagnosis of spinal tuberculosi, and also can detect rifampin resistance. The number of mycobacterium tuberculosis in pus specimens has a great influence in the sensitivity of Xpert MTB/RIF assay.

Xpert MTB/RIF Ultra for detection of Mycobacterium tuberculosis and rifampicin resistance: a prospective multicentre diagnostic accuracy study.

Science.gov (United States)

Dorman, Susan E; Schumacher, Samuel G; Alland, David; Nabeta, Pamela; Armstrong, Derek T; King, Bonnie; Hall, Sandra L; Chakravorty, Soumitesh; Cirillo, Daniela M; Tukvadze, Nestani; Bablishvili, Nino; Stevens, Wendy; Scott, Lesley; Rodrigues, Camilla; Kazi, Mubin I; Joloba, Moses; Nakiyingi, Lydia; Nicol, Mark P; Ghebrekristos, Yonas; Anyango, Irene; Murithi, Wilfred; Dietze, Reynaldo; Lyrio Peres, Renata; Skrahina, Alena; Auchynka, Vera; Chopra, Kamal Kishore; Hanif, Mahmud; Liu, Xin; Yuan, Xing; Boehme, Catharina C; Ellner, Jerrold J; Denkinger, Claudia M

2018-01-01

The Xpert MTB/RIF assay is an automated molecular test that has improved the detection of tuberculosis and rifampicin resistance, but its sensitivity is inadequate in patients with paucibacillary disease or HIV. Xpert MTB/RIF Ultra (Xpert Ultra) was developed to overcome this limitation. We compared the diagnostic performance of Xpert Ultra with that of Xpert for detection of tuberculosis and rifampicin resistance. In this prospective, multicentre, diagnostic accuracy study, we recruited adults with pulmonary tuberculosis symptoms presenting at primary health-care centres and hospitals in eight countries (South Africa, Uganda, Kenya, India, China, Georgia, Belarus, and Brazil). Participants were allocated to the case detection group if no drugs had been taken for tuberculosis in the past 6 months or to the multidrug-resistance risk group if drugs for tuberculosis had been taken in the past 6 months, but drug resistance was suspected. Demographic information, medical history, chest imaging results, and HIV test results were recorded at enrolment, and each participant gave at least three sputum specimen on 2 separate days. Xpert and Xpert Ultra diagnostic performance in the same sputum specimen was compared with culture tests and drug susceptibility testing as reference standards. The primary objectives were to estimate and compare the sensitivity of Xpert Ultra test with that of Xpert for detection of smear-negative tuberculosis and rifampicin resistance and to estimate and compare Xpert Ultra and Xpert specificities for detection of rifampicin resistance. Study participants in the case detection group were included in all analyses, whereas participants in the multidrug-resistance risk group were only included in analyses of rifampicin-resistance detection. Between Feb 18, and Dec 24, 2016, we enrolled 2368 participants for sputum sampling. 248 participants were excluded from the analysis, and 1753 participants were distributed to the case detection group (n=1439

Efficacy of the amino-acetonitrile derivative, monepantel, against experimental and natural adult stage gastro-intestinal nematode infections in sheep.

Science.gov (United States)

Sager, Heinz; Hosking, Barry; Bapst, Béatrice; Stein, Philip; Vanhoff, Kathleen; Kaminsky, Ronald

2009-01-22

Multiple drug resistance by nematodes, against anthelmintics has become an important economic problem in sheep farming worldwide. Here we describe the efficacy of monepantel, a developmental molecule from the recently discovered anthelmintic class, the amino-acetonitrile derivatives (AADs). Efficacy was tested against adult stage gastro-intestinal nematodes (GINs) in experimentally and naturally infected sheep at a dose of 2.5mg/kg body weight when administered as an oral solution. Some of the isolates used in experimental infection studies were known to be resistant to the benzimidazoles or levamisole anthelmintics; strains resistant to the macrocyclic lactones were not available for these tests. Worm count-based efficacies of >98% were determined in these studies. As an exception, Oesophagostomum venulosum was only reduced by 88% in one study, albeit with a low worm burden in the untreated controls (geometric mean 15.4 worms). Similar efficacies for monepantel were also confirmed in naturally infected sheep. While the efficacy against most species was >99%, the least susceptible species was identified as Nematodirus spathiger, and although efficacy was 92.4% in one study it was generally >99%. Several animals were infected with Trichuris ovis, which was not eliminated after the treatment. Monepantel demonstrated high activity against a broad range of the important GINs of sheep, which makes this molecule an interesting candidate for use in this species, particularly in regions with problems of anthelmintic resistance. Monepantel was well tolerated by the treated sheep, with no treatment related adverse events documented.

Comparative efficacy of different anthelmintics against fenbendazole-resistant nematodes of pashmina goats.

Science.gov (United States)

Ram, H; Rasool, T J; Sharma, A K; Meena, H R; Singh, S K

2007-08-01

A trial using albendazole, albendazole plus rafoxanide combination, ivermectin and doramectin was conducted in Pashmina goats having history of fenbendazole resistance to Haemonchus spp. and maintained at high altitude (>2350 m above sea level). Day 0 infection level was variable in different groups of animals and their larval cultures indicated Haemonchus, Trichostrongylus, Ostertagia and Oesophagostomum spp. infection, in addition to Nematodirus spp. as observed in egg counts. Efficacy of drugs was calculated on day 14 post treatment by faecal egg count reduction test (FECRT). Albendazole was least effective (14%) followed by its combination with rafoxanide (54%). However, ivermectin and doramectin were 96% and 94% effective against gastrointestinal nematodes of Pashmina goats. It was concluded that use of albendazole and its combination with rafoxanide are ineffective in controlling the nematodes of goats at this farm; hence, future use must be avoided. However, regular monitoring of the efficacy of ivermectin and doramectin is needed.

Detecting insulin resistance in polycystic ovary syndrome: purposes and pitfalls.

Science.gov (United States)

Legro, Richard S; Castracane, V Daniel; Kauffman, Robert P

2004-02-01

Approximately 50% to 70% of all women with polycystic ovary syndrome (PCOS) have some degree of insulin resistance, and this hormone insensitivity probably contributes to the hyperandrogenism that is responsible for the signs and symptoms of PCOS. Although uncertainty exists, early detection and treatment of insulin resistance in this population could ultimately reduce the incidence or severity of diabetes mellitus, dyslipidemia, hypertension, and cardiovascular disease. Even if that proves to be the case, there are still several problems with our current approach to insulin sensitivity assessment in PCOS, including the apparent lack of consensus on what defines PCOS and "normal" insulin sensitivity, ethnic and genetic variability, the presence of other factors contributing to insulin resistance such as obesity, stress, and aging, and concern about whether simplified models of insulin sensitivity have the precision to predict treatment needs, responses, and future morbidity. Although the hyperinsulinemic-euglycemic clamp technique is the gold standard for measuring insulin sensitivity, it is too expensive, time-consuming, and labor-intensive to be of practical use in an office setting. Homeostatic measurements (fasting glucose/insulin ratio or homeostatic model assessment [HOMA] value) and minimal model tests (particularly the oral glucose tolerance test [OGTT]) represent the easiest office-based assessments of insulin resistance in the PCOS patient. The OGTT is probably the best simple, office-based method to assess women with PCOS because it provides information about both insulin resistance and glucose intolerance. The diagnosis of glucose intolerance holds greater prognostic and treatment implications. All obese women with PCOS should be screened for the presence of insulin resistance by looking for other stigmata of the insulin resistance syndrome such as hypertension, dyslipidemia, central obesity, and glucose intolerance.

Detection of Antibiotic Resistant Staphylococcus aureus from Milk: A Public Health Implication

Science.gov (United States)

Akindolire, Muyiwa Ajoke; Babalola, Olubukola Oluranti; Ateba, Collins Njie

2015-01-01

The aim of this study was to investigate the occurrence, antibiotic susceptibility profiles, and virulence genes determinants of S. aureus isolated from milk obtained from retail outlets of the North-West Province, South Africa. To achieve this, 200 samples of raw, bulk and pasteurised milk were obtained randomly from supermarkets, shops and some farms in the North-West Province between May 2012 and April 2013. S. aureus was isolated and positively identified using morphological (Gram staining), biochemical (DNase, catalase, haemolysis and rapid slide agglutination) tests, protein profile analysis (MALDI-TOF mass spectrometry) and molecular (nuc specific PCR) methods. The antimicrobial resistance profiles of the isolates were determined using the phenotypic agar diffusion method. Genes encoding enterotoxins, exfoliative toxins and collagen adhesins were also screened using PCR. Among all the samples examined, 30 of 40 raw milk samples (75%), 25 of 85 bulk milk samples (29%) and 10 of 75 pasteurised milk samples (13%) were positive for S. aureus. One hundred and fifty-six PCR-confirmed S. aureus isolates were obtained from 75 contaminated milk samples. A large proportion (60%–100%) of the isolates was resistant to penicillin G, ampicillin, oxacillin, vancomycin, teicoplanin and erythromycin. On the contrary, low level resistance (8.3%–40%) was observed for gentamicin, kanamycin and sulphamethoxazole. Methicillin resistance was detected in 59% of the multidrug resistant isolates and this was a cause for concern. However, only a small proportion (20.6%) of these isolates possessed PBP2a which codes for Methicillin resistance in S. aureus. In addition, 32.7% of isolates possessed the sec gene whereas the sea, seb sed, see, cna, eta, etb genes were not detected. The findings of this study showed that raw, bulk and pasteurised milk in the North-West Province is contaminated with toxigenic and multi-drug resistant S. aureus strains. There is a need to implement

Comparison of Molecular and Phenotypic Methods for the Detection and Characterization of Carbapenem Resistant Enterobacteriaceae.

Science.gov (United States)

Somily, Ali M; Garaween, Ghada A; Abukhalid, Norah; Absar, Muhammad M; Senok, Abiola C

2016-03-01

In recent years, there has been a rapid dissemination of carbapenem resistant Enterobacteriaceae (CRE). This study aimed to compare phenotypic and molecular methods for detection and characterization of CRE isolates at a large tertiary care hospital in Saudi Arabia. This study was carried out between January 2011 and November 2013 at the King Khalid University Hospital (KKUH) in Saudi Arabia. Determination of presence of extended-spectrum beta-lactamases (ESBL) and carbapenem resistance was in accordance with Clinical and Laboratory Standards Institute (CLSI) guidelines. Phenotypic classification was done by the MASTDISCS(TM) ID inhibitor combination disk method. Genotypic characterization of ESBL and carbapenemase genes was performed by the Check-MDR CT102. Diversilab rep-PCR was used for the determination of clonal relationship. Of the 883 ESBL-positive Enterobacteriaceae detected during the study period, 14 (1.6%) isolates were carbapenem resistant. Both the molecular genotypic characterization and phenotypic testing were in agreement in the detection of all 8 metalo-beta-lactamases (MBL) producing isolates. Of these 8 MBL-producers, 5 were positive for blaNDM gene and 3 were positive for blaVIM gene. Molecular method identified additional blaOXA gene isolates while MASTDISCS(TM) ID detected one AmpC producer isolate. Both methods agreed in identifying 2 carbapenem resistant isolates which were negative for carbapenemase genes. Diversilab rep-PCR analysis of the 9 Klebsiella pneumoniae isolates revealed polyclonal distribution into eight clusters. MASTDISCS(TM) ID is a reliable simple cheap phenotypic method for detection of majority of carbapenemase genes with the exception of the blaOXA gene. We recommend to use such method in the clinical laboratory.

Geo electrical Resistivity Survey for Ancient Tunnel Detection at Bukit Tenggek, Setiu, Terengganu

International Nuclear Information System (INIS)

Siti Nazira Masrom; Mohd Hariri Arifin; Abd Rahim Harun; Abdul Rahim Samsudin

2011-01-01

Geo electrical resistivity survey was conducted in the Bukit Tenggek, Setiu, Terengganu to detect the possible existence of an ancient tunnel which is believed to be in the area. Geo electrical resistivity method was found very effective in searching for archaeological exploration and underground structures (tunnels and artifacts). Geo electrical resistivity survey was carried out using Terrameter ABEM SAS1000 and Wenner array electrode configuration. The survey area is located in a damp valley with a stream across the region. 2-D resistivity image showed the existence of anomalies in several areas that can be associated with the structure. Low resistivity value represents the estimated existence of the old tunnel, while isolated rounded anomalies are believed to be associated with barrels/artifacts. 3-D resistivity profiles, shows anomalies that may be caused by the existence of a horizontal and two vertical tunnels (shaft). However, the drillings work need to be done to figure out the exact cause of these anomalies. (author)

Au-nanoprobes for detection of SNPs associated with antibiotic resistance in Mycobacterium tuberculosis

International Nuclear Information System (INIS)

Veigas, Bruno; Baptista, Pedro V; Machado, Diana; Couto, Isabel; Viveiros, Miguel; Perdigao, Joao; Portugal, Isabel

2010-01-01

Tuberculosis (TB) is one of the leading causes of infection in humans, causing high morbility and mortality all over the world. The rate of new cases of multidrug resistant tuberculosis (MDRTB) continues to increase, and since these infections are very difficult to manage, they constitute a serious health problem. In most cases, drug resistance in Mycobacterium tuberculosis has been related to mutations in several loci within the pathogen's genome. The development of fast, cheap and simple screening methodologies would be of paramount relevance for the early detection of these mutations, essential for the timely and effective diagnosis and management of MDRTB patients. The use of gold nanoparticles derivatized with thiol-modified oligonucleotides (Au-nanoprobes) has led to new approaches in molecular diagnostics. Based on the differential non-cross-linking aggregation of Au-nanoprobes, we were able to develop a colorimetric method for the detection of specific sequences and to apply this approach to pathogen identification and single base mutations/single nucleotide polymorphisms (SNP) discrimination. Here we report on the development of Au-nanoprobes for the specific identification of SNPs within the beta subunit of the RNA polymerase (rpoB locus), responsible for resistance to rifampicin in over 95% of rifampicin resistant M. tuberculosis strains.

Au-nanoprobes for detection of SNPs associated with antibiotic resistance in Mycobacterium tuberculosis

Energy Technology Data Exchange (ETDEWEB)

Veigas, Bruno; Baptista, Pedro V [CIGMH, Departamento de Ciencias da Vida, Faculdade de Ciencias e Tecnologia, Universidade Nova de Lisboa, Caparica (Portugal); Machado, Diana; Couto, Isabel; Viveiros, Miguel [Unidade de Micobacterias, Instituto de Higiene e Medicina Tropical, Universidade Nova de Lisboa (IHMT/UNL) (Portugal); Perdigao, Joao; Portugal, Isabel, E-mail: pmvb@fct.unl.pt [Centro de Patogenese Molecular/URIA, Faculdade de Farmacia, Universidade de Lisboa, Lisboa (Portugal)

2010-10-15

Tuberculosis (TB) is one of the leading causes of infection in humans, causing high morbility and mortality all over the world. The rate of new cases of multidrug resistant tuberculosis (MDRTB) continues to increase, and since these infections are very difficult to manage, they constitute a serious health problem. In most cases, drug resistance in Mycobacterium tuberculosis has been related to mutations in several loci within the pathogen's genome. The development of fast, cheap and simple screening methodologies would be of paramount relevance for the early detection of these mutations, essential for the timely and effective diagnosis and management of MDRTB patients. The use of gold nanoparticles derivatized with thiol-modified oligonucleotides (Au-nanoprobes) has led to new approaches in molecular diagnostics. Based on the differential non-cross-linking aggregation of Au-nanoprobes, we were able to develop a colorimetric method for the detection of specific sequences and to apply this approach to pathogen identification and single base mutations/single nucleotide polymorphisms (SNP) discrimination. Here we report on the development of Au-nanoprobes for the specific identification of SNPs within the beta subunit of the RNA polymerase (rpoB locus), responsible for resistance to rifampicin in over 95% of rifampicin resistant M. tuberculosis strains.

Flow cytometry as a rapid test for detection of penicillin resistance directly in bacterial cells in Enterococcus faecalis and Staphylococcus aureus.

Science.gov (United States)

Jarzembowski, T; Wiśniewska, K; Józwik, A; Bryl, E; Witkowski, J

2008-08-01

We studied the usefulness of flow cytometry for detection of penicillin resistance in E. faecalis and S. aureus by direct binding of commercially available fluorescent penicillin, Bocillin FL, to cells obtained from culture. There were significantly lower percentages of fluorescent cells and median and mean fluorescence values per particle in penicillin-resistant than in penicillin-sensitive strains of both species observed. The method allows rapid detection of penicillin resistance in S. aureus and E. faecalis. The results encourage further investigations on the detection of antibiotic resistance in bacteria using flow cytometry.

An introductory survey of helminth control practices in South Africa and anthelmintic resistance on Thoroughbred stud farms in the Western Cape Province

Directory of Open Access Journals (Sweden)

S. Matthee

2002-07-01

Full Text Available Fifty-one per cent of 110 questionnaires, designed for obtaining information on helminth control practices and management on Thoroughbred stud farms in South Africa, were completed by farmers during 2000. The number of horses per farm included in the questionnaire survey ranged from 15 to 410. Foals, yearlings and adult horses were treated with anthelmintics at a mean of 7.3+ / -3.0, 6.6+ / -2.7 and 5.3+ / -2.3 times per year, respectively. An average of 3.4 different drugs were used annually, with ivermectin being used by most farmers during 1997-2000. On 43% of farms the weights of horses were estimated by weigh band and 45% of farmers estimated visually, while both were used on 7% of farms and scales on the remaining 5%. Doses were based on average group weight on 50% of the farms and on individual weights on 46%. Forty-three per cent of farmers performed faecal egg count reduction tests (FECRT. Most farmers rotated horses between pastures and treated new horses at introduction. Faecal removal was practiced on 61% of farms and less than 50% of farmers used alternate grazing with ruminants. Faecal egg count reduction tests were done on 283 horses, using oxibendazole, ivermectin and moxidectin on 10, 9 and 5 farms, respectively, in the Western Cape Province during 2001. While the efficacy of oxibendazole was estimated by FECRT to range from 0-88% and moxidectin from 99-100%, ivermectin resulted in a 100% reduction in egg counts. Only cyathostome larvae were recovered from post-treatment faecal cultures.

Detection of Panton-Valentine Leukocidin DNA from methicillin-resistant Staphylococcus aureus by resistive pulse sensing and loop-mediated isothermal amplification with gold nanoparticles

International Nuclear Information System (INIS)

Yang, Alice Kar Lai; Lu, Haifei; Wu, Shu Yuen; Kwok, Ho Chin; Ho, Ho Pui; Yu, Samuel; Cheung, Anthony Ka Lun; Kong, Siu Kai

2013-01-01

Graphical abstract: -- Highlights: •A novel diagnostic assay is developed to detect the MRSA's Panton-Valentine Leukocidin toxin. •Detection is based on target DNA amplification at one single temperature at 65 °C by LAMP. •Amplicons are then hybridized with 2 Au-nanoparticles with specific DNA probes for sensing. •The supra-assemblies are subsequently sensed by resistive pulse sensing. •Detection limit: ∼200 copies of DNA; time for detection: completed within 2 h. -- Abstract: This report describes a novel diagnostic assay for rapid detection of the Panton-Valentine Leukocidin (PVL) toxin of methicillin-resistant Staphylococcus aureus (MRSA) utilizing resistive pulse sensing (RPS), loop-mediated isothermal DNA amplification (LAMP) in combination with gold nanoparticles (AuNPs). The PVL DNA from MRSA was specifically amplified by LAMP using four primers at one temperature (65 °C). The DNA products with biotin were then conjugated to a first AuNP1 (55 ± 2 nm) through biotin–avidin binding. A second AuNP2 (30 ± 1.5 nm) coated with a specific DNA probe hybridized with the LAMP DNA products at the loop region to enhance assay sensitivity and specificity, to generate supra-AuNP1-DNA-AuNP2 assemblies. Scanning electron microscopy confirmed the presence of these supra-assemblies. Using RPS, detection and quantitation of the agglomerated AuNPs were performed by a tunable fluidic nanopore sensor. The results demonstrate that the LAMP-based RPS sensor is sensitive and rapid for detecting the PVL DNA. This technique could achieve a limit of detection (LOD) up to about 500 copies of genomic DNA from the bacteria MRSA MW2 and the detection can be completed within two hours with a straightforward signal-to-readout setup. It is anticipated that this LAMP-based AuNP RPS may become an effective tool for MRSA detection and a potential platform in clinical laboratory to report the presence or absence of other types of infectious agents

Development of an in vitro bioassay for measuring susceptibility to macrocyclic lactone anthelmintics in Dirofilaria immitis.

Science.gov (United States)

Evans, Christopher C; Moorhead, Andrew R; Storey, Bobby E; Wolstenholme, Adrian J; Kaplan, Ray M

2013-12-01

For more than 20 years, anthelmintics of the macrocyclic lactone (ML) drug class have been widely and effectively used as preventives against the canine heartworm, Dirofilaria immitis. However, in recent years an increased number of lack of efficacy (LOE) cases are being reported, in which dogs develop mature heartworm infections despite receiving monthly prophylactic doses of ML drugs. While this situation is raising concerns that heartworms may be developing resistance to MLs, compelling evidence for this is still lacking. Resolution of this dilemma requires validated biological or molecular diagnostic assays, but, unfortunately, no such tests currently exist. To address this need, we developed and optimized a larval migration inhibition assay (LMIA) for use with D. immitis third-stage larvae. The LMIA was used to measure the in vitro dose-response of two ML drugs (ivermectin and eprinomectin) on a known ML-susceptible laboratory strain of D. immitis. A nonlinear regression model was fit to the dose-response data, from which IC50 values were calculated; the mean IC50 and 95% confidence interval for IVM was 4.56 μM (1.26-16.4 μM), greater than that for EPR at 2.02 μM (1.68-2.42 μM), and this difference was significant (p = 0.0428). The R (2) value for EPR assays (0.90) was also greater than that for IVM treatment (0.71). The consistency and reproducibility of the dose-response data obtained with this assay suggests that it may be a useful technique for investigating the relative susceptibilities to ML drugs in other D. immitis populations.

Impact of chemical structure of flavanol monomers and condensed tannins on in vitro anthelmintic activity against bovine nematodes

DEFF Research Database (Denmark)

Desrues, Olivier; Fryganas, Christos; Ropiak, Honorata M.

2016-01-01

Plants containing condensed tannins (CT) may have potential to control gastrointestinal nematodes (GIN) of cattle. The aim was to investigate the anthelmintic activities of four flavan-3-ols, two galloyl derivatives and 14 purified CT fractions, and to define which structural features of CT...... susceptible to all CT fractions than C. oncophora L1. The mean degree of polymerization of CT (i.e. average size) was the most important structural parameter: large CT reduced larval feeding more than small CT. The flavan-3-ols of prodelphinidin (PD)-type tannins had a stronger negative influence on parasite...

Study of binding interaction between anthelmintic 2, 3-dihydroquinazolin-4-ones with bovine serum albumin by spectroscopic methods

Energy Technology Data Exchange (ETDEWEB)

Hemalatha, K.; Madhumitha, G., E-mail: madhumitha.g@vit.ac.in

2016-10-15

A new series of brominated derivatives of 2, 3-dihydroquinazolin-4(1H)-one were synthesized and their structures were confirmed using IR, NMR and mass spectra. The synthesized derivatives were screened for their in vitro anthelmintic activity. The investigations on interaction of the bioactive compound, 2i with bovine serum albumin (BSA) were evaluated. The quenching mechanism of the compound, 2i was deduced based on the results of Stern–Volmer equation. The number of binding site, prediction of binding site region and the changes in the secondary structure of protein were predicted using various spectroscopic studies.

Evaluation of a new test, genotype HelicoDR, for molecular detection of antibiotic resistance in Helicobacter pylori.

Science.gov (United States)

Cambau, Emmanuelle; Allerheiligen, Vera; Coulon, Céline; Corbel, Céline; Lascols, Christine; Deforges, Lionel; Soussy, Claude-James; Delchier, Jean-Charles; Megraud, Francis

2009-11-01

The eradication rate of Helicobacter pylori by standard therapy is decreasing due to antibiotic resistance, mainly to clarithromycin. Our aim was to provide a new molecular test to guide the treatment of new and relapsed cases. We first studied 126 H. pylori strains for phenotypic (MIC) and genotypic resistance to clarithromycin (rrl mutation) and levofloxacin (gyrA mutation) and then developed a DNA strip genotyping test on the basis of the correlation results and literature data. Clinical strains (n = 92) and gastric biopsy specimens containing H. pylori (n = 105) were tested blindly with the new molecular test GenoType HelicoDR. The presence of mutations or the absence of hybridization with wild-type sequences was predictive, in rrl for clarithromycin resistance in 91 cases (mostly the A2147G mutation) and in gyrA for levofloxacin resistance in 58 cases (mutations at codon 87 or 91). Genotyping revealed a mix of genotypes in 33% of the cases, reflecting a coinfection or selection for resistant mutants. The sensitivity and specificity of detecting resistance were 94% and 99% for clarithromycin and 87% and 98.5% for levofloxacin, respectively. The concordance scores were 0.96 for clarithromycin and 0.94 for levofloxacin. With global resistance rates of 46% for clarithromycin and 25% for levofloxacin, which were observed for consecutive positive biopsy specimens from 2007 and 2008, the positive and negative predictive values for detecting resistance were 99% and 94% for clarithromycin and 96% and 96% for fluoroquinolone. GenoType HelicoDR is efficient at detecting mutations predictive of antibiotic resistance in H. pylori when applied to strains or directly to gastric biopsy specimens.

Supplementary Material for: Detection of antibiotic resistance in probiotics of dietary supplements

KAUST Repository

Wong, Aloysius Tze

2015-01-01

Abstract Background Probiotics are live microorganisms that confer nutrition- and health-promoting benefits if consumed in adequate amounts. Concomitant with the demand for natural approaches to maintaining health is an increase in inclusion of probiotics in food and health products. Since probiotic bacteria act as reservoir for antibiotic resistant determinants, the transfer of these genes to pathogens sharing the same intestinal habitat is thus conceivable considering the fact that dietary supplements contain high amounts of often heterogeneous populations of probiotics. Such events can confer pathogens protection against commonly-used drugs. Despite numerous reports of antibiotic resistant probiotics in food and biological sources, the antibiogram of probiotics from dietary supplements remained elusive. Findings Here, we screened five commercially available dietary supplements for resistance towards antibiotics of different classes. Probiotics of all batches of products were resistant towards vancomycin while batch-dependent resistance towards streptomycin, aztreonam, gentamycin and/or ciprofloxacin antibiotics was detected for probiotics of brands Bi and Bn, Bg, and L. Isolates of brand Cn was also resistant towards gentamycin, streptomycin and ciprofloxacin antibiotics. Additionally, we also report a discrepancy between the enumerated viable bacteria amounts and the claims of the manufacturers. Conclusions This short report has highlighted the present of antibiotic resistance in probiotic bacteria from dietary supplements and therefore serves as a platform for further screenings and for in-depth characterization of the resistant determinants and the molecular machinery that confers the resistance.

A method for detection and location of high resistance earth faults

Energy Technology Data Exchange (ETDEWEB)

Haenninen, S; Lehtonen, M [VTT Energy, Espoo (Finland); Antila, E [ABB Transmit Oy (Finland)

1998-08-01

In the first part of this presentation, the theory of earth faults in unearthed and compensated power systems is briefly presented. The main factors affecting the high resistance fault detection are outlined and common practices for earth fault protection in present systems are summarized. The algorithms of the new method for high resistance fault detection and location are then presented. These are based on the change of neutral voltage and zero sequence currents, measured at the high voltage / medium voltage substation and also at the distribution line locations. The performance of the method is analyzed, and the possible error sources discussed. Among these are, for instance, switching actions, thunder storms and heavy snow fall. The feasibility of the method is then verified by an analysis based both on simulated data, which was derived using an EMTP-ATP simulator, and by real system data recorded during field tests at three substations. For the error source analysis, some real case data recorded during natural power system events, is also used

Detection of Individual Molecules and Ions by Carbon Nanotube-Based Differential Resistive Pulse Sensor.

Science.gov (United States)

Peng, Ran; Tang, Xiaowu Shirley; Li, Dongqing

2018-04-01

This paper presents a new method of sensing single molecules and cations by a carbon nanotube (CNT)-based differential resistive pulse sensing (RPS) technique on a nanofluidic chip. A mathematical model for multichannel RPS systems is developed to evaluate the CNT-based RPS signals. Individual cations, rhodamine B dye molecules, and ssDNAs are detected successfully with high resolution and high signal-to-noise ratio. Differentiating ssDNAs with 15 and 30 nucleotides are achieved. The experimental results also show that translocation of negatively charged ssDNAs through a CNT decreases the electrical resistance of the CNT channel, while translocation of positively charged cations and rhodamine B molecules increases the electrical resistance of the CNT. The CNT-based nanofluidic device developed in this work provides a new avenue for single-molecule/ion detection and offers a potential strategy for DNA sequencing. © 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Effect of anthelmintics on reproductive performance and first-lactation culling rate in Holstein heifers.

Science.gov (United States)

Mejía, M E; Perri, A F; Miglierina, M M; Formía, N; Becú-Villalobos, D; Lacau-Mengido, I M

2009-12-26

Female Holstein calves were treated with ivermectin from birth to first oestrus to study the effect of parasitic burden and anthelmintic treatment on reproductive and productive performance. First oestrus, age at first service and age at calving were advanced by 30, 70 and 110 days, respectively (P<0.05), in ivermectin-treated animals compared with controls. No significant differences were observed in the conception rate, the number of services and the characteristics of the newborn calves and any problems at calving between the two groups. Daily milk yield, fat content in milk during first lactation, and the concentrations of growth hormone, insulin-like growth factor type 1, insulin and prolactin in serum were similar in both groups of cows. Culling during the first lactation was more common in untreated (47 per cent) than in treated (11 per cent) cows (P<0.05).

The unique resistance and resilience of the Nigerian West African Dwarf goat to gastrointestinal nematode infections

Science.gov (United States)

2011-01-01

Background West African Dwarf (WAD) goats serve an important role in the rural village economy of West Africa, especially among small-holder livestock owners. They have been shown to be trypanotolerant and to resist infections with Haemonchus contortus more effectively than any other known breed of goat. Methods In this paper we review what is known about the origins of this goat breed, explain its economic importance in rural West Africa and review the current status of our knowledge about its ability to resist parasitic infections. Conclusions We suggest that its unique capacity to show both trypanotolerance and resistance to gastrointestinal (GI) nematode infections is immunologically based and genetically endowed, and that knowledge of the underlying genes could be exploited to improve the capacity of more productive wool and milk producing, but GI nematode susceptible, breeds of goats to resist infection, without recourse to anthelmintics. Either conventional breeding allowing introgression of resistance alleles into susceptible breeds, or transgenesis could be exploited for this purpose. Appropriate legal protection of the resistance alleles of WAD goats might provide a much needed source of revenue for the countries in West Africa where the WAD goats exist and where currently living standards among rural populations are among the lowest in the world. PMID:21291550

Detection of expression and modulation of multidrug-resistance (MDR) and establishment of a new bioassay

International Nuclear Information System (INIS)

Berger, W.

1993-08-01

The present thesis deals with the resistance of human malignant cells against cellular toxicity of anticancer drugs, a phenomenon representing one of the major obstacles to successful chemotherapy. One mechanism underlying a cross-resistance to different drugs called multidrug resistance (MDR) is characterized by the expression of an active transport protein (P-glycoprotein), causing decreased intracellular drug retention and cytotoxicity. The main subjects of the present work were to establish different detection methods for MDR and its modulation (by substances blocking activity of P-glycoprotein) including immunological methods (immunocytochemistry, radioimmunoassay), molecular biology (slot-blot analysis, in-situ hybridization) and functional assays (drug-accumulation analysis, drug-cytotoxicity analysis). The methods were evaluated and compared using human and mouse MDR control cell lines and human tumor cell lines established in our laboratory. In cell lines derived from human melanoma - a malignancy insensitive to chemotherapy - expression of P-glycoprotein of relatively low transporting activity was detected by different methods in 8 of 33 cases. Furthermore a new sensitive in vitro assay for the functional detection of MDR was established using the biological features of cytochalasins, a microfilament disrupting substance group. These compounds were shown to be substrates for the P-glycoprotein efflux pump and their effects on cell division (blockade of cytokinesis resulting in multinucleate cells) correlated with MDR-activity of the tested cells. With this new assay P-glycoprotein activity can be demonstrated and analysed over a wide range of resistance against different cytotoxic drugs. Therefore it may by a suitable tool for research and diagnosis in the field of drug resistance

A Novel Multiplex HRM Assay to Detect Clopidogrel Resistance.

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Zhang, Lichen; Ma, Xiaowei; You, Guoling; Zhang, Xiaoqing; Fu, Qihua

2017-11-22

Clopidogrel is an antiplatelet medicine used to prevent blood clots in patients who have had a heart attack, stroke, or other symptoms. Variability in the clinical response to clopidogrel treatment has been attributed to genetic factors. In particular, five SNPs of rs4244285, rs4986893, rs12248560, rs662 and rs1045642 have been associated with resistance to clopidogrel therapy in Chinese population. This work involves the development of a multiplex high-resolution melting (HRM) assay to genotype all five of these loci in 2 tubes. Amplicons corresponding to distinct SNPs in a common tube were designed with the aid of uMelt prediction software to have different melting temperatures Tm by addition of a GC-rich tail to the 5' end of the certain primers. Two kinds of commercial methods, Digital Fluorescence Molecular Hybridization (DFMH) and Sanger sequencing, were used as a control. Three hundred sixteen DFMH pretested samples from consecutive acute coronary syndrome patients were used for a blinded study of multiplex HRM. The sensitivity of HRM was 100% and the specificity was 99.93% reflecting detection of variants other than the known resistance SNPs. Multiplex HRM is an effective closed-tube, highly accurate, fast, and inexpensive method for genotyping the 5 clopidogrel resistance associated SNPs.

Detection of Anti-Hepatitis B Virus Drug Resistance Mutations Based on Multicolor Melting Curve Analysis.

Science.gov (United States)

Mou, Yi; Athar, Muhammad Ammar; Wu, Yuzhen; Xu, Ye; Wu, Jianhua; Xu, Zhenxing; Hayder, Zulfiqar; Khan, Saeed; Idrees, Muhammad; Nasir, Muhammad Israr; Liao, Yiqun; Li, Qingge

2016-11-01

Detection of anti-hepatitis B virus (HBV) drug resistance mutations is critical for therapeutic decisions for chronic hepatitis B virus infection. We describe a real-time PCR-based assay using multicolor melting curve analysis (MMCA) that could accurately detect 24 HBV nucleotide mutations at 10 amino acid positions in the reverse transcriptase region of the HBV polymerase gene. The two-reaction assay had a limit of detection of 5 copies per reaction and could detect a minor mutant population (5% of the total population) with the reverse transcriptase M204V amino acid mutation in the presence of the major wild-type population when the overall concentration was 10 4 copies/μl. The assay could be finished within 3 h, and the cost of materials for each sample was less than $10. Clinical validation studies using three groups of samples from both nucleos(t)ide analog-treated and -untreated patients showed that the results for 99.3% (840/846) of the samples and 99.9% (8,454/8,460) of the amino acids were concordant with those of Sanger sequencing of the PCR amplicon from the HBV reverse transcriptase region (PCR Sanger sequencing). HBV DNA in six samples with mixed infections consisting of minor mutant subpopulations was undetected by the PCR Sanger sequencing method but was detected by MMCA, and the results were confirmed by coamplification at a lower denaturation temperature-PCR Sanger sequencing. Among the treated patients, 48.6% (103/212) harbored viruses that displayed lamivudine monoresistance, adefovir monoresistance, entecavir resistance, or lamivudine and adefovir resistance. Among the untreated patients, the Chinese group had more mutation-containing samples than did the Pakistani group (3.3% versus 0.56%). Because of its accuracy, rapidness, wide-range coverage, and cost-effectiveness, the real-time PCR assay could be a robust tool for the detection if anti-HBV drug resistance mutations in resource-limited countries. Copyright © 2016, American Society for

Role of the Material Electrodes on Resistive Behaviour of Carbon Nanotube-Based Gas Sensors for H2S Detection

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M. Lucci

2012-01-01

Full Text Available Miniaturized gas-sensing devices that use single-walled carbon nanotubes as active material have been fabricated using two different electrode materials, namely, Au/Cr and NbN. The resistive sensors have been assembled aligning by dielectrophoresis the nanotube bundles between 40 μm spaced Au/Cr or NbN multifinger electrodes. The sensing devices have been tested for detection of the H2S gas, in the concentration range 10–100 ppm, using N2 as carrier gas. No resistance changes were detected using sensor fabricated with NbN electrodes, whereas the response of the sensor fabricated with Au/Cr electrodes was characterized by an increase of the resistance upon gas exposure. The main performances of this sensor are a detection limit for H2S of 10 ppm and a recovery time of few minutes. The present study suggests that the mechanism involved in H2S gas detection is not a direct charge transfer between molecules and nanotubes. The hypothesis is that detection occurs through passivation of the Au surfaces by H2S molecules and modification of the contact resistance at the Au/nanotube interface.

Evaluation of an expanded microarray for detecting antibiotic resistance genes in a broad range of gram-negative bacterial pathogens.

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Card, Roderick; Zhang, Jiancheng; Das, Priya; Cook, Charlotte; Woodford, Neil; Anjum, Muna F

2013-01-01

A microarray capable of detecting genes for resistance to 75 clinically relevant antibiotics encompassing 19 different antimicrobial classes was tested on 132 Gram-negative bacteria. Microarray-positive results correlated >91% with antimicrobial resistance phenotypes, assessed using British Society for Antimicrobial Chemotherapy clinical breakpoints; the overall test specificity was >83%. Microarray-positive results without a corresponding resistance phenotype matched 94% with PCR results, indicating accurate detection of genes present in the respective bacteria by microarray when expression was low or absent and, hence, undetectable by susceptibility testing. The low sensitivity and negative predictive values of the microarray results for identifying resistance to some antimicrobial resistance classes are likely due to the limited number of resistance genes present on the current microarray for those antimicrobial agents or to mutation-based resistance mechanisms. With regular updates, this microarray can be used for clinical diagnostics to help accurate therapeutic options to be taken following infection with multiple-antibiotic-resistant Gram-negative bacteria and prevent treatment failure.

Comparison of constitutive and thiabendazole-induced expression of five cytochrome P450 genes in fourth-stage larvae of Haemonchus contortus isolates with different drug susceptibility identifies one gene with high constitutive expression in a multi-resistant isolate

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Esra Yilmaz

2017-12-01

Full Text Available Benzimidazoles (BZs remain amongst the most widely used anthelmintic drug classes against gastro-intestinal nematode infections, although their efficacy is increasingly compromised by resistance. The primary underlying mechanisms for BZ resistance are single-nucleotide polymorphisms (SNPs in the isotype 1 β-tubulin gene causing the substitutions F167Y, E198A or F200Y. However, resistance is believed to be multi-genic and previous studies have shown that isolates carrying 90–100% F200Y can vary considerably in their resistance level in the egg hatch assay (EHA. Cytochrome P450 monooxygenases (CYPs are associated with drug resistance in mammals and arthropods and have been considered as mediators of anthelmintic resistance. In Caenorhabditis elegans, several members of the CYP34/35 and CYP31 families are BZ and/or xenobiotic inducible and thiabendazole (TBZ is metabolised by CYP35D1. Here, expression of all 5 CYPs closely related to the C. elegans CYP34/35 and CYP31 families was investigated in fourth-stage larvae of two susceptible and three BZ-resistant Haemonchus contortus isolates following in vitro exposure to TBZ for 3 and 6 h using real-time RT-PCR. The resistance status of all isolates was determined using EHAs and quantification of resistance-associated β-tubulin SNPs using pyrosequencing. While none of the CYPs was TBZ inducible, constitutive expression of CYP34/35 family member HCOI100383400 was significantly 2.4–3.7-fold higher in the multi-drug resistant WR isolate with the strongest BZ resistance phenotype compared to susceptible and intermediate-level BZ-resistant isolates. Although this increase is only moderate, HCOI100383400 might still be involved in high-level BZ resistance by further decreasing susceptibility in isolates already carrying 100% of a β-tubulin SNP causing BZ resistance. Lower transcript levels were observed for all CYPs in the intermediately resistant IRE isolate in comparison to the susceptible Hc

Efficacy and safety of anthelmintics tested against Taenia solium cysticercosis in pigs.

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Ernatus Martin Mkupasi

Full Text Available Porcine cysticercosis, an infection caused by Taenia solium metacestodes, is continuously being reported in low-income countries of Latin America, Asia, and sub-Saharan Africa. The disease was declared eradicable by the International Task Force for Diseases Eradication (ITFDE in 1993, and it is listed among the 17 WHO Neglected Tropical Diseases and Neglected Zoonoses that are potentially eradicable. In view of that, WHO has proposed a step-wise approach to its elimination, including chemotherapy of infected pigs. Different drugs have been tested on porcine cysticercosis with varying efficacies. These include flubendazole, fenbendazole, albendazole, albendazole sulphoxide, oxfendazole, praziquantel, and nitazoxanide. This review summarises available information on the efficacies and adverse effects shown by these drugs in pigs. Oxfendazole has shown to be effective for the control of porcine cysticercosis; however, it needs to be integrated with other control approaches. There is a need for standardised guidelines for evaluating the efficacy of anthelmintics against porcine cysticercosis, and more efficacy studies are needed since the conclusions so far are based on a limited number of studies using few infected pigs.

Detection of the acetylcholinesterase insecticide resistance mutation (G328A) in natural populations of Ceratitis capitata

International Nuclear Information System (INIS)

Elfekih, Samia; Haran, Julien; Shannon, Matthew; Vogler, Alfried P.

2015-01-01

Wild Mediterranean fruit fly specimens collected from various regions worldwide were screened for the glycine to alanine (Gly->Ala) point mutation (G328A) in the acetylcholinesterase enzyme, presumably causing resistance to organophosphates. We found that the single nucleotide polymorphism (SNP) responsible for this amino acid change is located at the beginning of exon 6 of the Ccace2 gene. The identification of the exact location of the SNP permitted PCR primer design around this site and direct sequencing of the corresponding genomic region. We detected the resistance allele in natural Mediterranean fruit fly populations from Brazil and Spain, but not from other sites in four continents. The known treatment history of sites suggests that the resistance build up is linked to organophosphate application in the held. The PCR-based detection provides a screening method useful for monitoring Mediterranean fruit fly insecticide resistance in local populations and improving pest management strategies accordingly. (author)

The value of microscopic-observation drug susceptibility assay in the diagnosis of tuberculosis and detection of multidrug resistance.

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Sertel Şelale, Denİz; Uzun, Meltem

2018-01-01

Inexpensive, rapid, and reliable tests for detecting the presence and drug susceptibility of Mycobacterium tuberculosis complex (MTBC) are urgently needed to control the transmission of tuberculosis. In this study, we aimed to assess the accuracy and speed of the microscopic-observation drug susceptibility (MODS) assay in the identification of MTBC and detection of multidrug resistance. Sputum samples from patients suspected to have tuberculosis were simultaneously tested with MODS and conventional culture [Löwenstein-Jensen (LJ) culture, BACTEC MGIT™ 960 (MGIT) system], and drug susceptibility testing (MGIT system) methods. A total of 331 sputum samples were analyzed. Sensitivity and specificity of MODS assay for detection of MTBC strains were 96% and 98.8%, respectively. MODS assay detected multidrug resistant MTBC isolates with 92.3% sensitivity and 96.6% specificity. Median time to culture positivity was similar for MGIT (8 days) and MODS culture (8 days), but was significantly longer with LJ culture (20 days) (p tuberculosis and detection of multidrug resistance. © 2017 APMIS. Published by John Wiley & Sons Ltd.

A randomized triple blind trial to assess the effect of an anthelmintic programme for working equids in Morocco

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Christley Robert M

2011-01-01

Full Text Available Abstract Background Gastro-intestinal parasitism has been identified as a significant cause of disease in working equids in many countries. This randomized triple-blind trial was designed to assess the impact of an anthelmintic treatment programme (using oral ivermectin and fenbendazole comparing treated and placebo control populations of working donkeys, mules and horses in field conditions in Morocco. In particular, we assessed animal body weight and condition score, together with a questionnaire-based owner evaluation of number of subjective animal health parameters. Faecal worm egg count was also measured. Results 239 animals completed the full study, 130 in the treatment group and 109 in the control group. Although the average animal weight increased during the study, this change was not significantly different between the two groups. Animals in the treatment group had a significantly lower strongyle worm egg count and increased in body condition score compared to animals in the control group at each examination during the study period. Owners of animals in the treatment group reported improvement in health and work ability and a beneficial effect on pruritus during the early period of the study. These differences in owner perception between treatment groups had disappeared in the latter stages of the study. Conclusion This study demonstrated that a routine anthelmintic treatment programme of three treatments annually can have a significant effect on faecal worm egg count. There may be beneficial consequences for the animal health and productivity. Further research on other populations of working equids in different environments would facilitate the objective planning of effective parasite control strategies for specific situations and provide better understanding of the likely clinical benefits of such programmes.

A randomized triple blind trial to assess the effect of an anthelmintic programme for working equids in Morocco

Science.gov (United States)

2011-01-01

Background Gastro-intestinal parasitism has been identified as a significant cause of disease in working equids in many countries. This randomized triple-blind trial was designed to assess the impact of an anthelmintic treatment programme (using oral ivermectin and fenbendazole) comparing treated and placebo control populations of working donkeys, mules and horses in field conditions in Morocco. In particular, we assessed animal body weight and condition score, together with a questionnaire-based owner evaluation of number of subjective animal health parameters. Faecal worm egg count was also measured. Results 239 animals completed the full study, 130 in the treatment group and 109 in the control group. Although the average animal weight increased during the study, this change was not significantly different between the two groups. Animals in the treatment group had a significantly lower strongyle worm egg count and increased in body condition score compared to animals in the control group at each examination during the study period. Owners of animals in the treatment group reported improvement in health and work ability and a beneficial effect on pruritus during the early period of the study. These differences in owner perception between treatment groups had disappeared in the latter stages of the study. Conclusion This study demonstrated that a routine anthelmintic treatment programme of three treatments annually can have a significant effect on faecal worm egg count. There may be beneficial consequences for the animal health and productivity. Further research on other populations of working equids in different environments would facilitate the objective planning of effective parasite control strategies for specific situations and provide better understanding of the likely clinical benefits of such programmes. PMID:21208398

Resistance Detection of Aedes aegypti Larvae to Cypermethrin from Endemic Area in Cimahi City West Java

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Endang Puji Astuti

2014-06-01

Full Text Available Vector control programs using chemical insecticide e.g organochlorin, organophosphate, carbamate, and pyrethroid (cypermethrin. When those insecticides were applied continuously, it may lead to vector resistance. The aim of this research was to detect any resistance of Ae. aegypti to cypermethrin in endemic areas of Cimahi. This research is a laboratory study that used biochemical test which referred to Lee’s method. Larva samples were collected from 8 villages, which are endemic area. Samples of larvae were collected from 15 villages belonged to dengue endemic areas in town of Cimahi, however, villages that meet the availability of larvae were only 8 villages. To detect the activity of monooxygenase enzyme, a biochemical assay was used in this research by created a reaction between larvae homogenate and sodium acetate substrate. The results of reaction were read using ELISA reader with spectrophotometer wave length of 595 nm. Overall, the results showed that most of the larvae in eight villages of Cimahi is still susceptible to cypermethrin. However, larvae from Cibabat village were 4% resistant, 2% tolerant, and 94% susceptible. On the other hand, Cigugur village showed that 12.7% larvae were tolerant and 87.3% still susceptible. Other villages like Cimahi, Cibeureum, Melong, Baros, Cipageran, and Pasirkaliki still remains susceptible. Resistance detection using biochemical assay of cypermethrin insecticide for Ae.aegypti resulting data stated that in 6 villages were still susceptible but in 3 other villages were already tolerant and 1 village was already resistance.

Detection of leaks in underground storage tanks using electrical resistance methods: 1996 results

International Nuclear Information System (INIS)

Ramirez, A.; Daily, W.

1996-10-01

This document provides a summary of a field experiment performed under a 15m diameter steel tank mockup located at the Hanford Reservation, Washington. The purpose of this test was to image a contaminant plume as it develops in soil under a tank already contaminated by previous leakage and to determine whether contaminant plumes can be detected without the benefit of background data. Measurements of electrical resistance were made before and during a salt water release. These measurements were made in soil which contained the remnants of salt water plumes released during previous tests in 1994 and in 1995. About 11,150 liters of saline solution were released along a portion of the tank's edge in 1996. Changes in electrical resistivity due to release of salt water conducted in 1996 were determined in two ways: (1) changes relative to the 1996 pre-spill data, and (2) changes relative to data collected near the middle of the 1996 spill after the release flow rate was increased. In both cases, the observed resistivity changes show clearly defined anomalies caused by the salt water release. These results indicate that when a plume develops over an existing plume and in a geologic environment similar to the test site environment, the resulting resistivity changes are easily detectable. Three dimensional tomographs of the resistivity of the soil under the tank show that the salt water release caused a region of low soil resistivity which can be observed directly without the benefit of comparing the tomograph to tomographs or data collected before the spill started. This means that it may be possible to infer the presence of pre-existing plumes if there is other data showing that the regions of low resistivity are correlated with the presence of contaminated soil. However, this approach does not appear reliable in defining the total extent of the plume due to the confounding effect that natural heterogeneity has on our ability to define the margins of the anomaly

Anthelmintic drug ivermectin inhibits angiogenesis, growth and survival of glioblastoma through inducing mitochondrial dysfunction and oxidative stress

International Nuclear Information System (INIS)

Liu, Yingying; Fang, Shanshan; Sun, Qiushi; Liu, Bo

2016-01-01

Glioblastoma is one of the most vascular brain tumour and highly resistant to current therapy. Targeting both glioblastoma cells and angiogenesis may present an effective therapeutic strategy for glioblastoma. In our work, we show that an anthelmintic drug, ivermectin, is active against glioblastoma cells in vitro and in vivo, and also targets angiogenesis. Ivermectin significantly inhibits growth and anchorage-independent colony formation in U87 and T98G glioblastoma cells. It induces apoptosis in these cells through a caspase-dependent manner. Ivermectin significantly suppresses the growth of two independent glioblastoma xenograft mouse models. In addition, ivermectin effectively targets angiogenesis through inhibiting capillary network formation, proliferation and survival in human brain microvascular endothelial cell (HBMEC). Mechanistically, ivermectin decreases mitochondrial respiration, membrane potential, ATP levels and increases mitochondrial superoxide in U87, T98G and HBMEC cells exposed to ivermectin. The inhibitory effects of ivermectin are significantly reversed in mitochondria-deficient cells or cells treated with antioxidants, further confirming that ivermectin acts through mitochondrial respiration inhibition and induction of oxidative stress. Importantly, we show that ivermectin suppresses phosphorylation of Akt, mTOR and ribosomal S6 in glioblastoma and HBMEC cells, suggesting its inhibitory role in deactivating Akt/mTOR pathway. Altogether, our work demonstrates that ivermectin is a useful addition to the treatment armamentarium for glioblastoma. Our work also highlights the therapeutic value of targeting mitochondrial metabolism in glioblastoma. - Highlights: • Ivermectin is effective in glioblastoma cells in vitro and in vivo. • Ivermectin inhibits angiogenesis. • Ivermectin induces mitochondrial dysfunction and oxidative stress. • Ivermectin deactivates Akt/mTOR signaling pathway.

Evaluation of a New Test, GenoType HelicoDR, for Molecular Detection of Antibiotic Resistance in Helicobacter pylori▿

Science.gov (United States)

Cambau, Emmanuelle; Allerheiligen, Vera; Coulon, Céline; Corbel, Céline; Lascols, Christine; Deforges, Lionel; Soussy, Claude-James; Delchier, Jean-Charles; Megraud, Francis

2009-01-01

The eradication rate of Helicobacter pylori by standard therapy is decreasing due to antibiotic resistance, mainly to clarithromycin. Our aim was to provide a new molecular test to guide the treatment of new and relapsed cases. We first studied 126 H. pylori strains for phenotypic (MIC) and genotypic resistance to clarithromycin (rrl mutation) and levofloxacin (gyrA mutation) and then developed a DNA strip genotyping test on the basis of the correlation results and literature data. Clinical strains (n = 92) and gastric biopsy specimens containing H. pylori (n = 105) were tested blindly with the new molecular test GenoType HelicoDR. The presence of mutations or the absence of hybridization with wild-type sequences was predictive, in rrl for clarithromycin resistance in 91 cases (mostly the A2147G mutation) and in gyrA for levofloxacin resistance in 58 cases (mutations at codon 87 or 91). Genotyping revealed a mix of genotypes in 33% of the cases, reflecting a coinfection or selection for resistant mutants. The sensitivity and specificity of detecting resistance were 94% and 99% for clarithromycin and 87% and 98.5% for levofloxacin, respectively. The concordance scores were 0.96 for clarithromycin and 0.94 for levofloxacin. With global resistance rates of 46% for clarithromycin and 25% for levofloxacin, which were observed for consecutive positive biopsy specimens from 2007 and 2008, the positive and negative predictive values for detecting resistance were 99% and 94% for clarithromycin and 96% and 96% for fluoroquinolone. GenoType HelicoDR is efficient at detecting mutations predictive of antibiotic resistance in H. pylori when applied to strains or directly to gastric biopsy specimens. PMID:19759218

Rapid detection of methicillin resistance in Staphylococcus aureus isolates by the MRSA-screen latex agglutination test

NARCIS (Netherlands)

W.B. van Leeuwen (Willem); C. van Pelt (Cindy); A. Luijendijk (Ad); H.A. Verbrugh (Henri); W.H.F. Goessens (Wil)

1999-01-01

textabstractThe slide agglutination test MRSA-Screen (Denka Seiken Co., Niigata, Japan) was compared with the mecA PCR ("gold standard") for the detection of methicillin resistance in Staphylococcus aureus. The MRSA-Screen test detected the penicillin-binding protein 2a

Method for rapid detection and identification of chaetomium and evaluation of resistance to peracetic acid.

Science.gov (United States)

Nakayama, Motokazu; Hosoya, Kouichi; Tomiyama, Daisuke; Tsugukuni, Takashi; Matsuzawa, Tetsuhiro; Imanishi, Yumi; Yaguchi, Takashi

2013-06-01

In the beverage industry, peracetic acid has been increasingly used as a disinfectant for the filling machinery and environment due to merits of leaving no residue, it is safe for humans, and its antiseptic effect against fungi and endospores of bacteria. Recently, Chaetomium globosum and Chaetomium funicola were reported resistant to peracetic acid; however, little is known concerning the detail of peracetic acid resistance. Therefore, we assessed the peracetic acid resistance of the species of Chaetomium and related genera under identical conditions and made a thorough observation of the microstructure of their ascospores by transmission electron microscopy. The results of analyses revealed that C. globosum and C. funicola showed the high resistance to peracetic acid (a 1-D antiseptic effect after 900 s and 3-D antiseptic effect after 900 s) and had thick cell walls of ascospores that can impede the action mechanism of peracetic acid. We also developed specific primers to detect the C. globosum clade and identify C. funicola by using PCR to amplify the β-tubulin gene. PCR with the primer sets designed for C. globosum (Chae 4F/4R) and C. funicola (Cfu 2F/2R) amplified PCR products specific for the C. globosum clade and C. funicola, respectively. PCR with these two primer sets did not detect other fungi involved in food spoilage and environmental contamination. This detection and identification method is rapid and simple, with extremely high specificity.

Rapid Aminoglycoside NP Test for Rapid Detection of Multiple Aminoglycoside Resistance in Enterobacteriaceae.

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Nordmann, Patrice; Jayol, Aurélie; Dobias, Jan; Poirel, Laurent

2017-04-01

The rapid aminoglycoside NP (Nordmann/Poirel) test was developed to rapidly identify multiple aminoglycoside (AG) resistance in Enterobacteriaceae It is based on the detection of the glucose metabolism related to enterobacterial growth in the presence of a defined concentration of amikacin plus gentamicin. Formation of acid metabolites was evidenced by a color change (orange to yellow) of the red phenol pH indicator. The rapid aminoglycoside NP test was evaluated by using bacterial colonies of 18 AG-resistant isolates producing 16S rRNA methylases, 20 AG-resistant isolates expressing AG-modifying enzymes (acetyl-, adenyl-, and phosphotransferases), and 10 isolates susceptible to AG. Its sensitivity and specificity were 100% and 97%, respectively, compared to the broth dilution method, which was taken as the gold standard for determining aminoglycoside resistance. The test is inexpensive, rapid (<2 h), and implementable worldwide. Copyright © 2017 American Society for Microbiology.

Multiplex TaqMan® detection of pathogenic and multi-drug resistant Salmonella.

Science.gov (United States)

Singh, Prashant; Mustapha, Azlin

2013-09-02

Overuse of antibiotics in the medical and animal industries is one of the major causes for the development of multi-drug-resistant (MDR) food pathogens that are often difficult to treat. In the past few years, higher incidences of outbreaks caused by MDR Salmonella have been increasingly documented. The objective of this study was to develop a rapid multiplex real-time polymerase chain reaction (PCR) assay for simultaneous detection of pathogenic and MDR Salmonella spp. A multiplex TaqMan®real-time PCR was designed by targeting the invasin virulence gene (invA), and four commonly found antibiotic resistance genes, viz. ampicillin, chloramphenicol, streptomycin and tetracycline. To avoid false negative results and to increase the reliability of the assay, an internal amplification control (IAC) was added which was detected using a locked nucleic acid (LNA) probe. In serially diluted (5 ng-50 fg) DNA samples, the assay was able to detect 100 genomic equivalents of Salmonella, while in a multiplex format, the sensitivity was 1000 genomic equivalents. The assay performed equally well on artificially contaminated samples of beef trim, ground beef of different fat contents (73:27, 80:20, 85:15 and 93:7), chicken rinse, ground chicken, ground turkey, egg, spinach and tomato. While the detection limit for un-enriched inoculated food samples was 10(4) CFU/g, this was improved to 10 CFU/g after a 12-h enrichment in buffered peptone water, with 100% reproducibility. The multiplex real-time assay developed in this study can be used as a valuable tool to detect MDR virulent Salmonella, thus enhancing the safety of food. © 2013.

Application of Electrical Resistivity Tomography for Detecting Root Biomass in Coffee Trees

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Carlos Mauricio Paglis

2013-01-01

Full Text Available Roots play an important role in plants and are responsible for several functions; among them are anchorage and nutrient and water absorption. Several methodologies are being tested and used to study plant root systems in order to avoid destructive root sampling. Electrical resistivity tomography is among these methodologies. The aim of this preliminary study was to use electrical resistivity for detecting root biomass in coffee trees. Measurements were performed in a soil transect with an ABM AL 48-b resistivimeter with a pole-dipole configuration. The tomograms indicated variability in soil resistivity values ranging from 120 to 1400 Ω·m−1. At the first 0.30 cm soil layer, these values were between 267 and 952 Ω·m−1. Oriented by this result, root samples were taken at 0.10, 0.20, and 0.30 m depths within 0.50 m intervals along the soil transect to compare soil resistivity with root mass density (RMD. RMD data, up to this depth, varied from 0.000019 to 0.009469 Mg·m−3, showing high spatial variability and significant relationship to the observed values of soil resistivity. These preliminary results showed that the electrical resistivity tomography can contribute to root biomass studies in coffee plants; however, more experiments are necessary to confirm the found results in Brazil coffee plantations.

Risk analysis and detection of thrombosis by measurement of electrical resistivity of blood.

Science.gov (United States)

Sapkota, Achyut; Asakura, Yuta; Maruyama, Osamu; Kosaka, Ryo; Yamane, Takashi; Takei, Masahiro

2013-01-01

Monitoring of thrombogenic process is very important in ventricular assistance devices (VADs) used as temporary or permanent measures in patients with advanced heart failure. Currently, there is a lack of a system which can perform a real-time monitoring of thrombogenic activity. Electrical signals vary according to the change in concentration of coagulation factors as well as the distribution of blood cells, and thus have potential to detect the thrombogenic process in an early stage. In the present work, we have made an assessment of an instrumentation system exploiting the electrical properties of blood. The experiments were conducted using bovine blood. Electrical resistance tomography with eight-electrode sensor was used to monitor the spatio-temporal change in electrical resistivity of blood in thrombogenic and non-thrombogenic condition. Under non-thrombogenic condition, the resistivity was uniform across the cross-section and average resistivity monotonically decreased with time before remaining almost flat. In contrary, under thrombogenic condition, there was non-uniform distribution across the cross-section, and average resistivity fluctuated with time.

Electrochemical and DFT study of an anticancer and active anthelmintic drug at carbon nanostructured modified electrode.

Science.gov (United States)

Ghalkhani, Masoumeh; Beheshtian, Javad; Salehi, Maryam

2016-12-01

The electrochemical response of mebendazole (Meb), an anticancer and effective anthelmintic drug, was investigated using two different carbon nanostructured modified glassy carbon electrodes (GCE). Although, compared to unmodified GCE, both prepared modified electrodes improved the voltammetric response of Meb, the carbon nanotubes (CNTs) modified GCE showed higher sensitivity and stability. Therefore, the CNTs-GCE was chosen as a promising candidate for the further studies. At first, the electrochemical behavior of Meb was studied by cyclic voltammetry and differential pulse and square wave voltammetry. A one step reversible, pH-dependent and adsorption-controlled process was revealed for electro-oxidation of Meb. A possible mechanism for the electrochemical oxidation of Meb was proposed. In addition, electronic structure, adsorption energy, band gap, type of interaction and stable configuration of Meb on the surface of functionalized carbon nanotubes were studied by using density functional theory (DFT). Obtained results revealed that Meb is weakly physisorbed on the CNTs and that the electronic properties of the CNTs are not significantly changed. Notably, CNTs could be considered as a suitable modifier for preparation of the modified electrode for Meb analysis. Then, the experimental parameters affecting the electrochemical response of Meb were optimized. Under optimal conditions, high sensitivity (b(Meb)=dIp,a(Meb)/d[Meb]=19.65μAμM(-1)), a low detection limit (LOD (Meb)=19nM) and a wide linear dynamic range (0.06-3μM) was resulted for the voltammetric quantification of Meb. Copyright © 2016 Elsevier B.V. All rights reserved.

New Real-Time PCR Assays for Detection of Inducible and Acquired Clarithromycin Resistance in the Mycobacterium abscessus Group.

Science.gov (United States)

Shallom, Shamira J; Moura, Natalia S; Olivier, Kenneth N; Sampaio, Elizabeth P; Holland, Steven M; Zelazny, Adrian M

2015-11-01

Members of the Mycobacterium abscessus group (MAG) cause lung, soft tissue, and disseminated infections. The oral macrolides clarithromycin and azithromycin are commonly used for treatment. MAG can display clarithromycin resistance through the inducible erm(41) gene or via acquired mutations in the rrl (23S rRNA) gene. Strains harboring a truncation or a T28C substitution in erm(41) lose the inducible resistance trait. Phenotypic detection of clarithromycin resistance requires extended incubation (14 days), highlighting the need for faster methods to detect resistance. Two real-time PCR-based assays were developed to assess inducible and acquired clarithromycin resistance and tested on a total of 90 clinical and reference strains. A SYBR green assay was designed to distinguish between a full-length and truncated erm(41) gene by temperature shift in melting curve analysis. Single nucleotide polymorphism (SNP) allele discrimination assays were developed to distinguish T or C at position 28 of erm(41) and 23S rRNA rrl gene mutations at position 2058 and/or 2059. Truncated and full-size erm(41) genes were detected in 21/90 and 69/90 strains, respectively, with 64/69 displaying T at nucleotide position 28 and 5/69 containing C at that position. Fifteen isolates showed rrl mutations conferring clarithromycin resistance, including A2058G (11 isolates), A2058C (3 isolates), and A2059G (1 isolate). Targeted sequencing and phenotypic assessment of resistance concurred with molecular assay results. Interestingly, we also noted cooccurring strains harboring an active erm(41), inactive erm(41), and/or acquired mutational resistance, as well as slowly growing MAG strains and also strains displaying an inducible resistance phenotype within 5 days, long before the recommended 14-day extended incubation. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

Detection of First-Line Drug Resistance Mutations and Drug-Protein Interaction Dynamics from Tuberculosis Patients in South India.

Science.gov (United States)

Nachappa, Somanna Ajjamada; Neelambike, Sumana M; Amruthavalli, Chokkanna; Ramachandra, Nallur B

2018-05-01

Diagnosis of drug-resistant tuberculosis predominantly relies on culture-based drug susceptibility testing, which take weeks to produce a result and a more time-efficient alternative method is multiplex allele-specific PCR (MAS-PCR). Also, understanding the role of mutations in causing resistance helps better drug designing. To evaluate the ability of MAS-PCR in the detection of drug resistance and to understand the mechanism of interaction of drugs with mutant proteins in Mycobacterium tuberculosis. Detection of drug-resistant mutations using MAS-PCR and validation through DNA sequencing. MAS-PCR targeted five loci on three genes, katG 315 and inhA -15 for the drug isoniazid (INH), and rpoB 516, 526, and 531 for rifampicin (RIF). Furthermore, the sequence data were analyzed to study the effect on interaction of the anti-TB drug molecule with the target protein using in silico docking. We identified drug-resistant mutations in 8 out of 114 isolates with 2 of them as multidrug-resistant TB using MAS-PCR. DNA sequencing confirmed only six of these, recording a sensitivity of 85.7% and specificity of 99.3% for MAS-PCR. Molecular docking showed estimated free energy of binding (ΔG) being higher for RIF binding with RpoB S531L mutant. Codon 315 in KatG does not directly interact with INH but blocks the drug access to active site. We propose DNA sequencing-based drug resistance detection for TB, which is more accurate than MAS-PCR. Understanding the action of resistant mutations in disrupting the normal drug-protein interaction aids in designing effective drug alternatives.

Detection of Soot Using a Resistivity Sensor Device Employing Thermophoretic Particle Deposition

Directory of Open Access Journals (Sweden)

Doina Lutic

2010-01-01

Full Text Available Results are reported for thermophoretic deposition of soot particles on resistivity sensors as a monitoring technique for diesel exhaust particles with the potential of improved detection limit and sensitivity. Soot with similar characteristics as from diesel exhausts was generated by a propane flame and diluted in stages. The soot in a gas flow at 240–270C∘ was collected on an interdigitated electrode structure held at a considerably lower temperature, 105–125C∘. The time delay for reaching measurable resistance values, the subsequent rate, and magnitude of resistance decrease were a function of the distance between the fingers in the electrodes and the degree of dilution of the soot containing flow. Soot deposition and subsequent removal by heating the sensor support was also performed in a real diesel exhaust. Good similarities between the behavior in our laboratory system and the real diesel exhaust were noticed.

Metabolic profiling for detection of Staphylococcus aureus infection and antibiotic resistance.

Directory of Open Access Journals (Sweden)

Henrik Antti

Full Text Available Due to slow diagnostics, physicians must optimize antibiotic therapies based on clinical evaluation of patients without specific information on causative bacteria. We have investigated metabolomic analysis of blood for the detection of acute bacterial infection and early differentiation between ineffective and effective antibiotic treatment. A vital and timely therapeutic difficulty was thereby addressed: the ability to rapidly detect treatment failures because of antibiotic-resistant bacteria. Methicillin-resistant Staphylococcus aureus (MRSA and methicillin-sensitive S. aureus (MSSA were used in vitro and for infecting mice, while natural MSSA infection was studied in humans. Samples of bacterial growth media, the blood of infected mice and of humans were analyzed with combined Gas Chromatography/Mass Spectrometry. Multivariate data analysis was used to reveal the metabolic profiles of infection and the responses to different antibiotic treatments. In vitro experiments resulted in the detection of 256 putative metabolites and mice infection experiments resulted in the detection of 474 putative metabolites. Importantly, ineffective and effective antibiotic treatments were differentiated already two hours after treatment start in both experimental systems. That is, the ineffective treatment of MRSA using cloxacillin and untreated controls produced one metabolic profile while all effective treatment combinations using cloxacillin or vancomycin for MSSA or MRSA produced another profile. For further evaluation of the concept, blood samples of humans admitted to intensive care with severe sepsis were analyzed. One hundred thirty-three putative metabolites differentiated severe MSSA sepsis (n = 6 from severe Escherichia coli sepsis (n = 10 and identified treatment responses over time. Combined analysis of human, in vitro, and mice samples identified 25 metabolites indicative of effective treatment of S. aureus sepsis. Taken together, this

Evaluation of conventional PCR for detection of Strongylus vulgaris on horse farms.

Science.gov (United States)

Bracken, M K; Wøhlk, C B M; Petersen, S L; Nielsen, M K

2012-03-23

Strongyle parasites are ubiquitous in grazing horses. Of these, the bloodworm Strongylus vulgaris is regarded as most pathogenic. Increasing levels of anthelmintic resistance in strongyle parasites has led to recommendations of decreased treatment intensities, and there is now a pronounced need for reliable tools for detection of parasite burdens in general and S. vulgaris in particular. The only method currently available for diagnosing S. vulgaris in practice is the larval culture, which is laborious and time-consuming, so veterinary practitioners most often pool samples from several horses together in one culture to save time. Recently, molecular tools have been developed to detect S. vulgaris in faecal samples. The aim of this study was to compare the performance of a conventional polymerase chain reaction (PCR) assay with the traditional larval culture and furthermore test the performance of pooled versus individual PCR for farm screening purposes. Faecal samples were obtained from 331 horses on 18 different farms. Farm size ranged from 6 to 56 horses, and horses aged between 2 months and 31 years. Larval cultures and PCR were performed individually on all horses. In addition, PCR was performed on 66 faecal pools consisting of 3-5 horses each. Species-specific PCR primers previously developed were used for the PCR. PCR and larval culture detected S. vulgaris in 12.1 and 4.5% of individual horses, respectively. On the farm level, eight farms tested positive with the larval culture, while 13 and 11 farms were positive with the individual and pooled PCRs, respectively. The individual PCR method was statistically superior to the larval culture, while no statistical difference could be detected between pooled and individual PCR for farm screening. In conclusion, pooled PCR appears to be a useful tool for farm screening for S. vulgaris. Copyright © 2011 Elsevier B.V. All rights reserved.

Association between large strongyle genera in larval cultures--using rare-event poisson regression.

Science.gov (United States)

Cao, X; Vidyashankar, A N; Nielsen, M K

2013-09-01

Decades of intensive anthelmintic treatment has caused equine large strongyles to become quite rare, while the cyathostomins have developed resistance to several drug classes. The larval culture has been associated with low to moderate negative predictive values for detecting Strongylus vulgaris infection. It is unknown whether detection of other large strongyle species can be statistically associated with presence of S. vulgaris. This remains a statistical challenge because of the rare occurrence of large strongyle species. This study used a modified Poisson regression to analyse a dataset for associations between S. vulgaris infection and simultaneous occurrence of Strongylus edentatus and Triodontophorus spp. In 663 horses on 42 Danish farms, the individual prevalences of S. vulgaris, S. edentatus and Triodontophorus spp. were 12%, 3% and 12%, respectively. Both S. edentatus and Triodontophorus spp. were significantly associated with S. vulgaris infection with relative risks above 1. Further, S. edentatus was associated with use of selective therapy on the farms, as well as negatively associated with anthelmintic treatment carried out within 6 months prior to the study. The findings illustrate that occurrence of S. vulgaris in larval cultures can be interpreted as indicative of other large strongyles being likely to be present.

Recent advances in biochemical and molecular diagnostics for the rapid detection of antibiotic-resistant Enterobacteriaceae: a focus on ß-lactam resistance.

Science.gov (United States)

Decousser, Jean-Winoc; Poirel, Laurent; Nordmann, Patrice

2017-04-01

The rapid detection of resistance is a challenge for clinical microbiologists who wish to prevent deleterious individual and collective consequences such as (i) delaying efficient antibiotic therapy, which worsens the survival rate of the most severely ill patients, or (ii) delaying the isolation of the carriers of multidrug-resistant bacteria and promoting outbreaks; this last consequence is of special concern, and there are an increasing number of approaches and market-based solutions in response. Areas covered: From simple, cheap biochemical tests to whole-genome sequencing, clinical microbiologists must select the most adequate phenotypic and genotypic tools to promptly detect and confirm β-lactam resistance from cultivated bacteria or from clinical specimens. Here, the authors review the published literature from the last 5 years about the primary technical approaches and commercial laboratory reagents for these purposes, including molecular, biochemical and immune assays. Furthermore, the authors discuss their intrinsic and relative performance, and we challenge their putative clinical impact. Expert commentary: Until the availability of fully automated wet and dry whole genome sequencing solutions, microbiologists should focus on inexpensive biochemical tests for cultured isolates or monomicrobial clinical specimen and on using the expensive molecular PCR-based strategies for the targeted screening of complex biological environments.

Effect of the surface film electric resistance on eddy current detectability of surface cracks in Alloy 600 tubes

International Nuclear Information System (INIS)

Saario, T.; Paine, J.P.N.

1995-01-01

The most widely used technique for NDE of steam generator tubing is eddy current. This technique can reliably detect cracks grown in sodium hydroxide environment only at depths greater than 50% through wall. However, cracking caused by thiosulphate solutions have been detected and sized at shallower depths. The disparity has been proposed to be caused by the different electric resistance of the crack wall surface films and corrosion products in the cracks formed in different environments. This work was undertaken to clarify the role of surface film electric resistance on the disparity found in eddy current detectability of surface cracks in alloy 600 tubes. The proposed model explaining the above mentioned disparity is the following. The detectability of tightly closed cracks by the eddy current technique depends on the electric resistance of the surface films of the crack walls. The nature and resistance of the films which form on the crack walls during operation depends on the composition of the solution inside the crack and close to the crack location. During cooling down of the steam generator, because of contraction and loss of internal pressurization, the cracks are rather tightly closed so that exchange of electrolyte and thus changes in the film properties become difficult. As a result, the surface condition prevailing at high temperature is preserved. If the environment is such that the films formed on the crack walls under operating conditions have low electric resistance, eddy current technique will fail to indicate these cracks or will underestimate the size of these cracks. However, if the electric resistance of the films is high, a tightly closed crack will resemble an open crack and will be easily indicated and correctly sized by eddy current technique

Detection of meca gene from methicillin resistant staphylococcus aureus isolates of north sumatera

Science.gov (United States)

Septiani Nasution, Gabriella; Suryanto, Dwi; Lia Kusumawati, R.

2018-03-01

Methicillin Resistant Staphylococcus aureus (MRSA) is a major pathogen associated with hospital-acquired infections (nosocomial infections). MRSA is a type of S. aureus resistant to the sub-group of beta-lactam antibiotics such as penicillin, cephalosporin, monobactam, and carbapenem. MRSA is resistant because of genetic changes caused by exposure to irrational antibiotic therapy. This study aimed to detect mecA gene in North Sumatra isolates of MRSA and to determine the pattern of antibiotic resistance in S.aureus isolates classified as MRSA by Vitek 2 Compact in the Central Public Hospital Haji Adam Malik, Medan. Samples were 40 isolates of S. aureus classified as MRSA obtained from clinical microbiology specimens. DNA isolation of the isolates was conducted by a method of freeze-thaw cycling. Amplification of mecA gene was done by PCR technique using specific primer for the gene. PCR products were visualized using mini-gel electrophoresis. The results showed that all MRSA isolates showed to have 533 bp band of mecA. Antibiotics test of Vitek 2 Compact showed that despite all isolates were resistant to beta-lactam antibiotics groups; the isolates showed multidrug resistant to other common antibiotics, such as aminoglycosides, macrolides, and fluoroquinolones. However, they were still sensitive to vancomycin (82.5% isolates), linezolid (97.5% isolates), and tigecycline (100% isolates).

Detection of gas in landfills using resistivity measurements; Detektering av gas i deponier med resistivitet

Energy Technology Data Exchange (ETDEWEB)

Rosqvist, Haakan; Leroux, Virginie; Lindsjoe, Magnus (NSR AB, Helsingborg (Sweden)); Dahlin, Torleif (Lund Univ., LTH (Sweden)); Svensson, Mats; Maansson, Carl-Henrik (Tyrens AB, Stockholm (Sweden))

2009-05-15

The main objective with the research project was to develop a methodology to improve the understanding of landfill gas migration in landfills, based on measurements with electrical resistivity. Consequently, the project aimed at an improvement of the utilisation of the energy potential in landfill gas, and to reduce the environmental impact to the atmosphere. Further more, the objective was to improve techniques for investigations of internal structures in landfills. The project also aimed at better understanding of gas migration in the waste body and the mitigation through a landfill cover. Measurements were performed at four landfills; the Biocell reactor (NSR, Helsingborg), the Filborna landfill (NSR, Helsingborg), the Hyllstofta landfill (Naarab, Klippan) and the Flishult landfill (Vetab, Vetlanda). Three dimensional (3D) measurements and analysis were performed. The measurements were repeated in time in order to study changes with time for the resistivity. Supplementary information was created by measurement of other parameters, such as, groundwater table and soil temperature. The results from the resistivity measurements agreed with previous measurements performed at landfills, and thus, the results are therefore regarded as reliable. The measurements showed large temporal and spatial variations, and all of the measurements showed the highest variability near the surface. The results show that the resistivity technique is a powerful tool for investigations of the internal of landfills. Water and gas migration are important features in landfill management and both processes can be detected by using resistivity. Degradation of organic waste results in process with high variability in time and space. Also the degradation rate varies in a landfill and high variability was registered during the resistivity measurements. The high variability in resistivity is likely to be explained by changes in gas pressure and thus indicating gas migration. Therefore, the project

Plasmonic Nanobubbles Rapidly Detect and Destroy Drug-Resistant Tumors

Science.gov (United States)

Lukianova-Hleb, Ekaterina Y.; Ren, Xiaoyang; Townley, Debra; Wu, Xiangwei; Kupferman, Michael E.; Lapotko, Dmitri O.

2012-01-01

The resistance of residual cancer cells after oncological resection to adjuvant chemoradiotherapies results in both high recurrence rates and high non-specific tissue toxicity, thus preventing the successful treatment of such cancers as head and neck squamous cell carcinoma (HNSCC). The patients' survival rate and quality of life therefore depend upon the efficacy, selectivity and low non-specific toxicity of the adjuvant treatment. We report a novel, theranostic in vivo technology that unites both the acoustic diagnostics and guided intracellular delivery of anti-tumor drug (liposome-encapsulated doxorubicin, Doxil) in one rapid process, namely a pulsed laser-activated plasmonic nanobubble (PNB). HNSCC-bearing mice were treated with gold nanoparticle conjugates, Doxil, and single near-infrared laser pulses of low energy. Tumor-specific clusters of gold nanoparticles (solid gold spheres) converted the optical pulses into localized PNBs. The acoustic signals of the PNB detected the tumor with high specificity and sensitivity. The mechanical impact of the PNB, co-localized with Doxil liposomes, selectively ejected the drug into the cytoplasm of cancer cells. Cancer cell-specific generation of PNBs and their intracellular co-localization with Doxil improved the in vivo therapeutic efficacy from 5-7% for administration of only Doxil or PNBs alone to 90% thus demonstrating the synergistic therapeutic effect of the PNB-based intracellular drug release. This mechanism also reduced the non-specific toxicity of Doxil below a detectable level and the treatment time to less than one minute. Thus PNBs combine highly sensitive diagnosis, overcome drug resistance and minimize non-specific toxicity in a single rapid theranostic procedure for intra-operative treatment. PMID:23139725

Development of a real-time PCR melt curve assay for simultaneous detection of virulent and antibiotic resistant Salmonella.

Science.gov (United States)

Singh, Prashant; Mustapha, Azlin

2014-12-01

Multiple drug resistance in Salmonella is an emerging problem in the area of food safety. Depending on the virulence and antibiotic resistance characteristics of the Salmonella strain, infections of varying severity could result. In this study, a multiplex melt curve real-time PCR assay for the detection of virulent and antibiotic resistance strains of Salmonella was developed with two primer sets. The first set targets the virulence gene, invasin (invA), and tetracycline (tetG), streptomycin (aadA2) and sulphonamide (sulI) antibiotic resistance genes, and the second set amplifies ampicillin (blaPSE,blaTEM) and chloramphenicol (floR) resistance genes. The multiplex assay was evaluated using 41 Salmonella strains and was further tested on eight different artificially inoculated food samples. The fluorescent DNA intercalating dye, SYTO9, generated high resolution melt curve peaks and, hence, was used for the development of the assay. This multiplex assay worked efficiently over a DNA concentration range of 20 ng-200 fg and showed a sensitivity of 290 CFU/mL with serially diluted broth cultures. The detection limit for un-enriched artificially inoculated food samples was 10(4) CFU/g, but an enrichment period of 6 h allowed for detection of 10 CFU/g of cells in the samples. Copyright © 2014 Elsevier Ltd. All rights reserved.

Evaluation of genotype MTBDRplus assay for rapid detection of isoniazid and rifampicin resistance in Mycobacterium tuberculosis clinical isolates from Pakistan

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Hasnain Javed

2016-01-01

Conclusions: As evidenced in this study, the major concern with the GenoType MTBDRplus assay were false negative results. In comparison to conventional drug susceptibility testing, the assay was unable to detect 30 (30/100; 30% strains resistant to INH and 23 (23/100; 23% strains resistant to RMP. The GenoType MTBDRplus failed to identify 38 MDR (38/100; 38% strains. Resistance in those strains probably originate from mutations in other codons and/or genes than those covered by the test. For detecting INH and RMP resistance in TB cases, especially in high TB incidence countries, such as Pakistan, molecular approaches should still be a complement rather than areplacement to conventional drug susceptibility testing.

Optimization of a Laboratory-Developed Test Utilizing Roche Analyte-Specific Reagents for Detection of Staphylococcus aureus, Methicillin-Resistant S. aureus, and Vancomycin-Resistant Enterococcus Species▿

OpenAIRE

Mehta, Maitry S.; Paule, Suzanne M.; Hacek, Donna M.; Thomson, Richard B.; Kaul, Karen L.; Peterson, Lance R.

2008-01-01

Nasal and perianal swab specimens were tested for detection of Staphylococcus aureus and vancomycin-resistant Enterococcus species (VRE) using a laboratory-developed real-time PCR test and microbiological cultures. The real-time PCR and culture results for S. aureus were similar. PCR had adequate sensitivity, but culture was more specific for the detection of VRE.

Synergistic interaction of ten essential oils against Haemonchus contortus in vitro

Science.gov (United States)

Anthelmintic resistance in sheep gastrointestinal nematodes is a worldwide problem. Multi-drug resistant haemonchosis is the most serious impediment for small ruminant systems, and there are no new drug candidates currently under development. Molecules from natural sources have demonstrated anthelmi...

Detection of emerging antibiotic resistance in bacteria isolated from subclinical mastitis in cattle in West Bengal

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Arnab Das

2017-05-01

Full Text Available Aim: The aim of this work was to detect antibiotic resistance in Gram-negative bacteria isolated from subclinical mastitis in cattle in West Bengal. Materials and Methods: The milk samples were collected from the cattle suffering with subclinical mastitis in West Bengal. The milk samples were inoculated into the nutrient broth and incubated at 37°C. On the next day, the growth was transferred into nutrient agar and MacConkey agar. All the pure cultures obtained from nutrient agar slant were subjected to Gram-staining and standard biochemical tests. All the bacterial isolates were tested in vitro for their sensitivity to different antibiotics commonly used in veterinary practices. All Gram-negative isolates including positive control were subjected to polymerase chain reaction (PCR for detection of blaCTX-M, blaTEM, blaSHV, blaVIM, tetA, tetB, tetC, and tetM genes considered for extended-spectrum β-lactamase (ESBL, metallo-β-lactamase, and tetracycline resistance. Results: In total, 50 Gram-negative organisms (Escherichia coli, Proteus, Pseudomonas, Klebsiella, and Enterobacter were isolated from milk samples of subclinical mastitis infected cattle. Among these Gram-negative isolates, 48% (24/50 were found either ESBL producing or tetracycline resistant. Out of total 50 Gram-negative isolates, blaCTX-M was detected in 18 (36% isolates, and 6 (12% harbored blaTEM genes in PCR. None of the isolates carried blaSHV genes. Further, in this study, 5 (10% isolates harbored tet(A gene, and 8 (16% isolates carried tet(B gene. No tet(C gene was detected from the isolates. Conclusion: This study showed emerging trend of antibiotic-resistant Gram-negative bacteria associated with subclinical mastitis in cattle in West Bengal, India.

Phenotypic Detection of Metallo-Beta-Lactamases in Carbapenem Resistant Acinetobacter baumannii Isolated from Pediatric Patients in Pakistan

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Muneeza Anwar

2016-01-01

Full Text Available Multidrug resistant A. baumannii has emerged as an important and problematic human pathogen as it is the causative agent of several types of infections especially in neonates and immunocompromised patients because they have least capacity to fight against infections. Carbapenems are used as last resort antibiotics for treating these infections but currently resistance against carbapenems due to MBL production is on the rise. The objective of this study was to determine the frequency of antibiotic resistance in A. baumannii and also to compare the efficacy of combined disk test and double disk synergy test for detection of metallo-beta-lactamases. A total of 112 A. baumannii were identified from various clinical samples and antibiotic susceptibility profile was determined by Kirby-Bauer Disk Diffusion method. Out of 112, 66 (58.9% isolates were resistant to both imipenem and meropenem (OXOID. These resistant isolates were tested for carbapenemase production, and 55 (83.3% were carbapenemase producers by Modified Hodge Test. These isolates were further tested for MBL production by combined disk test and double disk synergy test. Out of 66, 49 isolates were positive by both methods, CDT and DDST, and only one isolate was detected as negative (with kappa value = 0.038. All MBL producing strains showed remarkable resistance to cephalosporins, fluoroquinolones, aminoglycosides, and piperacillin/tazobactam (OXOID. The antibiotic resistance was very high in A. baumannii which were isolated from children in Pakistan specially attending a nephrology unit.

Detection of QTLs for traits associated with pre-harvest sprouting resistance in bread wheat (Triticum aestivum L.).

Science.gov (United States)

Cao, Liangzi; Hayashi, Kazuki; Tokui, Mayumi; Mori, Masahiko; Miura, Hideho; Onishi, Kazumitsu

2016-03-01

Pre-harvest sprouting (PHS) is one of the serious problems for wheat production, especially in rainy regions. Although seed dormancy is the most critical trait for PHS resistance, the control of heading time should also be considered to prevent seed maturation during unfavorable conditions. In addition, awning is known to enhance water absorption by the spike, causing PHS. In this study, we conducted QTL analysis for three PHS resistant related traits, seed dormancy, heading time and awn length, by using recombinant inbred lines from 'Zenkouji-komugi' (high PHS resistance) × 'Chinese Spring' (weak PHS resistance). QTLs for seed dormancy were detected on chromosomes 1B (QDor-1B) and 4A (QDor-4A), in addition to a QTL on chromosome 3A, which was recently cloned as TaMFT-3A. In addition, the accumulation of the QTLs and their epistatic interactions contributed significantly to a higher level of dormancy. QDor-4A is co-located with the Hooded locus for awn development. Furthermore, an effective QTL, which confers early heading by the Zenkouji-komugi allele, was detected on the short arm of chromosome 7B, where the Vrn-B3 locus is located. Understanding the genetic architecture of traits associated with PHS resistance will facilitate the marker assisted selection to breed new varieties with higher PHS resistance.

Determining resistance to mastitis in a bovine subject comprises detecting the presence or absence of a genetic marker that is linked to a trait indicative of mastitis resistance

DEFF Research Database (Denmark)

2007-01-01

The invention relates to a method for determining mastitis resistance in bovine subjects, wherein mastitis resistance comprise resistance to both sub-clinical and clinical mastitis. In particular, the method of the invention involves identification of genetic markers and/or Quantitative Trait Locus...... (QTL) for the determination of mastitis resistance in a bovine subject. The determination of mastitis resistance involves resolution of the specific microsatellite status. Furthermore, the invention relates to a diagnostic kit for detection of genetic marker(s) associated with mastitis resistance....... The method and kit of the present invention can be applied for selection of bovine subjects for breeding purposes. Thus, the invention provides a method of genetically selecting bovine subjects with mastitis resistance, thereby yielding cows less prone to mastitis...

Mechanisms of methicillin resistance in Staphylococcus aureus and methods for laboratory detection.

Science.gov (United States)

Jorgensen, J H

1991-01-01

Three distinctly different mechanisms of methicillin resistance have been described in Staphylococcus aureus. The best-documented and probably most important mechanism is production of a unique, low affinity penicillin-binding protein, PBP 2a. Strains possessing PBP 2a are resistant to methicillin, oxacillin, and probably all other currently available beta-lactam antibiotics. Two additional mechanisms of reduced susceptibility to methicillin have been described. Borderline resistance (BORSA) to the semi-synthetic penicillins has been attributed to the hyperproduction of normal staphylococcal beta-lactamase. A third mechanism has recently been advanced that describes an intermediate level of resistance to methicillin due to production of modified, normal PBPs with reduced affinity for beta-lactams (MODSA). Little is known regarding the prevalence or clinical significance of the BORSA and MODSA strains. The most reliable in vitro susceptibility test methods for detecting MRSA (strains possessing PBP 2a) include the microdilution minimum inhibitory concentration (MIC) test (with 2% NaCl supplemented broth), the oxacillin agar screen plate test (incorporating 6 micrograms/ml oxacillin in 4% NaCl supplemented agar), and the National Committee for Clinical Laboratory Standards (NCCLS) disk diffusion test with oxacillin. All three methods use direct inoculum preparation and incubation of tests at 35 degrees C for a full 24 hours.

Environmental determinants of total IgE among school children living in the rural Tropics: importance of geohelminth infections and effect of anthelmintic treatment

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Benitez Susana M

2008-06-01

Full Text Available Abstract Background The environmental factors that determine the elevated levels of polyclonal IgE observed in populations living in the Tropics are poorly understood but may include geohelminth infections. We investigated the association between geohelminth infections and total IgE levels in school children in rural tropical Ecuador, and assessed the effect on IgE of repeated anthelmintic treatments over a period of 12 months. The study was nested within a cluster-randomized study that randomized 68 schools to receive either 400 mg of albendazole every 2 months over a year or no treatment. We studied random samples of children completing follow-up and representing four groups stratified by the presence of geohelminth infection at baseline and treatment allocation. We measured levels of total IgE and anti-A. lumbricoides IgG (used as a measure of past and current geohelminth infectious exposure in blood samples collected at the start of the study and after 12 months. Results We observed elevated levels of total IgE (compared to standard reference values at the start of the study in this population of school children (geometric mean, 1,004 IU/mL, range 12 to 22,608 IU/mL and baseline IgE levels were strongly associated with parameters of geohelminth infection but not with age, nutritional and socioeconomic status. After 12 months, levels of IgE fell significantly in the treatment (by 35.1% and no treatment (by 10.4% groups, respectively, but the fall was significantly greater in the treatment group. Falls in IgE were independently associated with albendazole treatment, having a baseline geohelminth infection and with high baseline levels of anti-A. lumbricoides IgG. Increases in IgE at 12 months were associated with the presence of geohelminth infections and increasing levels of anti-A. lumbricoides IgG at 12 months independent of treatment allocation. Conclusion The data provide evidence that geohelminth infections are an important determinant

Molecular characterization of two high-level ceftriaxone-resistant Neisseria gonorrhoeae isolates detected in Catalonia, Spain.

Science.gov (United States)

Cámara, Jordi; Serra, Judit; Ayats, Josefina; Bastida, Teresa; Carnicer-Pont, Dolors; Andreu, Antònia; Ardanuy, Carmen

2012-08-01

The aim of this study was to characterize the first two extended-spectrum cephalosporin-resistant and multidrug-resistant (MDR) Neisseria gonorrhoeae isolates collected from two sexually related patients (men who have sex with men) in Spain. Antimicrobial susceptibility was studied by Etest. Genes involved in quinolone, ceftriaxone and multidrug resistance were amplified by PCR and sequenced in both directions. The isolates were typed by N. gonorrhoeae multi-antigen sequence typing (NG-MAST). The two isolates had the same MDR profile, showing resistance to penicillin (MIC 0.094 mg/L; β-lactamase negative), ceftriaxone (MIC 1.5 mg/L), cefixime (MIC 1.5 mg/L), cefotaxime (MIC 1 mg/L), ciprofloxacin (MIC >32 mg/L) and tetracycline (MIC 1.5 mg/L). NG-MAST showed that both isolates belonged to sequence type (ST) 1407 (porB-908 and tbpB-110). Ciprofloxacin resistance was due to amino acid substitutions in GyrA (S91F and D95G) and ParC (S87R). An A deletion in the promoter of the MtrCDE efflux pump (mtrR) was detected. No changes were detected in the pilQ gene. The outer membrane protein PorB showed two substitutions at G120K and A121N. An L421P substitution was observed in the PBP1A (ponA) sequence. The sequence of PBP2 (penA) showed a mosaic structure related to genotype XXXIV with a single additional amino acid substitution (A501P). This genotype was identical to a recently described French isolate (F89). This is the first reported case of high-level extended-spectrum cephalosporin-resistant N. gonorrhoeae transmission. The molecular typing and MDR genotype suggest possible European spread of this strain, highlighting the need for surveillance and the importance of testing the susceptibility of N. gonorrhoeae to extended-spectrum cephalosporins.

Nanoparticle mechanics: deformation detection via nanopore resistive pulse sensing

Science.gov (United States)

Darvish, Armin; Goyal, Gaurav; Aneja, Rachna; Sundaram, Ramalingam V. K.; Lee, Kidan; Ahn, Chi Won; Kim, Ki-Bum; Vlahovska, Petia M.; Kim, Min Jun

2016-07-01

liposomes inside nanopores. We observed a significant difference in resistive pulse characteristics between soft liposomes and rigid polystyrene nanoparticles especially at higher applied voltages. We used theoretical simulations to demonstrate that the difference can be explained by shape deformation of liposomes as they translocate through the nanopores. Comparing our results with the findings from electrodeformation experiments, we demonstrated that the rigidity of liposomes can be qualitatively compared using resistive pulse characteristics. This application of nanopores can provide new opportunities to study the mechanics at the nanoscale, to investigate properties of great value in fundamental biophysics and cellular mechanobiology, such as virus deformability and fusogenicity, and in applied sciences for designing novel drug/gene delivery systems. Electronic supplementary information (ESI) available: Supplementary figures; signal processing and event detection; multiphysics simulation in COMSOL. See DOI: 10.1039/c6nr03371g

Anthelmintic Resistance of Strongyle Nematodes to Ivermectin and Fenbendazole on Cart Horses in Gondar, Northwest Ethiopia

OpenAIRE

Seyoum, Zewdu; Zewdu, Alemu; Dagnachew, Shimelis; Bogale, Basazinew

2017-01-01

A study was conducted from November 2015 to April 2016 to determine fenbendazole and ivermectin resistance status of intestinal nematodes of cart horses in Gondar, Northwest Ethiopia. Forty-five strongyle infected animals were used for this study. The animals were randomly allocated into three groups (15 horses per group). Group I was treated with fenbendazole and Group II with ivermectin and Group III was left untreated. Faecal samples were collected from each cart horse before and after tre...

Detection of clarithromycin-resistant Helicobacter pylori by polymerase chain reaction using residual samples from rapid urease test

Directory of Open Access Journals (Sweden)

Jae-Sik Jeon

2017-01-01

Full Text Available Background: Approximately 50% of the world population is infected with Helicobacter pylori, which corresponds to a high infection rate. Furthermore, the incidence of antibiotic-resistant H. pylori has increased with the recent rise in use of antibiotics for H. pylori elimination, suggesting growing treatment failures. Aim: The study was aimed to assess the use of residual samples from rapid urease test (RUT for biomolecular testing as an effective and accurate method to detect antibiotic-resistant H. pylori. Settings and Design: This study was a retrospective study performed using data obtained from medical records of previously isolated H. pylori strains. Materials and Methods: RUT was conducted for 5440 biopsy samples from individuals who underwent health examination in South Korea. Subsequently, 469 RUT residual samples were randomly selected and subjected to polymerase chain reaction (PCR to detect antibiotic-resistant H. pylori. Statistical Analysis Used: The Chi-square test was used to analyse categorical data. P < 0.05 was considered statistically significant. Results: The results showed a concordance between the results of PCR and conventional RUT in 450 of 469 samples, suggesting that the H. pylori PCR test is a time- and cost-effective detection method. Conclusions: This study demonstrated that PCR test can aid physicians to prescribe the appropriate antibiotics at the time of diagnosis, thus preventing the reduction in H. pylori eradication due to antibiotic resistance, averting progression to serious diseases and increasing the treatment success rate.

Current Status for Gastrointestinal Nematode Diagnosis in Small Ruminants: Where Are We and Where Are We Going?

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Sarah Jane Margaret Preston

2014-01-01

Full Text Available Gastrointestinal nematode (GIN parasites pose a significant economic burden particularly in small ruminant production systems. Anthelmintic resistance is a serious concern to the effective control of GIN parasites and has fuelled the focus to design and promote sustainable control of practices of parasite control. Many facets of sustainable GIN parasite control programs rely on the ability to diagnose infection both qualitatively and quantitatively. Diagnostics are required to determine anthelmintic efficacies, for targeted treatment programs and selection of animals for parasite resistant breeding. This review describes much of the research investigated to date to improve the current diagnostic for the above practices which is based on counting the number of parasite eggs in faeces.

Multi-wall carbon nanotube networks as potential resistive gas sensors for organic vapor detection

Czech Academy of Sciences Publication Activity Database

Slobodian, P.; Říha, Pavel; Lengálová, A.; Svoboda, P.; Sáha, P.

2011-01-01

Roč. 49, č. 7 (2011), s. 2499-2507 ISSN 0008-6223 Institutional research plan: CEZ:AV0Z20600510 Keywords : carbon nanotube network * KMnO 4 oxidation * electrical resistance * organic vapor detection * adsorption /desorption cycles Subject RIV: JB - Sensors, Measurment, Regulation Impact factor: 5.378, year: 2011

Reprofiling a classical anthelmintic, pyrvinium pamoate, as an anti-cancer drug targeting mitochondrial respiration

Energy Technology Data Exchange (ETDEWEB)

Ishii, Isao [Department of Biochemistry, Keio University Graduate School of Pharmaceutical Sciences, Tokyo (Japan); Harada, Yasuo [Fujii Memorial Research Institute, Otsuka Pharmaceutical Co., Ltd., Shiga (Japan); Kasahara, Tadashi, E-mail: isao-ishii@umin.ac.jp [Department of Biochemistry, Keio University Graduate School of Pharmaceutical Sciences, Tokyo (Japan)

2012-10-02

Pyrvinium pamoate (PP) is an FDA-approved classical anthelmintic, but is now attracting particular attention as an anti-cancer drug after recent findings of its potent cytotoxicity against various cancer cell lines only during glucose starvation, as well as its anti-tumor activity against hypovascular pancreatic cancer cells transplanted in mice. The molecular mechanisms by which PP promotes such preferential toxicity against cancer cells are currently under extensive investigation. PP suppressed the NADH-fumarate reductase system that mediates a reverse reaction of the mitochondrial electron-transport chain complex II in anaerobic organisms such as parasitic helminthes or mammalian cells under tumor microenvironment-mimicking hypoglycemic/hypoxic conditions, thereby inhibiting efficient ATP production. PP also inhibited the unfolded protein response induced by glucose starvation, thereby inhibiting the proliferation of pancreatic cancer cells. Even under normoglycemic/normoxic conditions, PP suppressed the mitochondrial electron-transport chain complex I and thereby STAT3, inhibiting the proliferation of myeloma/erythroleukemia cells. Here, we review accumulating knowledge on its working mechanisms and evaluate PP as a novel anti-cancer drug that targets mitochondrial respiration.

Reprofiling a classical anthelmintic, pyrvinium pamoate, as an anti-cancer drug targeting mitochondrial respiration.

Directory of Open Access Journals (Sweden)

Isao eIshii

2012-10-01

Full Text Available Pyrvinium pamoate (PP is an FDA-approved classical anthelmintic, but is now attracting particular attention as an anti-cancer drug after recent findings of its potent cytotoxicity against various cancer cell lines only during glucose starvation, as well as its anti-tumor activity against hypovascular pancreatic cancer cells transplanted in mice. The molecular mechanisms by which PP promotes such preferential toxicity against cancer cells are currently under extensive investigation. PP suppressed the NADH-fumarate reductase system that mediates a reverse reaction of the mitochondrial electron-transport chain complex II in anaerobic organisms such as parasitic helminthes or mammalian cells under tumor microenvironment-mimicking hypoglycemic/hypoxic conditions, thereby inhibiting efficient ATP production. PP also inhibited the unfolded protein response induced by glucose starvation, thereby inhibiting the proliferation of pancreatic cancer cells. Even under normoglycemic/normoxic conditions, PP suppressed the mitochondrial electron-transport chain complex I and thereby STAT3, inhibiting the proliferation of myeloma/erythroleukemia cells. Here, we review accumulating knowledge on its working mechanisms and evaluate PP as a novel anti-cancer drug that targets mitochondrial respiration.

Reprofiling a classical anthelmintic, pyrvinium pamoate, as an anti-cancer drug targeting mitochondrial respiration

International Nuclear Information System (INIS)

Ishii, Isao; Harada, Yasuo; Kasahara, Tadashi

2012-01-01

Pyrvinium pamoate (PP) is an FDA-approved classical anthelmintic, but is now attracting particular attention as an anti-cancer drug after recent findings of its potent cytotoxicity against various cancer cell lines only during glucose starvation, as well as its anti-tumor activity against hypovascular pancreatic cancer cells transplanted in mice. The molecular mechanisms by which PP promotes such preferential toxicity against cancer cells are currently under extensive investigation. PP suppressed the NADH-fumarate reductase system that mediates a reverse reaction of the mitochondrial electron-transport chain complex II in anaerobic organisms such as parasitic helminthes or mammalian cells under tumor microenvironment-mimicking hypoglycemic/hypoxic conditions, thereby inhibiting efficient ATP production. PP also inhibited the unfolded protein response induced by glucose starvation, thereby inhibiting the proliferation of pancreatic cancer cells. Even under normoglycemic/normoxic conditions, PP suppressed the mitochondrial electron-transport chain complex I and thereby STAT3, inhibiting the proliferation of myeloma/erythroleukemia cells. Here, we review accumulating knowledge on its working mechanisms and evaluate PP as a novel anti-cancer drug that targets mitochondrial respiration.

Bioaerosol emissions and detection of airborne antibiotic resistance genes from a wastewater treatment plant

Science.gov (United States)

Li, Jing; Zhou, Liantong; Zhang, Xiangyu; Xu, Caijia; Dong, Liming; Yao, Maosheng

2016-01-01

Air samples from twelve sampling sites (including seven intra-plant sites, one upwind site and four downwind sites) from a wastewater treatment plant (WWTP) in Beijing were collected using a Reuter Centrifugal Sampler High Flow (RCS); and their microbial fractions were studied using culturing and high throughput gene sequence. In addition, the viable (fluorescent) bioaerosol concentrations for 7 intra-plant sites were also monitored for 30 min each using an ultraviolet aerodynamic particle sizer (UV-APS). Both air and water samples collected from the plant were investigated for possible bacterial antibiotic resistance genes and integrons using polymerase chain reaction (PCR) coupled with gel electrophoresis. The results showed that the air near sludge thickening basin was detected to have the highest level of culturable bacterial aerosols (up to 1697 CFU/m3) and fungal aerosols (up to 930 CFU/m3). For most sampling sites, fluorescent peaks were observed at around 3-4 μm, except the office building with a peak at 1.5 μm, with a number concentration level up to 1233-6533 Particles/m3. About 300 unique bacterial species, including human opportunistic pathogens, such as Comamonas Testosteroni and Moraxella Osloensis, were detected from the air samples collected over the biological reaction basin. In addition, we have detected the sul2 gene resistant to cotrimoxazole (also known as septra, bactrim and TMP-SMX) and class 1 integrase gene from the air samples collected from the screen room and the biological reaction basin. Overall, the screen room, sludge thickening basin and biological reaction basin imposed significant microbial exposure risks, including those from airborne antibiotic resistance genes.

Evaluation of efficiency of nested multiplex allele-specific PCR assay for detection of multidrug resistant tuberculosis directly from sputum samples.

Science.gov (United States)

Mistri, S K; Sultana, M; Kamal, S M M; Alam, M M; Irin, F; Nessa, J; Ahsan, C R; Yasmin, M

2016-05-01

For an effective control of tuberculosis, rapid detection of multidrug resistant tuberculosis (MDR-TB) is necessary. Therefore, we developed a modified nested multiplex allele-specific polymerase chain reaction (MAS-PCR) method that enables rapid MDR-TB detection directly from sputum samples. The efficacy of this method was evaluated using 79 sputum samples collected from suspected tuberculosis patients. The performance of nested MAS-PCR method was compared with other MDR-TB detection methods like drug susceptibility testing (DST) and DNA sequencing. As rifampicin (RIF) resistance conforms to MDR-TB in greater than 90% cases, only the presence of RIF-associated mutations in rpoB gene was determined by DNA sequencing and nested MAS-PCR to detect MDR-TB. The concordance between nested MAS-PCR and DNA sequencing results was found to be 96·3%. When compared with DST, the sensitivity and specificity of nested MAS-PCR for RIF-resistance detection were determined to be 92·9 and 100% respectively. For developing- and high-TB burden countries, molecular-based tests have been recommended by the World Health Organization for rapid detection of MDR-TB. The results of this study indicate that, nested MAS-PCR assay might be a practical and relatively cost effective molecular method for rapid detection of MDR-TB from suspected sputum samples in developing countries with resource poor settings. © 2016 The Society for Applied Microbiology.

Real-time polymerase chain reaction assay for the rapid detection and characterization of chloroquine-resistant Plasmodium falciparum malaria in returned travelers.

Science.gov (United States)

Farcas, Gabriella A; Soeller, Rainer; Zhong, Kathleen; Zahirieh, Alireza; Kain, Kevin C

2006-03-01

Imported drug-resistant malaria is a growing problem in industrialized countries. Rapid and accurate diagnosis is essential to prevent malaria-associated mortality in returned travelers. However, outside of a limited number of specialized centers, the microscopic diagnosis of malaria is slow, unreliable, and provides little information about drug resistance. Molecular diagnostics have the potential to overcome these limitations. We developed and evaluated a rapid, real-time polymerase chain reaction (PCR) assay to detect Plasmodium falciparum malaria and chloroquine (CQ)-resistance determinants in returned travelers who are febrile. A real-time PCR assay based on detection of the K76T mutation in PfCRT (K76T) of P. falciparum was developed on a LightCycler platform (Roche). The performance characteristics of the real-time assay were compared with those of the nested PCR-restriction fragment-length polymorphism (RFLP) and the sequence analyses of samples obtained from 200 febrile returned travelers, who included 125 infected with P. falciparum (48 of whom were infected CQ-susceptible [K76] and 77 of whom were CQ-resistant [T76] P. falciparum), 22 infected with Plasmodium vivax, 10 infected with Plasmodium ovale, 3 infected with Plasmodium malariae malaria, and 40 infected with other febrile syndromes. All patient samples were coded, and all analyses were performed blindly. The real-time PCR assay detected multiple pfcrt haplotypes associated with CQ resistance in geographically diverse malaria isolates acquired by travelers. Compared with nested-PCR RFLP (the reference standard), the real-time assay was 100% sensitive and 96.2% specific for detection of the P. falciparum K76T mutation. This assay is rapid, sensitive, and specific for the detection and characterization of CQ-resistant P. falciparum malaria in returned travelers. This assay is automated, standardized, and suitable for routine use in clinical diagnostic laboratories.

A locked nucleic acid (LNA-based real-time PCR assay for the rapid detection of multiple bacterial antibiotic resistance genes directly from positive blood culture.

Directory of Open Access Journals (Sweden)

Lingxiang Zhu

Full Text Available Bacterial strains resistant to various antibiotic drugs are frequently encountered in clinical infections, and the rapid identification of drug-resistant strains is highly essential for clinical treatment. We developed a locked nucleic acid (LNA-based quantitative real-time PCR (LNA-qPCR method for the rapid detection of 13 antibiotic resistance genes and successfully used it to distinguish drug-resistant bacterial strains from positive blood culture samples. A sequence-specific primer-probe set was designed, and the specificity of the assays was assessed using 27 ATCC bacterial strains and 77 negative blood culture samples. No cross-reaction was identified among bacterial strains and in negative samples, indicating 100% specificity. The sensitivity of the assays was determined by spiking each bacterial strain into negative blood samples, and the detection limit was 1-10 colony forming units (CFU per reaction. The LNA-qPCR assays were first applied to 72 clinical bacterial isolates for the identification of known drug resistance genes, and the results were verified by the direct sequencing of PCR products. Finally, the LNA-qPCR assays were used for the detection in 47 positive blood culture samples, 19 of which (40.4% were positive for antibiotic resistance genes, showing 91.5% consistency with phenotypic susceptibility results. In conclusion, LNA-qPCR is a reliable method for the rapid detection of bacterial antibiotic resistance genes and can be used as a supplement to phenotypic susceptibility testing for the early detection of antimicrobial resistance to allow the selection of appropriate antimicrobial treatment and to prevent the spread of resistant isolates.

ANTHELMINTIC ACTIVITY OF LAPACHOL, β-LAPACHONE AND ITS DERIVATIVES AGAINST Toxocara canis LARVAE

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Taís MATA-SANTOS

2015-06-01

Full Text Available Anthelmintics used for intestinal helminthiasis treatment are generally effective; however, their effectiveness in tissue parasitosis (i.e. visceral toxocariasis is moderate. The aim of this study was to evaluate the in vitro activity of lapachol, β-lapachone and phenazines in relation to the viability of Toxocara canis larvae. A concentration of 2 mg/mL (in duplicate of the compounds was tested using microculture plates containing Toxocara canis larvae in an RPMI-1640 environment, incubated at 37 °C in 5% CO2 tension for 48 hours. In the 2 mg/mL concentration, four phenazines, lapachol and three of its derivatives presented a larvicide/larvistatic activity of 100%. Then, the minimum larvicide/larvistatic concentration (MLC test was conducted. The compounds that presented the best results were nor-lapachol (MLC, 1 mg/mL, lapachol (MLC 0.5 mg/mL, β-lapachone, and β-C-allyl-lawsone (MLC, 0.25 mg/mL. The larvae exposed to the compounds, at best MLC with 100% in vitro activity larvicide, were inoculated into healthy BALB/c mice and were not capable of causing infection, confirming the larvicide potential in vitro of these compounds.

Phenotypic and genotypic detection of ESBL mediated cephalosporin resistance in Klebsiella pneumoniae: emergence of high resistance against cefepime, the fourth generation cephalosporin.

Science.gov (United States)

Grover, S S; Sharma, Meenakshi; Chattopadhya, D; Kapoor, Hema; Pasha, S T; Singh, Gajendra

2006-10-01

Cephalosporins belonging to second and third generation are commonly used in India for the treatment of Klebsiella pneumoniae. Report on resistance among K. pneumoniae strains to second and third generation cephalosporins are on rise in this country, which has been attributed to emergence of strains expressing extended-spectrum beta-lactamases (ESBLs). The aim of this study was to evaluate the in vitro susceptibility of K. pneumoniae to broad-spectrum cephalosporins particularly to cefepime, a recently introduced fourth generation cephalosporin in relation to ESBL production. This study has been carried out in two phases among K. pneumoniae strains isolated between October 2001 and September 2002 (phase I, before marketing of cefepime in India) and between August 2003 and July 2004 (phase II, after marketing of cefepime in India). Minimum Inhibitory Concentration (MIC) was determined by a commercial strip containing gradient of antimicrobials (Strip E-test). Detection for ESBL production was carried out by DDST, E-test ESBL and PCR. Antimicrobial resistance profile of K. pneumoniae strains to five cephalosporins as analyzed by WHONET 5 identified 15 different resistance profiles among the 108 phase I isolates, ranging from resistance to none (19.44%) to all the five cephalosporin (8.33%) and eight different resistance profiles among the 99 phase II isolates, ranging from resistance to none (9.1%) to all the five cephalosporins (36.4%). Among the 108 phase I isolates a total of 71 (65.72%) and out of 99 phase II isolates, a total of 87 (88.0%) could be identified as ESBL producers. Among the isolates, regardless of the phase of the isolation, those characterized by production of ESBL showed overall higher frequency of resistance to cephalosporins (range 19.7-85.9% and 51.7-100% in phase I and phase II, respectively) compared to those for ESBL non-producers (range 0-13.5% and 0-25% in phase I and phase II, respectively). Ten randomly selected isolates from the most

Contribution of katG, ahpC and inhA mutations to the detection of isoniazid-resistant Mycobacterium tuberculosis isolates from Lebanon and Syria

Directory of Open Access Journals (Sweden)

F Dabboussi

2015-01-01

Conclusions: This study showed that the pyrosequencing applied to katG, inhA promoter and ahpC-oxyR intergenic region was able to detect a relatively large proportion of Syrian INH-resistant MTB isolates (80.7% in Syria. This strategy may be inappropriate for Lebanese strains, as the genetic mechanisms of resistance remain unidentified for approximately half of the isolates, so it is quite possible to detect the presence of other mechanisms of resistance.

Development of a Real-Time Resistance Measurement for Vibrio parahaemolyticus Detection by the Lecithin-Dependent Hemolysin Gene

Science.gov (United States)

Xiang, Guiming; Pu, Xiaoyun; Jiang, Dongneng; Liu, Linlin; Liu, Chang; Liu, Xiaobo

2013-01-01

The marine bacterium Vibrio parahaemolyticus (V. parahaemolyticus) causes gastroenteritis in humans via the ingestion of raw or undercooked contaminated seafood, and early diagnosis and prompt treatment are important for the prevention of V. parahaemolyticus-related diseases. In this study, a real-time resistance measurement based on loop-mediated isothermal amplification (LAMP), electrochemical ion bonding (Crystal violet and Mg2+), real-time monitoring, and derivative analysis was developed. V. parahaemolyticus DNA was first amplified by LAMP, and the products (DNA and pyrophosphate) represented two types of negative ions that could combine with a positive dye (Crystal violet) and positive ions (Mg2+) to increase the resistance of the reaction liquid. This resistance was measured in real-time using a specially designed resistance electrode, thus permitting the quantitative detection of V. parahaemolyticus. The results were obtained in 1–2 hours, with a minimum bacterial density of 10 CFU.mL−1 and high levels of accuracy (97%), sensitivity (96.08%), and specificity (97.96%) when compared to cultivation methods. Therefore, this simple and rapid method has a potential application in the detection of V. parahaemolyticus on a gene chip or in point-of-care testing. PMID:23991096

Detection of resistance, cross-resistance, and stability of resistance to new chemistry insecticides in Bemisia tabaci (Homoptera: Aleyrodidae).

Science.gov (United States)

Basit, Muhammad; Saeed, Shafqat; Saleem, Mushtaq Ahmad; Denholm, Ian; Shah, Maqbool

2013-06-01

Resistance levels in whitefly, Bemisia tabaci (Gennadius) collections from cotton and sunflower (up to four districts) for five neonicotinoids and two insect growth regulators (IGRs) were investigated for two consecutive years. Based on the LC50(s), all collections showed slight to moderate levels of resistance for the tested insecticides compared with the laboratory susceptible population. The data also indicated that cotton and sunflower collections had similar resistance levels. In comparison (four collections), Vehari collections showed higher resistance for acetamiprid, thiacloprid, and nitenpyram compared with those of others. Average resistance ratios for acetamiprid, thiacloprid, and nitenpyram ranged from 5- to 13-, 4- to 8-, and 9- to 13-fold, respectively. Multan and Vehari collections also exhibited moderate levels (9- to 16-fold) of resistance to buprofezin. Furthermore, toxicity of neonicotinoids against immature stages was equal to that of insect growth regulators. The data also suggested that resistance in the field populations was stable. After selection for four generations with bifenthrin (G1 to G4), resistance to bifenthrin increased to 14-fold compared with the laboratory susceptible population. Selection also increased resistance to fenpropathrin, lambdacyhalothrin, imidacloprid, acetamiprid, and diafenthuron. Cross-resistance and stability of resistance in the field populations is of some concern. Rotation of insecticides having no cross-resistance and targeting the control against immature stages may control the resistant insects, simultaneously reducing the selection pressure imposed.

Determining resistance to mastitis in a bovine subject involves detecting presence or absence of genetic marker associated with trait indicative of mastitis resistance of the bovine subject and/or off-spring from it

DEFF Research Database (Denmark)

2010-01-01

NOVELTY - Determining (m1) resistance to mastitis in a bovine subject, involves detecting in a sample from the bovine subject the presence or absence of at least one genetic marker that is associated with at least one trait indicative of mastitis resistance of the bovine subject and/or off......-spring from it, where the genetic marker is located on the bovine chromosome BTA11 in the region flanked by and including the zeta-chain associated protein 70kD (ZAP70) and CD8B genes, where the presence or absence of the genetic marker is indicative of mastitis resistance. USE - For determining resistance...... to mastitis in a bovine subject for determining mastitis resistance in a bovine subject; for detecting the presence or absence in a bovine subject of at least one genetic marker associated with resistance to mastitis; and for estimating breeding value in respect of susceptibility to mastitis in a bovine...

Molecular detection and antimicrobial resistance of diarrheagenic Escherichia coli strains isolated from diarrheal cases

International Nuclear Information System (INIS)

Aslani, Mehdi M.; Salmanzadeh-Ahrabi, S.; Jafari, F.; Zali, Reza M.; Mani, M.; Alikhani, Yousef M.

2008-01-01

Objective was to identify and classify Iranian isolates of diarrheagenic Escherichia coli (E. coli) on the basis of presence of virulence genes and to determine antibiotic susceptibility of isolated strains. The current cross-sectional study was conducted in 2005 at the Pasteur Institute, Tehran, Iran. One hundred and ninety-three diarrheagenic E. coli isolated from diarrheal patients in different regions of Iran were included in current study. Virulence factors genees for diarrheagenic E. coli were detected by polymerase chain reaction. Of the 193 diarrheagenic E. coli detected by PCR, 86(44.5%) were Shiga toxin-producing E. coli (STEC), 74 (38.4%) enteropathogenic E. coli (EPEC), 19 (9.8%) enteroaggregative E. coli and 14 (7.3%) enterotoxigenic E. coli isolates. Susceptibility to 12 clinically important antimicrobial agents was determined for 193 strains of diarrhheagenic E. coli. A high incidence of resistance to tetracycline (63%), ampicillin (62%), streptomycin (56%), amoxicillin/clavulanic acid (44.5%), trimetoprim/sulphamethoxazole (39.5%) and cephalothin (37%) was observed. The STEC and EPEC strains with high resistance to tetracycline and ampicillin but highly susceptible to quinolones are among the most important causative agent of diarrhea in Iran. This study suggests that antimicrobial resistance is wide spread among E. coli strains colonizing Iranian patients. Guidelines for appropriate use of antibiotics in developing countries require updating. (author)

Simultaneous detection of hepatitis B virus genotypes and mutations associated with resistance to lamivudine, adefovir, and telbivudine by the polymerase chain reaction-ligase detection reaction

Directory of Open Access Journals (Sweden)

Yong-Zhong Wang

Full Text Available OBJECTIVES: Detection of mutations associated to nucleos(tide analogs and hepatitis B virus (HBV genotyping are essential for monitoring treatment of HBV infection. We developed a multiplex polymerase chain reaction-ligase detection reaction (PCR-LDR assay for the rapid detection of HBV genotypes and mutations associated with lamivudine, adefovir, and telbivudine resistance in HBV-infected patients. METHODS: HBV templates were amplified by PCR, followed by LDR and electrophoresis on a sequencer. The assay was evaluated using plasmids that contained wild-type or mutant HBV sequences and 216 clinical samples. RESULTS: The PCR-LDR assay and sequencing gave comparable results for 158 of the 216 samples (73.1% with respect to mutation detection and genotyping. Complete agreement between the two methods was observed for all the samples (100% at codon 180 and codon 204. Concordant results were observed for 99.4% of the 158 samples at codon 181 and 98.7% at codon 236. The genotyping results were completely concordant between the PCR-LDR assay and sequencing. The PCR-LDR assay could detect a proportion of 1% mutant plasmid in a background of wild-type plasmid. CONCLUSION: The PCR-LDR assay is sensitive and specific for detection of HBV genotypes and drug resistance mutations, and could be helpful for decision making in the treatment of HBV infection.

Autoradiographic detection of diphtheria toxin resistant mutants in human diploid fibroblasts

International Nuclear Information System (INIS)

Gupta, R.S.; Singh, B.

1985-01-01

An autoradiographic procedure for the detection of diphtheria toxin (DT) resistant (Dip/sub R/) mutants in human diploid fibroblast (HDF) cells has been developed. The assay is based on the observation that when HDFs from confluent cultures are seeded in medium containing 0.01 flocculating units/ml or higher concentration of DT, protein synthesis in sensitive cells is severely inhibited by 4-6 hr. If at this or later time, a radiolabeled protein precursor (eg, 3 H-leucine) is added to the culture, it is almost exclusively incorporated into the resistant cells, which are then readily identified by autoradiography. These studies provide strong evidence that the labeled cells identified by autoradiography are bona fide Dip/sub R/ mutants. The detection of Dip/sub R/ cells by autoradiography is apparently not affected by the presence of the sensitive cells in the mixtures. The spontaneous frequency of Dip/sub R/ cells in HDFs has been found to be in the range of 1-5 x 10 -6 , and this increases in a dose dependent manner upon treatment with the mutagen ethyl methanesulfonate. These results indicate that the autoradiographic assay could be used for quantitative mutagenesis. Since the autoradiographic assay does not depend on cell division, it may prove useful in estimating the incidence of pre-existing mutations in cell populations that either do not divide or have very limited growth potential (eg, lymphocytes, muscle cells, neurons, senescent fibroblasts, etc.)

Phenotypic and Genotypic Detection of Metallo-beta-lactamases among Imipenem-Resistant Gram Negative Isolates

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Mohammad Mohammadzadeh

2016-08-01

Full Text Available Background:   Imipenem-resistant gram negative bacteria, resulting from metallo-beta-lactamase (MBLs-producing strains have been reported to be among the important causes of nosocomial infections and of serious therapeutic problem worldwide. Because of their broad range, potent carbapenemase activity and resistance to inhibitors, these enzymes can confer resistance to almost all beta-lactams. The prevalence of metallo-beta-lactamase among imipenem-resistant Acinetobacter spp., Pseudomonas spp. and Enerobacteriaceae isolates is determined.Methods:   In this descriptive study 864 clinical isolates of Acinetobacter spp., Pseudomonas spp. and Enterobacteriaceae, were initially tested for imipenem susceptibility. The metallo-beta-lactamase production was detected using combined disk diffusion, double disk synergy test, and Hodge test. Then all imipenem resistant isolates were tested by PCR for imp, vim and ndm genes. Results:   Among 864 isolates, 62 (7.17 % were imipenem-resistant. Positive phonetypic test for metallo-beta-lactamase was 40 (64.5%, of which 24 (17.1% and 16 (9.2% isolates were Acinetobacter spp. and Pseudomonas spp., respectively. By PCR method 30 (48.4% of imipenem resistant Acinetobacter, and Pseudomonas isolates were positive for MBL-producing genes. None of the Enterobacteriaceae isolates were positive for metallo-beta-lactamase activity. Conclusion:   The results of this study are indicative of the growing number of nosocomial infections associated with multidrug-resistant gram negative bacteria in this region leading to difficulties in antibiotic therapy. Thereby, using of phenotypic methods can be helpful for management of this problem.

Detection and linkage to mobile genetic elements of tetracycline resistance gene tet(M) in Escherichia coli isolates from pigs

DEFF Research Database (Denmark)

Jurado-Rabadan, Sonia; de la Fuente, Ricardo; Ruiz-Santa-Quiteria, Jose A.

2014-01-01

Background: In Escherichia coli the genes involved in the acquisition of tetracycline resistance are mainly tet(A) and tet(B). In addition, tet(M) is the most common tetracycline resistance determinant in enterococci and it is associated with conjugative transposons and plasmids. Although tet......(M) has been identified in E. coli, to our knowledge, there are no previous reports studying the linkage of the tet(M) gene in E. coli to different mobile genetic elements. The aim of this study was to determine the occurrence of tet(A), tet(B), and tet(M) genes in doxycycline-resistant E. coli isolates...... from pigs, as well as the detection of mobile genetic elements linked to tet(M) in E. coli and its possible transfer from enterococci. Results: tet(A) was the most frequently detected gene (87.9%) in doxycycline-resistant isolates. tet(M) was found in 13.1% E. coli isolates. The tet(M) gene...

Worm control practice against gastro-intestinal parasites in Norwegian sheep and goat flocks

Science.gov (United States)

2011-01-01

Background Anthelmintic treatment is the most common way of controlling nematode infections in ruminants. However, several countries have reported anthelmintic resistance (AR), representing a limitation for sustainable small ruminant production. The knowledge regarding worm control management represents a baseline to develop a guideline for preventing AR. The aim of the present study was therefore to improve our knowledge about the worm control practices in small ruminant flocks in Norway. Methods A questionnaire survey regarding worm control practices was performed in small ruminant flocks in Norway. Flocks were selected from the three main areas of small ruminant farming, i.e. the coastal, inland and northern areas. A total of 825 questionnaires, comprising 587 sheep flocks (return rate of 51.3%) and 238 goat flocks (52.6%) were included. Results The results indicated that visual appraisal of individual weight was the most common means of estimating the anthelmintic dose used in sheep (78.6%) and goat (85.1%) flocks. The mean yearly drenching rate in lambs and ewes were 2.5 ± 1.7 and 1.9 ± 1.1, respectively, whereas it was 1.0 (once a year) in goats. However, these figures were higher in sheep in the coastal area with a rate of 3.4 and 2.2 in lambs and ewes, respectively. Benzimidazoles were the predominant anthelmintic class used in sheep flocks (64.9% in 2007), whereas benzimidazoles and macrocyclic lactones were both equally used in dairy goat flocks. In the period of 2005-2007, 46.3% of the sheep flocks never changed the anthelmintic class. The dose and move strategy was practiced in 33.2% of the sheep flocks. Conclusions The present study showed that inaccurate weight calculation gives a risk of under-dosing in over 90% of the sheep and goat flocks in Norway. Taken together with a high treatment frequency in lambs, a lack of anthelmintic class rotation and the common use of a dose-and-move strategy, a real danger for development of anthelmintic resistance

Worm control practice against gastro-intestinal parasites in Norwegian sheep and goat flocks

Directory of Open Access Journals (Sweden)

Vatn Synnøve

2011-05-01

Full Text Available Abstract Background Anthelmintic treatment is the most common way of controlling nematode infections in ruminants. However, several countries have reported anthelmintic resistance (AR, representing a limitation for sustainable small ruminant production. The knowledge regarding worm control management represents a baseline to develop a guideline for preventing AR. The aim of the present study was therefore to improve our knowledge about the worm control practices in small ruminant flocks in Norway. Methods A questionnaire survey regarding worm control practices was performed in small ruminant flocks in Norway. Flocks were selected from the three main areas of small ruminant farming, i.e. the coastal, inland and northern areas. A total of 825 questionnaires, comprising 587 sheep flocks (return rate of 51.3% and 238 goat flocks (52.6% were included. Results The results indicated that visual appraisal of individual weight was the most common means of estimating the anthelmintic dose used in sheep (78.6% and goat (85.1% flocks. The mean yearly drenching rate in lambs and ewes were 2.5 ± 1.7 and 1.9 ± 1.1, respectively, whereas it was 1.0 (once a year in goats. However, these figures were higher in sheep in the coastal area with a rate of 3.4 and 2.2 in lambs and ewes, respectively. Benzimidazoles were the predominant anthelmintic class used in sheep flocks (64.9% in 2007, whereas benzimidazoles and macrocyclic lactones were both equally used in dairy goat flocks. In the period of 2005-2007, 46.3% of the sheep flocks never changed the anthelmintic class. The dose and move strategy was practiced in 33.2% of the sheep flocks. Conclusions The present study showed that inaccurate weight calculation gives a risk of under-dosing in over 90% of the sheep and goat flocks in Norway. Taken together with a high treatment frequency in lambs, a lack of anthelmintic class rotation and the common use of a dose-and-move strategy, a real danger for development of

Electrochemical and DFT study of an anticancer and active anthelmintic drug at carbon nanostructured modified electrode

International Nuclear Information System (INIS)

Ghalkhani, Masoumeh; Beheshtian, Javad; Salehi, Maryam

2016-01-01

The electrochemical response of mebendazole (Meb), an anticancer and effective anthelmintic drug, was investigated using two different carbon nanostructured modified glassy carbon electrodes (GCE). Although, compared to unmodified GCE, both prepared modified electrodes improved the voltammetric response of Meb, the carbon nanotubes (CNTs) modified GCE showed higher sensitivity and stability. Therefore, the CNTs-GCE was chosen as a promising candidate for the further studies. At first, the electrochemical behavior of Meb was studied by cyclic voltammetry and differential pulse and square wave voltammetry. A one step reversible, pH-dependent and adsorption-controlled process was revealed for electro-oxidation of Meb. A possible mechanism for the electrochemical oxidation of Meb was proposed. In addition, electronic structure, adsorption energy, band gap, type of interaction and stable configuration of Meb on the surface of functionalized carbon nanotubes were studied by using density functional theory (DFT). Obtained results revealed that Meb is weakly physisorbed on the CNTs and that the electronic properties of the CNTs are not significantly changed. Notably, CNTs could be considered as a suitable modifier for preparation of the modified electrode for Meb analysis. Then, the experimental parameters affecting the electrochemical response of Meb were optimized. Under optimal conditions, high sensitivity (b(Meb) = dI p,a (Meb) / d[Meb] = 19.65 μA μM −1 ), a low detection limit (LOD (Meb) = 19 nM) and a wide linear dynamic range (0.06–3 μM) was resulted for the voltammetric quantification of Meb. - Highlights: • Electrochemical oxidation mechanism of Meb was investigated. • A carbon nanostructure modified electrode was developed for the determination of Meb. • The modified electrode surface was characterized by SEM and impedance studies. • This study provides an effective chemically modified electrode with satisfactory repeatability and reproducibility

Development of a TaqMan Allelic Discrimination Assay for detection of Single Nucleotides Polymorphisms associated with anti-malarial drug resistance

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Kamau Edwin

2012-01-01

Full Text Available Abstract Background Anti-malarial drug resistance poses a threat to current global efforts towards control and elimination of malaria. Several methods are used in monitoring anti-malarial drug resistance. Molecular markers such as single nucleotide polymorphism (SNP for example are increasingly being used to identify genetic mutations related to anti-malarial drug resistance. Several methods are currently being used in analysis of SNP associated with anti-malarial drug resistance and although each one of these methods has unique strengths and shortcoming, there is still need to improve and/or develop new methods that will close the gap found in the current methods. Methods TaqMan Allelic Discrimination assays for detection of SNPs associated with anti-malarial drug resistance were designed for analysis on Applied Biosystems PCR platform. These assays were designed by submitting SNP sequences associated with anti-malarial drug resistance to Applied Biosystems website. Eleven SNPs associated with resistance to anti-malarial drugs were selected and tested. The performance of each SNP assay was tested by creating plasmid DNAs carrying codons of interests and analysing them for analysis. To test the sensitivity and specificity of each SNP assay, 12 clinical samples were sequenced at codons of interest and used in the analysis. Plasmid DNAs were used to establish the Limit of Detection (LoD for each assay. Results Data from genetic profiles of the Plasmodium falciparum laboratory strains and sequence data from 12 clinical samples was used as the reference method with which the performance of the SNP assays were compared to. The sensitivity and specificity of each SNP assay was establish at 100%. LoD for each assay was established at 2 GE, equivalent to less than 1 parasite/μL. SNP assays performed well in detecting mixed infection and analysis of clinical samples. Conclusion TaqMan Allelic Discrimination assay provides a good alternative tool in

Simultaneous densitometric determination of anthelmintic drug albendazole and its metabolite albendazole sulfoxide by HPTLC in human plasma and pharmaceutical formulations.

Science.gov (United States)

Pandya, Jui J; Sanyal, Mallika; Shrivastav, Pranav S

2017-09-01

A new, simple, accurate and precise high-performance thin-layer chromatographic method has been developed and validated for simultaneous determination of an anthelmintic drug, albendazole, and its active metabolite albendazole, sulfoxide. Planar chromatographic separation was performed on aluminum-backed layer of silica gel 60G F 254 using a mixture of toluene-acetonitrile-glacial acetic acid (7.0:2.9:0.1, v/v/v) as the mobile phase. For quantitation, the separated spots were scanned densitometrically at 225 nm. The retention factors (R f ) obtained under the established conditions were 0.76 ± 0.01 and 0.50 ± 0.01 and the regression plots were linear (r 2  ≥ 0.9997) in the concentration ranges 50-350 and 100-700 ng/band for albendazole and albendazole sulfoxide, respectively. The method was validated for linearity, specificity, accuracy (recovery) and precision, repeatability, stability and robustness. The limit of detection and limit of quantitation found were 9.84 and 29.81 ng/band for albendazole and 21.60 and 65.45 ng/band for albendazole sulfoxide, respectively. For plasma samples, solid-phase extraction of analytes yielded mean extraction recoveries of 87.59 and 87.13% for albendazole and albendazole sulfoxide, respectively. The method was successfully applied for the analysis of albendazole in pharmaceutical formulations with accuracy ≥99.32%. Copyright © 2017 John Wiley & Sons, Ltd.

Electrical resistivity for detecting subsurface non-aqueous phase liquids: A progress report

International Nuclear Information System (INIS)

Lee, K.H.; Shan, C.; Javandel, I.

1995-06-01

Soils and groundwater have been contaminated by hazardous substances at many places in the United States and many other countries. The contaminants are commonly either petroleum products or industrial solvents with very low solubility in water. These contaminants are usually called non-aqueous phase liquids (NAPLs). The cost of cleaning up the affected sites in the United States is estimated to be of the order of 100 billion dollars. In spite of the expenditure of several billion dollars during the last 15 years, to date, very few, if any major contaminated site has been restored. The presence of NAPL pools in the subsurface is believed to be the main cause for the failure of previous cleanup activities. Due to their relatively low water solubility, and depending on their volume, it takes tens or even hundreds of years to deplete the NAPL sources if they are not removed from the subsurface. The intrinsic electrical resistivity of most NAPLs is typically in the range of 10 7 to 10 12 Ω-m, which is several orders of magnitude higher than that of groundwater containing dissolved solids (usually in the range of a few Ω-m to a few thousand Ω-m). Although a dry soil is very resistive, the electrical resistivity of a wet soil is on the order of 100 Ω-m and is dependent on the extent of water saturation. For a given soil, the electrical resistivity increases with decrease of water saturation. Therefore, if part of the pore water is replaced by a NAPL, the electrical resistivity will increase. At many NAPL sites, both the vadose and phreatic zones can be partially occupied by NAPL pools. It is the great contrast in electrical resistivity between the NAPLs and groundwater that may render the method to be effective in detecting subsurface NAPLs at contaminated sites. The following experiments were conducted to investigate the change of the electrical resistivity of porous media when diesel fuel (NAPL) replaces part of the water

Characterization of staphylococci in urban wastewater treatment plants in Spain, with detection of methicillin resistant Staphylococcus aureus ST398

International Nuclear Information System (INIS)

Gómez, Paula; Lozano, Carmen; Benito, Daniel; Estepa, Vanesa; Tenorio, Carmen; Zarazaga, Myriam; Torres, Carmen

2016-01-01

The objective of this study was to determine the prevalence of Staphylococcus in urban wastewater treatment plants (UWTP) of La Rioja (Spain), and to characterize de obtained isolates. 16 wastewater samples (8 influent, 8 effluent) of six UWTPs were seeded on mannitol-salt-agar and oxacillin-resistance-screening-agar-base for staphylococci and methicillin-resistant Staphylococcus aureus recovery. Antimicrobial susceptibility profile was determined for 16 antibiotics and the presence of 35 antimicrobial resistance genes and 14 virulence genes by PCR. S. aureus was typed by spa, agr, and multilocus-sequence-typing, and the presence of immune-evasion-genes cluster was analyzed. Staphylococcus spp. were detected in 13 of 16 tested wastewater samples (81%), although the number of CFU/mL decreased after treatment. 40 staphylococci were recovered (1–5/sample), and 8 of them were identified as S. aureus being typed as (number of strains): spa-t011/agr-II/ST398 (1), spa-t002/agr-II/ST5 (2), spa-t3262/agr-II/ST5 (1), spa-t605/agr-II/ST126 (3), and spa-t878/agr-III/ST2849 (1). S. aureus ST398 strain was methicillin-resistant and showed a multidrug resistance phenotype. Virulence genes tst, etd, sea, sec, seg, sei, sem, sen, seo, and seu, were detected among S. aureus and only ST5 strains showed genes of immune evasion cluster. Thirty-two coagulase-negative Staphylococcus of 12 different species were recovered (number of strains): Staphylococcus equorum (7), Staphylococcus vitulinus (4), Staphylococcus lentus (4), Staphylococcus sciuri (4), Staphylococcus fleurettii (2), Staphylococcus haemolyticus (2), Staphylococcus hominis (2), Staphylococcus saprophyticus (2), Staphylococcus succinus (2), Staphylococcus capitis (1), Staphylococcus cohnii (1), and Staphylococcus epidermidis (1). Five presented a multidrug resistance phenotype. The following resistance and virulence genes were found: mecA, lnu(A), vga(A), tet(K), erm(C), msr(A)/(B), mph(C), tst, and sem. We found that

Detection of multidrug resistance using molecular nuclear technique

International Nuclear Information System (INIS)

Lee, Jae Tae; Ahn, Byeong Cheol

2004-01-01

Although the outcome of cancer patients after cytotoxic chemotherapy is related diverse mechanisms, multidrug resistance (MDR) for chemotherapeutic drugs due to cellular P-glycoprotein (Pgp) or multidrug-resistance associated protein (MRP) is most important factor in the chemotherapy failure to cancer. A large number of pharmacologic compounds, including verapamil, quinidine, tamoxifen, cyclosporin A and quinolone derivatives have been reported to overcome MDR. Single photon emission computed tomography (SPECT) and positron emission tomography (PET) are available for the detection of Pgp and MRP-mediated transporter. 99 m-Tc-MIBI and other 99 m-Tc-radiopharmaceuticals are substrates for Pgp and MRP, and have been used in clinical studies for tumor imaging, and to visualize blockade of Pgp-mediated transport after modulation of Pgp pump. Colchicine, verapamil and daunorubicin labeled with 11 C have been evaluated for the quantification of Pgp-mediated transport with PET in vivo and reported to be feasible substrates with which to image Pgp function in tumors. Leukotrienes are specific substrates for MRP and N-( 11 C)acetyl-leukotriene E4 provides an opportunity to study MRP function non-invasively in vivo. SPECT and PET pharmaceuticals have successfully used to evaluate pharmacologic effects of MDR modulators. Imaging of MDR and reversal of MDR with bioluminescence in a living animal is also evaluated for future clinical trial. We have described recent advances in molecular imaging of MDR and reviewed recent publications regarding feasibility of SPECT and PET imaging to study the functionality of MDR transporters in vivo

Detection of Ampicillin Resistance Genes (bla in Clinical Isolates of Escherichia coli with Polymerase Chain Reaction Method

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Tiana Milanda

2014-09-01

Full Text Available Escherichia coli is a rod negative Gram which could be pathogenic, if its value increases or located in outer gastrointestinal tract. Pathogenic E. coli will produce enterotoxin which will cause diarrhoea or infection in urine tract. Ampicilin was one of particular antibiotics to overcome infection. Ampicilin nowadays is no longer used as primary medicine, because of its resistance case. The aim of this research is to detect the presence of gene which is responsible to ampicilin resistant E. coli. We used isolated midstream urine from cystitis object in Hasan Sadikin Hospital (RSHS as samples. Polymerase Chain Reaction (PCR method (colony-PCR and DNA-PCR were done to invenstigate the antibiotic resistency. Based on the result of antibiotic susceptibility testing to ampicillin, E. coli samples were resistant to ampicilin. Elektroforegram products of colony-PCR and DNA-PCR showed that the resistance case of ampicilin caused by bla gene (199 bp. Selective and rational antibiotic treatment is required to prevent ampicillin resistance in patients with symptoms

Anti-parasitic activity of pelleted sainfoin (Onobrychis viciifolia) against Ostertagia ostertagi and Cooperia oncophora in calves

DEFF Research Database (Denmark)

Desrues, O.; Pena-Espinoza, Miguel Angel; Hansen, T. V.

2016-01-01

BACKGROUND: Increasing anthelmintic-resistance in nematodes of ruminants emphasises the need for sustainable parasite control. Condensed tannin-containing legume forages such as sainfoin (Onobrychis viciifolia) have shown promising anthelmintic properties in small ruminants but this has never been......) or concentrate and grass-clover hay (Group CO; n = 6, two pens). After 16 days of adaptation, all animals were experimentally infected with 10,000 and 66,000 third-stage larvae of Ostertagia ostertagi and Cooperia oncophora, respectively. Egg excretion, blood parameters and bodyweights were recorded throughout...

Contribution of 3-D electrical resistivity tomography for landmines detection

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M. Metwaly

2008-12-01

Full Text Available Landmines are a type of inexpensive weapons widely used in the pre-conflicted areas in many countries worldwide. The two main types are the metallic and non-metallic (mostly plastic landmines. They are most commonly investigated by magnetic, ground penetrating radar (GPR, and metal detector (MD techniques. These geophysical techniques however have significant limitations in resolving the non-metallic landmines and wherever the host materials are conductive. In this work, the 3-D electric resistivity tomography (ERT technique is evaluated as an alternative and/or confirmation detection system for both landmine types, which are buried in different soil conditions and at different depths. This can be achieved using the capacitive resistivity imaging system, which does not need direct contact with the ground surface. Synthetic models for each case have been introduced using metallic and non-metallic bodies buried in wet and dry environments. The inversion results using the L₁ norm least-squares optimization method tend to produce robust blocky models of the landmine body. The dipole axial and the dipole equatorial arrays tend to have the most favorable geometry by applying dynamic capacitive electrode and they show significant signal strength for data sets with up to 5% noise. Increasing the burial depth relative to the electrode spacing as well as the noise percentage in the resistivity data is crucial in resolving the landmines at different environments. The landmine with dimension and burial depth of one electrode separation unit is over estimated while the spatial resolutions decrease as the burial depth and noise percentage increase.

Evaluation of the BACTEC MGIT 960 SL DST Kit and the GenoType MTBDRsl Test for Detecting Extensively Drug-resistant Tuberculosis Cases.

Science.gov (United States)

Tekin, Kemal; Albay, Ali; Simsek, Hulya; Sig, Ali Korhan; Guney, Mustafa

2017-10-01

The present study aimed to evaluate the performances of the BACTEC MGIT 960 SL DST kit and the GenoType MTBDRsl test for detecting second-line antituberculosis drug resistance in Multidrug-resistant TB (MDR-TB) cases. Forty-six MDR-TB strains were studied. Second-line antituberculosis drug resistances were detected using the BACTEC MGIT 960 SL DST kit and the GenoType MTBDRsl test. The Middlebrook 7H10 agar proportion method was used as the reference test. The sensitivity and specificity values for the BACTEC MGIT 960 SL DST kit were both 100% for amikacin, kanamycin, capreomycin (4 µg/mL), and ofloxacin; 100% and 95.3%, respectively, for capreomycin (10 µg/mL); and 85.7% and 100%, respectively, for moxifloxacin (0.5 µg/mL). The sensitivity and specificity values for the GenoType MTBDRsl test to detect fluoroquinolone and aminoglycoside/cyclic peptide resistance were 88.9% and 100%, respectively, for ofloxacin and 85.7% and 94.9%, respectively, for moxifloxacin (0.5 µg/mL). The accuracy of the GenoType MTBDRsl assay for kanamycin, capreomycin, ofloxacin, and moxifloxacin was lower than that of the BACTEC MGIT 960 SL DST. The BACTEC MGIT 960 SL DST kit and the GenoType MTBDRsl were successful in detecting second-line antituberculosis drug resistance. Preliminary results of the GenoType MTBDRsl are very valuable for early treatment decisions, but we still recommend additional BACTEC MGIT 960 SL DST kit usage in the routine evaluation of drug-resistant tuberculosis.

Evaluation of the BACTEC MGIT 960 SL DST Kit and the GenoType MTBDRsl Test for Detecting Extensively Drug-resistant Tuberculosis Cases

Science.gov (United States)

Tekin, Kemal; Albay, Ali; Simsek, Hulya; Sig, Ali Korhan; Guney, Mustafa

2017-01-01

Objective: The present study aimed to evaluate the performances of the BACTEC MGIT 960 SL DST kit and the GenoType MTBDRsl test for detecting second-line antituberculosis drug resistance in Multidrug-resistant TB (MDR-TB) cases. Materials and Methods: Forty-six MDR-TB strains were studied. Second-line antituberculosis drug resistances were detected using the BACTEC MGIT 960 SL DST kit and the GenoType MTBDRsl test. The Middlebrook 7H10 agar proportion method was used as the reference test. Results: The sensitivity and specificity values for the BACTEC MGIT 960 SL DST kit were both 100% for amikacin, kanamycin, capreomycin (4 µg/mL), and ofloxacin; 100% and 95.3%, respectively, for capreomycin (10 µg/mL); and 85.7% and 100%, respectively, for moxifloxacin (0.5 µg/mL). The sensitivity and specificity values for the GenoType MTBDRsl test to detect fluoroquinolone and aminoglycoside/cyclic peptide resistance were 88.9% and 100%, respectively, for ofloxacin and 85.7% and 94.9%, respectively, for moxifloxacin (0.5 µg/mL). The accuracy of the GenoType MTBDRsl assay for kanamycin, capreomycin, ofloxacin, and moxifloxacin was lower than that of the BACTEC MGIT 960 SL DST. Conclusion: The BACTEC MGIT 960 SL DST kit and the GenoType MTBDRsl were successful in detecting second-line antituberculosis drug resistance. Preliminary results of the GenoType MTBDRsl are very valuable for early treatment decisions, but we still recommend additional BACTEC MGIT 960 SL DST kit usage in the routine evaluation of drug-resistant tuberculosis. PMID:29123441

Suspension Array for Multiplex Detection of Eight Fungicide-Resistance Related Alleles in Botrytis cinerea

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Xin Zhang

2016-09-01

Full Text Available A simple and high-throughput assay to detect fungicide resistance is required for large-scale monitoring of the emergence of resistant strains of Botrytis cinerea. Using suspension array technology performed on a Bio-Plex 200 System, we developed a single-tube allele-specific primer extension (ASPE assay that can simultaneously detect eight alleles in one reaction. These eight alleles include E198 and 198A of the β-Tubulin gene (BenA, H272 and 272Y of the Succinate dehydrogenase iron–sulfur subunit gene (SdhB, I365 and 365S of the putative osmosensor histidine kinase gene (BcOS1, and F412 and 412S of the 3-ketoreductase gene (erg27. This assay was first established and optimized with eight plasmid templates containing the DNA sequence variants BenA-E198, BenA-198A, SdhB-H272, SdhB-272Y, BcOS1-I365, BcOS1-365S, erg27-F412, and erg27-412S. Results indicated that none of the probes showed cross-reactivity with one another. The minimum limit of detection for these genotypes was one copy per test. Four mutant plasmids were mixed with 10 ng/μL wild-type genomic DNA in different ratios. Detection sensitivity of mutant loci was 0.45% for BenA-E198A, BcOS1-I365S, and erg27-F412S, and was 4.5% for SdhB-H272Y. A minimum quantity of 0.1 ng of genomic DNA was necessary to obtain reliable results. This is the first reported assay that can simultaneously detect mutations in BenA, SdhB, BcOS1, and erg27.

Suspension Array for Multiplex Detection of Eight Fungicide-Resistance Related Alleles in Botrytis cinerea.

Science.gov (United States)

Zhang, Xin; Xie, Fei; Lv, Baobei; Zhao, Pengxiang; Ma, Xuemei

2016-01-01

A simple and high-throughput assay to detect fungicide resistance is required for large-scale monitoring of the emergence of resistant strains of Botrytis cinerea . Using suspension array technology performed on a Bio-Plex 200 System, we developed a single-tube allele-specific primer extension assay that can simultaneously detect eight alleles in one reaction. These eight alleles include E198 and 198A of the β-Tubulin gene ( BenA ), H272 and 272Y of the Succinate dehydrogenase iron-sulfur subunit gene ( SdhB) , I365 and 365S of the putative osmosensor histidine kinase gene ( BcOS1 ), and F412 and 412S of the 3-ketoreductase gene ( erg27 ). This assay was first established and optimized with eight plasmid templates containing the DNA sequence variants BenA- E198, BenA- 198A, SdhB- H272, SdhB- 272Y, BcOS1- I365, BcOS1- 365S, erg27 -F412, and erg27 -412S. Results indicated that none of the probes showed cross-reactivity with one another. The minimum limit of detection for these genotypes was one copy per test. Four mutant plasmids were mixed with 10 ng/μL wild-type genomic DNA in different ratios. Detection sensitivity of mutant loci was 0.45% for BenA- E198A, BcOS1- I365S, and erg27 -F412S, and was 4.5% for SdhB- H272Y. A minimum quantity of 0.1 ng of genomic DNA was necessary to obtain reliable results. This is the first reported assay that can simultaneously detect mutations in BenA , SdhB , BcOS1 , and erg27 .

Determination of the influence of dispersion pattern of pesticide-resistant individuals on the reliability of resistance estimates using different sampling plans.

Science.gov (United States)

Shah, R; Worner, S P; Chapman, R B

2012-10-01

Pesticide resistance monitoring includes resistance detection and subsequent documentation/ measurement. Resistance detection would require at least one (≥1) resistant individual(s) to be present in a sample to initiate management strategies. Resistance documentation, on the other hand, would attempt to get an estimate of the entire population (≥90%) of the resistant individuals. A computer simulation model was used to compare the efficiency of simple random and systematic sampling plans to detect resistant individuals and to document their frequencies when the resistant individuals were randomly or patchily distributed. A patchy dispersion pattern of resistant individuals influenced the sampling efficiency of systematic sampling plans while the efficiency of random sampling was independent of such patchiness. When resistant individuals were randomly distributed, sample sizes required to detect at least one resistant individual (resistance detection) with a probability of 0.95 were 300 (1%) and 50 (10% and 20%); whereas, when resistant individuals were patchily distributed, using systematic sampling, sample sizes required for such detection were 6000 (1%), 600 (10%) and 300 (20%). Sample sizes of 900 and 400 would be required to detect ≥90% of resistant individuals (resistance documentation) with a probability of 0.95 when resistant individuals were randomly dispersed and present at a frequency of 10% and 20%, respectively; whereas, when resistant individuals were patchily distributed, using systematic sampling, a sample size of 3000 and 1500, respectively, was necessary. Small sample sizes either underestimated or overestimated the resistance frequency. A simple random sampling plan is, therefore, recommended for insecticide resistance detection and subsequent documentation.

Variability in faecal egg counts – a statistical model to achieve reliable determination of anthelmintic resistance in livestock

DEFF Research Database (Denmark)

Nielsen, Martin Krarup; Vidyashankar, Anand N.; Hanlon, Bret

statistical model was therefore developed for analysis of FECRT data from multiple farms. Horse age, gender, zip code and pre-treatment egg count were incorporated into the model. Horses and farms were kept as random effects. Resistance classifications were based on model-based 95% lower confidence limit (LCL...

Questionnaire study on parasite control practices on Thoroughbred and Standardbred breeding farms in New Zealand.

Science.gov (United States)

Bolwell, Charlotte F; Rosanowski, Sarah M; Scott, Ian; Sells, Patrick D; Rogers, Chris W

2015-04-15

Against a global background of increasing anthelmintic resistance in parasites, little is known about the current parasite control strategies adopted within the equine industry in New Zealand. The aim of the study was to describe and compare the current parasite management and control practices used on Thoroughbred and Standardbred stud farms in New Zealand. An online questionnaire was used to collect data on the demographics of respondents, parasite control methods, grazing management, and use of faecal egg counts. Questions regarding parasite control strategy, how often horses were dewormed, number of treatments per year and stocking density were stratified by horse type: young stock (foals/weanlings/yearlings), wet mares (nursing a foal) or dry mares, and industry (Thoroughbred and Standardbred). Questions on grazing management were stratified by horse type and the breeding and non-breeding season. In total, 136 respondents completed the survey, of which most (66%; 90/136) were involved in the Thoroughbred breeding industry. Most (98%; 134/136) respondents used anthelmintic products to treat the horses on their property, and regardless of industry type most respondents were using interval drenching for young stock (86/129; 53%), dry mares (51/124; 41%) or wet mares (50/126; 40%). Of those respondents treating on regular interval, 55% (68/123), 42% (50/119) and 38% (46/122) were treating young stock, wet mares and dry mares every 6-8 weeks. The median number of treatments per year for young stock, dry mares and wet mares was 6 (IQR 4-8), 4 (IQR 3-6) and 4 (IQR 3-6), respectively; there was no difference in frequency of treatments by industry type. In the last 12 months respondents used a median of 2 (IQR 2-4) and 3 (IQR 2-4) different anthelmintic products to treat horses on Thoroughbred and Standardbred breeding farms, respectively. Of the respondents reporting the anthelmintic products used in the last 12 months, 95% used at least one product containing

[Molecular characterization of resistance mechanisms: methicillin resistance Staphylococcus aureus, extended spectrum β-lactamases and carbapenemases].

Science.gov (United States)

Oteo, Jesús; Belén Aracil, María

2015-07-01

Multi-drug resistance in bacterial pathogens increases morbidity and mortality in infected patients and it is a threat to public health concern by their high capacity to spread. For both reasons, the rapid detection of multi-drug resistant bacteria is critical. Standard microbiological procedures require 48-72 h to provide the antimicrobial susceptibility results, thus there is emerging interest in the development of rapid detection techniques. In recent years, the use of selective and differential culture-based methods has widely spread. However, the capacity for detecting antibiotic resistance genes and their low turnaround times has made molecular methods a reference for diagnosis of multidrug resistance. This review focusses on the molecular methods for detecting some mechanisms of antibiotic resistance with a high clinical and epidemiological impact: a) Enzymatic resistance to broad spectrum β-lactam antibiotics in Enterobacteriaceae, mainly extended spectrum β-lactamases (ESBL) and carbapenemases; and b) methicillin resistance in Staphylococcus aureus. Copyright © 2015 Elsevier España, S.L.U. All rights reserved.

Sinkhole detection using electrical resistivity tomography in Saudi Arabia

International Nuclear Information System (INIS)

Youssef, Ahmed M; Zabramawi, Yasser A; El-Kaliouby, Hesham

2012-01-01

Karst phenomena exist in different areas in the Kingdom of Saudi Arabia, causing serious environmental problems that affect urban development and infrastructure (buildings, roads and highways). One of the most important problems are sinkholes, which most of the time consist of unfilled voids. These sinkholes are formed as a result of the chemical leaching of carbonate and evaporite formations by percolating water. Field investigations show that there are many surface expressions of sinkholes in the area; some appear on the ground surface and others are hidden in the subsurface. Geophysical data were collected at the study area using two-dimensional electrical resistivity tomography (ERT) with different electrode spacings to delineate buried sinkholes and associated subsurface cavities. Our findings indicated that the dipole–dipole method using an electrode spacing of 1 m was successful in detecting a known subsurface sinkhole. According to the ERT method the detected sinkhole depth ranges from 2 to 4 m, its height ranges from 2 to 4 m, and its width ranges from 5 to 7 m. Field observation has verified the geophysical data, especially along the profile A-A. Finally, closely spaced ERT profiles were successful in determining the three-dimensional volume of the subsurface sinkhole. (paper)

Detection of macrolide resistance genes in culture-negative specimens from Bangladeshi children with invasive pneumococcal diseases.

Science.gov (United States)

Hasanuzzaman, Md; Malaker, Roly; Islam, Maksuda; Baqui, Abdullah H; Darmstadt, Gary L; Whitney, Cynthia G; Saha, Samir K

2017-03-01

In recent years, an increasing prevalence of macrolide resistance among pneumococci in Bangladesh has been observed. However, the scenario remains incomplete, as few isolates (80%) are culture-negative. This study optimised a triplex PCR method to detect macrolide resistance genes (MRGs) (mefA and ermB) and cpsA from culture-negative pneumococcal cases to predict the prevalence and level of macrolide resistance. The presence of MRGs among pneumococcal strains (n=153) with a wide range of erythromycin MICs (culture-negative clinical specimens and corresponding isolates. The known impact of the presence of specific MRG(s) on MICs of strains was used to predict the MICs of non-culturable strains based on the presence/absence of MRG(s) in the specimens. None of the erythromycin-susceptible isolates possessed any of the MRGs, and all non-susceptible strains had ≥1 MRG. MICs were 2-16mg/L and ≥256mg/L for 93% of strains with mefA and ermB, respectively, whereas 100% of isolates with both genes had MICs≥256mg/L. PCR for body fluids showed 100% concordance with corresponding isolates when tested for MRG(s) in parallel. Erythromycin MICs can be predicted for non-culturable strains with 93-100% precision based on detection of ermB and/or mefA. This method will be useful for establishing comprehensive surveillance for macrolide resistance among pneumococci, specifically in the population with prior antibiotic use. Copyright © 2017. Published by Elsevier Ltd.

Detection and genetic distance of resistant populations of Pseudosuccinea columella (Mollusca: Lymnaeidae) to Fasciola hepatica (Trematoda: Digenea) using RAPD markers.

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Calienes, Aymé Fernandez; Fraga, Jorge; Pointier, Jean-Pierre; Yong, Mary; Sanchez, Jorge; Coustau, Christine; Gutiérrez, Alfredo; Théron, André

2004-09-01

Twelve natural populations of Pseudosuccinea columella snails, sampled in the western and central regions of Cuba, were analyzed using the RAPD-PCR technique to screen for resistance to Fasciola hepatica. Ten OPA primers previously shown to produce marker bands for resistance and susceptibility were tested. A new population of P. columella (El Azufre, Pinar del Río) exhibited the amplification patterns of resistant snails, and its resistant status was confirmed after experimental exposure to miracidia. No genetic variability was detected across or within the susceptible isolates. Similarly, the novel resistant isolate displayed an RAPD profile identical to the profile of two other isolates previously identified as resistant to F. hepatica. However, clear differences in RAPD banding patterns and genetic distance were observed between resistant and susceptible isolates.

Production trials involving use of the FAMACHA© system for haemonchosis in sheep : preliminary results

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J.A. Van Wyk

2008-09-01

Full Text Available In three trials conducted on two separate farms the production of sheep treated for naturally acquired haemonchosis using the FAMACHA© system of targeted selective treatment (TST (i.e. to treat only those animals unable to manage unaided in the face of heavy Haemonchus challenge was compared to that of suppressively drenched sheep in the same flock. As expected by the research team who developed and evaluated the FAMACHA© system, TST did result in some loss in production. However, despite high levels of worm challenge in two of the trials and the fact that the comparison was with suppressive drenching which is not sustainable, the total effect was relatively small in relation to the important advantage of using the TST as regards reduced selection for anthelmintic resistance (AR. Concerning the sustainability of worm control, it is concluded that the development of drug resistance to anthelmintics leaves sheep and goat farmers in South Africa no choice but to use methods of TST such as FAMACHA©. The FAMACHA© system can also be a useful clinical aid for early on-farm detection of AR by farmers; the degree of improvement in the colour of the ocular mucous membrane from pale to red in individually drenched anaemic animals over a period of 7-14 days can give a good indication of the efficacy of the compound(s used.

Detection of methicillin resistant Staphylococcus aureus (MRSA) from recreational beach using the mecA gene