Reinemeyer Craig R
Full Text Available Abstract Since 2002, macrocyclic lactone resistance has been reported in populations of Parascaris equorum from several countries. It is apparent that macrocyclic lactone resistance developed in response to exclusive and/or excessively frequent use of ivermectin or moxidectin in foals during the first year of life. The development of anthelmintic resistance was virtually inevitable, given certain biological features of Parascaris and unique pharmacologic characteristics of the macrocyclic lactones. Practitioners can utilize the Fecal Egg Count Reduction Test to detect anthelmintic resistance in Parascaris, and the same technique can be applied regularly to confirm the continued efficacy of those drugs currently in use. In the face of macrocyclic lactone resistance, piperazine or anthelmintics of the benzimidazole or pyrimidine classes can be used to control ascarid infections, but Parascaris populations that are concurrently resistant to macrocyclic lactones and pyrimidine drugs have been reported recently from Texas and Kentucky. Compared to traditional practices, future recommendations for ascarid control should feature: 1 use of only those anthelmintics known to be effective against indigenous populations, 2 initiation of anthelmintic treatment no earlier than 60 days of age, and 3 repetition of treatments at the longest intervals which prevent serious environmental contamination with Parascaris eggs. In the interest of decreasing selection pressure for anthelmintic resistance, horse owners and veterinarians must become more tolerant of the passage of modest numbers of ascarid eggs by some foals. Anthelmintic resistance is only one of several potential responses to genetic selection. Although still only theoretical, changes in the immunogenicity of ascarid isolates or reduction of their prepatent or egg reappearance periods could pose far greater challenges to effective control than resistance to a single class of anthelmintics.
Larsen, Mette L.; Ritz, Christian; Petersen, Stig L.
cyathostomins and P. equorum in Danish horses. A total of 196 animals were selected from 52 farms, all of which were using a selective anthelmintic treatment strategy. ERP was investigated with weekly samples from 96 horses from nine farms. Horses were treated with ivermectin oral paste by their owners...
Hatem A Shalaby
Full Text Available Many parasitic helminthes of veterinary importance have genetic features that favor development of anthelmintic resistance, this becoming a major worldwide constrain in livestock production. The development of anthelmintic resistance poses a large threat to future production and welfare of grazing animals. Development of variable degrees of resistance among different species of gastrointestinal nematodes has been reported for all the major groups of anthelmintic drugs. It has been observed that frequent usage of the same group of anthelmintic; use of anthelmintics in sub-optimal doses, prophylactic mass treatment of domestic animals and frequent and continuous use of a single drug have contributed to the widespread development of anthelmintic resistance in helminthes. The degree and extent of this problem especially with respect to multidrug resistance in nematode populations is likely to increase. Maintaining parasites in refugia and not exposed to anthelmintics, seems to be a key point in controlling and delaying the development of resistance, because the susceptible genes are preserved. Targeted selective treatments attract the interest of scientists towards this direction. Additionally, adoption of strict quarantine measures and a combination drug strategy are two important methods of preventing of anthelmintic resistance. Experience from the development of anthelmintic resistance suggests that modern control schemes should not rely on sole use of anthelmintics, but employ other, more complex and sustainable recipes, including parasite resistant breeds, nutrition, pasture management, nematode-trapping fungi, antiparasitic vaccines and botanical dewormers. Most of them reduce reliance on the use of chemicals and are environmental friendly. Finally, if new anthelmintic products are released, an important question will be raised about how they should be used. It is suggested that slowing the development of resistance to a new
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 ...
Borgsteede, F.H.M.; Pekelder, J.J.; Dercksen, D.P.
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
A cross-sectional study was carried out from November 2013 to April 2014 in pastoral area, Yabello districts, to estimate the prevalence and to identify risk factors associated with gastrointestinal strongyle infection in sheep and goats. Moreover, to assess the anthelmintic resistance in goats gastrointestinal nematodes a total ...
Galvan, Noe; Middleton, John R.; Nagy, Dusty W.; Schultz, Loren G.; Schaeffer, Josh W.
A herd of alpacas was examined because of a history of severe endoparasitism, anemia, hypoproteinemia, and weight loss. Resistance of gastrointestinal nematodes to albendazole, fenbendazole, and doramectin was documented. This report suggests that anthelmintic resistance may be an emerging problem in South American camelids in North America. PMID:23729829
Kaplan, Ray M; Vidyashankar, Anand N
Over the past 10-15 years, we have witnessed a rapid increase in both the prevalence and magnitude of anthelmintic resistance, and this increase appears to be a worldwide phenomenon. Reports of anthelmintic resistance to multiple drugs in individual parasite species, and in multiple parasite species across virtually all livestock hosts, are increasingly common. In addition, since the introduction of ivermectin in 1981, no novel anthelmintic classes were developed and introduced for use in livestock until recently with the launch of monepantel in New Zealand. Thus, livestock producers are often left with few options for effective treatment against many important parasite species. While new anthelmintic classes with novel mechanisms of action could potentially solve this problem, new drugs are extremely expensive to develop, and can be expected to be more expensive than older drugs. Thus, it seems clear that the "Global Worming" approach that has taken hold over the past 40-50 years must change, and livestock producers must develop a new vision for parasite control and sustainability of production. Furthermore, parasitologists must improve methods for study design and data analysis that are used for diagnosing anthelmintic resistance, especially for the fecal egg count reduction test (FECRT). Currently, standards for diagnosis of anthelmintic resistance using FECRT exist only for sheep. Lack of standards in horses and cattle and arbitrarily defined cutoffs for defining resistance, combined with inadequate analysis of the data, mean that errors in assigning resistance status are common. Similarly, the lack of standards makes it difficult to compare data among different studies. This problem needs to be addressed, because as new drugs are introduced now and in the future, the lack of alternative treatments will make early and accurate diagnosis of anthelmintic resistance increasingly important. Copyright © 2011 Elsevier B.V. All rights reserved.
Holm, Signe A.; Sørensen, Camilla; Thamsborg, Stig M.
The prevalence of gastrointestinal parasites in Danish goats and the presence of anthelmintic resistance (AR) in 10 selected herds were investigated during April-September 2012. All Danish herds (n = 137) with 10 or more adult goats were invited to participate, and of these 27 herds met the inclu......The prevalence of gastrointestinal parasites in Danish goats and the presence of anthelmintic resistance (AR) in 10 selected herds were investigated during April-September 2012. All Danish herds (n = 137) with 10 or more adult goats were invited to participate, and of these 27 herds met...... the inclusion criterion of more than 10 young kids never treated with anthelmintics. Questionnaire data on management were collected, and faecal samples from 252 kids were analysed by the McMaster technique. From all herds with a mean faecal egg count (FEC) above 300 eggs per g of faeces, pooled samples were...
Knubben-Schweizer, Gabriela; Pfister, Kurt
Anthelmintic resistance of gastrointestinal nematodes in small ruminants, but also in cattle and horses, is now found worldwide. The reason for increasing anthelmintic resistance is, in particular, the extensive use of all the anthelmintic agents available on the market. A non-targeted use leads to the selection of naturally occurring resistance genes within parasite populations. The most practical method for evaluating the efficacy of an anthelmintic is the fecal egg-count reduction test. To reduce the rate of anthelmintic resistance development, the available active substances must be applied less and in a targeted manner. When applying targeted (selective) treatment, part of the herd is left untreated. Therefore, it is necessary to identify the animals that require treatment for health or economic reasons. To decide on anthelmintic treatment, findings can be collected from single animals or from a group of animals in a herd. To determine which groups of animals are to be treated within a herd (targeted treatment), pooled fecal samples (cattle and small ruminants), serum pepsinogen concentration (cattle), or Ostertagia ostertagi antibodies in the bulk milk (cattle) can be analyzed. For individual animal (targeted selective) treatment, criteria including fecal egg count (cattle and small ruminants), conjuctival color as an indicator for infection with Haemonchus contortus (FAMACHA ® , small ruminants), body condition in adult animals (small ruminants), weight gain in juvenile animals (cattle and small ruminants), and the consistency of the feces (small ruminants) are used. These decision criteria can also be combined to enhance the informative value. Furthermore, an efficacy test of the anthelmintics used should be performed regularly at the beginning of the pasture season. During the pasture season, a low infection pressure should be maintained by pasture management strategies. The goal of sustainable parasite management is the reduction of anthelmintic
Gasbarre, Louis C
The first documented case of macrocyclic lactone resistance in gastrointestinal (GI) nematodes of cattle was seen in the US approximately 10 years ago. Since that time the increase incidence of anthelmintic resistance has continued at an alarming rate. Currently parasites of the genera Cooperia and/or Haemonchus resistant to generic or brand-name macrocyclic lactones have be demonstrated in more than half of all operations examined. Both of these parasite genera are capable of causing economic losses by decreasing food intake and subsequently animal productivity. Currently, there are no easy and quick means to detect anthelmintic resistant GI nematodes. Definitive identification requires killing of cattle. The most commonly used field detection method is the fecal egg count reduction test (FECRT). This method can be adapted for use as a screening agent for Veterinarians and producers to identify less than desired clearance of the parasites after anthelmintic treatment. Further studies can then define the reasons for persistence of the egg counts. The appearance of anthelmintic resistance is largely due to the development of very effective nematode control programs that have significantly improved the productivity of the US cattle industry, but at the same time has placed a high level of selective pressure on the parasite genome. The challenges ahead include the development of programs that control the anthelmintic resistant nematodes but at the same time result in more sustainable parasite control. The goal is to maintain high levels of productivity but to exert less selective pressures on the parasites. One of the most effective means to slow the development of drug resistance is through the simultaneous use of multiple classes of anthelmintics, each of which has a different mode of action. Reduction of the selective pressure on the parasites can be attained through a more targeted approach to drug treatments where the producer's needs are met by selective
Young, K E; Garza, V; Snowden, K; Dobson, R J; Powell, D; Craig, T M
Diversity of parasite populations was compared between two herds of horses, one a regularly treated herd the other a feral herd which has bad no anthelmintic treatment for at least 25 years. Eggs obtained from fecal samples of both herds were tested for anthelmintic resistance by use of an in-vitro larval hatch/development assay (LDA), DrenchRite. A fecal egg reduction test was also performed with the domesticated herd using fenbendazole, pyrantel pamoate and ivermectin. Cyathostomes were the predominant group of worms present in both herds. Trichostrongylus axei was seen in both herds, but Strongylus equinus, Strongylus vulgaris, Gyalocephalus capitatus, Poteriostomum spp. and Strongyloides westeri were only found in the feral horses. Larvae of Strongylus edentatus were found in a single domesticated horse. Fecal egg reduction tests with the domesticated herd showed a 32% egg count reduction for fenbendazole, a 93% reduction with pyrantel, and a 99% reduction with ivermectin. From the LDA, anthelmintic resistance was evaluated by determining the resistance ratio of the domesticated herd compared with the feral herd. For benzimidazoles in the domesticated herd, 45% of the cyathostome population was 9.4 times more tolerant than the feral herd's parasite population. The parasite population in the domesticated herd was 1.5 times more tolerant to Levamisole, and 1.7 times more tolerant to the benzimidazole/levamisole combination than the parasite population within the feral herd. 9% of the parasite population in the domesticated herd was 90 times more tolerant to avermectins than the feral herd's parasite population, even though a subpopulation of worms in the feral herd were tolerant to low concentrations of avermectins despite never being previously exposed to this class of anthelmintic.
Full Text Available Abstract Anthelmintic resistance has been reported in most sheep producing countries. Prior to the mid 1990s, reports of anthelmintic resistance in Ireland were sparse and focused on benzimidazole, one of the three classes of anthelmintic available during this period. This evidence for efficacy issues on Irish farms combined with awareness that anthelmintic resistance was increasingly being reported in other countries prompted the need for more comprehensive investigations on Irish farms. Faecal egg count reduction and micro-agar larval development tests were employed to investigate resistance to benzimidazole, levamisole and macrocyclic lactone. There is compelling evidence for resistance to both benzimidazole (>88% of flocks and levamisole (>39% of flocks. Resistance of nematode populations to macrocyclic lactone was suspected on a small number of farms (11% but needs to be confirmed. The recent introduction of two new classes of anthelmintics, after over a 25 year interval, together with the evidence that anthelmintic resistance is reported within a relatively short time following the introduction of a new anthelmintic compound means that the challenge to the industry is immediate. Actions are urgently required to manage anthelmintic resistance so as to prolong the lifespan of anthelmintics.
Maroto, R.; Jiménez, A. E.; Romero, J. J.; Alvarez, V.; De Oliveira, J. B.; Hernández, J.
As the prevalence and severity of anthelmintic resistance continue to rise, nematode infections in sheep correspondingly reduce the profitability of the sheep industry. In Costa Rica, sheep production systems are increasing in both number and importance. A field trial study was carried out to detect the level of anthelmintic resistance to albendazole and ivermectin in gastrointestinal nematodes (GIN) of sheep from seven farms in Costa Rica. Resistance was determined using the fecal egg count reduction test (FECRT). Three treatment groups were assessed on each farm: control, albendazole, and ivermectin. Haemonchus spp. (71%), Strongyloides sp. (57%), and Trichostrongylus spp. (43%) presented resistance levels to albendazole, whereas Strongyloides sp. (43%), Haemonchus spp. (29%), and Trichostrongylus spp. (29%) were resistant to ivermectin. Haemonchus spp., Strongyloides sp., and Trichostrongylus spp. were the most resistant GIN to both products. This study suggests that frequency of treatment, exclusive chemical control, and visual estimation of animal weight to calculate dosage may contribute to the high levels of anthelmintic resistance that were observed on the farms analyzed herein. PMID:21772962
Full Text Available The anthelmintic resistance in small ruminants is a common problem and concern worldwide. The aim of this study was to verify anthelmintic treatment efficacy in naturally infected sheep. This study was conducted on nine herds that used the same anthelmintic management for over a year. In each farm, the animals were divided into two groups: untreated control group (n = 5 and treated (n = 10 according to the number of eggs per gram of feces (EPG. The treatment effect was checked based on EPG results and larval culture performed before treatment and 10 days after treatment. Significant differences were not observed (P> 0.05 on EPG results between untreated and treated groups. The coproculture showed that the animals were infected primarily byHaemonchus spp., Trichostrongylus spp.,Teladorsagia spp., Cooperia spp. andOesophagostomum spp. In all farms, anthelmintic resistance by genera Haemonchus and Trichostrongylus was found, but this resistance varied greatly between farms.Haemonchus spp. showed resistance to closantel, levamisole, and albendazole. Trichostrongylus spp. was shown to be resistant to closantel, levamisole, and albendazole. The drugs tested showed to be efficient against the genera Teladorsagia,Cooperia, and Oesophagostomum. Based on these results, we conclude that the anthelmintic resistance to the tested drugs is a problem present in the farms evaluated.
Full Text Available As the prevalence and severity of anthelmintic resistance continue to rise, nematode infections in sheep correspondingly reduce the profitability of the sheep industry. In Costa Rica, sheep production systems are increasing in both number and importance. A field trial study was carried out to detect the level of anthelmintic resistance to albendazole and ivermectin in gastrointestinal nematodes (GIN of sheep from seven farms in Costa Rica. Resistance was determined using the fecal egg count reduction test (FECRT. Three treatment groups were assessed on each farm: control, albendazole, and ivermectin. Haemonchus spp. (71%, Strongyloides sp. (57%, and Trichostrongylus spp. (43% presented resistance levels to albendazole, whereas Strongyloides sp. (43%, Haemonchus spp. (29%, and Trichostrongylus spp. (29% were resistant to ivermectin. Haemonchus spp., Strongyloides sp., and Trichostrongylus spp. were the most resistant GIN to both products. This study suggests that frequency of treatment, exclusive chemical control, and visual estimation of animal weight to calculate dosage may contribute to the high levels of anthelmintic resistance that were observed on the farms analyzed herein.
Jordana Andrioli Salgado
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.
Full Text Available A survey to detect anthelmintic resistance in nematode parasites of sheep was conducted on 10 randomly-distributed farms in the Chivhu District, Mashonaland East Province, Zimbabwe. Before the survey, a questionnaire was circulated to the farmers concerning nematode parasite control. Results showed that parasite control using anthelmintic treatment was the only method practised and that the benzimidazoles were the most frequently used anthelmintic drugs. The faecal egg count reduction test was used to detect resistance. The anthelmintic groups tested were benzimidazoles, levamisole and ivermectin. Resistance to benzimidazoles was detected on 6 of 10 farms and levamisole resistance on 2 of 3 farms. Ivermectin resistance was not observed on the farms surveyed. Post-treatment larval cultures indicated that Haemonchus contortus survived administration of fenbendazole, albendazole, oxfendazole and levamisole. A Cooperia sp. strain resistant to albendazole was detected and this is the first report in Zimbabwe of a resistant parasite in this genus.
Full Text Available The intensive usage of anthelmintic in most of farms led to resistances of livestock gastrointestinal nematodes against anthelmintic. Many reports of resistance that increased every year happen following the continuing helminth control programmes. The succesful implementation of helminth control programmes that designed to minimize the development of resistance in nematode populations depends on the availability of effective and sensitive method for its detection and monitoring. A variety of in vivo and in vitro tests have been developed for detecting nematode population resistance to the main anthelmintic groups. This paper will discuss the development of detection method of anthelmintic resistance based on conventional and molecular approach according to their strengths and weakness.
Craven, J.; Bjørn, H.; Henriksen, S.A.
This study reports on the prevalence of anthelmintic resistance in strongyles of horses in Denmark; Of 5 methods used for the calculation of faecal egg count reduction (FECR) the method recommended by the World Association for the Advancement of Veterinary Parasitology, for the detection of resis......This study reports on the prevalence of anthelmintic resistance in strongyles of horses in Denmark; Of 5 methods used for the calculation of faecal egg count reduction (FECR) the method recommended by the World Association for the Advancement of Veterinary Parasitology, for the detection...... of resistance in sheep was the most sensitive procedure for detecting resistance. Using this method benzimidazole resistance was detected on 33 of 42 farms (79%) examined. Pyrantel was tested on 15 farms and FECR tests indicate resistance on 3 (30%) farms. On 2 farms on which resistance to pyrantel was detected...... resistance to benzimidazoles was also detected. On one of 16 farms examined ivermectin resistance was indicated at Day 14 but not at Day 19. On the 15 remaining farms ivermectin was effective. Due to the high prevalence of anthelmintic resistance in Danish horse herds it is recommended that tests...
Garretson, Pamela D; Hammond, Elizabeth E; Craig, Thomas M; Holman, Patricia J
A young male giraffe (Giraffa camelopardalis) recently acquired by the Lion Country Safari in Loxahatchee, Florida, was diagnosed and successfully treated for Haemonchus infection while in quarantine. Seven weeks after introduction into a group of resident giraffes, this giraffe presented with diarrhea. Fecal evaluation revealed an extremely high count of 16,700 eggs/g, with larval identification of the parasite as Haemonchus. A larval development assay showed resistance to the three classes of anthelmintics currently used to treat Haemonchus contortus: the benzimidazoles, imidazothiazoles, and macrocyclic lactones. The giraffe was treated with a combination of moxidectin topically and fenbendazole orally, and follow-up fecal examination 2 wk later showed a marked reduction in strongyle-type eggs. However, within 2 mo the giraffe had a packed cell volume of 22% and an eggs per gram count of 11,900. The animal was then treated with moxidectin topically and copper oxide wire particles orally and removed from the contaminated area. Because of the unusual host, molecular analysis of the parasite was employed, which confirmed the nematode as H. contortus. It is likely that the monthly rotational deworming schedule first implemented more than 5 yr earlier contributed to the development of multiple anthelmintic resistance in this H. contortus population. The proper use of anthelmintics and good pasture management are crucial to reducing the parasite burden in captive giraffe.
Carlos M.B. Gárcia
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.
Víctor Humberto Suarez
Full Text Available Risk factors for anthelmintic resistance (AR on bovine ranches were studied. Data were derived from a survey made to 50 ranch owners, who had conducted a faecal egg-count-reduction test. The questionnaire contained descriptors of bovine ranch management and nematode control. A case-control design study was undertaken and AR cases were present in 26 herds. Associations between the binary outcome variable (AR versus not AR and risk factors recorded in the questionnaire were evaluated. Variables associated with the presence of AR at P 2 were subjected to a multivariable logistic regression model. The main effects contributing to general AR (avermectin AVM and/or benzimidazole in the final model were total number of annual treatments (OR 7.68; 95% CI 2.4 to 28.3 and use of more than 75% of AVM in the past (OR= 18.6; 95% CI 1.3 to 97.3, whereas for AVM resistance alone were total number of AVM annual treatments (OR= 11.5; 95% CI 2.9 to 45.5 and number of AVM Nov-Jan treatments (OR= 5.8; 95% CI 1.71 to 47.9. The results showed that treatment frequency, date of treatment and frequency of treatment in the past with a single drug were the main risk factors involved in AR development.
Han, Tianlong; Wang, Min; Zhang, Guanghe; Han, Dongsheng; Li, Xinwei; Liu, Guowen; Li, Xiaobing; Wang, Zhe
Gastrointestinal nematodes (GIN) are a crucial restraint to grazing sheep production worldwide. This study was conducted to determine the infections and anthelmintic resistance (AR) of GIN in pasture-based sheep in the Eastern Inner Mongolia, China. GIN eggs were tested from 600 grazing sheep feces of 10 farms using saturated saline flotation method and McMaster's method. The egg hatch test (EHT) and the faecal egg count reduction test (FECRT) were used to evaluate resistance of GIN to anthelmintics. We found that the average infection rate was 79.2% (range: 45%-100%). The grand mean faecal egg count (FEC) was 1813.2 eggs per gram (EPG) (range: 0-32400 EPG). There were significant differences in GIN infection among different breeds of sheep. The sequence of infection intensity and infection rate were Small fat tail > Ujimqin > Ju Ud (pgrazing sheep were very common. AR, especially in Haemonchus, was a serious problem in these sheep flocks. Thus, actions are urgently required to taken to mitigate the worsening situation.
Andrew C. Kotze
Full Text Available Anthelmintic resistance has a great impact on livestock production systems worldwide, is an emerging concern in companion animal medicine, and represents a threat to our ongoing ability to control human soil-transmitted helminths. The Consortium for Anthelmintic Resistance and Susceptibility (CARS provides a forum for scientists to meet and discuss the latest developments in the search for molecular markers of anthelmintic resistance. Such markers are important for detecting drug resistant worm populations, and indicating the likely impact of the resistance on drug efficacy. The molecular basis of resistance is also important for understanding how anthelmintics work, and how drug resistant populations arise. Changes to target receptors, drug efflux and other biological processes can be involved. This paper reports on the CARS group meeting held in August 2013 in Perth, Australia. The latest knowledge on the development of molecular markers for resistance to each of the principal classes of anthelmintics is reviewed. The molecular basis of resistance is best understood for the benzimidazole group of compounds, and we examine recent work to translate this knowledge into useful diagnostics for field use. We examine recent candidate-gene and whole-genome approaches to understanding anthelmintic resistance and identify markers. We also look at drug transporters in terms of providing both useful markers for resistance, as well as opportunities to overcome resistance through the targeting of the transporters themselves with inhibitors. Finally, we describe the tools available for the application of the newest high-throughput sequencing technologies to the study of anthelmintic resistance.
Kotze, Andrew C.; Hunt, Peter W.; Skuce, Philip; von Samson-Himmelstjerna, Georg; Martin, Richard J.; Sager, Heinz; Krücken, Jürgen; Hodgkinson, Jane; Lespine, Anne; Jex, Aaron R.; Gilleard, John S.; Beech, Robin N.; Wolstenholme, Adrian J.; Demeler, Janina; Robertson, Alan P.; Charvet, Claude L.; Neveu, Cedric; Kaminsky, Ronald; Rufener, Lucien; Alberich, Melanie; Menez, Cecile; Prichard, Roger K.
Anthelmintic resistance has a great impact on livestock production systems worldwide, is an emerging concern in companion animal medicine, and represents a threat to our ongoing ability to control human soil-transmitted helminths. The Consortium for Anthelmintic Resistance and Susceptibility (CARS) provides a forum for scientists to meet and discuss the latest developments in the search for molecular markers of anthelmintic resistance. Such markers are important for detecting drug resistant worm populations, and indicating the likely impact of the resistance on drug efficacy. The molecular basis of resistance is also important for understanding how anthelmintics work, and how drug resistant populations arise. Changes to target receptors, drug efflux and other biological processes can be involved. This paper reports on the CARS group meeting held in August 2013 in Perth, Australia. The latest knowledge on the development of molecular markers for resistance to each of the principal classes of anthelmintics is reviewed. The molecular basis of resistance is best understood for the benzimidazole group of compounds, and we examine recent work to translate this knowledge into useful diagnostics for field use. We examine recent candidate-gene and whole-genome approaches to understanding anthelmintic resistance and identify markers. We also look at drug transporters in terms of providing both useful markers for resistance, as well as opportunities to overcome resistance through the targeting of the transporters themselves with inhibitors. Finally, we describe the tools available for the application of the newest high-throughput sequencing technologies to the study of anthelmintic resistance. PMID:25516826
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.
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
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
George, Melissa M; Paras, Kelsey L; Howell, Sue B; Kaplan, Ray M
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
Leathwick, Dave M; Luo, Dongwen
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.
Muchiut, Sebastián Manuel; Fernández, Alicia Silvina; Steffan, Pedro Eduardo; Riva, Eliana; Fiel, César Alberto
Sheep production in tropical and temperate regions is hampered by the presence of Haemonchus contortus, the blood-sucking nematode that is the major cause of economic losses in small ruminant enterprises. The most limiting factor in the control of this parasitic disease is the steady progress of anthelmintic resistance worldwide. The search for control strategies that minimise the use of anthelmintics is therefore central to various efforts worldwide. One strategy is the introduction of susceptible parasites in refugia when these refugia are at low levels. This strategy could lead to a renewed possibility anthelmintics being effective. At farm level, this management practice could recover the use of anthelmintics in flocks with high levels of resistance. This review explores the possibility of replacing resistant H. contortus populations with susceptible ones through refugia management and. highlights the experiences of on-farm research attempts carried out in different geographical areas, reaching various degrees of success. Copyright © 2018 Elsevier B.V. All rights reserved.
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.
Funato, Michinori; Kaneko, Hideo; Ohkusu, Kiyofumi; Sasai, Hideo; Kubota, Kazuo; Ohnishi, Hidenori; Kato, Zenichiro; Fukao, Toshiyuki; Kondo, Naomi
We describe a 35-year-old man with X-linked agammaglobulinemia who had refractory chronic pleurisy caused by a Helicobacter equorum-like bacterium. Broad-range bacterial PCR targeting the 16S and 23S rRNA genes and in situ hybridization targeting the 16S rRNA gene of H. equorum confirmed the presence of this pathogen in a human for the first time. PMID:21677071
Jeong, Do-Won; Han, Seulhwa; Lee, Jong-Hoon
To select starters for jeotgal, a traditional Korean high-salt-fermented seafood, the safety and technological properties of its predominant bacteria isolates, which were identified as Staphylococcus equorum, were assessed. Among the 185 S. equorum isolates from jeotgal, 126 ampicillin-sensitive strains were subjected to assessments for antibiotic susceptibility and safety hazards. Sixty-six out of the 126 S. equorum strains exhibited phenotypic resistances to at least one antibiotic, and their prevailing resistances were to penicillin G (34.1%), erythromycin (9.5%) and trimethoprim (9.5%). Twenty-four S. equorum strains expressed resistance to at least two antibiotics. The lnuA for lincomycin (four strains) and pbp for β-lactam (three strains) were amplified by PCR. α-Hemolytic activity was not detected from the 126 strains, and 87 strains presented δ-hemolytic activity. Among the 87 strains, three strains exhibited β-hemolytic activity. Thirty-seven strains formed a biofilm. A hemolysin gene homologous to that of Staphylococcus epidermidis was amplified from an S. equorum strain with β-hemolytic activity by PCR; however, no PCR product homologous to the previously known staphylococcal enterotoxin genes was amplified. Thirty-nine S. equorum strains cleared all of the tested safety hazards and were adopted for technological property assessments. Among these strains, 16 strains exhibited protease, lipase and nitrate reductase activities, and seven strains did not produce four types of biogenic amines. Five biogenic amine non-producers exhibited three enzyme activities. Most of the strains could grow on the agar with 20% NaCl, and 13 strains maintained growth at the 25% NaCl condition. S. equorum KS1039, which is the most applicable strain that covers the safety and technological requirements for jeotgal, can grow at the 25% NaCl condition. Through this research study, we reconfirmed the necessity of characterization in the functionality and safety of S. equorum
Dever, M L; Kahn, L P
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.
Robles-Pérez, D; Martínez-Pérez, J M; Rojo-Vázquez, F A; Martínez-Valladares, M
The aim of this study was to develop a PCR for the diagnosis of Fasciola hepatica infection in feces of sheep based on the ribosomal internal transcribed spacer. Detection of infection was possible from the second week post-infection in experimentally infected sheep by amplification of a 292bp fragment. This PCR was employed for the detection of anthelmintic resistance (AR) in naturally infected sheep flocks, and results were compared with techniques such as the fecal egg count reduction test (FECRT) and the copro-antigen reduction test (CRT). The FECRT was carried out in two flocks, Santillan de la Vega (SV) and Corullón (CR), with sheep treated with albendazole (ABZ), clorsulon (CL), or triclabendazole (TCBZ). Feces were collected from individuals on days 0, 7, 15, and 30 post-treatment (pt). The FECRT showed adult F. hepatica to be resistant to ABZ and CL in both flocks. All parasite stages in the SV flock were susceptible to TCBZ, while in the CR flock, adult flukes showed resistance and immature forms were susceptible to the treatment. To compare FECRT and the PCR results, we calculated the percent of positive sheep on day 1 pt. In both flocks, the percent positive sheep was consistently higher by PCR than by sedimentation, confirming that the PCR is a more sensitive method of diagnosing infection and therefore to detect the resistance in infected animals. The CRT was carried out in the SV flock using a sandwich ELISA kit. The percent of sheep found positive by PCR was higher than with ELISA. Comparison of FECRT, CRT, and PCR for the detection of AR showed PCR to be the most sensitive. Copyright © 2013 Elsevier B.V. All rights reserved.
Eurico A. Sczesny-Moraes
, 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%.
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
Ingrid Kelly Zanchet
Full Text Available
Para conhecer a situação da resistência anti-hel-míntica em ovinos de propriedades localizadas nos mu-nicípios da Associação dos Municípios do Alto Irani (AMAI, oeste de Santa Catarina, Brasil, foram avalia-dos nove rebanhos pelo teste de redução da OPG (ovos por grama de fezes. Este teste consiste na comparação da média da OPG de um grupo de animais quatorze dias após o tratamento com a média de um grupo controle não-medicado. Consideraram-se efetivas as drogas capazes de reduzir a OPG em 95%. Os princípios ativos utilizados foram: levamisol (7,5 mg/kg, closantel (7,5 mg/Kg, al-bendazol (10 e 5 mg/Kg, ivermectin e moxidectin (0,2 mg/Kg. Detectou-se resistência dos nematódeos gastrin-testinais a todos os grupos anti-helmínticos testados, sen-do que 100% das propriedades apresentam resistência ao ivermectin; 66,7% ao moxidectin, 44,4% ao levamisol e 75% aos benzimidazóis. Para as lactonas macrocíclicas e benzimidazóis, tanto o gênero Haemonchus sp. quanto Trichostrongylus sp. apresentaram resistência. Para o le-vamisol, a resistência está restrita a Trichostrongylus sp. Também foi detectada a presença de uma população de Haemonchus sp. resistente ao closantel e uma de Nema-todirus sp. resistente ao albendazol. Estes dados mostram a urgência de difundir medidas de controle integrado de parasitoses, visando prolongar a vida útil dos princípios ativos ainda disponíveis para uso.
PALAVRAS-CHAVES: Ovinos, resistência anti-helmíntica, Santa Catarina.
In order to know the situation of the anthelmintic resistance in sheep farms in the municipalities of Asso-ciation of the municipalities of the High Irani Region - AMAI, West of Santa Catarina State, Brazil, nine flocks were submitted to the faecal egg counts reduction test (FECRT. This test consists in comparing the mean FEC of a group of sheep, 14 days after treatment with the mean FEC of a non-treated control group. Only drugs that could
Nematode parasites are known to pose a challenge to small ruminant production in Tanzania due to their fast development of resistance to the commonly used anthelmintics. The objective of this study was to determine the resistance of anthelmintics in small ruminants. A total of 30 sheep and 30 goats aged between 6 and ...
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
Ana Carolina Fonseca Lindoso Melo
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
Nielsen, M K; Pfister, K; von Samson-Himmelstjerna, G
Since the 1960s equine parasite control has relied heavily on frequent anthelmintic treatments often applied with frequent intervals year-round. However, increasing levels of anthelmintic resistance in cyathostomins and Parascaris equorum are now forcing the equine industry to change to a more surveillance-based treatment approach to facilitate a reduction in treatment intensity. The principle of selective therapy has been implemented with success in small ruminant parasite control, and has also found use in horse populations. Typically, egg counts are performed from all individuals in the population, and those exceeding a predetermined cutoff threshold are treated. Several studies document the applicability of this method in populations of adult horses, where the overall cyathostomin egg shedding can be controlled by only treating about half the horses. However, selective therapy has not been evaluated in foals and young horses, and it remains unknown whether the principle is adequate to also provide control over other important parasites such as tapeworms, ascarids, and large strongyles. One recent study associated selective therapy with increased occurrence of Strongylus vulgaris. Studies are needed to evaluate potential health risks associated with selective therapy, and to assess to which extent development of anthelmintic resistance can be delayed with this approach. The choice of strongyle egg count cutoff value for anthelmintic treatment is currently based more on tradition than science, and a recent publication illustrated that apparently healthy horses with egg counts below 100 eggs per gram (EPG) can harbor cyathostomin burdens in the range of 100,000 luminal worms. It remains unknown whether leaving such horses untreated constitutes a potential threat to equine health. The concept of selective therapy has merit for equine strongyle control, but several questions remain as it has not been fully scientifically evaluated. There is a great need for new and
Coprological examination revealed the existence of six species and five genera of helminths namely, Strongylus species 39.5 %, Cyathostomes 35.1 %, Trichostrongylus axei 29.0 %, Fasciola hepatica 23.1 %, Triodonthophorus species 13.9%, Parascaris equorum 11.7 %, Oxyuris equi 1.8 %, Strongyloides westeri 0.7 % ...
Lätti, Reet, 1983-
Hobustel esinevad parasiitide - askariidid e varsasolkmed (Parascaris equorum), väikesed ümarussid (Cyathostominae), suured strongüliidid (Strongylus vulgaris, S. edentatus), väikesed ümarussid (Strongyloides westeri), paeluss (Anoplocephala perfoliata) - tõrjest ning nakatumise kontrollimisest
Sczesny-Moraes, Eurico A.; Bianchin, Ivo; Silva, Karina F. da; Catto, João Batista; Honer, Michael Robin; Paiva, Fernando
Entre os métodos de controle da verminose gastrintestinal em ovinos, a utilização de produtos químicos é o mais empregado. Porém, o uso indiscriminado e continuado desses produtos tem selecionado populações de helmintos resistentes aos anti-helmínticos, fenômeno relatado no mundo todo. Este trabalho teve como objetivo identificar as espécies de parasitos gastrintestinais e diagnosticar a situação da resistência anti-helmíntica em ovinos no Estado de Mato Grosso do Sul. Foram realizados testes...
Based on these findings, it is necessary to educate farmers on adapting integrated approach to helminth control with appropriate use of anthelmintics. Les cas de résistance anthelminthique chez les nématodes gastro-intestinaux dans une ferme ovine à Kabete au Kenya ont fait l\\'objet d\\'enquête entre octobre 2005 et mars ...
Nielsen, Martin Krarup; Vidyashankar, Anand N.; Hanlon, Bret
contribute to cause this high variability and these must be taken into account to accurately identify a reduction in anthelmintic efficacy. To address this problem, we developed a hierarchical statistical model for analysis of FECRT data from multiple farms. The model includes animal effect and farm clusters...... = 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...
Díez-Baños, P; Pedreira, J; Sánchez-Andrade, R; Francisco, I; Suárez, J L; Díaz, P; Panadero, R; Arias, M; Painceira, A; Paz-Silva, A; Morrondo, P
A coprological survey to analyze the presence of flock resistance to benzimidazoles (BZ) and macrocyclic lactones (ML) was performed in sheep under field conditions. Fecal samples were collected from 2,625 sheep in 72 commercial farms from Galicia (NW Spain). The in vitro (FECRT, fecal egg count reduction test) and in vivo (EHA, egg hatch assay, and LFIA, larval feeding inhibition assay) tests were used to assess the efficacy of these anthelmintics. Coprocultures were also developed to obtain knowledge on the main genera of trichostrongylid nematoda prior to, and after, the administration of the anthelmintics. By using the FECRT, BZ resistance was observed in 13 (18%) flocks, whereas ML resistance was only detected in 2 (3%) farms. The number of resistant flocks to BZ was 21 (29%) by using the EHA and 7 (10%) by means of the LFIA. None of the flocks used in this study showed simultaneous resistance to both employed anthelmintics. The results from the in vitro and in vivo tests revealed that 92% of the flocks FECRT resistant to BZ were also resistant with the EHA. The LFIA confirmed all the farms resistant to ML by using the in vivo test. After the administration of BZ, nematode larvae belonging to Teladorsagia circumcincta (32.2%), Trichostrongylus spp. (29%), Nematodirus spp. (6.5%), and Chabertia ovis (3.2%) were identified. In the flocks receiving ML, only T. circumcincta was identified (57%). We recommend the use of in vitro tests because they are more efficient. As the use of macrocyclic lactones is increasing in this region, further investigation is needed for detecting resistance to the anthelmintic family compounds by the LFIA.
Pena-Espinoza, Miguel Angel; Enemark, Heidi L.; Thansborg, Stig M.
kids were selected for faecal egg count (FEC) reduction tests. Animals were allocated into one of 5 treatment groups, or 1 untreated control group, for each species. Lambs were treated with 5 mg/kg fenbendazole (FBZ), 0.2 mg/kg moxidectin (MOX), 7 mg/kg levamisole (LEV), 0.2 mg/kg ivermectin (IVM...
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...
Os eqüinos apresentam uma grande variedade de parasitas em sua fauna helmíntica, e algumas espécies/gêneros são de relevada importância, como: Parascaris equorum, Anoplocephala perfoliata, Oxyuris equi, Cyathostomum spp. e Strongylus spp. O controle destas infecções depende principalmente da utilização de produtos antiparasitários de forma supressiva ou estratégica e, em menor escala, de forma curativa. O tratamento supressivo é o fator mais importante na promoção da seleção de organismos res...
Full Text Available Within the practical training of students of the fifth year of studies of veterinary medicine, a demonstration was performed of stallion castration. The owner of the horse decided to take this step because of the unpredictable temperament of the stallion. The castration was carried out under general anaesthesia. The stallion was laid down and immobilized for castration in keeping with the so-called in-the-field conditions. The castration proceeded without any complications, and the postoperative course was in order. Three days after castration, the horse died with symptoms of colic. The autopsy showed obturation of the ileum and ileocecal valve by Parascaris equorum parasites.
Saleh Mohammed Jajere
Full Text Available Aim. This survey study was conducted from April 2014 through March 2015 in Bauchi, Yobe, and Gombe states, northeastern Nigeria, to explore the risk factors associated with the occurrence of gastrointestinal helminthosis among indigenous donkeys (Equus asinus. Materials and Methods. A total of six hundred fresh faecal samples were randomly collected from indigenous donkeys of varying age, sex, and settlements. Simple flotation and sedimentation techniques were used for the detection of helminths eggs. Results. Three gastrointestinal nematode parasites were encountered including Strongyle, Parascaris equorum, and Oxyuris equi. An overall prevalence of 98.3% was obtained, of which 78.3%, 40.3%, and 17.5% were, respectively, from Strongyle, Parascaris equorum, and Oxyuris equi. Age, sex, and season were not statistically associated with the risk of helminth infection as were the different study areas (p>0.05. However, body condition score, settlement, anthelminthic medication history, and management practices were significantly associated with the risk of gastrointestinal helminthosis. Statistically high prevalence of helminthic infections was observed in donkeys, with poor (thin body condition, from rural settlements, that were not dewormed and raised under poor management systems (p<0.001. Conclusion. It is concluded from the study that gastrointestinal helminths particularly Strongyle were endemic among the indigenous donkeys in northeastern Nigeria. Further control and preventive measures were discussed.
Mohammed Jajere, Saleh; Rabana Lawal, Jallailudeen; Mohammed Bello, Amina; Wakil, Yakaka; Aliyu Turaki, Usman; Waziri, Ibrahim
Aim. This survey study was conducted from April 2014 through March 2015 in Bauchi, Yobe, and Gombe states, northeastern Nigeria, to explore the risk factors associated with the occurrence of gastrointestinal helminthosis among indigenous donkeys (Equus asinus). Materials and Methods. A total of six hundred fresh faecal samples were randomly collected from indigenous donkeys of varying age, sex, and settlements. Simple flotation and sedimentation techniques were used for the detection of helminths eggs. Results. Three gastrointestinal nematode parasites were encountered including Strongyle, Parascaris equorum, and Oxyuris equi. An overall prevalence of 98.3% was obtained, of which 78.3%, 40.3%, and 17.5% were, respectively, from Strongyle, Parascaris equorum, and Oxyuris equi. Age, sex, and season were not statistically associated with the risk of helminth infection as were the different study areas (p > 0.05). However, body condition score, settlement, anthelminthic medication history, and management practices were significantly associated with the risk of gastrointestinal helminthosis. Statistically high prevalence of helminthic infections was observed in donkeys, with poor (thin) body condition, from rural settlements, that were not dewormed and raised under poor management systems (p < 0.001). Conclusion. It is concluded from the study that gastrointestinal helminths particularly Strongyle were endemic among the indigenous donkeys in northeastern Nigeria. Further control and preventive measures were discussed.
Determining the Degree of Anthelmintic Resistance against Macrocyclic Lactones in Small Strongylus, Based on the Larvae Development Analysis (LDA in Horses from Grasslands in the Department of Casanare
Germán Alonso Prada
Full Text Available This study was conducted in the grasslands of the Department of Casanare, seeking to determine the degree of anthelmintic susceptibility or resistance against macrocyclic lactones in small populations of Strongylus in this region of the country. Samples were taken from four municipalities in the department: Aguazul, Paz de Ariporo, Maní and El Yopal, between June, 2006 and April, 2007, where ten fresh fecal samples were collected in each municipality, directly from the field and in a completely random way. The samples were processed using the MacMaster coprological technique, thus determining the highest count of fecal epg by municipality. L3 larvae were extracted from each of the six samples with the highest count of fecal epg through the coprological and Baermann-Wetzel tests with which the Larvae Development Analysis (LDA test was run, detecting Small Strongylus highly susceptible to the action of macrocyclic lactones (ivermectin.
Fernández, V; Estein, S; Ortiz, P; Luchessi, P; Solana, V; Solana, H
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.
Antonio Pereira de Souza
Full Text Available Com o objetivo de avaliar focos de resistência de helmintos gastrintestinais parasitos de bovinos, com aptidão para corte, à ivermectina, ao fosfato de levamisole e ao sulfóxido de albendazole foram realizadas avaliações em 39 propriedades localizadas no Planalto Catarinense. Em cada uma foram coletadas, em média, 60 amostras de fezes de animais, com idade entre sete e 18 meses, para avaliar o número de ovos por grama de fezes (OPG, o cultivo e a identificação de larvas. Foram formados três grupos de animais com média de OPG semelhante. A média de OPG de todos os animais coletados, antes do tratamento, foi considerada como testemunha. Cada grupo recebeu um tratamento diferente por via subcutânea (ivermectina na dose de 0,2mg kg-1, fosfato de levamisole na dose de 5mg kg-1 e sulfóxido de albendazole na dose de 2,5mg kg-1. Sete dias após foram repetidas as avaliações por grupo. Considerou-se resistência quando a eficácia da droga foi menor que 95% e o limite inferior do intervalo de confiança IC95% menor que 90%. Entre as propriedades avaliadas, 82,1% apresentaram animais com helmintos resistentes à ivermectina, 15,4% ao fosfato de levamisole e 7,8% ao sulfóxido de albendazole. Em apenas 10,3% das propriedades, a eficácia de todos os anti-helmínticos foi superior a 95%. Com base no diagnóstico genérico, verificou-se que os gêneros predominantes resistentes à ivermectina foram Cooperia spp e Haemonchus spp, ao fosfato de levamisole, Ostertagia spp, Cooperia spp e Trichostrongylus spp, e ao sulfóxido de albendazole, Cooperia spp.In order to estimate possible focus of resistance of bovine gastrointestinal helminths to ivermectin, levamisole phosphate and albendazole sulphoxide, 39 properties in the Santa Catarina Plateau were evaluated. In each location a mean number of 60 samples of feces was collected from animals between seven and 18 months of age, for the evaluation of the number of eggs per gram of feces (EPG, for culture and for larvae identification. Three groups with similar EPG mean were comprised, with the mean EPG being considered the group control. Animal from each group received a different subcutaneous treatment: a 0.2mg kg-1 ivermectin, b 5mg kg-1 levamisole phosphate, and c 2.5mg kg-1 albendazole sulphoxide. The fecal evaluation, per group, was repeated seven days later. Resistance was considered when the efficacy of the drug was lesser than 95% and when the inferior limit of the confidence interval CI95% was lesser than 90%. From the properties examined, 82.1% presented animals with helminths resistant to ivermectin, 15.4% to levamisole phosphate, and 7.8% to albendazole sulphoxide. In only 10.3% of the properties the efficacy of all the anthelmintics was superior to 95%. Based on the generic diagnosis, through the larvae identification, Cooperia spp and Haemonchus spp were the prevailing resistant genus to ivermectin, Ostertagia spp, Cooperia spp and Trichostrongylus spp were associated with resistance to levamisole phosphate, and Cooperia spp. to albendazole sulphoxide.
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.
Weslen Fabricio Pires Teixeira
Full Text Available With the aim of studying the endoparasite fauna of horses from the Formiga city, located in center-west region of the state of Minas Gerais, 25 animals that were naturally infected with helminths were evaluated. By means of parasitological necropsies, different endoparasites were found. The subfamily Cyathostominae presented the highest incidence, followed by Trichostrongylus axei, Oxyuris equi, Triodontophorus serratus, Strongyloides westeri, Strongylus edentatus, Habronema muscae, Parascaris equorum, Probstmayria vivipara, Strongylus vulgaris, Gasterophilus nasalis, Anoplocephala magna and Anoplocephala perfoliata. In the present study, if the species Probstmayria vivipara was not considered in the prevalence, the frequency of Cyathostominae was equivalent to 94.85%. The results obtained in this study allowed us to detect and identify different species of helminths in horses, and confirmed the high incidence of nematodes belonging to the subfamily Cyathostominae in the center-west region of Minas Gerais.
Full Text Available Acetylcholine receptors are pentameric ligand-gated channels involved in excitatory neuro-transmission in both vertebrates and invertebrates. In nematodes, they represent major targets for cholinergic agonist or antagonist anthelmintic drugs. Despite the large diversity of acetylcholine-receptor subunit genes present in nematodes, only a few receptor subtypes have been characterized so far. Interestingly, parasitic nematodes affecting human or animal health possess two closely related members of this gene family, acr-26 and acr-27 that are essentially absent in free-living or plant parasitic species. Using the pathogenic parasitic nematode of ruminants, Haemonchus contortus, as a model, we found that Hco-ACR-26 and Hco-ACR-27 are co-expressed in body muscle cells. We demonstrated that co-expression of Hco-ACR-26 and Hco-ACR-27 in Xenopus laevis oocytes led to the functional expression of an acetylcholine-receptor highly sensitive to the anthelmintics morantel and pyrantel. Importantly we also reported that ACR-26 and ACR-27, from the distantly related parasitic nematode of horses, Parascaris equorum, also formed a functional acetylcholine-receptor highly sensitive to these two drugs. In Caenorhabditis elegans, a free-living model nematode, we demonstrated that heterologous expression of the H. contortus and P. equorum receptors drastically increased its sensitivity to morantel and pyrantel, mirroring the pharmacological properties observed in Xenopus oocytes. Our results are the first to describe significant molecular determinants of a novel class of nematode body wall muscle AChR.
Luiz Silva Vieira
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 treatment and 7 days later, to provide material for egg counts and larval cultures. Among 34 surveyed herds 20.6% showed levamisole resistance, 17.6% showed resistance to benzimidazole, and 35.3% had multiple resistance. At the time of the assessment 52.9% of the farmers were using broad spectrum anthelmintics. Only 26.5% of the surveyed herds had nematode populations susceptible to the anthelmintics assessed. The results of larval cultures showed that larvae surviving the treatment with oxfendazole were mainly Haemonchus sp and, to a lesser extent, Oesophagostomum sp; those surviving levamisole treatment were Haemonchus sp, Oesophagostomum sp and Trichostrongylus sp.
Maria do Socorro Veloso Leite Ferraz da Costa
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 redução significativa (p > 0,05 nas contagens de OPG em nenhum dos grupos após os tratamentos anti-helmínticos, e as larvas encontradas na coprocultura foram Cooperia, Haemonchus, Oesophagostomum e poucos Trichostrongylus, com predominância de Cooperia nos quatro meses iniciais e Haemonchus nos meses seguintes. No ano de 2003, bezerros traçadores foram alocados mensalmente nos pastos, apresentando altas cargas parasitárias na maioria dos meses do ano, sendo as principais espécies identificadas: Cooperia punctata e Haemonchus contortus. Os resultados indicam que os nematódeos da fazenda apresentam resistência anti-helmíntica, principalmente às lactonas macrocíclicas, e o desenvolvimento de imunidade foi primordial para reduzir a carga parasitária dos animais mestiços.
Marcelo Beltrão Molento
Full Text Available Os eqüinos apresentam uma grande variedade de parasitas em sua fauna helmíntica, e algumas espécies/gêneros são de relevada importância, como: Parascaris equorum, Anoplocephala perfoliata, Oxyuris equi, Cyathostomum spp. e Strongylus spp. O controle destas infecções depende principalmente da utilização de produtos antiparasitários de forma supressiva ou estratégica e, em menor escala, de forma curativa. O tratamento supressivo é o fator mais importante na promoção da seleção de organismos resistentes, prejudicando a sustentabilidade de qualquer programa sanitário. As formas de detecção da resistência parasitária são onerosas e as mais comuns expressam resultados imprecisos. Entretanto, estas técnicas servem para monitorar a evolução e determinar os organismos envolvidos. A combinação de drogas é uma ferramenta que deve ser utilizada com muita cautela, pois esta alternativa não garante uma redução significativa de organismos resistentes aos compostos envolvidos. O objetivo deste artigo é apresentar formas de planejamento que auxiliem a melhorar a condição sanitária, o bem-estar dos animais e preserve o efeito tóxico dos produtos antiparasitários.Equines harbour a variety of parasitic organisms on their helminth fauna and there are a few species/genus of interest, such as: Parascaris equorum, Anoplocephala perfoliata, Oxyuris equi, Cyathostomum spp. and Strongylus spp. The control of these infections relies mostly on the suppressive or strategic usage of antiparasitic compounds, and to a less extent on curative/salvage treatments. Suppressive treatment is the most important factor regarding the selection of resistant organisms, causing the impairment of sanitary programs. Detection methods of parasite resistance are expensive and the most common ones express variable results. Although, these techniques allow monitoring the evolution and the determination of which organisms are involved. Drug combination is a
Durant, Jean-Francois; Irenge, Leonid M; Fogt-Wyrwas, Renata; Dumont, Catherine; Doucet, Jean-Pierre; Mignon, Bernard; Losson, Bertrand; Gala, Jean-Luc
Toxocarosis is a zoonotic disease caused by Toxocara canis (T. canis) and/or Toxocara cati (T. cati), two worldwide distributed roundworms which are parasites of canids and felids, respectively. Infections of humans occur through ingestion of embryonated eggs of T. canis or T. cati, when playing with soils contaminated with dogs or cats feces. Accordingly, the assessment of potential contamination of these areas with these roundworms eggs is paramount. A duplex quantitative real-time PCR (2qPCR) targeting the ribosomal RNA gene internal transcribed spacer (ITS2) has been developed and used for rapid and specific identification of T. canis and T. cati eggs in fecal and soil samples. The assay was set up on DNA samples extracted from 53 adult worms including T. canis, T. cati, T. leonina, Ascaris suum (A. suum) and Parascaris equorum (P. equorum). The assay was used to assess the presence of T. cati eggs in several samples, including 12 clean soil samples spiked with eggs of either T. cati or A. suum, 10 actual soil samples randomly collected from playgrounds in Brussels, and fecal samples from cats, dogs, and other animals. 2qPCR results on dogs and cats fecal samples were compared with results from microscopic examination. 2qPCR assay allowed specific detection of T. canis and T. cati, whether adult worms, eggs spiked in soil or fecal samples. The 2qPCR limit of detection (LOD) in spiked soil samples was 2 eggs per g of soil for a turnaround time of 3 hours. A perfect concordance was observed between 2qPCR assay and microscopic examination on dogs and cats feces. The newly developed 2qPCR assay can be useful for high throughput prospective or retrospective detection of T.canis and/or T. cati eggs in fecal samples as well as in soil samples from playgrounds, parks and sandpits.
Pérez Cordón, G; Hitos Prados, A; Romero, D; Sánchez Moreno, M; Pontes, A; Osuna, A; Rosales, M J
Gastrointestinal parasites cause serious diarrhoea in captive animals. Therefore, we have undertaken this study to establish programmes to prevent, control, and treat intestinal parasitism in the animals of the zoological garden "Peña Escrita" of Almuñecar (Granada). An annual survey was conduced to estimate the occurrence of gastrointestinal parasites and the seasonality of this parasitism. Between June 2006 and May 2007, 432 samples were collected from primates, carnivores, perissoodactyla, artiodactyla, rodentia, diprotodontia, galliformes, anseriformes and struthioniformes. One or more intestinal parasites were identified in 72.5% of the animals. The most frequent pathogenic endoparasites were Eimeria spp. (17.3%), Trichuris spp. (5.1%), Strongyloides spp. (4.5%), Cyclospora spp. (4.5%), Cryptosporidium spp. (3.2%) and Isospora spp. (2.6%). Iodamoeba butschlii, Parascaris equorum and Trichuris spp. did not vary with season and Cryptosporidium spp., Dicrocoelium dendriticum, Metastrongylus spp. and Cylicospirura spp. appeared exclusively in Artiodactyla. Multiple parasitic infections were common, 70% of animals presented with at least two parasites (maximum=6). The most frequent cases of multiple parasitism were Eimeria spp. plus Blastocystis spp. and Eimeria spp. plus Nematodirus spp., in the last case the animals presented explosive diarrhoea. In accord with our results, after each sampling, some of the affected animals were treated and the corresponding programmes of prevention and control were designed.
Hernández, José A; Vázquez-Ruiz, Rosa A; Cazapal-Monteiro, Cristiana F; Valderrábano, Esther; Arroyo, Fabián L; Francisco, Iván; Miguélez, Silvia; Sánchez-Andrade, Rita; Paz-Silva, Adolfo; Arias, María S
Abstract : There are certain saprophytic fungi in the soil able to develop an antagonistic effect against eggs of parasites. Some of these fungal species are ingested by animals during grazing, and survive in their feces after passing through the digestive tract. To identify and isolate ovicidal fungi in the feces of wild captive animals, a total of 60 fecal samples were taken from different wild animals kept captive in the Marcelle Natureza Zoological Park (Lugo, Spain). After the serial culture of the feces onto Petri dishes with different media, their parasicitide activity was assayed against eggs of trematodes ( Calicophoron daubneyi ) and ascarids ( Parascaris equorum ). Seven fungal genera were identified in the feces. Isolates from Fusarium , Lecanicillium , Mucor , Trichoderma , and Verticillium showed an ovicidal effect classified as type 3, because of their ability to adhere to the eggshell, penetrate, and damage permanently the inner embryo. Penicillium and Gliocladium developed a type 1 effect (hyphae attach to the eggshell but morphological damage was not provoked). These results provide very interesting and useful information about fungi susceptible for being used in biological control procedures against parasites.
Vercruysse, Jozef; Levecke, Bruno; Prichard, Roger
With the London Declaration on neglected tropical disease (NTD), we are entering a new era of combating NTDs. However, the worldwide prospects of increased mass drug administration (MDA) treatments warrant caution on the development of anthelmintic resistance. In this review, we discuss the practical implications of MDA programs on the development of anthelmintic resistance in human soil-transmitted helminths (STH). There is poor evidence of anthelmintic resistance in human STH. Moreover, there is presumptive evidence that the refugia in MDA programs to control human STH is currently large, suggesting that the development of anthelmintic resistance in STH will be slow or may not occur. It remains unclear whether the current MDA strategy to control STH will sufficiently delay or prevent the development of anthelmintic resistance. First, differences in efficacy across and within STH species, and seasonal transmission of STH have not yet been considered. Second, any surveillance system to monitor drug efficacy is lacking. Finally, there is still no agreed strategy on how to deal with anthelmintic resistance once it emerges. Although anthelmintic resistance in human STH is currently of limited concern, various actions should be put in place for its delay and monitoring, and strategies should be developed in case anthelmintic resistance occurs.
André Antonio Cutolo
Full Text Available Twenty horses naturally infected with nematodes were included in a blind, controlled field study on efficacy and safety of an oral 2% ivermectin formulation at a dose of 0.2 mg.kg-1. Horses were divided into treated and non-treated (control groups with ten animals each based on preliminary counts of eggs per gram of feces (EPG. Stool samples were collected after treatment for identification of nematode species. Clinical evaluations and EPG counts were performed on days 0, +5, +14 and +19. Nineteen nematode species were identified: Coronocyclus ulambajari, Craterostomum acuticaudatum, Cyathostomum catinatum, Cyathostomum pateratum, Cylicocyclus brevicapsulatus, Cylicocyclus insigne, Cylicocyclus leptostomum, Cylicocyclus nassatus, Cylicocyclus ultrajectinus, Cylicocyclus spp., Cylicostephanus calicatus, Cylicostephanus longibursatus, Cylicostephanus poculatus, Habronema muscae, Habronema spp., Parascaris equorum, Poteriostomum imparidentatum, Oxyuris equi and Triodontophorus spp. The mean EPG counts of treated and non-treated (control groups on Days -15, 0, +5, +14 and +19 were 1925, 1340, 0, 12.5, 0, 1470, 790, 875, 1605 and 1240 respectively. The efficacy of treatment on Days +5, +14 and +19 was 100, 99.2 and 100% respectively, with a significant difference compared to the control group (p Vinte equinos naturalmente infectados com nematódeos foram utilizados em estudo cego, controlado, de eficácia e segurança clínica a campo de uma formulação oral de ivermectina a 2%, na dosagem de 0,2 mg.kg-1. Foram distribuídos em grupos: tratado e sem tratamento, de dez animais cada, baseados na contagem prévia de ovos por grama de fezes (OPG. Amostras de fezes foram colhidas pós-tratamento para identificação da helmintofauna. Avaliações clínicas e OPG foram realizados nos dias 0, +5, +14 e +19. Identificou-se dezenove espécies de nematódeos: Coronocyclus ulambajari, Craterostomum acuticaudatum, Cyathostomum catinatum, Cyathostomum pateratum
Güiris, A D M; Rojas, H N M; Berovides, A V; Sosa, P J; Pérez, E M E; Cruz, A E; Chávez, H C; Moguel, A J A; Jimenez-Coello, M; Ortega-Pacheco, A
A cross sectional survey was performed to identify gastrointestinal helminths and protozoans in naturally infected horses from the biosphere reserve known as "La Sierra Madre de Chiapas", Mexico (El Triunfo and La Sepultura). During a three-year survey, fecal samples from 90 horses and parasites from 2 necropsied animals were collected. Five families from the Nematoda class: Ascaridae, Kathlanidae, Oxyuridae, Strongylidae and Trichostrongylidae were found, whereas, only one family from the class Cestoda, was observed: Anoplocephalidae. One family from the class Insecta, was observed: Gasterophiilidae. The number of species of parasites ranged from 13 to 18 with an average of 15 per animal. Adult parasites were recovered from the large intestine luminal contents at necropsy. Species recovered included: Strongylus vulgaris, S. equinus, S. edentatus, Oxyuris equi, Parascaris equorum, Coronocyclus coronatum, C. labiatus, C. labratus, Cyathostomum tetracanthum, Cylicocyclus insigne, C. leptostomus, Cylicodontophorus bicoronatus, Cylicostephanus asymetricus, C. bidentatus, C. minutus, C. longibursatus, Petrovinema poculatum, Poteriostomum imparidentatum, Cylicostephanus goldi, Tridentoinfundibulum gobi, Triodontophorus serratus and T. tenuicollis. One species of Diptera were recovered from stomach and identified: Gasterophilus intestinalis. Furthermore, different species of protozoa were recovered from fresh horse-dung and identified in four classes: Sporozoa, Litostomatea, Ciliasida and Suctoria. Nine families: Cryptosporidiidae, Eimeriidae, Balantidiidae, Buetschliidae, Blepharocorythidae, Cycloposthiidae, Spirodiniididae, Ditoxidae, Acinetidae; and 31 ciliates species were recorded: Allantosoma dicorniger, A. intestinalis, Alloiozona trizona, Blepharosphaera intestinalis, Blepharoprosthium pireum, Blepharoconus benbrooki, Bundleia postciliata, Didesmis ovalis, D. quadrata, Sulcoarcus pellucidulus, Blepharocorys angusta, B. cardionucleata, B. curvigula, B. juvata, B
Abongwa, Melanie; Baber, Katherine E; Martin, Richard J; Robertson, Alan P
Nematode parasite infections pose a significant threat in human and veterinary medicine. At least a third of the world's population is at risk from nematode parasite infections. These infections not only cause health problems, but also cause loss of livestock production and hence, economic losses. Anthelmintic drugs are the mainstay by which control of nematode parasite infections is achieved. Many of the currently available anthelmintics act on nicotinic acetylcholine receptors (nAChRs). However, the detailed mode of action (MOA) of these anthelmintics is not clearly understood. Elucidation of the MOA of anthelmintics is highly desirable; an in-depth knowledge of the MOA will better inform on mechanisms of resistance development and on ways to slow down or overcome resistance. The cholinomimetic anthelmintic, morantel, has a complex MOA involving the activation and block of levamisole-sensitive single nAChR channels (L-type nAChR or L-nAChR). More recently, morantel has been demonstrated to activate Haemonchus contortus and Parascaris equorum ACR-26/ACR-27 nAChRs expressed in Xenopus laevis oocytes. Previous studies in our laboratory, however, have shown morantel does not activate the nicotine-sensitive nAChR (N-type nAChR or N-nAChR), Ascaris suum ACR-16 (Asu-ACR-16). In this study, we used two-electrode voltage-clamp (TEVC) electrophysiology to investigate the inhibitory effects of morantel, on expressed Asu-ACR-16 nAChRs in X. laevis oocytes. Our results show that morantel acts as a non-competitive antagonist on Asu-ACR-16. This non-competitive antagonism by morantel was further demonstrated to be voltage-sensitive. We conclude based on our findings that morantel is a non-competitive voltage-sensitive open channel blocker of Asu-ACR-16.
Full Text Available Thirty-one Polish primitive horses (Equus caballus from three herds (two from the reserve and onefrom the stable were dewormed with ivermectin+praziquantel and examined for the gastrointestinalparasite fauna. A total of 21.231 parasites were collected from the faeces at 24, 36 and 48 hoursposttreatment. There were 35 nematode species, one cestode and one botfl y larva. Strongyloideswesteri infection was confirmed pretreatment by faecal sample examination and no threadwormspecimens were found after deworming. Large and small strongyle prevalence was 90 % – 100 % and represented by 31 species. Among a total of 25 cyathostome species recovered (from 19 to 24in each group, five species (C. catinatum, C. minutus, C. longibursatus, C. nassatus and C. ashworthihad a prevalence of 100 % in three groups of horses. Meanwhile 14 species were 100 % prevalent in one herd. A total of six large strongyle species were found in adult horses. Oxyuris equiwas recorded in 60 – 100 % of the horses while Parascaris equorum was detected in 100 % of foalsand 16.7 % – 30 % of adult mares. Habronema muscae was found in 30 % of the horses from onefree-ranging herd. Tapeworms (Anoplocephala perfoliata were found in 90 % of the horses from onefree-ranging group, whereas botfl y larvae (Gasterophilus intestinalis were found in 50 – 80 % of allsurveyed horses. The present results are compared with earlier studies of Polish primitive wild horsesfrom similar reserves in Poland. A total of 36 gastrointestinal parasite species were recorded fromwild and stabled horses from the Biebrza National Park. This is in comparison with 35 such speciesin free-ranging and stabled horses from the Roztocze National Park and with 28 such species offree-ranginghorses from the Popielno forest reserve.
..., Strongylus equinus), small strongyles (Trichonema spp., Triodontophorus), pinworms (Oxyuris), and large...., Triodontophorus), pinworms (Oxyuris), and large roundworms (Parascaris). (ii) It is administered as a single dose...
Charlier, J.; Levecke, B.; Devleesschauwer, B.; Vercruysse, J.; Hogeveen, Henk
Current control practices against gastrointestinal nematodes in dairy cows rely strongly on anthelmintic use. To reduce the development of anthelmintic resistance or disposition of drug residues in the environment, novel control approaches are currently proposed that target anthelmintic treatment to
Full Text Available 50764_1( AY350764 |pid:none) Parascaris univalens ribosomal pro... 95 2e-18 AE016817_243( AE016817 |pid:none...5e-17 EU125004_1( EU125004 |pid:none) Eurythoe complanata ribosomal prot... 91 5e-17 AY350763_1( AY350763 |pid:none) Parascaris
Full Text Available ntromere-associated protein OS=Parascaris univalens Align length 142 Score (bit) 48.1 E-value 2.0e-05 Report...nd centromere-associated protein OS=Parascaris univalens GN=PUMA1 PE=2 SV=1 Lengt
Full Text Available t : Swiss-Prot sp_hit_id O61308 Definition sp|O61308|PUMA_PARUN 227 kDa spindle- and centromere-associated protein OS=Parascaris...dle- and centromere-associated protein OS=Parascaris univalens GN=PUMA1 PE=2 SV=1 Length = 1955 Score = 44.7
Otsen, M.; Plas, M. E.; Lenstra, J. A.; Roos, M. H.; Hoekstra, R.
The alarming development of anthelmintic resistance in important gastrointestinal nematode parasites of man and live-stock is caused by selection for specific genotypes. In order to provide genetic tools to study the nematode populations and the consequences of anthelmintic treatment, we isolated
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...
Co-administration of Albendazole and Levamisole to control multiple anthelmintic resistant nematodes in a sheep farm in Kabete Kenya · EMAIL FULL TEXT EMAIL FULL TEXT DOWNLOAD FULL TEXT DOWNLOAD FULL TEXT. CJ Nganga, DW Gakuya, RO Otieno, RW Githinji, 275-278 ...
Maingi, N.; Bjørn, H.; Gichohi, V.M.
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...
Githinji, RW. Vol 62, No 3 (2014) - Articles Co-administration of Albendazole and Levamisole to control multiple anthelmintic resistant nematodes in a sheep farm in Kabete Kenya Abstract. ISSN: 0378-9721. AJOL African Journals Online. HOW TO USE AJOL... for Researchers · for Librarians · for Authors · FAQ's · More ...
The prevalence of anthelmintic resistance and the consumer demand for alternative farming systems that limit the use of chemical anthelmintics has made the search for alternative gastrointestinal nematode parasites control methods crucial. Traditional medicinal/herbal plants can offer an alternative to the reliance on ...
Magwisha, H.B.. Vol 32, No 1 (2017) - Articles Determination of anthelmintic resistance in goats and sheep using faecal egg count reduction test at Luguruni farm, Dar es Salaam, Tanzania Abstract · Vol 32, No 1 (2017) - Articles Molecular diversity of Theileria parva: a case study of Kilosa district, Morogoro, Tanzania
Nielsen, Martin Krarup; Peterson, David S.; Monrad, Jesper
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 s...
Because of a high prevalence of anthelmintic resistance and consumer demand for chemical free meat products, management tools to minimize the need for deworming are needed. The objective was to examine the effectiveness of grazing sericea lespedeza (SL) in a mixed grass or a pure forage system for ...
O'Grady, M. R.; Slocombe, J. O. D.
Several variables in a standard vial fecal gravitational flotation technique were investigated. These were the specific gravity of the sodium nitrate flotation solution, duration of flotation and mesh sizes of strainers. The number of eggs which floated and adhered to a coverslip were counted and estimates of the number of eggs remaining in the strained fecal suspension and in the feces trapped on the strainer were made. Eggs from hookworms, Trichuris vulpis and Toxocara canis in feces from dogs, Nematodirus spp. from sheep and Parascaris equorum from horses floated equally well in solutions with specific gravities (SpGr) ranging from 1.22-1.38. Taenia spp. from dogs had a slightly narrower range (SpGr 1.27-1.38) for best recovery. Eggs from Haemonchus contortus from sheep appeared to float best between SpGr 1.22- 1.32. Strongyles from one horse floated best with SpGr 1.27-1.32 and from another with SpGr 1.11-1.38. Coccidial oocysts from sheep floated best in a narrow range of SpGr from 1.22-1.27. However, as the SpGr of the solution was increased the recognition of eggs under the coverslip was increasingly difficult and especially so at SpGr 1.38 with sheep feces. This was due to the increasing amount of debris and the more rapid formation of crystals with evaporation with solutions of higher SpGr. It appeared, therefore, that solutions with SpGr of 1.22-1.35 would be best for routine laboratory use. At specific gravity 1.27, there appeared to be no difference in the number of eggs recovered for a four, eight and 12 min flotation period. Only 3-7% of the eggs in 4 g of feces were counted under the coverslip. This poor efficacy resulted first because approximately 50% of the eggs were trapped in the feces and retained on the strainer. Secondly, only one half of the strained fecal suspension, containing approximately 25% of the eggs, was placed in the vial for examination. Thirdly, of those eggs in the vial only 16-29% were counted under the coverslip. When the
This conference covered areas like, technology delivery, compared the precision of experiments in biometrics, the role of agro-veterinary shops in animal health services,Preliminary economic evaluation,anthelmintic resistance survey,Impact of AIDS/HIV, improvement of dairy cattle productivity, dry season feeding for smallholder dairy farmers, screening of tree species, diagnosis of cowdriosis in sheep in Kenya, Camel diseases, effect of growth environment, and many others
Andersen, Ulla Vestergaard; Howe, Daniel K.; Olsen, Susanne Nautrup
Strongyle parasites are ubiquitous in grazing horses, with cyathostomins being the most prevalent, but the large strongyles having larger clinical impact. Strongylus vulgaris is considered most pathogenic nematode, with migrating larvae causing verminous endarteritis and potentially ischaemic...... infarction of intestinal segments. Developing anthelmintic resistance in equine parasites has put emphasis on less intensive treatment regimens to maintain efficacy of current anthelmintics. This has been associated with apparent re-emergence of S. vulgaris. Currently there are no methods for diagnosing...
Hördegen, P.; Cabaret, J.; Hertzberg, H.; Langhans, W.; Maurer, V.
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...
Silvestre, A; Cabaret, J
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.
Full Text Available Intestinal strongyles are the most problematic endoparasites of equids as a result of their wide distribution and the spread of resistant isolates throughout the world. While abundant literature can be found on the extent of anthelmintic resistance across continents, empirical knowledge about associated risk factors is missing. This study brought together results from anthelmintic efficacy testing and risk factor analysis to provide evidence-based guidelines in the field. It involved 688 horses from 39 French horse farms and riding schools to both estimate Faecal Egg Count Reduction (FECR after anthelmintic treatment and to interview farm and riding school managers about their practices. Risk factors associated with reduced anthelmintic efficacy in equine strongyles were estimated across drugs using a marginal modelling approach. Results demonstrated ivermectin efficacy (96.3%Â Â±Â 14.5% FECR, the inefficacy of fenbendazole (42.8%Â Â±Â 33.4% FECR and an intermediate profile for pyrantel (90.3%Â Â±Â 19.6% FECR. Risk factor analysis provided support to advocate for FEC-based treatment regimens combined with individual anthelmintic dosage and the enforcement of tighter biosecurity around horse introduction. The combination of these measures resulted in a decreased risk of drug resistance (relative risk of 0.57, pÂ =Â 0.02. Premises falling under this typology also relied more on their veterinarians suggesting practitionners play an important role in the sustainability of anthelmintic usage. Similarly, drug resistance risk was halved in premises with frequent pasture rotation and with stocking rate below five horses/ha (relative risk of 0.53, pÂ <Â 0.01. This is the first empirical risk factor analysis for anthelmintic resistance in equids. Our findings should guide the implementation of more sustained strongyle management in the field. Keywords: Horse, Nematode, Anthelmintic resistance, Strongyle, Cyathostomin
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.
Nielsen, Martin Krarup
in 1966. The province of Quebec in Canada, and an increasing number of European countries, have implemented prescription-only restrictions on anthelmintic drugs. Denmark introduced this legislation ten years ago, and some evidence has been generated describing potential consequences. It is without dispute...... 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...
Desrues, O.; Pena-Espinoza, Miguel Angel; Hansen, T. V.
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...... difference in total worm burdens of C. oncophora was found between the groups. Weight gains were lower for Group SF (P phosphorus levels in the SF diet, despite similar feed intake at pen-level. CONCLUSIONS: Overall, the effect of sainfoin on abomasal...
Andersen, Ulla Vestergaard; Howe, Daniel K.; Dangoudoubiyam, Sriveny
Strongyle parasites are ubiquitous in grazing horses. Strongylus vulgaris, the most pathogenic of the large strongyles, is known for its extensive migration in the mesenteric arterial system. The lifecycle of S. vulgaris is characterised by a long prepatent period where the migrating larvae...... are virtually undetectable as there currently is no test available for diagnosing prepatent S. vulgaris infection. Presence of S. vulgaris larvae in the arterial system causes endarteritis and thrombosis with a risk of non-strangulating intestinal infarctions. Emergence of anthelmintic resistance among...
Preston, Sarah Jane Margaret; Sandeman, Mark; Gonzalez, Jorge; Piedrafita, David
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.
Gori, Klaus; Jespersen, Lene
). Corynebacterium casei, Microbacterium barkeri, Microbacterium gubbeenense and S. equorum subsp. linens (all isolated from the smear of surface ripened cheeses) using the AI-2 bioluminescence assay. This indicates that AI-2 signaling could take place between bacteria found in the smear of surface ripened cheeses....
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
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.
Mahmmod, Yasser; Svennesen, Line; Pedersen, Karl
identified in milk samples. Staphylococcus chromogenes was detected in both milk (n= 2) and teat skin (n= 1) samples. Data collection will be finished in April 2017. The final results will give new insights into herd specific CNS species patterns and the microbial ecology and epidemiology of common CNS...... on Staphylococcus selective medium (SA Select) and calf blood agar. Colonies from quarters suspect of having CNS in milk and/or teat skin samples (cut-off five CFU) are subjected for MALDI-TOF assay for species identification. Only isolates from the right hind and left front quarters are analyzed by MALDI-TOF assay...... swabs and one milk sample) were harboring more than one type of the CNS species. Staphylococcus epidermidis and Staphylococcus equorum were the most frequently isolated CNS species from milk samples (7/17) and (5/17), respectively. Staphylococcus equorum, Staphylococcus haemolyticus and Staphylococcus...
Gori, Klaus; Moslehi Jenabian, Saloomeh; Purrotti, Micol
Bacterial strains of the species Arthrobacter nicotianae, Corynebacterium ammoniagenes, Corynebacterium casei, Microbacterium barkeri, Microbacterium gubbeenense and Staphylococcus equorum subsp. linens, all isolated from the smear of surface ripened cheeses, were found to possess autoinducer-2 (...... increased by dairy-relevant stress conditions, indicates that AI-2 signalling might be important in regulation of microbial succession during ripening of surface ripened cheeses.......Bacterial strains of the species Arthrobacter nicotianae, Corynebacterium ammoniagenes, Corynebacterium casei, Microbacterium barkeri, Microbacterium gubbeenense and Staphylococcus equorum subsp. linens, all isolated from the smear of surface ripened cheeses, were found to possess autoinducer-2 (AI......-2) activity using the Vibrio harveyi (BB170) bioluminescence assay. In contrast, Brevibacterium casei and Brevibacterium linens strains were not found to have AI-2 activity. When exposed to low pH and high NaCl concentrations, AI-2 activities increased between 5.0 and 11.6× for C. casei 44701, M...
Alan D. Fisher
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.
Verschave, Sien H; Charlier, Johannes; Rose, Hannah; Claerebout, Edwin; Morgan, Eric R
Nematode infections are an important economic constraint to cattle farming. Future risk levels and transmission dynamics will be affected by changes in climate and farm management. The prospect of altered parasite epidemiology in combination with anthelmintic resistance requires the adaptation of current control approaches. Mathematical models that simulate disease dynamics under changing climate and farm management can help to guide the optimization of helminth control strategies. Recent efforts have increasingly employed such models to assess the impact of predicted climate scenarios on future infection pressure for gastrointestinal nematodes (GINs) in cattle, and to evaluate possible adaptive control measures. This review aims to consolidate progress in this field to facilitate further modeling and application. Copyright © 2016 Elsevier Ltd. All rights reserved.
Sörensen, C.; Holm, S. A.; Thamsborg, S. M.
The aims were to examine prevalence of gastrointestinal parasites in Danish goats, based on faecal examination, in relation to geographical distribution and risk factors, and to investigate the occurrence of anthelmintic resistance in nematodes in selected farms. In April 2012 all Danish goat farms......, and herds with a mean EPG>150 were offered a faecal egg count reduction test (FECRT). All herds were asked to complete a questionnaire about management and risk factors concerning parasites, particularly nematodes. Faecal egg counts were generally low; 2 out of 25 herds had a mean EPG>150. Herd prevalence...... of Nematodirus battus was 16%; likewise PNA‐staining revealed H. contortus in 16% of the herds (4 of 5 herds with individual EPG > 500). The overall prevalence of parasites detected by faecal examination were: Eimeria spp.100%, gastrointestinal trichostrongyles (excl. Nematodirus spp.) 66%, Trichuris ovis 36...
Peña-Espinoza, Miguel; Drag, Markus; Hansen, Tina Vicky Alstrup
efficacy of IVM was confirmed by FECRT in 5 out of 6 farms at day 14 p.t. and in all six farms at day 21 p.t. The qPCR was able to identify O. ostertagi and C. oncophora populations surviving IVM treatment. This highlights the need of monitoring the efficacy of anthelmintic treatments in cattle using......Field reports of anthelmintic resistance against the widely-used macrocyclic lactones (ML) in gastrointestinal nematodes (GIN) of cattle have appeared in NW-Europe in recent years. Objective: The aim of this study was to assess the efficacy of ivermectin (IVM) against field infections with GIN...... in Danish cattle. In addition, we evaluated a novel quantitative (q) real-time PCR assay for accurate identification of surviving nematode species after treatment. Methods: Six farms were selected based on mean faecal egg counts (FEC) of ≥ 100 nematode eggs per gram in first season grazing heifers. All...
Hördegen, P; Cabaret, J; Hertzberg, H; Langhans, W; Maurer, V
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.
Alan D. Fisher
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.
DE Lara, Ana Paula DE Souza Stori; Lorenzon, Lucas Bigolin; Vianna, Ana Muñoz; Santos, Francisco Denis Souza; Pinto, Luciano Silva; Aires Berne, Maria Elisabeth; Leite, Fábio Pereira Leivas
Effective control of gastrointestinal parasites is necessary in sheep production. The development of anthelmintics resistance is causing the available chemically based anthelmintics to become less effective. Biological control strategies present an alternative to this problem. In the current study, we tested the larvicidal effects of Bacillus thuringiensis var. israelensis Cry11Aa toxin against Haemonchus contortus larvae. Bacterial suspensions [2 × 108 colony-forming units (CFU) g-1 of the feces] of B. thuringiensis var. israelensis and recombinant Escherichia coli expressing Cry11Aa toxin were added to naturally H. contortus egg-contaminated feces. The larvae were quantified, and significant reductions of 62 and 81% (P var. israelensis and recombinant E. coli expressing Cry11Aa toxin were then orally administered to lambs naturally infected with H. contortus. Twelve hours after administration, feces were collected and submitted to coprocultures. Significant larvae reductions (P var. israelensis is a promising new class of biological anthelmintics for treating sheep against H. contortus.
Aggarwal, Rama; Bagai, Upma
Increasing anthelmintic resistance and the impact of conventional anthelmintics on the environment, it is important to look for alternative strategies against helminth parasite in sheep. Important lipogenic enzymes like glucose-6-phosphate dehydrogenase (G-6-PDH) and malate dehydrogenase (MDH) show subcellular distribution pattern. Activity of G-6-PDH was largely restricted to cytosolic fraction while MDH was found in both cytosolic and mitochondrial fraction in Gastrothylax indicus. Following in vitro treatment with ethanolic and aqueous extracts of Punica granatum fruit peel and commercial anthelmintic, albendazole G-6-PDH activity was decreased by 19-32 %, whereas MDH was suppressed by 24-41 %, compared to the respective control. Albendazole was quite effective when compared with negative control and both the extracts. The results indicate that phytochemicals of plant may act as potential vermifuge or vermicide.
Sarah Jane Margaret Preston
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.
Sallé, G; Cortet, J; Bois, I; Dubès, C; Guyot-Sionest, Q; Larrieu, C; Landrin, V; Majorel, G; Wittreck, S; Woringer, E; Couroucé, A; Guillot, J; Jacquiet, P; Guégnard, F; Blanchard, A; Leblond, A
Intestinal strongyles are the most problematic endoparasites of equids as a result of their wide distribution and the spread of resistant isolates throughout the world. While abundant literature can be found on the extent of anthelmintic resistance across continents, empirical knowledge about associated risk factors is missing. This study brought together results from anthelmintic efficacy testing and risk factor analysis to provide evidence-based guidelines in the field. It involved 688 horses from 39 French horse farms and riding schools to both estimate Faecal Egg Count Reduction (FECR) after anthelmintic treatment and to interview farm and riding school managers about their practices. Risk factors associated with reduced anthelmintic efficacy in equine strongyles were estimated across drugs using a marginal modelling approach. Results demonstrated ivermectin efficacy (96.3% ± 14.5% FECR), the inefficacy of fenbendazole (42.8% ± 33.4% FECR) and an intermediate profile for pyrantel (90.3% ± 19.6% FECR). Risk factor analysis provided support to advocate for FEC-based treatment regimens combined with individual anthelmintic dosage and the enforcement of tighter biosecurity around horse introduction. The combination of these measures resulted in a decreased risk of drug resistance (relative risk of 0.57, p = 0.02). Premises falling under this typology also relied more on their veterinarians suggesting practitionners play an important role in the sustainability of anthelmintic usage. Similarly, drug resistance risk was halved in premises with frequent pasture rotation and with stocking rate below five horses/ha (relative risk of 0.53, p risk factor analysis for anthelmintic resistance in equids. Our findings should guide the implementation of more sustained strongyle management in the field. Copyright © 2017 The Authors. Published by Elsevier Ltd.. All rights reserved.
Doyle, Stephen R; Laing, Roz; Bartley, David J; Britton, Collette; Chaudhry, Umer; Gilleard, John S; Holroyd, Nancy; Mable, Barbara K; Maitland, Kirsty; Morrison, Alison A; Tait, Andy; Tracey, Alan; Berriman, Matthew; Devaney, Eileen; Cotton, James A; Sargison, Neil D
The parasitic nematode Haemonchus contortus is an economically and clinically important pathogen of small ruminants, and a model system for understanding the mechanisms and evolution of traits such as anthelmintic resistance. Anthelmintic resistance is widespread and is a major threat to the sustainability of livestock agriculture globally; however, little is known about the genome architecture and parameters such as recombination that will ultimately influence the rate at which resistance may evolve and spread. Here, we performed a genetic cross between two divergent strains of H. contortus, and subsequently used whole-genome resequencing of a female worm and her brood to identify the distribution of genome-wide variation that characterizes these strains. Using a novel bioinformatic approach to identify variants that segregate as expected in a pseudotestcross, we characterized linkage groups and estimated genetic distances between markers to generate a chromosome-scale F1 genetic map. We exploited this map to reveal the recombination landscape, the first for any helminth species, demonstrating extensive variation in recombination rate within and between chromosomes. Analyses of these data also revealed the extent of polyandry, whereby at least eight males were found to have contributed to the genetic variation of the progeny analyzed. Triploid offspring were also identified, which we hypothesize are the result of nondisjunction during female meiosis or polyspermy. These results expand our knowledge of the genetics of parasitic helminths and the unusual life-history of H. contortus, and enhance ongoing efforts to understand the genetic basis of resistance to the drugs used to control these worms and for related species that infect livestock and humans throughout the world. This study also demonstrates the feasibility of using whole-genome resequencing data to directly construct a genetic map in a single generation cross from a noninbred nonmodel organism with a
Wanyangu, S.W; Rugut, M.K; Nginyi, J.M; Onyango-Abuye, J.A; Mugambi, J.M; Bain, A.M; Monteiro, F; Jackson, Mackellar F
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
Full Text Available Echinostome metacercariae are the infective stage for humans and animals. The identification of echinostomes has been based until recently on morphology but molecular techniques using sequences of ribosomal RNA and mitochondrial DNA have indicated major clades within the group. In this study we have used the ITS2 region of ribosomal RNA and the ND1 region of mitochondrial DNA to identify metacercariae from snails collected from eight well-separated sites from an area of 4000 km2 in Lamphun Province, Thailand. The derived sequences have been compared to those collected from elsewhere and have been deposited in the nucleotide databases. There were two aims of this study; firstly, to determine the species of echinostome present in an endemic area, and secondly, to assess the intra-specific genetic diversity, as this may be informative with regard to the potential for the development of anthelmintic resistance and with regard to the spread of infection by the definitive hosts. Our results indicate that the most prevalent species are most closely related to E. revolutum, E. trivolvis, E. robustum, E. malayanum and Euparyphium albuferensis. Some sites harbour several species and within a site there could be considerable intra-species genetic diversity. There is no significant geographical structuring within this area. Although the molecular techniques used in this study allowed the assignment of the samples to clades within defined species, however, within these groupings there were significant differences indicating that cryptic speciation may have occurred. The degree of genetic diversity present would suggest the use of targeted regimes designed to minimise the selection of anthelmintic resistance. The apparent lack of geographic structuring is consistent with the transmission of the parasites by the avian hosts.
Kerboeuf, Dominique; Blackhall, William; Kaminsky, Ronald; von Samson-Himmelstjerna, Georg
Infestation with parasitic helminths is a common problem in human populations of third world countries and is ubiquitous in livestock and other domestic animals. The cell-membrane efflux pump, P-glycoprotein (Pgp), appears to contribute to anthelmintic resistance. Pgp have been identified from both phyla of parasitic helminths, Platyhelmintha and Nematoda, and alterations in expression levels and allele frequencies of Pgp in anthelmintic-resistant populations have been observed in nematodes. Localisation of Pgp has been studied in the free-living nematode Caenorhabditis elegans and in the sheep parasite Haemonchus contortus using specific monoclonal antibodies or lectins. Reversing agents used in human studies, such as the calcium-channel blocker verapamil (VPL), appear to have similar effects in helminths as they do in human cancer cells: the efficacy of drug treatment is increased in drug-resistant parasites when reversing agents are co-administered with the anthelmintic. The functional role of the Pgp glycosylation was also studied using a lectin specific for the alpha-mannosyl residues and showed that resistance can be associated with a decreased affinity of the lectin for Pgp sites and that up to 50% reversion in the resistance to benzimidazoles (BZ) can be obtained using this lectin. Furthermore, the current knowledge on the role of Pgp in molecular mechanisms of drug resistance in the parasitic protozoan genus Trypanosoma is discussed. In some Trypanosoma species it was shown that drug resistance was associated with reduced uptake and in other ones with increased efflux. Several trypanosome Pgp-coding sequences have been described. In contrast to earlier data, most recent observations, based on experimentally overexpressed Pgp in Trypanosoma brucei, indicate a possible involvement in the mechanism of drug resistance in this parasite.
Torres-Acosta, J F J; Molento, M; Mendoza de Gives, P
The widespread presence of anthelmintic resistant gastrointestinal parasitic nematodes in outdoor ruminant production systems has driven the need to identify and develop novel approaches for the control of helminths with the intention to reduce the dependence on commercial anthelmintic drugs. This paper identifies what has been done in Latin America (LA) in terms of estimating the prevalence of anthelmintic resistance (AR) in ruminant production systems and the application of different novel approaches for the control of helminths in those systems, including research and extension activities. Firstly, the paucity of knowledge of AR is discussed in the context of different countries, as well as, the available economic resources for research, the technical infrastructure available and the practical difficulties of the production systems. It is then proposed that the search for novel approaches is not only driven by AR but also by the need for techniques that are feasible for application by resource-poor farmers in non-commercial subsistence farming systems. However, the commercial benefits of these approaches are often limited and so are funding inputs in most countries. The workers participating in the research into different novel approaches are identified as well as the different methods being studied in the different areas of LA according to their published results. In addition, the difficulties experienced during extension efforts to reach farmers and help them to adopt novel approaches for the control of parasitic nematodes in LA are discussed. The role of regulatory authorities in these countries is discussed as some methods of control might need an official confirmation of their efficacy as well as authorization prior to application as they may affect animal products (i.e. residues) and/or impose a hazard for animal welfare. The role of the pharmaceutical companies is also discussed. Copyright © 2011. Published by Elsevier B.V.
Besier, R B; Kahn, L P; Sargison, N D; Van Wyk, J A
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.
Full Text Available The current level of anthelmintic resistance in the horse-breeding industry is extremely high and therefore more emphasis is being placed on studies that focus on the judicious use of anthelmintic products. The aims of the study were to: 1 establish if there is variation in the egg excretion pattern of strongyles between the different age classes of Thoroughbred horses in the Western Cape Province (WCP, 2 test if a selective treatment approach successfully reduces the number of anthelmintic treatments and maintains acceptably low helminth burdens in adult Thoroughbred horses, and 3 evaluate the efficacy of subsampling large horse herds for faecal egg counts (FECs to monitor the strongyle burden. In 2001 the FECs of 4 adult mare, 5 yearling and 3 weanling herds from 8 different farms were compared in the WCP. Within the mare herds there were generally fewer eggexcreting individuals with lower mean FECs compared with the younger age classes. Individual faecal samples were collected every 3-4 weeks from 52 adult Thoroughbred mares from 1 farm in the WCP during a 12-month period (2002/2003. Animals with strongyle FECs > 100 eggs per gram (epg were treated with an ivermectin-praziquantel combination drug (Equimax oral paste, Virbac. The mean monthly strongyle FEC for the entire group was < 300 epg throughout the study and the number of treatments was reduced by 50 %. Resampling methods showed that an asymptote to mean FEC was reached at 55 animals for each of the pooled weanling, yearling and mare egg counts. Resampling within 4 different mare herds recorded asymptotes of between 24 and 28 animals. Subsampling entire herds for FECs therefore provided an effective approach to treatment management. This study demonstrates that selective treatment is both a practical and an effective approach to the management of anthelmintic resistance.
Sampimon, O C; Zadoks, R N; De Vliegher, S; Supré, K; Haesebrouck, F; Barkema, H W; Sol, J; Lam, T J G M
In this study, the accuracy of two phenotypic tests, API Staph ID 32 and Staph-Zym, was determined for identification of coagulase-negative staphylococci (CNS) from bovine milk samples in comparison with identification based on DNA-sequencing. A total of 172 CNS isolated from bovine milk were classified into 17 species. The most frequently isolated species based on rpoB sequencing were Staphylococcus chromogenes and Staphylococcus epidermidis, followed by Staphylococcus xylosus, Staphylococcus warneri and Staphylococcus equorum (37, 13, 9, 8 and 6% of isolates, respectively). The API Staph ID 32 correctly identified 41% of the CNS isolates. Best agreement with rpoB sequence based species identification was found for S. epidermidis, Staphylococcus hyicus and S. xylosus (100, 89 and 87%, respectively). The positive predictive value was 89, 100 and 52%, respectively. Poor sensitivity was observed for 3 of the 5 most frequently found species, S. chromogenes (37%), Staphylococcus warneri (15%) and S. equorum (0%) albeit with specificity of 100%. The Staph-Zym needed additional tests for 66% of the isolates and identified 31% of the CNS isolates correctly. Good sensitivity was found for S. epidermidis, S. simulans and S. xyloxus (100, 78 and 73%, respectively). The positive predictive value was 89, 78 and 98%, respectively. Poor sensitivity was observed for S. chromogenes, S. warneri and S. equorum (0, 54 and 0%, respectively) but with a specificity of 100, 99 and 100%, respectively. Both phenotypic tests misidentified a large proportion of CNS isolates and were thus unsuitable for identification of CNS species from bovine milk samples.
Gori, Klaus; Jespersen, Lene
A large number of Gram-positive and Gram-negative bacteria have been found to produce the signaling molecule autoinducer two (AI-2), which is used for interspecies communication. In this study, AI-2 activity was for the first time determined in Arthrobacter nicotianae, Brevibacterium linens (BL2......). Corynebacterium casei, Microbacterium barkeri, Microbacterium gubbeenense and S. equorum subsp. linens (all isolated from the smear of surface ripened cheeses) using the AI-2 bioluminescence assay. This indicates that AI-2 signaling could take place between bacteria found in the smear of surface ripened cheeses....
Liébano-Hernández, E; González-Olvera, M; Vázquez-Peláez, C; Mendoza-de-Gives, P; Ramírez-Vargas, G; Peralta-Lailson, M; Reyes-García, M E; Osorio, J; Sánchez-Pineda, H; López-Arellano, M E
Because of the natural adaptation of Mexican sheep, the aim of the present study was to identify the presence or absence of gastrointestinal parasitic nematodes (GIN) resistant to benzimidazole (BZ) in both Chiapas and Pelibuey sheep breeds on local farms. Both male and female GIN-infected grazing sheep of the two breeds were selected. Sheep faecal samples were collected to obtain infective larvae (L3). This evolving stage of the parasite was used for taxonomic identification of the genus, based on its morphological characteristics. BZ anthelmintic resistance was evaluated using a nematode-compound in vitro interaction bioassay and the allele-specific polymerase chain reaction technique to detect mutations of residues 198 and 200 on isotype 1 of the β-tubulin gene. Three BZ-based compounds (febendazole (FBZ), tiabendazole (TBZ) and albendazole (ABZ)) at concentrations of 1, 0.5, 0.25, 0.125, 0.062 and 0.03 mg/ml were used to estimate the anthelmintic efficacy and lethal dose (LD50, LD90 and LD99) of the drugs. Two parasitic nematodes, Haemonchus and Teladorsagia, were identified in both isolates. Also, the proportions of anthelmintic resistance identified in GIN of the two sheep breeds were 68% in isolates from the Chiapas breed and 71.8% in the Pelibuey breed. The specific lethal activity obtained with FBZ was higher than 90%. However, TBZ and ABZ showed a lethal activity lower than 50%. High variability in the discriminating dose values was found among the BZ drugs. For example, FBZ LD ranged from 0.01 to 1.20 mg/ml; on the other hand, TBZ and ABZ required a dose ranging from 0.178 to 759 mg/ml. In addition, amino acid changes of Phe (TTC) to Tyr (TAC) at codon 200 of the β-tubulin gene, showing resistance to BZ, and no changes at codon 198 Glu (GAA) to Ala (GCA) were observed for both isolates. These results confirmed the presence of a genetic mutation associated with BZ in both Chiapas and Pelibuey nematode isolates.
Bolwell, Charlotte F; Rosanowski, Sarah M; Scott, Ian; Sells, Patrick D; Rogers, Chris W
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
Peña-Espinoza, Miguel; Thamsborg, Stig M; Demeler, Janina; Enemark, Heidi L
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
Candy, Paul M; Waghorn, Tania S; Miller, Chris M; Ganesh, Siva; Leathwick, Dave M
A replicated field trial was conducted to measure the effect on liveweight gain of failing to adequately control anthelmintic resistant populations of Cooperia oncophora and to determine whether populations, and hence production losses, increased with time. Eight mobs of 10 Friesian-Hereford calves were run on independent farmlets from January to December, over each of two years. All mobs were routinely treated with a pour-on formulation of eprinomectin every six weeks, which controlled parasites other than Cooperia. Four mobs also received six weekly treatments with an oral levamisole plus albendazole combination anthelmintic to control Cooperia. Liveweights, condition scores, faecal egg counts and larval numbers on pasture were measured throughout. In the first year animals treated with eprinomectin alone were 12.9 kg lighter in November than those treated with eprinomectin plus albendazole and levamisole, however, in the second year there was no difference between the treatment groups. The data, therefore, support the view that while C. oncophora is less pathogenic than other cattle parasite species it can still cause production losses when present in sufficient numbers. In the first year of the study, parasite load, as measured by faecal nematode egg count and larval numbers on herbage, tended to be higher and calf growth rates lower than in the second year. In both years, counts of infective larvae on herbage declined over winter-spring to be at low levels before mid-summer. This suggests that the carry-over of infection from one crop of calves to the next was relatively small and hence that the level of challenge to the young calves at the start of each year was largely due to the effectiveness of the quarantine treatments administered when the animals arrived on the trial site. Low survival of larvae on pasture between grazing seasons, resulting in small larval populations on pasture when drenching programmes start each summer, might help to explain the
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
Wilson, L; Rhodes, A P; Dodunski, G
The drivers for anthelmintic use today are substantial and anthelmintic use has become an embedded normalised behaviour. The cheapness and easy availability of anthelmintic products has meant that New Zealand farmers have had access to easy "solutions" for dealing with parasites and minimal forward planning or system redesign has been required. Despite 30 years of messaging about the emerging issue of anthelmintic resistance, management to reduce parasitism and the need to change behaviour, farmer practice has largely remained unchanged. Traditional approaches to extension, particularly around parasite management, appear to have been quite ineffective, apart from encouraging change in anthelmintic products and a switch to use of anthelmintics in combination. More effective approaches are required. The evolving nature of anthelmintic resistance and sustainable management of parasitism require attitudes, knowledge and behaviour to change. This is a challenge for all players in the industry; researchers, manufacturers and sellers, advisors and farmers. Looking beyond agriculture to the health sector provides some insight into models of decision making and behaviour change that can inform future strategies. Features in the health belief model including concepts of self-efficacy and cues to action appear to align with the issues, challenges and culture prevailing in farming, and parasite management in particular. Programmes through which farmers have made substantial beneficial behaviour change and the lessons learnt are discussed. Effecting consistent behaviour change around parasite management will involve new approaches by all participants in the process. And the process itself also needs to change. It requires an understanding of whole-farm systems, and the consideration of all the sources of influence on the farmer and the other participants in the process. The process of knowledge sharing involving the farmer should be based on equality; each person in the process
Full Text Available Histidine, lysine, ornithine and tyrosine decarboxylase activities were tested in 38 strains of Staphylococcus (15 of Staph. equorum, 11 of Staph. epidermidis, 7 of Staph. saprophyticus, and 5 of Staph. pasteuri and 19 strains of Bacillus (13 of B. subtilis and 6 of B. amyloliquefaciens isolated from two Spanish traditional sausage varieties.The four decarboxylase activities were present in most of the strains studied, but some variability was observed between strains within each microbial species.Accumulation of putrescine and cadaverine was assessed in the culture media of the strains that displayed ornithine and lysine decarboxylase activities. The aminogenic potential of the strains was low, with amounts accumulated lower than 25 mg/L for the putrescine and than 5 mg/L for the cadaverine, with the exception of a strain of Staph. equorum that produced 1415 mg/L of putrescine, and of a strain of Staph. epidermidis that accumulated 977 mg/L of putrescine and 36 mg/L of cadaverine.
Fonseca, Sonia; Cachaldora, Aida; Gómez, María; Franco, Inmaculada; Carballo, Javier
The dynamics of the bacterial population throughout the ripening of Galician chorizo, a traditional dry fermented sausage produced in the north-west of Spain, were investigated by using classical and molecular approaches. Fermented sausages are certainly complex matrices with PCR inhibitors and background microbiota. Therefore, two different DNA preparation methods were performed to elaborate each standard curve and no significant differences were found, showing a very good correlation between both methods (R(2)>0.994). The combination of the results obtained from microbial counts, species and genus-specific PCR as well as real-time quantitative PCR (qPCR) allowed the identification for the dominant bacterial species and the study of the variation in the community composition over the ripening period. According to the data obtained both by identification of plate isolates and by real-time PCR, the dominant species among staphylococci and lactobacilli were Staphylococcus equorum and Lactobacillus sakei, respectively. However, only real-time PCR assay showed enough sensitivity to detect and quantify staphylococci in meat batter before stuffing, showing values of 5.28logCFU/g when quantifying Staphylococcus spp. and 2.87logCFU/g when quantifying S. equorum. In conclusion, real-time PCR was shown to be an efficient tool for the study of the complex associations developed in meat fermentations and for the characterization of dominant populations. Copyright © 2012. Published by Elsevier Ltd.
Tadesse, Dereje; Eguale, Tadesse; Giday, Mirutse; Mussa, Abiy
The widespread development of anthelmintic resistance and high cost of the conventional anthelmintic drugs, has limited the control of gastrointestinal nematode parasites of sheep and goats and hence led to evaluation of medicinal plants as an alternative source of anthelmintics. In the current study, in vitro ovicidal and larvicidal activity of the leaves and fruits of the aqueous and hydro-alcoholic extracts of Maesa lanceolata and aerial parts of Plectranthus punctatus were evaluated on the egg and larvae of Haemonchus contortus using egg hatch assay and larval development test. All extracts of plants tested have shown complete inhibition of egg hatching at or below 1 mg/ml. ED50 for egg hatch inhibition ranged from 0.11 to 0.29 mg/ml, for both the aqueous and hydro-alcoholic extracts of Plectranthus punctatus and Maesa lanceolata. All extracts have shown dose dependent inhibition of larval development with variable results. The complete inhibition (100%) at the maximum concentration tested (50 mg/ml) was obtained only for hydro-alcoholic extract of the fruits of Maesa lanceolata and the lowest inhibition (50.33%) was recorded for the hydro-alcoholic extract of the leaves of the same plant. The overall findings of the present study has shown that Plectranthus punctatus and Maesa lanceolata contain possible anthelmintic compounds and further evaluation of different extracts and fractions of these plants should be carried out.
Slocombe, J. Owen D.; Coté, John F.; de Gannes, Rolph V.G.
Three clinical trials with fecal egg count reduction tests and coproculture were conducted on 2 standardbred farms in Ontario. On Farm A, the treatment groups were mebendazole and ivermectin in trial 1, and fenbendazole and moxidectin in another. On Farm B, treatment groups were mebendazole and ivermectin. All horses treated with mebendazole or fenbendazole were subsequently treated with ivermectin or moxidectin. Strongyle eggs/g feces were estimated pre- and post-treatment using the Cornell-McMaster dilution and Cornell-Wisconsin centrifugal flotation techniques. After treatment, there was no change in the arithmetic mean eggs/g feces for horses given mebendazole, and a reduction of only 49.1% for those given fenbendazole. All horses receiving ivermectin or moxidectin had their egg counts reduced to 0. Only cyathostomes were found on culture. On both farms the benzimidazole resistant strains appeared to have persisted for at least 10 years. Development of and monitoring for anthelmintic resistance are briefly discussed. PMID:18320979
J.A. Van Wyk
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.
Solana, María Victoria; Mera y Sierra, Roberto; Scarcella, Silvana; Neira, Gisela; Solana, Hugo Daniel
Anthelmintic resistance in livestock parasites is currently a worldwide problem. Fasciola hepatica is a cosmopolitan parasite which causes considerable loss in sheep and cattle production systems all over the world. Chemotherapy is currently the main tool available for its control. The intensive use of triclabendazole, the drug of choice for more than 20 years, has resulted in the development of resistant strains. The therapeutic options are adulticides such as closantel (salicylanilide anthelmintic that binds extensively to plasma albumin) to treat chronic fascioliasis in sheep, and cattle. In the present work, an Egg Hatch Assay (EHA) and morphometric studies were used to evaluate in vivo the ovicidal activity and morphology F. hepatica eggs, recovered from closantel treated sheep collected at different time intervals post treatment. Statistically significant differences (p < 0.0001) were observed in egg morphometry between the control and the treated groups in all the parameters studied. Eggs recovered from treated animals tend to be narrower and longer. Significant differences were found in the embryonation and hatching of eggs between 36 h post treatment (32, 5%) vs. approximately 85% in control, 12 h and 24 h post treatment. Our results confirm that closantel affects in vivo the normal development of the eggs. As one of the first effects, this drug affects the performance of the trematode's reproductive physiology. Even though closantel treated animals may still eliminate eggs in the first days post treatment, these are not viable. Copyright © 2015 Elsevier Inc. All rights reserved.
Gasser, R B; Schwarz, E M; Korhonen, P K; Young, N D
Parasitic roundworms (nematodes) cause substantial mortality and morbidity in animals globally. The barber's pole worm, Haemonchus contortus, is one of the most economically significant parasitic nematodes of small ruminants worldwide. Although this and related nematodes can be controlled relatively well using anthelmintics, resistance against most drugs in common use has become a major problem. Until recently, almost nothing was known about the molecular biology of H. contortus on a global scale. This chapter gives a brief background on H. contortus and haemonchosis, immune responses, vaccine research, chemotherapeutics and current problems associated with drug resistance. It also describes progress in transcriptomics before the availability of H. contortus genomes and the challenges associated with such work. It then reviews major progress on the two draft genomes and developmental transcriptomes of H. contortus, and summarizes their implications for the molecular biology of this worm in both the free-living and the parasitic stages of its life cycle. The chapter concludes by considering how genomics and transcriptomics can accelerate research on Haemonchus and related parasites, and can enable the development of new interventions against haemonchosis. Copyright © 2016 Elsevier Ltd. All rights reserved.
P. T. Costa
Full Text Available The objective of this study was to evaluate the efficacy of different drugs in the control of parasites of sheep belonging to the Centro Agropecuário da Palma, Universidade Federal de Pelotas, municipality of Capão do Leão, RS. Seventy-eight female Corriedale and Ideal sheep with an initial body weight of 48.35 ± 4.71 kg were randomly selected and divided into six groups submitted to the following treatments: control treatment and treated with 34%nitroxynil, 18.8% levamisole hydrochloride, 10% closantel, 1% moxidectin, and 10% fenbendazole. The drugs were administered according to the recommendations of the manufacturers. Fecal samples were collected before (day 0 and after treatment (days 7, 4, 21 and 28 and were used for the determination of fecal egg count (FEC in the different groups. Efficacy was evaluated based on the percentage reduction in FEC and percent efficacy of the drugs. The fecal samples were processed for coproculture to identify the parasite genera present in the herd. The percentages of efficacy observed on day 28 post-treatment were 96.93% for nitroxynil, 95.8% for levamisole hydrochloride, 95.5% for closantel, 80.2% for moxidectin, and 27.5% for fenbendazole. The nematode species present in the herd was Haemonchus spp. (100%. Nitroxynil, closantel and levamisole hydrochloride were effective in eliminating gastrointestinal nematodes. Anthelmintic resistance was observed to moxidectin and fenbendazole.
Luciana Laitano Dias de Castro
Full Text Available Due to anthelmintic resistance in nematodes, several research studies have been developed seeking control alternatives to these parasites. This study evaluated the in vitro action of Origanum vulgare on gastrointestinal nematode eggs of cattle. In order to evaluate the ability to inhibit egg hatch, different dried leaves extracts of this plant were tested, such as dye, hydroalcoholic and aqueous extracts at concentrations varying from 0.62 to 80 mg/mL. Each assay was accompanied by control containing levamisole hydrochloride (0.2 mg/mL, distilled water and 70 ºGL grain alcohol at the same concentration of the extracts. Test results showed that the different O. vulgare extracts inhibited egg hatch of cattle gastrointestinal nematodes at a percentage that varied from 8.8 to 100%; dye and hydroalcoholic extract were the most promising inhibitors. In view of this ovicidal property, O. vulgare may be an important source of viable antiparasitic compounds for nematodiosis control in ruminants.
Zhao, Jianguo; Williams, Andrew R; Hansen, Tina Vicky Alstrup; Thamsborg, Stig M; Cai, Jianping; Song, Shuaibao; Chen, Gang; Kang, Ming; Zhang, Zhuangzhi; Liu, Qun; Han, Qian
In vitro methods have been developed for the detection of anthelmintic resistance in a range of nematode species. However, the life cycle of Ascaris suum renders the commonly used egg hatch assay and larval development assay unusable. In this study we developed a combined multi-well culture 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 of larvae that migrated to the surface of each treated and control well were used to calculate the drug concentration which inhibits 50% of the larvae migration (EC 50 ). The values of EC 50 of thiabendazole, fenbendazole, mebendazole, levamisole, and pyrantel against A. suum isolates ranged 74-150, 4.9-13.9, 2.3-4.3, 358-1150 and 1100-4000nM, respectively. This combined multi-well culture and agar gel larval migration assay was a sensitive bioassay for anthelmintic activity and could serve as an in vitro method to detect for lowered drug efficacy against A. suum or possibly to screen for anthelmintic drug candidates. Copyright © 2017 Elsevier B.V. All rights reserved.
Sallé, Guillaume; Cabaret, Jacques
In-depth knowledge of the use of anthelminthics in the field, especially by veterinarians, is required to design more sustainable parasite control strategies. An online survey was sent by e-mail to 940 equine veterinary practitioners to describe their equine practice, their awareness about parasites and the management strategies they apply. Gastrointestinal parasites were generally considered (68%) as an issue of moderate importance. Drug efficacy failure was a minor or moderate issue for 47% and 48% of responders, respectively. Parasite management mostly relied on the use of systematic calendar treatments across a wide variety of horse owners (ie, riding schools, studs or hobby horse owners). Almost half of the practitioners (42%) never performed Faecal Egg Count (FEC) before drenching. Horse owners or their employees in charge of equines were reported to be the only person managing drenching in 59% of the collected answers. This was associated with the report of many off-label uses of anthelmintics and the frequent buying of drugs using the internet. Given the critical situation regarding anthelmintic resistance, it seems necessary for veterinarians to reclaim parasite management and prevention as a specific topic. Implementation of stricter regulations for use of anthelmintics, like the one applied in Denmark, may make parasitic management in equids more sustainable.
Eduardo Robson Duarte
Full Text Available In the North of Minas Gerais the equineculture is an important activity because it corroborates the success in the breeding of beef cattle. The equine verminosis control in this region has not been applied considering the anthelmintic resistances of nematodes and this resistance has been observed in some equine herds in Brazil. The present study has the objective to evaluate the effectiveness of pyrantel pamoate and ivermectin association in mare verminosis in the North of Minas Gerais, during the peripatum. After fourteen days of the first faecal egg count (FEC and treatment of the animals with these respective bases, the FEC reduction test indicated 98.1% effectiveness and the coprocultures were negative. After the worm identification from the control group, was observed 30% of filariod worms of Strongyloides spp., 30% of the genus Haemonchus, 20% were worms of Cyathosminae, 10% of the genus Trichostrongylgys and 10% of the genus Oesophagodontus. The results observed suggest that the use of pyrantel pamoate associated with ivermectin was safe during the peripartum and efficient in the FEC reduction of naturally infected mares raised in pastures of the North of Minas Gerais.
Adriane Holtz Tirabassi
Full Text Available The widespread occurrence of anthelmintic-resistant gastrointestinal nematodes (GINs, particularly Haemonchus contortus, in sheep production systems has magnified the need to identify and develop alternative control strategies. Strategies include the selection of genetically GIN-resistant sheep and the implementation of biological parasite control to reduce dependence on anthelmintic drugs. In this study, we aimed to establish the molecular identity of bacterial communities present in the abomasum of sheep classified as resistant or susceptible to H. contortus, an abomasal parasite. Thirty-eight sheep were experimentally infected with L3 Haemonchus contortus and analyzed for fecal egg count (FEC and hematocrit (Ht to establish haemonchosis resistance or susceptibility. Four resistant sheep (RS and four susceptible sheep (SS were selected for microbial sampling and subsequent phylogenetic analysis. Molecular identification of the bacteria was based on amplification of the bacterial 16S rRNA gene, construction of a 16S rDNA clone library, and subsequent gene sequencing. Significant differences (p = 0.05 were observed in the occurrence of different phyla identified in RS and SS libraries: Firmicutes (61.4% and 37.2%, respectively, Proteobacteria (10.2% and 37.2%, respectively, Bacteroidetes (12.8% and 5.8%, respectively, and unclassified bacteria (12.8% and 17%, respectively. Differences between the proportions of bacterial communities present in the RS and SS pool samples were observed, contributing as a first step toward the assessment of the association between the gastrointestinal tract microbiota and nematode resistance in sheep.
Charlier, J; Morgan, E R; Rinaldi, L; van Dijk, J; Demeler, J; Höglund, J; Hertzberg, H; Van Ranst, B; Hendrickx, G; Vercruysse, J; Kenyon, F
Due to the development of anthelmintic resistance, there have been calls for more sustainable nematode control practices. Two important concepts were introduced to study and promote the sustainable use of anthelmintics: targeted treatments (TT), where the whole flock/herd is treated based on knowledge of the risk, or parameters that quantify the severity of infection; and targeted selective treatments (TST), where only individual animals within the grazing group are treated. The aim of the TT and TST approaches is to effectively control nematode-induced production impacts while preserving anthelmintic efficacy by maintaining a pool of untreated parasites in refugia. Here, we provide an overview of recent studies that assess the use of TT/TST against gastrointestinal nematodes in ruminants and investigate the economic consequences, feasibility and knowledge gaps associated with TST. We conclude that TT/TST approaches are ready to be used and provide practical benefits today. However, a major shift in mentality will be required to make these approaches common practice in parasite control. British Veterinary Association.
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.
Tzelos, Thomas; Matthews, Jacqueline B; Buck, Amy H; Simbari, Fabio; Frew, David; Inglis, Neil F; McLean, Kevin; Nisbet, Alasdair J; Whitelaw, C Bruce A; Knox, David P; McNeilly, Tom N
Teladorsagia circumcincta is a major cause of ovine parasitic gastroenteritis in temperate climatic regions. The development of high levels of anthelmintic resistance in this nematode species challenges its future control. Recent research indicates that many parasite species release extracellular vesicles into their environment, many of which have been classified as endocytic in origin, termed exosomes. These vesicles are considered to play important roles in the intercellular communication between parasites and their hosts, and thus represent potentially useful targets for novel control strategies. Here, we demonstrate that exosome-like extracellular vesicles can be isolated from excretory-secretory (ES) products released by T. circumcincta fourth stage larvae (Tci-L4ES). Furthermore, we perform a comparative proteomic analysis of vesicle-enriched and vesicle-free Tci-L4ES. Approximately 73% of the proteins identified in the vesicle-enriched fraction were unique to this fraction, whilst the remaining 27% were present in both vesicle-enriched and vesicle-free fraction. These unique proteins included structural proteins, nuclear proteins, metabolic proteins, proteolytic enzymes and activation-associated secreted proteins. Finally, we demonstrate that molecules present within the vesicles-enriched material are targets of the IgA and IgG response in T. circumcincta infected sheep, and could potentially represent useful targets for future vaccine intervention studies. Copyright © 2016 Elsevier B.V. All rights reserved.
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.
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.
Storey, Bobby E; Williamson, Lisa H; Howell, Sue B; Terrill, Thomas H; Berghaus, Roy; Vidyashankar, Anand N; Kaplan, Ray M
Haemonchus contortus resistant to multiple anthelmintics threaten the viability of the small ruminant industry in areas where this parasite is prevalent. In response to this situation, the FAMACHA© system was developed and validated for use with small ruminants as a way to detect clinical anemia associated with haemonchosis. Given that H. contortus and multiple anthelmintic resistance is a similar problem in camelids, the FAMACHA© system might also provide the same benefits. To address this need, a validation study of the FAMACHA© system was conducted on 21 alpaca and llama farms over a 2-year period. H. contortus was the predominant nematode parasite on 17 of the 21 farms (10 alpaca and 7 llama farms) enrolled in the study, based on fecal culture results. The FAMACHA© card was used to score the color of the lower palpebral (lower eye lid) conjunctiva on a 1-5 scale. Packed cell volume (PCV) values were measured and compared to FAMACHA© scores using FAMACHA© score cutoffs of ≥3 or ≥4 and with anemia defined as a PCV ≤15%, ≤17%, or≤20%. PCV was significantly associated with FAMACHA© score, fecal egg count (FEC), and body condition score (BCS), regardless of the FAMACHA© cutoff score or the PCV% chosen to define clinical anemia (pcamelids suffering from haemonchosis. Parameters for making treatment decisions based on FAMACHA© score in camelids should mirror those established for small ruminants. Copyright © 2017 Elsevier B.V. All rights reserved.
Chandrawathani, Panchacharam; Jamnah, Omar; Waller, Peter John; Höglund, Johan; Larsen, Michael; Zahari, Wan Mohammed
Approximately 2,800 fresh dung samples from animals, mainly ruminant livestock, were screened for the presence of nematophagous fungi in Malaysia. Arthrobotrys spp. was noted on numerous occasions, but only one isolate of Duddingtonia flagrans was made. For the purposes of producing sufficient quantities of this fungus for feeding trials in sheep, various, commonly available, cheap plant materials were tested as possible growth substrates. This showed that cereal grains (wheat, millet and rice) were the best media for fungal growth. Pen feeding trials were carried out using sheep, both naturally and experimentally infected with nematode parasites (predominantely Haemonchus contortus), to test the efficiency of D. flagrans when administered either in a grain supplement, or incorporated into a feed block. These showed that the fungus survived gut passage in sheep and that dose rates of approximately 1 x 10(6) D. flagrans spores / animal / day, reduced the percentage of infective larvae developing in faecal cultures by more than 90%. These results indicate that using D. flagrans as a biological control agent of nematode parasites, is a promising alternative to nematode parasite control of small ruminants in Malaysia, where anthelmintic resistance is now a major problem.
Full Text Available The egg reduction rate (ERR is the current standard mean to assess the efficacy of drugs against human soil-transmitted helminths (STHs; Ascaris lumbricoides, Trichuris trichiura and hookworm. Although the timing of post-treatment sampling is pivotal for a readily interpretation of drug efficacy, there is lack empirical data that allows recommending the optimal time point for a follow-up egg counting. In the present study, we re-analyzed both the kinetics of worm expulsion and egg output for Ascaris lumbricoides following a single oral dose of albendazole in a series of studies previously conducted in Kenyan communities. The results indicate that it takes up to 10 days post-treatment before the expulsion of both adult male and female Ascaris worms is completed, approximately 20% of the worms being expelled between day 7 and 10 post-treatment. The sequential analysis of the egg out put, indicated a poor ERR (89.4% at day 7 post-treatment, but a 100% ERR at day 14 and 21 post-treatment. Based on our findings we recommend to wait at least 14 days after an albendazole treatment before conducting the follow-up egg count. Any sampling before this time point may result in biased ERR estimates, due the release of residual eggs from moribund or degenerating worms. Keywords: Ascaris lumbricoides, Worm expulsion, Egg reduction rate, Anthelmintic resistance, Albendazole
Full Text Available Totally, 47 strains of S. aureus and 578 coagulase negative staphylococci were detected in samples from raw ewe milk. The 35 out 47 isolates of S. aureus from ewe milk were positive for the presence of staphylococcal enterotoxin genes: sea(4 %, sec (48 % a sed (48 %. Staphylococcus epidermis (33.04%, Staphylococcus caprae (21.28% were more prevalent. Staphylococcus chromogenes (7.44 %, Staphylococcus hominis (7.09%, Staphylococcus xylosus (6,92 %, a Staphylococcus warneri (6.40 % were isolated also in ewes milk. Staphylococcus haemolyticus (3.11 %, Staphylococcus capitis (2.94 %, Staphylococcus simulans (2.08 % and Staphylococcus saprophyticus (1.73 % were isolated very rarely from the taken individual milk ewe samples. Sporadically, only in few cases, the others coagulase negative staphylococci were isolated (Staphylococcus cohnii cohnii, Staphylococcus sciuri, Staphylococcus closii, Staphylococcus lugdunensis, Staphylococcus auricularis and Staphylococcus equorum. doi:10.5219/24
Ana Maria Simion Ciuciu
Full Text Available Selected starter cultures were used to produce the traditional Romanian dry cured sausage, Dacia. A control sausage was produced without starter culture (sausage A, one with L. sakei CECT 5964 and S. equorum SA25 (sausage B and one with L. sakei CECT 5964, S. equorum SA25 and L. acidophilus CECT 903 (sausage C. Samples from each batch of sausages were taken at 0 (mix before stuffing, and after 2, 4, 7, 14, 21 and 28 days of ripening. Counts of total aerobic mesophilic flora, lactic acid bacteria, salt tolerant flora, and Enterobacteriaceae and some physical-chemical parameters (moisture, NaCl, pH and aw values were determined. High microbial counts (log CFU were observed with values at the end of ripening period: for lactic acid bacteria 9.77 (A, 11.47 (B and 11.19 (C; for total aerobic mesophilic flora 9.89 (A, 11.38 (B and 11.30 (C; for salt tolerant flora 4.45 (A, 5.31 (B and 5.27 (C. The starter cultures had a significant inhibitory effect on Enterobacteriaceae counts (log CFU, values at the end of ripening period being 1.32 (A, 0.33 (B and not detected (C. A significant decrease in the pH values is observed until the seventh day of ripening, showing a slight, but progressive increase after the 14th day of ripening. Results show that the production and ripening process in a pilot scale chamber under controlled conditions contributes in obtaining safe and homogeneous products.
De Visscher, A; Piepers, S; Haesebrouck, F; De Vliegher, S
Coagulase-negative staphylococci (CNS) are the main cause of bovine intramammary infections and are also abundantly present in extramammary habitats such as teat apices. Teat apex colonization (TAC) with CNS has already been explored in lactating dairy cows at the species level, whereas this is not true for dry cows and end-term heifers. Therefore, the aim of this observational study was to describe CNS TAC in nonlactating dairy cows and end-term heifers in Flemish dairy herds and to identify associated risk factors at the herd, cow, and quarter level. All CNS were molecularly identified to the species level using transfer RNA intergenic spacer PCR (tDNA-PCR) and sequencing of the 16S rRNA gene, allowing for species-specific statistical analyses using multivariable, multilevel logistic regression. Staphylococcus devriesei, Staphylococcus chromogenes, Staphylococcus haemolyticus, and Staphylococcus equorum were the most frequently isolated species. Staphylococcus chromogenes was the sole species colonizing teat apices of cows and heifers in all herds, whereas large between-herd differences were observed for the other species. Teat apices of red and white Holstein Friesians, of quarters dried off without an internal teat sealer, and swabbed in months with lower precipitation and higher ambient temperature were significantly more likely to be colonized by S. devriesei. Slightly dirty teat apices and teat apices swabbed in months with lower precipitation had higher odds of being colonized by S. chromogenes, whereas teat apices sampled in months with lower precipitation and higher ambient temperature were more likely to be colonized by S. haemolyticus. Dirty teat apices and teat apices swabbed in months with lower ambient temperature in combination with low precipitation had higher odds of being colonized by S. equorum. Diverse factors explaining CNS TAC, yet mostly related to humidity, ambient temperature, and hygiene, substantiate differences in epidemiological
Ye, K; Liu, M; Liu, J; Jiang, J; Guo, C
The objective of this study was to investigate the effect of spices and packaging films on the bacterial dynamics of pot-stewed duck wings (PSDW) during storage at 8°C. The results showed that spices added at a 1 : 1 ratio (weight ratio of pungent spices and fragrant spices) could delay the lag period of bacterial growth and extend the shelf-life of PSDW. Denaturing Gel Gradient Electrophoresis (DGGE) analysis indicated that the predominant bacteria in PSDW at the end of storage were Weisella hellenica, Weisella diestrammenae, Staphylococcus equorum and Streptococcus parauberis, and spices added at a 1 : 1 ratio showed better inhibition for bacteria at the end of storage, except for W. hellenica. Thus, general-purpose and high barrier films in modified atmosphere packaging, had a good barrier effect for the isolation of air, and showed no effect on the bacteria. This study will help PSDW processing companies to understand the biodiversity of PSDW, and provide a scientific basis for the extension of shelf-life. This study confirmed that high barrier cover film, general barrier base film and spices added at a 1 : 1 ratio (weight ratio of pungent and fragrant spices), had a better inhibition effect on bacteria in pot-stewed duck wings products, which could be used for processing and storage conditions of this product. Simultaneously, the change in the bacterial community of this product during 8°C storage was analysed, where the predominant bacteria of modified atmosphere packaging pot-stewed duck wings at the end of 8°C storage included Weisella hellenica, Weisella diestrammenae, Staphylococcus equorum and Streptococcus parauberis. © 2016 The Society for Applied Microbiology.
Rajkowska, Katarzyna; Koziróg, Anna; Otlewska, Anna; Piotrowska, Małgorzata; Nowicka-Krawczyk, Paulina; Brycki, Bogumił; Kunicka-Styczyńska, Alina; Gutarowska, Beata
Quaternary ammonium compounds (QACs) are widely used in disinfection of water, surfaces and instruments as well as in textile, leather and food industries because of their relatively low toxicity, broad antimicrobial spectrum, non-volatility and chemical stability. Due to these advantages, QACs are also used in restoration and can be applied on historical material. The aim of this study was to determine the usefulness of biocides based on quaternary ammonium salts and containing various excipients in the protection of historical materials against microbial growth. The study determined the antimicrobial activity of three biocides against bacteria: Pseudomonas fluorescens, Staphylococcus equorum, Bacillus cereus, Bacillus muralis, Sporosarcina aquimarina and Rhodococcus fascians, and moulds: Chaetomium globosum, Penicillium citreonigrum, Cladosporium cladosporioides I, Acremonium strictum, Aspergillus fumigatus and Cladosporium cladosporioides II, all isolated from historical wood and brick. Staphylococcus equorum, Bacillus cereus, Sporosarcina aquimarina and Rhodococcus fascians bacteria, and Cladosporium cladosporioides I and Acremonium strictum moulds showed high sensitivity to quaternary ammonium biocides. Historical wood can be effectively disinfected by three applications of biocide A (30% v/v) containing dodecyl dimethyl ammonium chloride (DDAC), citric acid, propiconazole and propanol. Disinfection of historical brick can be carried out by three applications of 6% v/v solutions of biocide B (based on DDAC and ethylenediaminetetraacetic acid - EDTA) or biocide C (containing a non-ionic surfactant, DDAC and EDTA). Effective protection of historical building materials against microbial growth for a period of seven days can be achieved by the application of biocide A (30% v/v) on the wood surface and biocide B (6% v/v) on the brick surface.
Boubaker Elandalousi, Ramzi; Akkari, Hafidh; B'chir, Fatma; Gharbi, Mohamed; Mhadhbi, Moez; Awadi, Soufia; Darghouth, Mohamed Aziz
The increasing prevalence of anthelmintic resistant strains of helminths, the drug residues in animal products and the high cost of conventional anthelmintics has created an interest in studying medicinal plants as an alternative source of anthelmintics. Thymus capitatus (Lamiales: Lamiaceae) is used traditionally by people as spices and reported to possess some biological effects. The objective of this study is to evaluate the anthelmintic efficacy of T. capitatus in comparison to albendazole against the gastrointestinal nematodes of sheep. To fulfil the objectives, in vitro anthelmintic activities of crude aqueous and crude ethanolic extracts of aerial parts of T. capitatus were investigated on the eggs and adults of the nematode parasite Haemonchus contortus. Both extract types of T. capitatus completely inhibited egg hatching at a concentration close to 2 mg/ml. LC₅₀ of ethanolic extract of T. capitatus was 0.368 mg/ml while that of aqueous extract was 6.344 mg/ml (p<0.05). The ethanolic extract showed higher in vitro activity against adult parasites than the aqueous one in terms of the paralysis and/or death of the worms at different hours post-treatment. Dose dependent effect was observed for both extracts. Chemical analyses revealed that the overall profile of both extracts was dominated by oxygenated constituents. In addition, ethanolic extract is mainly composed of phenols among which thymol (71.22%) and camphor (17.18%). As far as the literature could be ascertained, this is the first publication on anthelmintic activity of T. capitatus. The results of the present study suggest that T. capitatus extracts are a promising alternative to the commercially available anthelmintics like albendazole for the treatment of small ruminants' gastrointestinal nematodes. Copyright © 2013 Elsevier B.V. All rights reserved.
Krücken, Jürgen; Fraundorfer, Kira; Mugisha, Jean Claude; Ramünke, Sabrina; Sifft, Kevin C; Geus, Dominik; Habarugira, Felix; Ndoli, Jules; Sendegeya, Augustin; Mukampunga, Caritas; Bayingana, Claude; Aebischer, Toni; Demeler, Janina; Gahutu, Jean Bosco; Mockenhaupt, Frank P; von Samson-Himmelstjerna, Georg
Control of human soil-transmitted helminths (STHs) relies on preventive chemotherapy of schoolchildren applying the benzimidazoles (BZ) albendazole or mebendazole. Anthelmintic resistance (AR) is a common problem in nematodes of veterinary importance but for human STHs, information on drug efficacy is limited and routine monitoring is rarely implemented. Herein, the efficacy of single dose albendazole (400 mg) was evaluated in 12 schools in the Huye district of Rwanda where Ascaris is the predominant STH. Ascaris eggs were detected by wet mount microscopy and the Mini-FLOTAC method to assess cure rate (CR) and faecal egg count reduction (FECR). Blood and faecal samples were analysed for co-infections with Plasmodium sp. and Giardia duodenalis, respectively. Ascaris positive samples collected before and after treatment were analysed for putatively BZ-resistance associated β-tubulin gene single nucleotide polymorphisms. The overall CR was 69.9% by Mini-FLOTAC and 88.6% by wet mount microscopy. The FECR was 75.4% and the 95% calculated confidence intervals were 50.4-87.8% using sample variance, 55.4-88.8% by bootstrapping, and 75.0-75.7% applying a Markov Chain Monte Carlo Bayesian approach. FECR varied widely between 0 and 96.8% for individual schools. No putative BZ-resistance associated polymorphisms were found in the four Ascaris β-tubulin isotype genes examined. Since FECRs Ascaris populations cannot be formally proven. However, since FECRs <95% indicate reduced efficacy, BZ resistance may be suspected which would be alarming and calls for further analyses and routine monitoring in preventive chemotherapy programs. Copyright © 2017 The Authors. Published by Elsevier Ltd.. All rights reserved.
Walker, Martin; Hall, Andrew; Basáñez, María-Gloria
Studying the distribution of parasitic helminth body size across a population of definitive hosts can advance our understanding of parasite population biology. Body size is typically correlated with egg production. Consequently, inequalities in body size have been frequently measured to infer variation in reproductive success (VRS). Body size is also related to parasite age (time since entering the definitive host) and potentially provides valuable information on the mode of acquisition and establishment of immature (larval) parasites within the host: whether parasites tend to establish singly or in aggregates. The mode of acquisition of soil-transmitted helminths has been a theoretical consideration in the parasitological literature but has eluded data-driven investigation. In this paper, we analyse individual Ascaris lumbricoides weight data collected from a cohort of human hosts before and after re-infection following curative treatment, and explore its distribution within and among individuals in the population. Lorenz curves and Gini coefficients indicate that levels of weight inequality (a proxy for VRS) in A.lumbricoides are lower than other published estimates from animal-helminth systems. We explore levels of intra-host weight aggregation using statistical models to estimate the intraclass correlation coefficient (ICC) while adjusting for covariates using a flexible fractional polynomial transformation approach capable of handling non-linear functional relationships. The estimated ICCs indicate that weights are aggregated within hosts both at equilibrium and after re-infection, suggesting that parasites may establish within the host in clumps. The implications of a clumped infection process are discussed in terms of ascariasis transmission dynamics, control and anthelmintic resistance. Copyright © 2010 Australian Society for Parasitology Inc. Published by Elsevier Ltd. All rights reserved.
Walker, Martin; Hall, Andrew; Basáñez, María-Gloria
The importance of the mode of acquisition of infectious stages of directly-transmitted parasitic helminths has been acknowledged in population dynamics models; hosts may acquire eggs/larvae singly in a "trickle" type manner or in "clumps". Such models have shown that the mode of acquisition influences the distribution and dynamics of parasite loads, the stability of host-parasite systems and the rate of emergence of anthelmintic resistance, yet very few field studies have allowed these questions to be explored with empirical data. We have analysed individual worm weight data for the parasitic roundworm of humans, Ascaris lumbricoides, collected from a three-round chemo-expulsion study in Dhaka, Bangladesh, with the aim of discerning whether a trickle or a clumped infection process predominates. We found that hosts tend to harbour female worms of a similar weight, indicative of a clumped infection process, but acknowledged that unmeasured host heterogeneities (random effects) could not be completely excluded as a cause. Here, we complement our previous statistical analyses using a stochastic infection model to simulate sizes of individual A. lumbricoides infecting a population of humans. We use the intraclass correlation coefficient (ICC) as a quantitative measure of similarity among simulated worm sizes and explore the behaviour of this statistic under assumptions corresponding to trickle or clumped infections and unmeasured host heterogeneities. We confirm that both mechanisms are capable of generating aggregates of similar-sized worms, but that the particular pattern of ICCs described pre- and post-anthelmintic treatment in the data is more consistent with aggregation generated by clumped infections than by host heterogeneities alone. This provides support to the notion that worms may be acquired in clumps. We discuss our results in terms of the population biology of A. lumbricoides and highlight the significance of our modelling approach for the study of the
Desrues, Olivier; Peña-Espinoza, Miguel; Hansen, Tina V A; Enemark, Heidi L; Thamsborg, Stig M
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 explored in cattle. Therefore, our aim was to examine the efficacy of sainfoin against cattle nematodes in vivo. Fifteen Jersey male calves (2-4 month-old) were allocated into two groups and fed isoproteic and isoenergetic diets mainly composed of sainfoin pellets (Group SF; n = 9, three pens) 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 the study. Worms were harvested by sieving for quantification and scanning electron microscopy (SEM) 42 days post-infection (dpi) when the calves were necropsied. The number of O. ostertagi adults in the abomasum was reduced by 50 % in Group SF compared with Group CO (P 0.05). Likewise, no statistical difference in total worm burdens of C. oncophora was found between the groups. Weight gains were lower for Group SF (P < 0.05), which may reflect lower digestibility and phosphorus levels in the SF diet, despite similar feed intake at pen-level. Overall, the effect of sainfoin on abomasal nematodes corroborates results from studies with small ruminants and encourages further investigations of the use of this crop for control of cattle nematodes.
Novobilský, Adam; Amaya Solis, Natalia; Skarin, Moa; Höglund, Johan
Anthelmintic resistance (AR) to Fasciola hepatica is emerging worldwide. Recently, AR to the adulticide compound albendazole (ABZ) was shown in Argentina and Spain. In Sweden, ABZ treatment failure against F. hepatica was first reported in sheep in 2012. The present study tested the efficacy of ABZ and triclabendazole (TCBZ) in sheep naturally infected with F. hepatica using a combination of three different diagnostic methods: faecal egg counts (FEC), coproantigen ELISA (cELISA) and Fasciola egg hatch test (FEHT). Two deworming trials, in November 2014 and January 2015, were performed on two sheep farms (farms A and B) in south-western Sweden. Except ABZ in November, treatment with ABZ or TCBZ achieved sufficient efficacy (97-100%) against adult F. hepatica on farm A. In contrast, ABZ treatment failed in the sheep flock on farm B, despite low initial faecal egg output. On farm B, ABZ efficacy based on FEC was 67% (95% CI: 35-84) and four of eight ewes tested were coproantigen-positive 21 days post-treatment. Ovicidal activity of ABZ against Fasciola eggs in isolates from both farms and one additional bovine isolate were tested by FEHT to exclude the presence of juvenile flukes and other factors such as dosing failure and poor quality of drug product. Irrespective of drug trial, data from FEHT showed significantly lower ovicidal activity of ABZ for the ovine farm B isolate than for the isolate from farm A. This confirms that the low efficacy of ABZ in sheep flock B was associated with ABZ resistance. Overall, the usefulness of three complementary methods for detection of ABZ resistance in the field was demonstrated. Copyright © 2016 The Authors. Published by Elsevier Ltd.. All rights reserved.
Beesley, Nicola J; Williams, Diana J L; Paterson, Steve; Hodgkinson, Jane
Fasciola hepatica, the liver fluke, is a trematode parasite of considerable economic importance to the livestock industry and is a re-emerging zoonosis that poses a risk to human health in F. hepatica-endemic areas worldwide. Drug resistance is a substantial threat to the current and future control of F. hepatica, yet little is known about how the biology of the parasite influences the development and spread of resistance. Given that F. hepatica can self-fertilise and therefore inbreed, there is the potential for greater population differentiation and an increased likelihood of recessive alleles, such as drug resistance genes, coming together. This could be compounded by clonal expansion within the snail intermediate host and aggregation of parasites of the same genotype on pasture. Alternatively, widespread movement of animals that typically occurs in the UK could promote high levels of gene flow and prevent population differentiation. We identified clonal parasites with identical multilocus genotypes in 61% of hosts. Despite this, 84% of 1579 adult parasites had unique multilocus genotypes, which supports high levels of genotypic diversity within F. hepatica populations. Our analyses indicate a selfing rate no greater than 2%, suggesting that this diversity is in part due to the propensity for F. hepatica to cross-fertilise. Finally, although we identified high genetic diversity within a given host, there was little evidence for differentiation between populations from different hosts, indicating a single panmictic population. This implies that, once those emerge, anthelmintic resistance genes have the potential to spread rapidly through liver fluke populations. Copyright © 2016 The Authors. Published by Elsevier Ltd.. All rights reserved.
Full Text Available Infections with parasitic helminths (nematodes and trematodes represent a significant economic and welfare burden to the global ruminant livestock industry. The increasing prevalence of anthelmintic resistance means that current control programmes are costly and unsustainable in the long term. Recent changes in the epidemiology, seasonality and geographic distribution of helminth infections have been attributed to climate change. However, other changes in environment (e.g., land use and in livestock farming, such as intensification and altered management practices, will also have an impact on helminth infections. Sustainable control of helminth infections in a changing world requires detailed knowledge of these interactions. In particular, there is a need to devise new, sustainable strategies for the effective control of ruminant helminthoses in the face of global change. In this paper, we consider the impact of helminth infections in grazing ruminants, taking a European perspective, and identify scientific and applied priorities to mitigate these impacts. These include the development and deployment of efficient, high-throughput diagnostic tests to support targeted intervention, modelling of geographic and seasonal trends in infection, more thorough economic data and analysis of the impact of helminth infections and greater translation and involvement of end-users in devising and disseminating best practices. Complex changes in helminth epidemiology will require innovative solutions. By developing and using new technologies and models, the use of anthelmintics can be optimised to limit the development and spread of drug resistance and to reduce the overall economic impact of helminth infections. This will be essential to the continued productivity and profitability of livestock farming in Europe and its contribution to regional and global food security.
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
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.
Cornelius, M P; Jacobson, C; Besier, R B
Sheep nematode control utilising refugia-based strategies have been shown to delay anthelmintic resistance, but the optimal indices to select individuals to be left untreated under extensive sheep grazing conditions are not clear. This experiment tested the hypothesis that high body condition can indicate ability of mature sheep to better cope with worms and therefore remain untreated in a targeted treatment programme. Adult Merino ewes from flocks on two private farms located in south-west Western Australia (Farm A, n = 271, and Farm B, n = 258) were measured for body condition score (BCS), body weight and worm egg counts (WEC) on four occasions between May and December (pre-lambing, lamb marking, lamb weaning and post-weaning). Half of the ewes in each flock received anthelmintic treatments to suppress WEC over the experimental period and half remained untreated (unless critical limits were reached). Response to treatment was analysed in terms of BCS change and percentage live weight change. No effect of high or low initial WEC groups was shown for BCS response, and liveweight responses were inconsistent. A relatively greater BCS response to treatment was observed in ewes in low BCS pre-lambing compared to better-conditioned ewes on one farm where nutrition was sub-optimal and worm burdens were high. Sheep in low body condition pre-lambing were more than three times more likely to fall into a critically low BCS (<2.0) if left untreated. Recommendations can be made to treat ewes in lower BCS and leave a proportion of the higher body condition sheep untreated in a targeted selective treatment programme, to provide a population of non-resistant worms to delay the development of resistance. Copyright © 2014 Elsevier B.V. All rights reserved.
Bredtmann, Christina M; Krücken, Jürgen; Murugaiyan, Jayaseelan; Kuzmina, Tetiana; von Samson-Himmelstjerna, Georg
Human and animal health is globally affected by a variety of parasitic helminths. The impact of co-infections and development of anthelmintic resistance requires improved diagnostic tools, especially for parasitic nematodes e.g., to identify resistant species or attribute pathological effects to individual species or particular species combinations. In horses, co-infection with cyathostomins is rather a rule than an exception with typically 5 to 15 species (out of more than 40 described) per individual host. In cyathostomins, reliable morphological species differentiation is currently limited to adults and requires highly specialized expertize while precise morphological identification of eggs and early stage larvae is impossible. The situation is further complicated by a questionable validity of some cyathostomins while others might actually represent cryptic species complexes. Several molecular methods using different target sequences were established to overcome these limitations. For adult worms, PCR followed by sequencing of mitochondrial genes or external or internal ribosomal RNA spacers is suitable to genetically confirm morphological identifications. The most commonly used method to differentiate eggs or larvae is the reverse-line-blot hybridization assay. However, both methods suffer from the fact that target sequences are not available for many species or even that GenBank® entries are unreliable regarding the cyathostomin species. Recent advances in proteomic tools for identification of metazoans including insects and nematodes of the genus Trichinella will be evaluated for suitability to diagnose cyathostomins. Future research should focus on the comparative analysis of morphological, molecular and proteomic data from the same cyathostomin specimen to optimize tools for species-specific identification.
Christina M. Bredtmann
Full Text Available Human and animal health is globally affected by a variety of parasitic helminths. The impact of co-infections and development of anthelmintic resistance requires improved diagnostic tools, especially for parasitic nematodes e.g., to identify resistant species or attribute pathological effects to individual species or particular species combinations. In horses, co-infection with cyathostomins is rather a rule than an exception with typically 5 to 15 species (out of more than 40 described per individual host. In cyathostomins, reliable morphological species differentiation is currently limited to adults and requires highly specialized expertize while precise morphological identification of eggs and early stage larvae is impossible. The situation is further complicated by a questionable validity of some cyathostomins while others might actually represent cryptic species complexes. Several molecular methods using different target sequences were established to overcome these limitations. For adult worms, PCR followed by sequencing of mitochondrial genes or external or internal ribosomal RNA spacers is suitable to genetically confirm morphological identifications. The most commonly used method to differentiate eggs or larvae is the reverse-line-blot hybridization assay. However, both methods suffer from the fact that target sequences are not available for many species or even that GenBank® entries are unreliable regarding the cyathostomin species. Recent advances in proteomic tools for identification of metazoans including insects and nematodes of the genus Trichinella will be evaluated for suitability to diagnose cyathostomins. Future research should focus on the comparative analysis of morphological, molecular and proteomic data from the same cyathostomin specimen to optimize tools for species-specific identification.
Alison J Dicker
reduce the need for anthelmintic treatment and the resultant selection for anthelmintic resistant parasites.
Lean, Ij; Westwood, Ct; Playford, Mc
This paper provides an overview of the changes in the pasture-based dairy systems of New Zealand and Australia that may influence the health of cattle. There are relatively few available data that can be used to quantify the effects of increased intensification of milk production on the health of cattle. There is evidence that increased production increases the risk of mastitis and culling for udder health. Increased risks of mastitis with treatment with somatotropin support these findings; however, the risk of mastitis may decrease with increased milking frequency. Larger herds with greater stocking density should increase the risk for infectious disease, but evidence to support this contention is sparse. Very intensive grazing patterns associated with higher grass yields achieved using better cultivars and greater use of fertilisers favour nematode parasites. There is some evidence of anthelmintic resistance in both nematodes and liver fluke. Veterinarians will need to be aware of the potential for these to reduce the productivity of cattle. There have been benefits of improved nutrition on the efficiency of energy use for dairy production. Diseases such as bloat and ketosis appear to be of lower prevalence. It also appears that mineral nutrition of pasture-fed cattle is being better addressed, with gains in the control of milk fever, hypomagnesaemia and trace-element deficiencies. However, acidosis is a condition with a high point prevalence in pasture-based dairy systems where cows are fed supplements; one study in Australia found a point prevalence of approximately 11% of cows with acidosis. There is evidence from this study that the neutral detergent fibre (NDF) in pasture-based diets may need to be higher than 30% of the diet to maintain rumen stability. Laminitis and acidosis are different conditions with a similar pathogenesis, specifically highly fermentable diets. The prevalence of lameness was 28% in herds in Australia, suggesting that this condition
Nielsen, M K; Reist, M; Kaplan, R M; Pfister, K; van Doorn, D C K; Becher, A
Due to widespread development of anthelmintic resistance in equine parasites, recommendations for their control are currently undergoing marked changes with a shift of emphasis toward more coprological surveillance and reduced treatment intensity. Denmark was the first nation to introduce prescription-only restrictions of anthelmintic drugs in 1999, but other European countries have implemented similar legislations over recent years. A questionnaire survey was performed in 2008 among Danish horse owners to provide a current status of practices and perceptions with relation to parasite control. Questions aimed at describing the current use of coprological surveillance and resulting anthelmintic treatment intensities, evaluating knowledge and perceptions about the importance of various attributes of parasite control, and assessing respondents' willingness to pay for advice and parasite surveillance services from their veterinarians. A total of 1060 respondents completed the questionnaire. A large majority of respondents (71.9%) were familiar with the concept of selective therapy. Results illustrated that the respondents' self-evaluation of their knowledge about parasites and their control associated significantly with their level of interest in the topic and their type of education (Pequine parasites and the concept of selective therapy, although there was some confusion over the terms small and large strongyles. They used a large degree of fecal surveillance in all age groups, with a majority of respondents sampling and/or treating around twice a year. Finally, respondents appeared willing to spend money on parasite control for their horses. It is of concern that the survey suggested that foals and young horses are treated in a manner very similar to adult horses, which is against current recommendations. Thus, the survey illustrates the importance of clear communication of guidelines for equine parasite control. Copyright © 2013 Elsevier B.V. All rights reserved.
Schneider, Stephanie; Pfister, Kurt; Becher, Anne M; Scheuerle, Miriam C
As a consequence of the increasing levels of anthelmintic resistance in cyathostomes, new strategies for equine parasite control are being implemented. To assess the potential risks of these, the occurrence of strongyles was evaluated in a group of 1887 horses. The distribution of fecal egg counts (FECs), the frequency of anthelmintic drug use, and the deworming intervals were also analyzed. Between June 2012 and May 2013, 1887 fecal samples from either selectively or strategically dewormed horses were collected at 195 horse farms all over Germany and analyzed quantitatively with a modified McMaster technique. All samples with FEC ≥20 eggs per gram (EPG) were subjected to coproculture to generate third-stage larvae (LIII) for species differentiation. Egg counts were below the limit of detection (20 EPG) in 1046 (55.4%) samples and above it in 841 (44.6%) samples. Strongylus vulgaris larvae were identified in two of the 841 positive samples. Infections with cyathostomes were found on every farm. The most frequently applied anthelmintic was ivermectin (788/50.8%), followed by pyrantel (336/21.6%). The mean time since last treatment was 6.3 months. High-egg-shedding (>500 EPG) strategically dewormed horses (183/1357) were treated, on average, three times/year. The planned treatment date was already exceeded by 72.5% of the high egg-shedders and by 58.1% of the moderate (200-500 EPG) and low egg-shedders (20-199 EPG). S. vulgaris seems to be rare in Germany and no difference in its frequency has yet been found between selectively treated horses and horses receiving treatment in strategic intervals. However, inconsistent parasite control has been observed. Therefore, to minimize the risks for disease, consistent and efficient parasite control should be implemented.
Habarugira, Gervais; Mbasinga, Gloria; Mushonga, Borden; Chitura, Teedzai; Kandiwa, Erick; Ojok, Lonzy
There are no published abattoir bovine hepatic lesion prevalence studies in cattle in Rwanda. This study estimated that 12.3% of the livers (n=4751) examined at Nyabugogo slaughterhouse in Kigali were condemned. Condemnation prejudiced the nation of 3492.00kg of meat with attendant economic losses of US$8932.40 during the study period. Risk factors for these lesions were also assessed. Male and female animals from 11 districts were used in this study. Hepatic lesions were higher in females (14.6%; n=1494) than in males (11.1%; n=3257). About 78.7% of the condemnations were due to fascioliasis, followed by abscesses (5.7%), hepatitis (5.3%), cirrhosis (4%) and other lesions (6.3%). Female animal livers showed more fascioliasis and abscesses (82.2% and 9.5%) than male animal livers (73.3% and 3.3%). The highest rate of condemnation was observed from Kayonza (40.2%; n=413) and the least was from Gakenke district (0.9%; n=1031). Cattle from the Eastern Province showed significantly (P<0.05) higher prevalence of condemnations (26.8%) than the rest of the provinces. Liver specimens of animals below 3 years and above 6 years of age had a significantly higher (P<0.05) condemnation rate (14.4%) (n=3000 and n=769) than the 3-6year age-group at 4.1% (n=982). We conclude that fascioliasis was responsible for a significant proportion of the liver condemnations at Nyabugogo slaughterhouse. Being a zoonosis, we recommend an epidemio-surveillance, implementation of control measures and anthelmintic resistance investigation for fascioliasis in Rwanda. Copyright © 2016 Elsevier B.V. All rights reserved.
Charlier, J; Thamsborg, S M; Bartley, D J; Skuce, P J; Kenyon, F; Geurden, T; Hoste, H; Williams, A R; Sotiraki, S; Höglund, J; Chartier, C; Geldhof, P; van Dijk, J; Rinaldi, L; Morgan, E R; von Samson-Himmelstjerna, G; Vercruysse, J; Claerebout, E
Gastrointestinal (GI) nematode control has an important role to play in increasing livestock production from a limited natural resource base and to improve animal health and welfare. In this synthetic review, we identify key research priorities for GI nematode control in farmed ruminants and pigs, to support the development of roadmaps and strategic research agendas by governments, industry and policymakers. These priorities were derived from the DISCONTOOLS gap analysis for nematodes and follow-up discussions within the recently formed Livestock Helminth Research Alliance (LiHRA). In the face of ongoing spread of anthelmintic resistance (AR), we are increasingly faced with a failure of existing control methods against GI nematodes. Effective vaccines against GI nematodes are generally not available, and anthelmintic treatment will therefore remain a cornerstone for their effective control. At the same time, consumers and producers are increasingly concerned with environmental issues associated with chemical parasite control. To address current challenges in GI nematode control, it is crucial to deepen our insights into diverse aspects of epidemiology, AR, host immune mechanisms and the socio-psychological aspects of nematode control. This will enhance the development, and subsequent uptake, of the new diagnostics, vaccines, pharma-/nutraceuticals, control methods and decision support tools required to respond to the spread of AR and the shifting epidemiology of GI nematodes in response to climatic, land-use and farm husbandry changes. More emphasis needs to be placed on the upfront evaluation of the economic value of these innovations as well as the socio-psychological aspects to prioritize research and facilitate uptake of innovations in practice. Finally, targeted regulatory guidance is needed to create an innovation-supportive environment for industries and to accelerate the access to market of new control tools. © 2017 Blackwell Verlag GmbH.
Fernanda S. Fortes
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.
Waller, P J
Because parasites are more abundant, small ruminants in the tropical/subtropical regions of the world experience much greater ravages from internal parasitic disease than those in the temperate regions. In the tropics/subtropics, the limiting ecological factor influencing the severity of parasitism is rainfall, as temperatures almost always favour hatching and development of the free-living stages. Attempts to expand sheep and goat production by replacing traditional village production systems, which rarely involve anthelmintic treatment, with large-scale intensive commercial enterprises invariably induce complete reliance on anthelmintics to control nematode parasites. This has led to the widespread development of high level, multiple anthelmintic resistance throughout the tropics/subtropics, and in certain regions this has reached the ultimate disastrous scenario of total chemotherapeutic failure. Immediate concerted efforts are needed to resolve this crisis. Significant benefits are likely to emerge from research into non-chemotherapeutic approaches to nematode parasite control, such as grazing management, worm vaccines, breed selection and biological control. However, it is likely that none, in isolation or collectively, will completely replace the need for effective anthelmintics. What is needed is the integration of all methods of parasite control as they come to hand, with the underlying aim of reducing the use and thus preserving the effectiveness of anthelmintics. Although cheap and simple procedures, based on sound epidemiological principles, can achieve dramatic benefits in worm control, they have been poorly adopted by livestock owners. Clearly then, the greatest need is for technology transfer and education programmes, but these activities are generally found to be chronically under-resourced.
Turnbull, Frank; Jonsson, Nicholas N; Kenyon, Fiona; Skuce, Philip J; Bisset, Stewart A
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.
Mihaela M Martis
Full Text Available The nematode Ascaridia galli (order Ascaridida is an economically important intestinal parasite responsible for increased food consumption, reduced performance and elevated mortality in commercial poultry production. This roundworm is an emerging problem in several European countries on farms with laying hens, as a consequence of the recent European Union (EU ban on conventional battery cages. As infection is associated with slow development of low levels of acquired protective immunity, parasite control relies on repeated use of dewormers (anthelmintics. Benzimidazoles (BZ are currently the only anthelmintic registered in the EU for use in controlling A. galli and there is an obvious risk of overuse of one drug class, selecting for resistance. Thus we developed a reference transcriptome of A. galli to investigate the response in gene expression before and after exposure to the BZ drug flubendazole (FLBZ. Transcriptional variations between treated and untreated A. galli showed that transcripts annotated as mitochondrial glutamate dehydrogenase and cytochrome P450 were significantly down-regulated in treated worms, whereas transcripts homologous to heat shock proteins (HSP, catalase, phosphofructokinase, and a multidrug resistance P-glycoprotein (PGP1 were significantly up-regulated in treated worms. Investigation of candidate transcripts responsible for anthelmintic resistance in livestock nematodes led to identification of several tubulins, including six new isoforms of beta-tubulin, and several ligand-gated ionotropic receptors and ABC-transporters. We discovered several transcripts associated with drug binding and processing genes, but further characterisation using a larger set of worms exposed to BZs in functional assays is required to determine how these are involved in drug binding and metabolism.
De Visscher, A; Piepers, S; Haesebrouck, F; Supré, K; De Vliegher, S
Coagulase-negative staphylococci (CNS) have become the main pathogens causing bovine mastitis in recent years. A huge variation in species distribution among herds has been observed in several studies, emphasizing the need to identify subgroup- and species-specific herd-level factors to improve our understanding of the differences in ecological and epidemiological nature between species. The use of bulk milk samples enables the inclusion of a large(r) number of herds needed to identify herd-level risk factors and increases the likelihood of recovering enough isolates per species needed for conducting subgroup- and, eventually, species-specific analyses at the same time. This study aimed to describe the prevalence and distribution of CNS species in bulk milk samples and to identify associated subgroup- and species-specific herd-level factors. Ninety percent of all bulk milk samples yielded CNS. Staphylococcus equorum was the predominant species, followed by Staphylococcus haemolyticus and Staphylococcus epidermidis. A seasonal effect was observed for several CNS species. Bulk milk samples from herds with a loose-pack or a tiestall housing system were more likely to yield CNS species compared with herds with a freestall barn, except for S. epidermidis, Staphylococcus simulans, and Staphylococcus cohnii. In September, herds in which udders were clipped had lower odds of yielding Staphylococcus chromogenes, S. simulans, and Staphylococcus xylosus, the CNS species assumed to be most relevant for udder health, in their bulk milk than herds in which udder clipping was not practiced. Bulk milk of herds participating in a monthly veterinary udder health-monitoring program was more likely to yield these 3 CNS species. Herds always receiving their milk quality premium or predisinfecting teats before attachment of the milking cluster had lower odds of having S. equorum in their bulk milk. Herds not using a single dry cotton or paper towel for each cow during premilking udder
Kastman, Erik K; Kamelamela, Noelani; Norville, Josh W; Cosetta, Casey M; Dutton, Rachel J; Wolfe, Benjamin E
Many metagenomic sequencing studies have observed the presence of closely related bacterial species or genotypes in the same microbiome. Previous attempts to explain these patterns of microdiversity have focused on the abiotic environment, but few have considered how biotic interactions could drive patterns of microbiome diversity. We dissected the patterns, processes, and mechanisms shaping the ecological distributions of three closely related Staphylococcus species in cheese rind biofilms. Paradoxically, the most abundant species (S. equorum) is the slowest colonizer and weakest competitor based on growth and competition assays in the laboratory. Through in vitro community reconstructions, we determined that biotic interactions with neighboring fungi help resolve this paradox. Species-specific stimulation of the poor competitor by fungi of the genus Scopulariopsis allows S. equorum to dominate communities in vitro as it does in situ Results of comparative genomic and transcriptomic experiments indicate that iron utilization pathways, including a homolog of the S. aureus staphyloferrin B siderophore operon pathway, are potential molecular mechanisms underlying Staphylococcus-Scopulariopsis interactions. Our integrated approach demonstrates that fungi can structure the ecological distributions of closely related bacterial species, and the data highlight the importance of bacterium-fungus interactions in attempts to design and manipulate microbiomes. Decades of culture-based studies and more recent metagenomic studies have demonstrated that bacterial species in agriculture, medicine, industry, and nature are unevenly distributed across time and space. The ecological processes and molecular mechanisms that shape these distributions are not well understood because it is challenging to connect in situ patterns of diversity with mechanistic in vitro studies in the laboratory. Using tractable cheese rind biofilms and a focus on coagulase-negative staphylococcus (CNS
Madsen, Anne Mette; Moslehi-Jenabian, Saloomeh; Islam, Md Zohorul; Frankel, Mika; Spilak, Michal; Frederiksen, Margit W
The aim of this study was to obtain knowledge about concentrations of Staphylococcus aureus, MRSA (methicillin-resistant S. aureus), and other Staphylococcus species in indoor air in Greater Copenhagen and about factors affecting the concentrations. The effects of season, temperature, relative humidity, air change rate (ACR), other bacterial genera, area per occupant, and presence of S. aureus-positive occupants were studied. In samples from 67 living rooms, S. hominis, S. warneri, S. epidermidis, and S. capitis were found in 13-25%; S. saprophyticus, S. cohnii, and S. pasteuri in 5-10%; and S. lugdunensis, S. haemolyticus, S. caprae, S. equorum, S. kloosii, S. pettenkoferi, S. simulans, and S. xylosus in less than 3%. Staphylococcus aureus were found in two of 67 living rooms: spa type t034 (an MRSA) was recovered from a farmhouse, while spa type t509 was found in an urban home. Two species, S. equorum and S. kloosii, were found only in the farmhouse. Staphylococcus was significantly associated with season with lowest concentration and richness in winter. Genera composition was associated with ACR with smaller fractions of Staphylococcus at higher ACR, while richness was significantly and negatively associated with area per occupant. Concentration of Staphylococcus correlated positively with the total concentration of bacteria, but negatively with the total concentration of other bacteria. The concentration of Staphylococcus was not significantly associated with concentrations of the other abundant genera Bacillus, Kocuria, and Micrococcus. In offices with S. aureus-positive occupants, airborne S. aureus was not found. In conclusion, Staphylococcus species constitute a considerable proportion of the airborne bacteria in the studied homes and offices. However, both S. aureus and MRSA had very low prevalence during all seasons. Thus, transmission of S. aureus and MRSA through the air in living rooms in Copenhagen is expected to be limited. The negative associations
Fijałkowski, Karol; Peitler, Dorota; Karakulska, Jolanta
The aim of this study was to analyse the staphylococci isolated from ready-to-eat meat products, including pork ham, chicken cold cuts, pork sausage, salami and pork luncheon meat, sliced in the store to the consumer's specifications, along with species identification and determination of antibiotic resistance. Genes encoding staphylococcal enterotoxins, staphylococcal enterotoxin-like proteins, exfoliative toxins, and toxic shock syndrome toxin 1 were also investigated. From the 41 samples, 75 different staphylococcal isolates were obtained. Based on PCR-RFLP analysis of the gap gene using AluI and HpyCH4V restriction enzymes, the isolates were identified as Staphylococcus equorum (28%), S. vitulinus (16%), S. carnosus (14%), S. succinus (11%), S. xylosus (11%), S. saprophyticus (9%), S. warneri (9%), S. haemolyticus (1%) and S. pasteuri (1%). The incidence and number of resistances to antimicrobials was found to be species but not source of isolation dependent. All S. xylosus, S. saprophyticus, S. haemolyticus and S. pasteuri isolates showed antibiotic resistance. A lower percentage of resistance was recorded for S. warneri (71%) and S. vitulinus (58%), followed by S. equorum (57%), S. carnosus (50%) and S. succinus (50%). The most frequent resistance was observed to fusidic acid (43%). The mecA gene was amplified in 4% of the staphylococci. However, phenotypic resistance to methicillin was not confirmed in any of these isolates. On the other hand, the mecA gene was not detected in any of 9% of the isolates resistant to cefoxitin. It was also found that among 75 isolates, 60 (80%) harbored from 1 to 10 out of 21 analyzed superantigenic toxin genes. The most prevalent genes were: sei (36% isolates) among enterotoxins, seln (32% isolates) among enterotoxin-like proteins and eta encoding exfoliative toxin A (37% isolates). The findings of this study further extend previous observations that, when present in food, not only S. aureus but also other species of
Mickiewicz, Marcin; Czopowicz, Michał; Górski, Paweł; Kaba, Jarosław
Fecal egg count reduction (FECR) test with albendazole and egg hatch test (EHT) with thiabendazole (TBZ) were performed in a dairy goat herd suspected of anthelmintic resistance to benzimidazoles. The herd had been regularly dewormed with fenbendazole for 5 previous years and despite that it remained infected with several species of gastrointestinal nematodes (Trichostrongylus colubriformis, Teladorsagia circumcincta, and Haemonchus contortus). Albendazole was administered per os at dose of 20 mg/kg to 10 goats (treated group), while 10 other goats remained untreated (control group). Fecal egg count (FEC) was determined using McMaster egg counting method before and 7 days after the treatment in the treated group, and once (at the latter moment) in the control group. EHT was performed on the pooled rectal sample collected from treated goats. EHT comprised the negative control and 7 consecutive concentrations of TBZ (0.05, 0.1, 0.2, 0.3, 0.5, 1.0, 2.0 μg/ml) according to the standard procedure. Two hundred eggs/larvae were counted to determine percentage of unhatched eggs, which was adjusted by the natural mortality. TBZ dose effective in preventing hatching of 50% of eggs (ED50) was determined using the log-probit transformation. Median FEC (range) before the treatment was 1000 (250–3450) epg in the treated group and dropped to 150 (50–500) epg after the treatment (p=0.005). Median FEC (range) after the treatment was also significantly lower in the treated than in control group (p=0.009), where it was 725 (0–5050) epg. FECR between the treated and control group was 81% (95% CI: 49%, 93%). FECR in the treated group was 83% and 74% based on average and individual approach, respectively. ED50 value of TBZ was 0.78 μg/ml. Only H. contortus persisted in the treated group after treatment. The results indicate resistance of H. contortus to a benzimidazole anthelmintic, which is the first such case reported in Polish goats.
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. .
Paolini, V; De La Farge, F; Prevot, F; Dorchies, Ph; Hoste, H
Due to the high prevalence of anthelmintic resistance in goats, the need to explore novel approaches to control nematodes and to reduce the exclusive reliance on chemotherapy is strongly demanded in this host species. In sheep, several studies have shown that the consumption of tannin-rich legume forages was associated with positive effects on host resilience and resistance to parasite infection. In goats, studies on such interactions between tanniferous plants and nematode infections remain few. The objectives of the current study were to examine under natural conditions the effects of consumption of sainfoin hay by goats on the parasite populations and on host resilience. Eighteen adult cull goats naturally infected with Haemonchus contortus, Teladorsagia circumcincta and Trichostrongylus colubriformis were used in the study. At the start of the assay, the goats were allocated into two groups, balanced according to weight and the levels of egg excretion. The two groups grazed separate pastures for 3 months with similar stocking rates. Goats from group S received each month indoors, for 7 days, sainfoin hay and control goats (group C) received hay of ryegrass. The diets in both groups were made isoenergetic and isoproteic and the refusals measured. Individual parasitological and pathophysiological measurements were performed fortnightly in order to compare host resistance and resilience. At the end of the study, five goats per group were necropsied. The distribution of sainfoin was associated with: (1) a higher consumption of hay; (2) significant, lower levels of nematode egg excretion which was associated with a decrease in worm fertility but no change in worm population; however, the number of intestinal worms was reduced by 50% in group S; (3) a better host resilience. In particular, after 2 months of grazing, two control goats died and half of the remaining animals needed to be treated whereas this was not the case in group S. These differences were related to
Collas, C; Sallé, G; Dumont, B; Cabaret, J; Cortet, J; Martin-Rosset, W; Wimel, L; Fleurance, G
The spread of anthelmintic resistance in equine strongyle nematodes has become a major problem, advocating for the development of alternative control for strongyles. Our study consisted of both in vivo and in vitro experiments. We investigate for the first time the efficacy of a short-term consumption of tannin-rich sainfoin (Onobrychis viciifolia) or extra proteins in naturally infected horses. We used 30 horses allocated into three groups of 10 individuals that received for 18 days either (i) a tannin-rich diet with 70% DM sainfoin pellets (Sd), (ii) a protein-rich diet with 52% DM Italian rye-grass pellets and 18% DM grinded linseed expeller (Pd), or (iii) a control diet with 45% DM barley and 25% DM cereal-based pellets (Cd). The three diets were isoenergetic, covering 94% of animal energy requirements on average, and the Sd and Pd diets were isoproteic and provided extra proteins (227% of protein requirements v. 93% for the Cd diet). Pd and Cd were compared to test for benefits of receiving extra proteins, while Sd and Pd were compared to account for the effect of sainfoin secondary metabolites. There were no between-diet differences in faecal egg counts (FEC) or in worm burden evaluated from worm counts in faeces of drenched horses at the end of the experiment. However, coprocultures from the faeces collected in each group at the beginning and at the end of the experiment suggested a lower rate of strongyle larval development in the Sd group at the end of the experiment (Sd=8.1%, Pd=30.5%, Cd=22.6%). In vitro tests using sainfoin solutions evidenced the influence of sainfoin on strongyle larval development: adding 29% of sainfoin pellets to faeces reduced the strongyle egg development into infective larvae by 82% (P<0.001) and using solutions with sainfoin concentrations higher than 7.5 mg/ml reduced egg hatching by 37% (P<0.05). The short-term use of tannin-rich plants in horse diet could thus constitute a promising strategy to reduce the risk of infection
Scantlebury, Claire E; Peachey, Laura; Hodgkinson, Jane; Matthews, Jacqui B; Trawford, Andrew; Mulugeta, Getachew; Tefera, Gebre; Pinchbeck, Gina L
Gastrointestinal nematode infections constitute a threat to the health and welfare of donkeys worldwide. Their primary means of control is via anthelmintic treatments; however, use of these drugs has constraints in developing countries, including cost, limited availability, access to cheaper generic forms of variable quality and potential anthelmintic resistance. As an alternative, bioactive plants have been proposed as an option to treat and control gastrointestinal helminths in donkeys. This study aimed to use participatory methodology to explore donkey owner knowledge, attitudes and beliefs relating to the use of plant-based treatments for gastrointestinal parasites of donkeys in Ethiopia. In focus groups, 22/29 groups stated they knew of plants used for the treatment of gastrointestinal parasites in donkeys. All groups volunteered plants that were used in cattle and/or small ruminants. In total, 21 plants were named by participants. 'Koso' (Hagenia abyssinica) 'Grawa' (Vernonia amygdalina) and a mixed roots and leaves preparation were the most frequently named plant preparations. 'Enkoko' (Embelia shimperi) and 'a mixture of roots and leaves' were ranked highly for effectiveness in donkeys. However, 'Grawa' and 'Koso' were the highest ranked when taking into account both the rank position and the number of groups ranking the plant.Thematic analysis of participants' current attitudes and beliefs surrounding traditional plant-based remedies for gastrointestinal parasites revealed that anthelmintics obtained from clinics were generally favoured due to their ease of administration and perceived higher effectiveness. There was doubt surrounding the effectiveness of some plant-based treatments, but there were also perceived advantages including their low cost, ease of cultivation and availability. However, plant-based treatments were considered a "past trend" and people favoured "modern" medicine, particularly among the younger generation. There was extensive
Full Text Available Fasciola spp. liver fluke cause pernicious disease in humans and animals. Whilst current control is unsustainable due to anthelmintic resistance, gene silencing (RNA interference, RNAi has the potential to contribute to functional validation of new therapeutic targets. The susceptibility of juvenile Fasciola hepatica to double stranded (dsRNA-induced RNAi has been reported. To exploit this we probe RNAi dynamics, penetrance and persistence with the aim of building a robust platform for reverse genetics in liver fluke. We describe development of standardised RNAi protocols for a commercially-available liver fluke strain (the US Pacific North West Wild Strain, validated via robust transcriptional silencing of seven virulence genes, with in-depth experimental optimisation of three: cathepsin L (FheCatL and B (FheCatB cysteine proteases, and a σ-class glutathione transferase (FheσGST.Robust transcriptional silencing of targets in both F. hepatica and Fasciola gigantica juveniles is achievable following exposure to long (200-320 nt dsRNAs or 27 nt short interfering (siRNAs. Although juveniles are highly RNAi-susceptible, they display slower transcript and protein knockdown dynamics than those reported previously. Knockdown was detectable following as little as 4h exposure to trigger (target-dependent and in all cases silencing persisted for ≥25 days following long dsRNA exposure. Combinatorial silencing of three targets by mixing multiple long dsRNAs was similarly efficient. Despite profound transcriptional suppression, we found a significant time-lag before the occurrence of protein suppression; FheσGST and FheCatL protein suppression were only detectable after 9 and 21 days, respectively.In spite of marked variation in knockdown dynamics, we find that a transient exposure to long dsRNA or siRNA triggers robust RNAi penetrance and persistence in liver fluke NEJs supporting the development of multiple-throughput phenotypic screens for control
Hernández-Villegas, M M; Borges-Argáez, R; Rodriguez-Vivas, R I; Torres-Acosta, J F J; Méndez-Gonzalez, M; Cáceres-Farfan, M
The development of anthelmintic resistance has impacted on the success of conventional anthelmintics (AH) for the control of gastrointestinal nematodes in grazing/browsing sheep and goats. Medicinal plants from the traditional herbolary in Mexico may provide new candidates that can be explored as alternative sources of AHs for ruminants. This study evaluated the leaf extracts derived from Phytolacca icosandra against infective L(3) larvae and eggs from Haemonchus contortus collected from sheep. Three extracts of different polarities were obtained from the leaf plants using ethanol, n-hexane and dichloromethane as the solvents. The effectiveness of the in vitro AH activity of the plant extracts was evaluated using larval migration inhibition (LMI) and egg hatch (EHA) assays. For the LMI assays, the ethanolic extract of P. icosandra showed 55.4% inhibition of larval migration at 2mg/mL (p<0.05). The dichloromethane extract of P. icosandra showed 67.1% inhibition of migration at 3mg/mL (p<0.05) and a dose-dependent response with an LD(50) of 0.90 mg/mL. The n-hexane extract failed to show inhibition of larval migration at any concentration explored. In the EHA for the ethanol extract, the lowest concentration tested (0.15 mg/mL) resulted in inhibition of egg hatching greater than 72.6%. Therefore, the LD(50) could not be calculated for this extract. The LD(50) of the dichloromethane extract of P. icosandra was 0. 28 mg/mL. An egg hatch inhibition greater than 90% was observed with both the ethanolic and dichloromethane extracts when using a concentration of 0.90 mg/mL or higher. The n-hexane extract failed to show egg hatch inhibition at any concentration tested. The AH activity reported for P. icosandra could be attributable to the flavonoids, steroids, terpenoids, coumarins and/or saponins that were present in the ethanolic and dichloromethane extracts. A combination of more than one component may also explain the observed AH activity against the H. contortus life
Cornelius, M P; Jacobson, C; Besier, R B
individual BCS assessment would be of no benefit under these circumstances except for identifying low BCS sheep at risk of falling below critical limits associated with health or welfare risks. No consistent relationship between WEC and BCS or bodyweight was observed, indicating that BCS selection would have no lesser or greater impact on worm pasture contamination compared to random selection. Summer treatments based on a random selection index (with a minimum BCS limit), with up to 30% of adult sheep untreated can be expected to delay the development of anthelmintic resistance, with minimal adverse effect on sheep health or production. Copyright © 2015 Elsevier B.V. All rights reserved.
Full Text Available El monepantel es un nuevo antihelmíntico registrado en nuestro país exclusivamente para el control de los nematodes gastrointestinales de los ovinos y su uso ha sido direccionado mayormente hacia el control de parásitos resistentes a las clases de antihelmínticos disponibles actualmente. Estos mismos nematodes también parasitan a los caprinos, pero en estos rumiantes la pato-fisiología y la respuesta a los antihelmínticos es diferente, lo cual resulta en un mayor parasitismo y complejidad en el manejo de la resistencia. La presente comunicación informa sobre la eficacia del monepantel en dos hatos caprinos mantenidos bajo condiciones de campo y parasitados naturalmente por los géneros de nematodes gastrointestinales más comunes del área central de la Argentina (Haemonchus y Trichostrongylus y portando alelos de resistencia múltiple (ivermectina y febendazole. El test de reducción en el conteo de huevos post tratamiento comparando diversas fórmulas, indicaron que en todos los hatos el monepantel por vía oral y a la dosis de 3,75 mg/kg de peso (1,5 veces mayor a la dosis ovina, resultó en eficacias del 99% al 100 %. Se realizan breves consideraciones sobre el uso potencial de esta droga en caprinos. SUMMARY. Monepantel effectiveness against Haemonchus contortus and Trichostrongylus spp. with multiple resistance (ivermectin and febendazole in goats. Monepantel is a new anthelmintic registered in Argentina exclusively for control of gastrointestinal nematodes of sheep and mostly directed toward controlling resistant parasites to current available classes of anthelmintics. The same nematodes also parasitize goats but pathophysiology and response are different in these hosts resulting in more severe parasitism and complexity in handling anthelmintic resistance. This report assess the efficacy of monepantel in goats maintained under field conditions and naturally parasitized by most common gastrointestinal nematodes from central
Vasileiou, N G C; Fthenakis, G C; Papadopoulos, E
The present paper discusses the spread of parasites by animal movements in small ruminant farms; it focuses in dissemination of parasitic forms that would lead to subsequent infection of sheep or goats. Systems of small ruminant production involve a component of animal movement (e.g., grazing) as part of routine husbandry, which favors spread of parasitic forms; that refers mainly to parasites of the digestive system (nematodes, trematodes, cestodes, protozoa), as well as helminthes of the respiratory system, although dissemination of the various parasitic forms in the environment would not always result to subsequent infection; external parasites may also be disseminated during movements, e.g., to inhabit wooden poles used in fencing. New livestock into a farm constitutes a biosecurity hazard and the most common means to introducing new parasitic pathogens into a farm; in contemporary small ruminant health management, this contributes in dissemination of anthelmintic resistant parasitic strains; other parasitic disease agents (e.g., mange mites, ticks) may also be spread into a farm that way. Often, especially in small scale farming, visits of rams or bucks take place from one farm to another during the mating season; in such cases, ectoparasites (e.g., mange mites) can be disseminated through direct contact of animals, as well other pathogens (e.g., Toxoplasma gondii, Neospora caninum) via the semen. During transportation of sheep/goats, parasitic forms can also spread, as well as during movement of sheep or goats to slaughterhouses, in which case dogs present in these places would contribute to their dissemination. Spread of life forms of various parasites can also occur from animal species present in the environment of sheep or goats; these include animals present within a farm, stray dogs roaming around a farm (e.g., for spread of Multiceps multiceps, Echinococcus granulosus, Taenia hydatigena, N. caninum), cats commanding the environment of a farm (e.g., for
Scare, J A; Slusarewicz, P; Noel, M L; Wielgus, K M; Nielsen, M K
Fecal egg counts are emphasized for guiding equine helminth parasite control regimens due to the rise of anthelmintic resistance. This, however, poses further challenges, since egg counting results are prone to issues such as operator dependency, method variability, equipment requirements, and time commitment. The use of image analysis software for performing fecal egg counts is promoted in recent studies to reduce the operator dependency associated with manual counts. In an attempt to remove operator dependency associated with current methods, we developed a diagnostic system that utilizes a smartphone and employs image analysis to generate automated egg counts. The aims of this study were (1) to determine precision of the first smartphone prototype, the modified McMaster and ImageJ; (2) to determine precision, accuracy, sensitivity, and specificity of the second smartphone prototype, the modified McMaster, and Mini-FLOTAC techniques. Repeated counts on fecal samples naturally infected with equine strongyle eggs were performed using each technique to evaluate precision. Triplicate counts on 36 egg count negative samples and 36 samples spiked with strongyle eggs at 5, 50, 500, and 1000 eggs per gram were performed using a second smartphone system prototype, Mini-FLOTAC, and McMaster to determine technique accuracy. Precision across the techniques was evaluated using the coefficient of variation. In regards to the first aim of the study, the McMaster technique performed with significantly less variance than the first smartphone prototype and ImageJ (psmartphone and ImageJ performed with equal variance. In regards to the second aim of the study, the second smartphone system prototype had significantly better precision than the McMaster (psmartphone system were 64.51%, 21.67%, and 32.53%, respectively. The Mini-FLOTAC was significantly more accurate than the McMaster (psmartphone system (psmartphone and McMaster counts did not have statistically different accuracies
Knapp-Lawitzke, Friederike; von Samson-Himmelstjerna, Georg; Demeler, Janina
of anthelmintic resistance and the presence of hot/dry weather conditions. Copyright © 2016 Australian Society for Parasitology Inc. Published by Elsevier Ltd. All rights reserved.
Kenyon, F; Jackson, F
In Europe, most nematodoses are subclinical involving morbid rather than mortal effects and control is largely achieved using anthelmintics. In cattle, the genera most associated with sub-optimal performance are Ostertagia and Cooperia whereas in sheep and goats, subclinical losses are most often caused by Teladorsagia and Trichostrongylus. In some regions, at certain times, other species such as Nematodirus and Haemonchus also cause disease in sheep and goats. Unfortunately, anthelmintic resistance has now become an issue for European small ruminant producers. One of the key aims of the EU-funded PARASOL project was to identify low input and sustainable approaches to control nematode parasites in ruminants using refugia-based strategies. Two approaches to optimise anthelmintic treatments in sheep and cattle were studied; targeted treatments (TT) - whole-group treatments optimised on the basis of a marker of infection e.g. faecal egg count (FEC), and targeted selected treatment (TST) - treatments given to identified individuals to provide epidemiological and/or production benefits. A number of indicators for TT and TST were assessed to define parasitological and production-system specific indicators for treatment that best suited the regions where the PARASOL studies were conducted. These included liveweight gain, production efficiency, FEC, body condition score and diarrhoea score in small ruminants, and pepsinogen levels and Ostertagia bulk milk tank ELISA in cattle. The PARASOL studies confirmed the value of monitoring FEC as a means of targeting whole-flock treatments in small ruminants. In cattle, bulk milk tank ELISA and serum pepsinogen assays could be used retrospectively to determine the levels of exposure and hence, in the next season to optimise anthelmintic usage. TST approaches in sheep and goats examined production efficiency and liveweight gain as indicators for treatment and confirmed the value of this approach in maintaining performance and
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.
Yilmaz, Esra; Ramünke, Sabrina; Demeler, Janina; Krücken, Jürgen
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 HcH isolate, which, except
Full Text Available Infestations of gastrointestinal parasitic nematodes in sheep result in significant costs to farmers. These infestations are controlled using synthetic anthelmintic treatments, which can result in the development of anthelmintic resistance in nematodes. The use of plants rich in condensed tannins (CTs is a promising alternative for controlling infestations of harmful parasites in sheep, and could allow reduction of the chemical products used. This study investigated the in vitro effect of CTs from Acacia mearnsii extract (AE on egg hatching and motility of third-stage larvae. Egg-hatching rate was measured after incubation with extracts for 48 h at 27 °C. The egg hatch test was performed with dilutions of 0.09, 0.19, 0.39, 0.78, 1.56, 3.12, 6.25, 12.5, 25, 50, and 100 mg mL-1. Distilled water was used as the negative control. The corresponding egg hatching inhibition percentages were 22.3, 32.3, 39.2, 49.1, 56.7, 59.0, 62.3, 77.3, 92.7, 98.3, and 100%. The concentration required to inhibit egg hatching in 50% of eggs (LC50 was 2.85 mg mL-1. The inhibition achieved with the negative control was 7.06%. A larval migration inhibition test was carried out after incubation with the extracts for 48 h at 27 oC, with AE and distilled water used in dilutions of 3.12, 6.25, 12.5, 25, 50, and 100 mg mL-1. The corresponding percentages of migration inhibition were 16.5, 37.0, 56.3, 79.4, 91.8, and 97.1%. The concentration required to inhibit migration of 50% of larvae (LC50 was 12.45 mg mL-1. The inhibition achieved with the negative control was 8.53%. The in vitro ovicidal and larvicidal activity of CTs from AE indicate the anthelmintic effect of AE, suggesting the potential of CT extracts to be used as alternatives for controlling gastrointestinal nematode infestations in small ruminants.
McMahon, Connor; Edgar, Hillary W J; Barley, Jason P; Hanna, Robert E B; Brennan, Gerard P; Fairweather, Ian
To address a lack of information on the control of ovine helminth parasites in Northern Ireland (NI), a number of research projects have been undertaken, dealing with gastrointestinal nematodes, tapeworms and liver fluke. This investigation concerns Nematodirus and concentrates on three aspects of disease: farm management strategies for its control, derived from the results of a Questionnaire; the efficacy of treatment used by farmers, as determined by a coprological survey; and the hatching requirements of Nematodirus eggs, that is, whether prolonged chilling is a pre-requisite for hatching. A Questionnaire was sent to 252 sheep farmers in NI in March 2012 (covering the years 2009-2012) and replies were received from 228 farmers. Under-dosing, inaccurate calibration of equipment and inappropriate product choice were poor practices identified. Following this survey, the efficacy of treatment of Nematodirus spp. in sheep flocks was evaluated in April and May 2012. Sampling kits were sent to 51 flock owners, all of whom returned pre- and post-anthelmintic dosing faecal samples to the laboratory for analysis. At the time of treatment, 41 flocks were positive for Nematodirus (as diagnosed by the presence of eggs). Reduced benzimidazole efficacy was detected in 35.7% of flocks tested ( n = 28). Although only involving a small number of flocks, reduced efficacy of levamisole treatment was detected in 50%, of avermectins in 33% and of moxidectin in 75% of flocks tested (n = 2, 6 and 4, respectively). In the egg hatch experiment, carried out under "chilled" and "non-chilled" conditions, 43% of the eggs in the "non-chilled" group were able to hatch, compared to 100% in the "chilled" group. The identification of inefficient control strategies argues for continued education of stockholders, in order to improve their management programmes. This is particularly important where the practices might impact on the development of anthelmintic resistance, which has been shown to
Vande Velde, F; Charlier, J; Hudders, L; Cauberghe, V; Claerebout, E
Emerging anthelmintic resistance emphasizes the need for sustainable control approaches against gastrointestinal nematode (GIN) infections in cattle. The uptake of diagnostic methods for sustainable control could enable more informed treatments and reduce excessive anthelmintic use. Unfortunately, the adoption of such methods remains relatively poor. A better understanding of farmers' motivations and behaviour would help to develop applicable advises and communication strategies for sustainable worm control strategies. A previous study created a general model for adoption intention of GIN diagnostics on dairy farms and measured the most important factors driving this intention (Vande Velde et al., 2015). The current research aimed to dig deeper into this model for the beliefs underlying these factors, and to identify additional factors impelling this specific behaviour. Data were collected through 22 semi-structured interviews with dairy farmers. Using analytic induction analysis, data were moved between deduction and induction. Results show that the adoption process of diagnostic methods for GIN occurs through three different phases: adoption intention, actual adoption and maintenance. Low infection awareness and low priority ('top of mind') of the disease are important barriers for adopting sustainable GIN control. Secondly, farmer behaviour is guided by two important social norms: the opinion of their veterinarian and their fellow farmers. However, farmers hold an incongruent relationship with both norms throughout different stages of behaviour: they do not value other farmers' opinions as a positive reference (intention phase), but follow and mimic their behaviour as a group (action phase). The veterinarian is seen as the most important positive reference, but also the responsible actor for GIN control. As such, the farmers do not hold themselves responsible for implementing sustainable control strategies. Thirdly, different types of motivations influence
Taylor, M A
Infections with gastrointestinal roundworms are an important cause of production losses in sheep and cattle. Worm control is a vital part of health and production management in sheep flocks and cattle herds in the UK, and good control is highly dependent on effective anthelmintics. Unfortunately, a direct and unavoidable consequence of using anthelmintics to control worm populations is selection for individuals that are resistant to the chemicals used. If left unchecked, anthelmintic resistance (AR) could prove to be one of the biggest challenges to sheep and cattle production and animal welfare within the UK. As a consequence of increasing reports of AR in sheep, a working group, "SCOPS" (sustainable control of parasites in sheep) was formed in 2003 with representatives from the UK sheep industry to promote practical guidelines for sheep farmers and their advisors. This led to the production of guidelines for 'sustainable worm control strategies for sheep' intended for veterinarians and sheep advisors, plus ongoing promotional literature aimed at farmers. Whilst there is some evidence of emerging resistance in roundworms of cattle, it appears to still be at a very low level in the UK. However the potential presence of such AR in cattle worms has been seen as a timely warning, which if ignored, could lead to a not dissimilar AR situation to that seen in sheep, and in other cattle areas around the world. Reports of AR in UK cattle nematodes have generally been limited to a small number of anecdotal reports of treatment failure with some macrocyclic lactone (ML) products, especially those formulated as pour-on preparations, and invariably involving the dose-limiting species, Cooperia oncophora. As a consequence of these observations, guidelines have been produced similar to those for sheep, for sustainable worm control strategies for cattle "COWS" (control of worms sustainably), and were launched in May 2010. Uptake and effectiveness of SCOPS recommendations are
Sadaf Bukhari and Prabir Kumar Sanyal
Full Text Available Because of the environmental and consumer concerns arising out of exponential growth in human population the world over, a term Sustainable Development has become an integral international concept, which is defined as one which meets the needs of the present without compromising the ability of future generations to meet their own needs. Ruminant animals appear sustainable as they do not compete with man for food, play a crucial role in the conversion of low quality plant material and crop residues to high quality human food as well as return valuable plant nutrients to the soil. Parasite control in ruminant livestock is a first-order input in any sustainable animal production system. As sustainable development is a compromise between reducing environmental degradation and positive economic growth, sustainable parasite control should aim towards less intensive, lower input, lesser risk of parasite induced losses with greater opportunities for integration of all available control resources. The compound scenario of rising anthelmintic resistance, food and environmental security and apathy of the pharmaceutical industry to go for the invention of new anthelmintic compounds has triggered the need for optimising the use of available anthelmintics with integration of all other alternative means for sustainable worm control. The “Sustainable Control of Parasitic Gastroenteritis in Ruminants” is thus encompasses a multidisciplinary approach involving integration of chemotherapy, grazing management, biological control, worm vaccines, genetic resistance of hosts, mathematical model based decision support and other strategies, if any. There is no single requirement more crucial to the rational and sustainable control of helminth parasites in grazing animals than a comprehensive knowledge of the epidemiology of the parasite as it interacts with the host in a specific climatic, management and production environment. In its absence, anthelmintic treatment
Semedo-Lemsaddek, Teresa; Carvalho, Laura; Tempera, Carolina; Fernandes, Maria H; Fernandes, Maria J; Elias, Miguel; Barreto, António S; Fraqueza, Maria J
The manufacture of dry fermented sausages is an important part of the meat industry in Southern European countries. These products are usually produced in small shops from a mixture of pork, fat, salt, and condiments and are stuffed into natural casings. Meat sausages are slowly cured through spontaneous fermentation by autochthonous microbiota present in the raw materials or introduced during manufacturing. The aim of this work was to evaluate the technological and safety features of coagulase-negative staphylococci (CNS) isolated from Portuguese dry fermented meat sausages in order to select autochthonous starters. Isolates (n = 104) obtained from 2 small manufacturers were identified as Staphylococcus xylosus, Staphylococcus equorum, Staphylococcus saprophyticus, and Staphylococcus carnosus. Genomically diverse isolates (n = 82) were selected for further analysis to determine the ability to produce enzymes (for example, nitrate-reductases, proteases, lipases) and antibiotic susceptibility. Autochthonous CNS producing a wide range of enzymes and showing low antibioresistance were selected as potential starters for future use in the production of dry fermented meat sausages. © 2016 Institute of Food Technologists®
León-Galván, Ma Fabiola; Barboza-Corona, José E; Lechuga-Arana, A Arianna; Valencia-Posadas, Mauricio; Aguayo, Daniel D; Cedillo-Pelaez, Carlos; Martínez-Ortega, Erika A; Gutierrez-Chavez, Abner J
Thirty-two farms (n = 535 cows) located in the state of Guanajuato, Mexico, were sampled. Pathogens from bovine subclinical mastitis (SCM) and clinical mastitis (CLM) were identified by 16S rDNA and the sensitivity to both antibiotics and bacteriocins of Bacillus thuringiensis was tested. Forty-six milk samples were selected for their positive California Mastitis Test (CMT) (≥3) and any abnormality in the udder or milk. The frequency of SCM and CLM was 39.1% and 9.3%, respectively. Averages for test day milk yield (MY), lactation number (LN), herd size (HS), and number of days in milk (DM) were 20.6 kg, 2.8 lactations, 16.7 animals, and 164.1 days, respectively. MY was dependent on dairy herd (DH), LN, HS, and DM (P resistant to penicillin, clindamycin, ampicillin, and cefotaxime. Bacteriocins synthesized by Bacillus thuringiensis inhibited the growth of multiantibiotic resistance bacteria such as S. agnetis, S. equorum, Streptococcus uberis, Brevibacterium stationis, and Brachybacterium conglomeratum, but they were not active against S. sciuri, a microorganism that showed an 84% resistance to antibiotics tested in this study.
Yang, Hang; Wang, Mengyue; Yu, Junping; Wei, Hongping
Biofilm formation renders Staphylococcus aureus highly resistant to conventional antibiotics and host defenses. Four D-amino acids (D-Leu, D-Met, D-Trp and D-Tyr) have been reported to be able to inhibit biofilm formation and disassemble established S. aureus biofilms. We report here for the first time that both D- and L-isoforms of aspartate (Asp) inhibited S. aureus biofilm formation on tissue culture plates. Similar biofilm inhibition effects were also observed against other staphylococcal strains, including S. saprophyticus, S. equorum, S. chromogenes and S. haemolyticus. It was found that Asp at high concentrations (>10 mM) inhibited the growth of planktonic N315 cells, but at subinhibitory concentrations decreased the cellular metabolic activity without influencing cell growth. The decreased cellular metabolic activity might be the reason for the production of less protein and DNA in the matrix of the biofilms formed in the presence of Asp. However, varied inhibition efficacies of Asp were observed for biofilms formed by clinical staphylococcal isolates. There might be mechanisms other than decreasing the metabolic activity, e.g. the biofilm phenotypes, affecting biofilm formation in the presence of Asp. © FEMS 2015. All rights reserved. For permissions, please e-mail: firstname.lastname@example.org.
Zhang, He; Li, Yan; Xu, Kunhua; Wu, Jiajia; Dai, Zhiyuan
Sanbao large yellow croaker is a kind of Chinese salted and fermented seafood processed by adding 50% of salt and fermenting at high temperature [around 30 °C] over 20 d. To get a comprehensive understanding of cultivable microorganism's diversity present in its fermentation, the chemical and microbial properties of this product, were detected initially, followed by identification of bacteria recovered from different fermentation periods using PCR-RFLP and 16S rRNA gene sequencing analysis. In total, 105 indigenous isolates were recovered with 3 different medium and majority of the isolates picked up from both MRS and MSA medium were finally clustered into the genus of Staphylococcus (S.). Among the 90 Staphylococci, S. xylosus, S. saprophyticus, and S. nepalensis were the most prevailing cultivable species recovered throughout the whole production process (70 isolates, 77.8%), while 5 other species, namely, S. aureus, S. vitulinus, S. sciuri, S. equorum, and S. succinus formed a minor fraction (20 isolates, 22.2%) of the Staphylococcus communities. Lactic acid bacteria, Pseudomonas, Proteus, and Bacillus constituted trivial populations in the initial period of the fermentation and then gave the way to the Staphylococcus immediately. High salt concentration used during the processing is like to have a pronounced influence on the microbial populations involved. Data obtained in this work could be referred in control and optimization of the fermentation process and selecting suitable strains for aquatic product fermentation. © 2015 Institute of Food Technologists®
León-Galván, Ma. Fabiola; Barboza-Corona, José E.; Lechuga-Arana, A. Arianna; Valencia-Posadas, Mauricio; Aguayo, Daniel D.; Cedillo-Pelaez, Carlos; Martínez-Ortega, Erika A.; Gutierrez-Chavez, Abner J.
Thirty-two farms (n = 535 cows) located in the state of Guanajuato, Mexico, were sampled. Pathogens from bovine subclinical mastitis (SCM) and clinical mastitis (CLM) were identified by 16S rDNA and the sensitivity to both antibiotics and bacteriocins of Bacillus thuringiensis was tested. Forty-six milk samples were selected for their positive California Mastitis Test (CMT) (≥3) and any abnormality in the udder or milk. The frequency of SCM and CLM was 39.1% and 9.3%, respectively. Averages for test day milk yield (MY), lactation number (LN), herd size (HS), and number of days in milk (DM) were 20.6 kg, 2.8 lactations, 16.7 animals, and 164.1 days, respectively. MY was dependent on dairy herd (DH), LN, HS, and DM (P < 0.01), and correlations between udder quarters from the CMT were around 0.49 (P < 0.01). Coagulase-negative staphylococci were mainly identified, as well as Staphylococcus aureus, Streptococcus uberis, Brevibacterium stationis, B. conglomeratum, and Staphylococcus agnetis. Bacterial isolates were resistant to penicillin, clindamycin, ampicillin, and cefotaxime. Bacteriocins synthesized by Bacillus thuringiensis inhibited the growth of multiantibiotic resistance bacteria such as S. agnetis, S. equorum, Streptococcus uberis, Brevibacterium stationis, and Brachybacterium conglomeratum, but they were not active against S. sciuri, a microorganism that showed an 84% resistance to antibiotics tested in this study. PMID:25815326
Martínez-Onandi, N; Castioni, A; San Martín, E; Rivas-Cañedo, A; Nuñez, M; Torriani, S; Picon, A
The microbiota of Serrano dry-cured ham of different chemical composition, subjected or not to high-pressure processing (HPP), was investigated using culture-dependent and culture-independent methods. Microbial counts were submitted to analysis of variance with physicochemical parameters (a w , NaCl concentration, salt-in-lean ratio and intramuscular fat content) or HPP as main effects. In untreated hams, physicochemical parameters significantly affected counts of aerobic mesophiles, psychrotrophs, and moulds and yeasts. NaCl concentration and fat content influenced the levels of four and three of the five studied microbial groups, respectively, whereas no influence of a w was stated. The HPP treatment had a significant effect on counts of all investigated microbial groups. Culture-independent methods showed the presence of bacteria such as Staphylococcus equorum, Staphylococcus succinus, Bacillus subtilis and Cellulosimicrobium sp., moulds like Penicillium commune, Aspergillus fumigatus, Sclerotinia sclerotiorum, Eurotium athecium and Moniliella mellis, and yeasts like Debaryomyces hansenii and Candida glucosophila. Absence of B. subtilis bands and weaker bands of E. athecium were recorded for HPP-treated hams. The higher microbial levels found in lean ham might result in a quicker deterioration. HPP treatment confirmed its suitability as a procedure to control spoilage microorganisms. DGGE did not seem to be sensitive enough to highlight changes caused by HPP treatment in the microbiota of ham, but contributed to the detection of microbial species not previously found in ham. Copyright Â© 2016 Elsevier B.V. All rights reserved.
Rieder, Gabriele; Krisch, Linda; Fischer, Harald; Kaufmann, Maria; Maringer, Adolf; Wessler, Silja
Nonspoiled food that nevertheless contains bacterial pathogens constitutes a much more serious health problem than spoiled food, as the consumer is not warned beforehand. However, data on the diversity of bacterial species in meat juice are rare. To study the bacterial load of fresh pork from ten different distributors, we applied a combination of the conventional culture-based and molecular methods for detecting and quantifying the microbial spectrum of fresh pork meat juice samples. Altogether, we identified 23 bacterial species of ten different families analyzed by 16S rRNA gene sequencing. The majority of isolates were belonging to the typical spoilage bacterial population of lactic acid bacteria (LAB), Enterococcaceae, and Pseudomonadaceae. Several additional isolates were identified as Staphylococcus spp. and Bacillus spp. originating from human and animal skin and other environmental niches including plants, soil, and water. Carnobacterium divergens, a LAB contributing to the spoilage of raw meat even at refrigeration temperature, was the most frequently isolated species in our study (5/10) with a bacterial load of 10(3) - 10(7) CFU mL(-1). In several of the analyzed pork meat juice samples, two bacterial faecal indicators, Serratia grimesii and Serratia proteamaculans, were identified together with another opportunistic food-borne pathogen, Staphylococcus equorum. Our data reveal a high bacterial load of fresh pork meat supporting the potential health risk of meat juice for the end consumer even under refrigerated conditions. © 2012 Federation of European Microbiological Societies. Published by Blackwell Publishing Ltd. All rights reserved.
Quijada, Narciso M; Mann, Evelyne; Wagner, Martin; Rodríguez-Lázaro, David; Hernández, Marta; Schmitz-Esser, Stephan
Cheese ripening involves the succession of complex microbial communities that are responsible for the organoleptic properties of the final products. The food processing environment can act as a source of natural microbial inoculation, especially in traditionally manufactured products. Austrian Vorarlberger Bergkäse (VB) is an artisanal washed-rind hard cheese produced in the western part of Austria without the addition of external ripening cultures. Here, the composition of the bacterial communities present on VB rinds and on different processing surfaces from two ripening cellars was assessed by near full length 16S rRNA gene amplification, cloning and sequencing. Non-inoculated aerobic bacteria dominated all surfaces in this study. VB production conditions (long ripening time, high salt concentration and low temperatures) favor the growth of psychro- and halotolerant bacteria. Several bacterial groups, such as coryneforms, Staphylococcus equorum and Halomonas dominated VB and were also found on most environmental surfaces. Analysis of OTUs shared between different surfaces suggests that VB rind bacteria are inoculated naturally during the ripening from the processing environment and that cheese surfaces exert selective pressure on these communities, as only those bacteria better adapted flourished on VB rinds. This study analyzed VB processing environment microbiota and its relationship with VB rinds for the first time, elucidating that the processing environment and the cheese microbiota should be considered as microbiologically linked ecosystems with the goal of better defining the events that take place during cheese maturation. Copyright © 2018 Elsevier B.V. All rights reserved.
Adouard, Nadège; Magne, Laurent; Cattenoz, Thomas; Guillemin, Hervé; Foligné, Benoît; Picque, Daniel; Bonnarme, Pascal
A mixture of nine microorganisms (six bacteria and three yeasts) from the microflora of surface-ripened cheeses were subjected to in vitro digestive stress in a three-compartment "dynamic gastrointestinal digester" (DIDGI). We studied the microorganisms (i) grown separately in culture medium only (ii) grown separately in culture medium and then mixed, (iii) grown separately in culture medium and then included in a rennet gel and (iv) grown together in smear-ripened cheese. The yeasts Geotrichum candidum, Kluyveromyces lactis and Debaryomyces hansenii, were strongly resistant to the whole DIDGI process (with a drop in viable cell counts of less than cheese-grown cultures. Ripening bacteria such as Hafnia alvei survived gastric stress less well when grown in cheese (with no viable cells after 90 min of exposure of the cheese matrix, compared with 6 CFU mL(-1) in lab cultures). The ability of Corynebacterium casei and Staphylococcus equorum to withstand digestive stress was similar for cheese and pure culture conditions. When grow in a cheese matrix, Brevibacterium aurantiacum and Arthrobacter arilaitensis were clearly more sensitive to the overall digestive process than when grown in pure cultures. Lactococcus lactis displayed poorer survival in gastric and duodenal compartments when it had been grown in cheese. In vivo experiments in BALB/c mice agreed with the DIDGI experiments and confirmed the latter's reliability. Copyright © 2015. Published by Elsevier Ltd.
Xia, Qian Hua; Zheng, Li Ping; Zhao, Pei Fei; Wang, Jian Wen
A biological method for synthesising silver nanoparticles (AgNPs) was developed using the callus extracts from Artemisia annua L. under sunlight at 25,000 lx. The AgNPs were characterised using transmission electron microscopy, atomic force microscope, X-ray diffraction and Fourier transform infrared spectroscopy. The AgNPs were mostly spherical with the size of 2.1 to 45.2 nm (average 10.9 nm). Pulse treatments of AgNPs at 125, 250 and 500 mg/l for 1 h extended vase life of cut carnation ( Dianthus caryophyllus cv. Green Land) flowers. Four dominant bacteria strains Arthrobacter arilaitensis, Kocuria sp., Staphylococcus equorum and Microbacterium oxydans were isolated from the stem-ends of cut D. caryophyllus flowers. AgNP pulse inhibited significantly bacterial growth in vase solution and cut stem ends during all of the vase period. The bacteria related blockage in the stem-ends was significantly alleviated by AgNP pulse because of its higher antibacterial efficacy against the dominant bacteria. In addition, ethylene release of cut carnation flowers was inhibited in response to AgNP pulse. This is the first time that the biologically synthesised AgNPs could be applied as a promising preservative agent for cut carnation flowers.
Full Text Available Rodents are important intermediate and paratenic hosts for carnivore parasites, including the important zoonotic agents Toxoplasma, Echinococcus and Toxocara. Monitoring of such parasites in rodents can be used to detect increasing risks for human and veterinary public health. Rodents were trapped at four sites in Berlin, two near the city center, two at the periphery. PCRs were conducted to detect Coccidia (target ITS-1 and specifically Toxoplasma gondii (repetitive element in brain and ascarids (ITS-2 in muscle or brain tissue. During necropsies, metacestodes were collected and identified using ITS-2 and 12S rRNA PCRs. An ELISA to detect antibodies against Toxocara canis ES antigens was performed. Within the 257 examined rodents, the most frequently observed parasite was Frenkelia glareoli predominantly found in Myodes glareolus. T. gondii was only detected in 12 rodents and Microtus spp. (although strongly underrepresented had a significantly increased chance of being positive. Neither Echinococcus nor typical Taenia parasites of dogs and cats were found but Mesocestoides litteratus and Taenia martis metacestodes were identified which can cause severe peritoneal or ocular cysticercosis in dogs, primates and humans. Using PCR, the ascarids T. canis (n = 8, Toxocara cati (4 and Parascaris sp. (1 were detected predominantly in muscles. Seroprevalence of T. canis was 14.2% and ELISA was thus more sensitive than PCR to detect infection with this parasite. Non-parametric multidimensional scaling and cluster analysis revealed that parasite communities could be grouped into an urban and a peri-urban cluster with high frequency of ascarid-positive rodents in urban and high frequency of F. glareoli in peri-urban sites. Prevalence rates of parasites in rodents with potential impact for human or veterinary public health are considerable and the monitoring of transmission cycles of carnivore parasites in intermediate rodent hosts is recommended to
Nemetschke, Linda; Eberhardt, Alexander G; Hertzberg, Hubertus; Streit, Adrian
When chromatin diminution occurs during a cell division a portion of the chromatin is eliminated, resulting in daughter cells with a smaller amount of genetic material. In the parasitic roundworms Ascaris and Parascaris, chromatin diminution creates a genetic difference between the soma and the germline. However, the function of chromatin diminution remains a mystery, because the vast majority of the eliminated DNA is noncoding. Within the parasitic roundworm genus Strongyloides, S. stercoralis (in man) and S. ratti (in rat) employ XX/XO sex determination, but the situation in S. papillosus (in sheep) is different but controversial. We demonstrate genetically that S. papillosus employs sex-specific chromatin diminution to eliminate an internal portion of one of the two homologs of one chromosome pair in males. Contrary to ascarids, the eliminated DNA in S. papillosus contains a large number of genes. We demonstrate that the region undergoing diminution is homologous to the X chromosome of the closely related S. ratti. The flanking regions, which are not diminished, are homologous to the S. ratti autosome number I. Furthermore, we found that the diminished chromosome is not incorporated into sperm, resulting in a male-specific transmission ratio distortion. Our data indicate that on the evolutionary path to S. papillosus, the X chromosome fused with an autosome. Chromatin diminution serves to functionally restore an XX/XO sex-determining system. A consequence of the fusion and the process that copes with it is a transmission ratio distortion in males for certain loci. Copyright © 2010 Elsevier Ltd. All rights reserved.
Kaspar, A; Pfister, K; Nielsen, M K; Silaghi, C; Fink, H; Scheuerle, M C
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.
Piñar, G; Kraková, L; Pangallo, D; Piombino-Mascali, D; Maixner, F; Zink, A; Sterflinger, K
The Capuchin Catacombs of Palermo, Italy, contain over 1800 mummies dating from the 16th to 20th centuries AD. Their environment is not conducive to the conservation of the remains due to, among other factors, water infiltration, which is producing salt efflorescences on the walls. A multiphasic approach was applied to investigate the halophilic microbiota present in the Catacombs. Enrichment cultures were conducted on media containing different NaCl concentrations, ranging from 3 to 20 %. For screening of the strains, the following two PCR-based methods were used and compared: fluorescence internal transcribed spacer PCR (f-ITS) and random amplification of polymorphic DNA (RAPD) analyses. Results derived from RAPD profiles were shown to be slightly more discriminative than those derived from f-ITS. In addition, the proteolytic and cellulolytic abilities were screened through the use of plate assays, gelatin agar and Ostazin Brilliant Red H-3B (OBR-HEC), respectively. Many of the strains isolated from the wall samples displayed proteolytic activities, such as all strains belonging to the genera Bacillus, Virgibacillus and Arthrobacter, as well as some strains related to the genera Oceanobacillus, Halobacillus and Idiomarina. In addition, many of the strains isolated from materials employed to stuff the mummies showed cellulolytic activities, such as those related to species of the genera Chromohalobacter and Nesterenkonia, as well as those identified as Staphylococcus equorum and Halomonas sp. Furthermore, many of the strains were pigmented ranging from yellow to a strong pink color, being directly related to the discoloration displayed by the materials.
Schirmer, B C T; Heir, E; Møretrø, T; Skaar, I; Langsrud, S
The background microbiota of 5 Norwegian small-scale cheese production sites was examined and the effect of the isolated strains on the growth and survival of Listeria monocytogenes was investigated. Samples were taken from the air, food contact surfaces (storage surfaces, cheese molds, and brine) and noncontact surfaces (floor, drains, and doors) and all isolates were identified by sequencing and morphology (mold). A total of 1,314 isolates were identified and found to belong to 55 bacterial genera, 1 species of yeast, and 6 species of mold. Lactococcus spp. (all of which were Lactococcus lactis), Staphylococcus spp., Microbacterium spp., and Psychrobacter sp. were isolated from all 5 sites and Rhodococcus spp. and Chryseobacterium spp. from 4 sites. Thirty-two genera were only found in 1 out of 5 facilities each. Great variations were observed in the microbial background flora both between the 5 producers, and also within the various production sites. The greatest diversity of bacteria was found in drains and on rubber seals of doors. The flora on cheese storage shelves and in salt brines was less varied. A total of 62 bacterial isolates and 1 yeast isolate were tested for antilisterial activity in an overlay assay and a spot-on-lawn assay, but none showed significant inhibitory effects. Listeria monocytogenes was also co-cultured on ceramic tiles with bacteria dominating in the cheese production plants: Lactococcus lactis, Pseudomonas putida, Staphylococcus equorum, Rhodococcus spp., or Psychrobacter spp. None of the tested isolates altered the survival of L. monocytogenes on ceramic tiles. The conclusion of the study was that no common background flora exists in cheese production environments. None of the tested isolates inhibited the growth of L. monocytogenes. Hence, this study does not support the hypothesis that the natural background flora in cheese production environments inhibits the growth or survival of L. monocytogenes. Copyright © 2013 American
Nathalie L. van der Mee-Marquet
Full Text Available We isolated from aerobic and anaerobic blood culture bottles from a febrile patient, a Helicobacter-like Gram negative, rod-shaped bacterium that MALDI-TOF MS failed to identify. Blood agar cultures incubated in a microaerobic atmosphere revealed a motile Gram negative rod, which was oxidase, catalase, nitrate reductase, esterase, and alkaline phosphatase positive. It grew at 42°C with no detectable urease activity. Antimicrobial susceptibility testing showed that the organism was susceptible to beta-lactams, gentamicin, erythromycin, and tetracycline but resistant to ciprofloxacin. Electronic microscopy analysis revealed a 3 × 0.5 μm curved rod bacterium harboring two sheathed amphitrichous flagella. Whole genome sequencing revealed a genome 1,708,265 base-pairs long with a GC content of 37.80% and a total of 1,697 coding sequences. The genomic analyses using the nucleotide sequences of the 16S rRNA gene, hsp60 and gyrB genes, as well as the GyrA protein sequence, and the results of Average Nucleotide Identity and in silico DNA-DNA hybridization suggest evidence for a novel Helicobacter species close to Helicobacter equorum and belonging to the group of enterohepatic Helicobacter species. As soon as the particular peptide mass fingerprint of this pathogen is added to the spectral databases, MALDI-TOF MS technology will improve its identification from clinical specimens, especially in case of “sterile infection”. We propose to associate the present strain with the Latin name of the place of isolation; Caesarodunum (Tours, France and suggest “Helicobacter caesarodunensis” for further description of this new bacterium.
De Visscher, A; Piepers, S; Haesebrouck, F; De Vliegher, S
Coagulase-negative staphylococci (CNS) are the main cause of bovine intramammary infections (IMI) in many countries. Despite a high prevalence of CNS IMI at parturition, species-specific risk factor studies, relying on accurate identification methods, are lacking. Therefore, this observational study aimed at determining the prevalence and distribution of different CNS species causing IMI in fresh heifers and dairy cows in Flemish dairy herds and identifying associated species- and subgroup-specific risk factors at the herd, cow, and quarter level. The effect on udder health was investigated as well. Staphylococcus chromogenes, S. sciuri, and S. cohnii were the most frequently isolated species. The only CNS species causing IMI in fresh heifers and dairy cows in all herds was Staphylococcus chromogenes, whereas large between-herd differences in distribution were observed for the other species. Quarters from heifers and quarters with an inverted teat end had higher odds of being infected with S. chromogenes, S. simulans, or S. xylosus as well as with S. chromogenes solely. Prepartum teat apex colonization with S. chromogenes increased the likelihood of S. chromogenes IMI in the corresponding quarters at parturition. Quarters with dirty teat apices before calving were more likely to be infected with S. cohnii, S. equorum, S. saprophyticus, or S. sciuri, supporting the environmental nature of these CNS species. Three species (S. chromogenes, S. simulans, and S. xylosus) were associated with a higher quarter somatic cell count at parturition as compared with uninfected quarters. Copyright © 2016 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.
Piessens, V; De Vliegher, S; Verbist, B; Braem, G; Van Nuffel, A; De Vuyst, L; Heyndrickx, M; Van Coillie, E
The aim of this study was to investigate whether the main coagulase-negative staphylococci (CNS) species involved in bovine intramammary infections (IMI) possess specific characteristics that promote colonization of the udder. Virulence markers associated with biofilm formation, antimicrobial resistance, and biocide tolerance were compared between typically contagious CNS species (Staphylococcus chromogenes, Staphylococcus epidermidis, Staphylococcus haemolyticus, and Staphylococcus simulans) and those rarely causing IMI (Staphylococcus sciuri, Staphylococcus equorum, and others) to find possible associations with pathogenicity. Coagulase-negative staphylococci isolates (n=366) belonging to 22 different species were analyzed by PCR for the presence of the biofilm-associated genes bap and icaA, and the methicillin resistance gene mecA. A selection of 82 isolates was additionally tested for their susceptibility to 5 antibiotics and 2 commercial teat dip products. Minimum inhibitory concentrations of antimicrobials were determined by Etest (AB bioMérieux, Marcy l'Etoile, France), and a microdilution method was optimized to determine minimum biocidal concentrations of teat dips. The bap, icaA, and mecA genes were detected significantly more in isolates from CNS species typically living in the cows' environment than in isolates from IMI-causing species. Antimicrobial resistance was mainly against erythromycin (23%) or oxacillin (16%), and was detected more often in the environmental species. The isolates least susceptible to the teat dips belonged to the IMI-causing species Staph. chromogenes and Staph. simulans. We concluded that carriage of biofilm genes and antimicrobial resistance were not associated with the ability to colonize the mammary gland because free-living CNS species constituted a more significant reservoir of biofilm and resistance determinants than did IMI-causing species. In contrast, increased tolerance to biocides may favor the establishment of
Mushtaq, Saleem; Rather, Muzafar Ahmad; Qazi, Parvaiz H; Aga, Mushtaq A; Shah, Aabid Manzoor; Shah, Aiyatullah; Ali, Md Niamat
The roots of Thalictrum minus are traditionally used in the treatment of inflammation and infectious diseases such as bovine mastitis. However, there are no reports available in literature till date regarding the antibacterial studies of T. minus against bovine mastitis. The present study was undertaken to evaluate the antibacterial potential of crude extract of T. minus (root) and some of its isolated constituents against bovine mastitis in order to scientifically validate its traditional use. A total of three alkaloid compounds were isolated from the DCM: MeOH extract of roots of T. minus using silica gel column chromatography. Structural elucidation of the isolated compounds was done by using spectroscopic techniques like mass spectrometry and NMR spectroscopy. Pathogens were isolated from cases of bovine mastitis and identified by using 16S rRNA gene sequencing. The broth micro-dilution method was used to evaluate the antibacterial activities of DCM: MeOH extract and isolated compounds against mastitis pathogens. The three isolated compounds were identified as benzylisoquinoline alkaloids (1) 5'-Hydroxythalidasine, (2) Thalrugosaminine and (3) O-Methylthalicberine. Compounds (2) and (3) are reported for the first time from the roots of T. minus. Five mastitis pathogens viz., Staphylococcus xylosus, Staphylococcus lentus, Staphylococcus equorum, Enterococcus faecalis and Pantoea agglomerans were identified on the basis of sequence analysis of isolates using the nucleotide BLAST algorithm. This study reports for the first time the isolation and molecular characterization of mastitis pathogens from Kashmir valley, India. The DCM: MeOH extract exhibited broad spectrum antibacterial activities that varied between the bacterial species (MIC=250-500µg/ml). 5'-Hydroxythalidasine and Thalrugosaminine showed promising antibacterial activity with MIC values of 64-128µg/ml while Staphylococcus species were found to be the most sensitive strains. The antibacterial
Storms, Goedele; Meersschaert, Carole; Farnir, Frédéric; Grauwels, Magda
To describe the bacterial flora of the normal conjunctiva of Huacaya alpacas (Vicugna pacos) and to determine the effect of age and gender on this flora. Fifty Huacaya alpacas. After a complete ophthalmic examination, conjunctival swabs were obtained from both eyes and cultured for aerobic and anaerobic bacteria. Logistic and Poisson regression analyses were used to evaluate the effect of age and gender on bacterial isolation. Four animals were excluded because of signs of external ocular disease. Of the remaining 46 alpacas, bacteria were recovered from 96.7% (89/92) of the eyes. A total of 190 bacterial isolates were cultured with a mean of 2.1 bacterial isolates per eye. The majority of isolates (70%) were Gram-positive. Staphylococcus xylosus (44/190: 23.2%) predominated, followed by viridans streptococci (32/190: 16.8%) and Pantoea agglomerans (24/190: 12.6%). Other frequently isolated bacteria included Rothia mucilaginosa (12/190: 6.3%), Staphylococcus equorum (12/190: 6.3%), Bacillus species (9/190: 4.7%), Moraxella ovis (9/190: 4.7%), and Moraxella catarrhalis (6/190: 3.2%). Statistical analysis showed that alpacas harboring viridans streptococci and Moraxella species were significantly younger. Gender did not significantly affect type of bacterial isolation. There appeared to be no significant effect of age or gender on number of bacteria isolated. Gram-positive aerobes were most commonly cultured, with S. xylosus and viridans streptococci predominating. To the best of the authors' knowledge, this is the first report describing the presence of Moraxella species in the healthy conjunctival sac of alpacas. Alpacas harboring viridans streptococci and Moraxella species were significantly younger. © 2015 American College of Veterinary Ophthalmologists.
Janssens, M; Van der Mijnsbrugge, A; Sánchez Mainar, M; Balzarini, T; De Vuyst, L; Leroy, F
The ability of coagulase-negative staphylococci (CNS) to use alternative energy sources in meat may partially explain their occurrence in fermented meats. Of 61 CNS strains tested, all metabolized adenosine and inosine in a meat simulation medium (MSM). The ability to catabolize arginine via the arginine deiminase (ADI) pathway varied between strains. All tested strains of Staphylococcus carnosus and Staphylococcus epidermidis possessed an arcA gene and showed ADI activity, whereas other species, such as Staphylococcus equorum and Staphylococcus succinus, did not. Arginine catabolic mobile elements (ACME), as in the positive control S. epidermidis ATCC 12228, were uncommon and only found in Staphylococcus xylosus 3PA6 (sausage isolate) and Staphylococcus chromogenes G222 (teat apex isolate). Monoculture experiments were performed in MSM with S. carnosus 833 and SS3-4, S. xylosus G211, and S. epidermidis ATCC 12228 and 2S7-4. At all pH values tested (5.3, 5.8, and 6.5), the strains of S. carnosus catabolized arginine faster than the strains of S. xylosus and S. epidermidis. Only at pH 6.5 could a low ADI activity be found for S. xylosus G211. Increased ADI activity occurred in the case of the ACME-positive S. epidermidis ATCC 12228, when compared to the ACME-negative S. epidermidis 2S7-4. Copyright © 2013 Elsevier Ltd. All rights reserved.
Fonseca, Sonia; Ivette Ouoba, Labia Irène; Franco, Inmaculada; Carballo, Javier
The development of Lactobacillus and Staphylococcus strains used as starter cultures throughout the ripening of Galician chorizo, a traditional dry fermented sausage from the north-west of Spain, was monitored combining different molecular-based techniques. The bacterial diversity occurring in the inoculated sausages at the beginning and the end of the ripening was also studied and compared to the indigenous population in an uninoculated control batch. Real-time PCR was used to monitor the Lactobacillus and Staphylococcus community using genus and species-specific primer to quantify the occurring microbiota. The identification of isolates at genus or species level was achieved by specific PCR and 16S rRNA gene sequencing. rep-PCR using (GTG)5-PCR primer was used to characterize this bacterial community at strain level. According to the data obtained, the strains Lactobacillus sakei LS131, Staphylococcus equorum SA25 and Staphylococcus saprophyticus SB12 were dominant during the ripening process, whereas the strain Staphylococcus epidermidis SA49, that was added in order to study its behaviour with a merely scientific purpose, did not succeed in dominating ripening, since it seemed to be outcompeted by autochthonous microbiota. In conclusion, the combination of a quantitative method such as real-time PCR with the identification and typing techniques used in this study (genus and species-specific PCR, 16S rRNA gene sequencing and (GTG)5-PCR) provided accurate and complete information about the starter cultures development, assessing their growth and survival over the ripening process. Copyright © 2013 Elsevier Ltd. All rights reserved.
Carta, A; Scala, A
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
Hoberg, E P; Zarlenga, D S
developmental thresholds defined by temperature and humidity for larval stages) will be substantial determinants in the potential outcomes for widespread geographical and host colonization which are predicted to unfold over the coming century. Insights about deeper evolutionary events, ecology and biogeography are critical as understanding history informs us about the possible range of responses in complex systems under new regimes of environmental forcing, especially, in this case, ecological perturbation linked to climate change. A deeper history of perturbation is relevant in understanding contemporary systems that are now strongly structured by events of invasion and colonization. The relaxation of abiotic and biotic controls on the occurrence of H. contortus, coincidental with inception and dissemination of anthelmintic resistance may be synergistic, serving to exacerbate challenges to control parasites or to limit the socioeconomic impacts of infection that can influence food security and availability. Studies of haemonchine nematodes contribute directly to an expanding model about the nature of diversity and the evolutionary trajectories for faunal assembly among complex host-parasite systems across considerable spatial and temporal scales. Copyright © 2016 Elsevier Ltd. All rights reserved.
Piessens, V; Van Coillie, E; Verbist, B; Supré, K; Braem, G; Van Nuffel, A; De Vuyst, L; Heyndrickx, M; De Vliegher, S
In many parts of the world, coagulase-negative staphylococci (CNS) are the predominant pathogens causing intramammary infections (IMI) in dairy cows. The cows' environment is thought to be a possible source for CNS mastitis and this was investigated in the present paper. A longitudinal field study was carried out in 6 well-managed dairy herds to determine the distribution and epidemiology of various CNS species isolated from milk, causing IMI and living freely in the cows' environment, respectively. In each herd, quarter milk samples from a cohort of 10 lactating cows and environmental samples from stall air, slatted floor, sawdust from cubicles, and sawdust stock were collected monthly (n=13). Isolates from quarter milk samples (n=134) and the environment (n=637) were identified to species level using amplified fragment length polymorphism (AFLP) genotyping. Staphylococcus chromogenes, S. haemolyticus, S. epidermidis, and S. simulans accounted for 81.3% of all CNS milk isolates. Quarters were considered infected with CNS (positive IMI status) only when 2 out of 3 consecutive milk samples yielded the same CNS AFLP type. The species causing IMI were S. chromogenes (n=35 samples with positive IMI status), S. haemolyticus (n=29), S. simulans (n=14), and S. epidermidis (n=6). The observed persistent IMI cases (n=17) had a mean duration of 149.4 d (range 63.0 to 329.8 d). The CNS species predominating in the environment were S. equorum, S. sciuri, S. haemolyticus, and S. fleurettii. Herd-to-herd differences in distribution of CNS species were observed in both milk and the environment, suggesting that herd-level factors are involved in the establishment of particular species in a dairy herd. Primary reservoirs of the species causing IMI varied. Staphylococcus chromogenes and S. epidermidis were rarely found in the environment, indicating that other reservoirs were more important in their epidemiology. For S. haemolyticus and S. simulans, the environment was found as a
Even, Sergine; Leroy, Sabine; Charlier, Cathy; Zakour, Nouri Ben; Chacornac, Jean-Paul; Lebert, Isabelle; Jamet, Emmanuel; Desmonts, Marie-Hélène; Coton, Emmanuel; Pochet, Sylvie; Donnio, Pierre-Yves; Gautier, Michel; Talon, Régine; Le Loir, Yves
Some coagulase negative staphylococci (CNS) species play an important role in the fermentation of meat and milk products and are considered as food-grade. However, the increasing clinical significance of CNS and the presence of undesirable and unsafe properties in CNS question their presence or use in food. Our goal was to assess the safety of CNS by developing a diagnostic microarray targeting 268 genes corresponding to safety hazards in a food context i.e. toxins (especially enterotoxins) and determinants of antibiotic resistance and biogenic amine production. Target genes were selected among staphylococci and Gram-positive species that may be in contact with CNS in foodstuffs. The diagnostic microarray was used to screen 129 strains belonging to the 2 dominant species isolated from foodstuffs (S. equorum and S. xylosus) and the 2 main species isolated both in foodstuffs and clinical samples (S. epidermidis and S. saprophyticus). Microarray data were further completed by antibiograms and measurement of biogenic amine production. Safety hazards associated with CNS were mostly limited to the presence of antibiotic resistance. Seventy-one percent of the strains possessed at least one gene encoding antibiotic resistance, while only one strain carried an enterotoxin gene. Most strains did not carry any genes encoding staphylococcal toxins (68%), non-staphylococcal toxins (95%) or decarboxylases involved in biogenic amine production (78%). Food safety hazards were more pronounced in S. epidermidis than in the three other species regardless the food or clinical origin of the strains. Seventy-six percent of the strains carrying genes encoding staphylococcal toxin and 69% of strains carrying 5 or more antibiotic determinants belonged to S. epidermidis species. The dominant antibiotic resistance targeted erythromycin, tetracycline and penicillin and were generally traced back to the presence of tetK and blaZ in the two latest cases. Six percent of the food-related strains
Ferrocino, Ilario; Bellio, Alberto; Romano, Angelo; Macori, Guerrino; Rantsiou, Kalliopi; Decastelli, Lucia; Cocolin, Luca
Valle d'Aosta Lard d'Arnad is a protected designation of origin (PDO) product produced from fat of the shoulder and back of heavy pigs. Its manufacturing process can be very diverse, especially regarding the maturation temperature and the NaCl concentration used for the brine; thereby, the main goal of this study was to investigate the impact of those parameters on the microbiota developed during curing and ripening. Three farms producing Lard d'Arnad were selected. Two plants, reflecting the industrial process characterized either by low maturation temperature (plant A [10% NaCl, 2°C]) or by using a low NaCl concentration (plant B [2.5% NaCl, 4°C]), were selected, while the third was characterized by an artisanal process (plant C [30% NaCl, 8°C]). Lard samples were obtained at time 0 and after 7, 15, 30, 60, and 90 days of maturation. From each plant, 3 independent lots were analyzed. The diversity of live microbiota was evaluated by using classical plate counts and amplicon target sequencing of small subunit (SSU) rRNA. The main taxa identified by sequencing were Acinetobacter johnsonii , Psychrobacter , Staphylococcus equorum , Staphylococcus sciuri , Pseudomonas fragi , Brochothrix , Halomonas , and Vibrio , and differences in their relative abundances distinguished samples from the individual plants. The composition of the microbiota was more similar among plants A and B, and it was characterized by the higher presence of taxa recognized as undesired bacteria in food-processing environments. Oligotype analysis of Halomonas and Acinetobacter revealed the presence of several characteristic oligotypes associated with A and B samples. IMPORTANCE Changes in the food production process can drastically affect the microbial community structure, with a possible impact on the final characteristics of the products. The industrial processes of Lard d'Arnad production are characterized by a reduction in the salt concentration in the brines to address a consumer demand
Sánchez Mainar, María; Leroy, Frédéric
The cured colour of European raw fermented meats is usually achieved by nitrate-into-nitrite reduction by coagulase-negative staphylococci (CNS), subsequently generating nitric oxide to form the relatively stable nitrosomyoglobin pigment. The present study aimed at comparing this classical curing procedure, based on nitrate reductase activity, with a potential alternative colour formation mechanism, based on nitric oxide synthase (NOS) activity, under different acidification profiles. To this end, meat models with and without added nitrate were fermented with cultures of an acidifying strain (Lactobacillus sakei CTC 494) and either a nitrate-reducing Staphylococcus carnosus strain or a rare NOS-positive CNS strain (Staphylococcus haemolyticus G110), or by relying on the background microbiota. Satisfactory colour was obtained in the models prepared with added nitrate and S. carnosus. In the presence of nitrate but absence of added CNS, however, cured colour was only obtained when L. sakei CTC 494 was also omitted. This was ascribed to the pH dependency of the emerging CNS background microbiota, selecting for nitrate-reducing Staphylococcus equorum strains at mild acidification conditions but for Staphylococcus saprophyticus strains with poor colour formation capability when the pH decrease was more rapid. This reliance of colour formation on the composition of the background microbiota was further explored by a side experiment, demonstrating the heterogeneity in nitrate reduction of a set of 88 CNS strains from different species. Finally, in all batches prepared with S. haemolyticus G110, colour generation failed as the strain was systematically outcompeted by the background microbiota, even when imposing milder acidification profiles. Thus, when aiming at colour formation through CNS metabolism, technological processing can severely interfere with the composition and functionality of the meat-associated CNS communities, for both nitrate reductase and NOS activities
Schmidt, Vanessa M; Williams, Nicola J; Pinchbeck, Gina; Corless, Caroline E; Shaw, Stephen; McEwan, Neil; Dawson, Susan; Nuttall, Tim
Coagulase-positive (CoPS) and coagulase-negative (CoNS) staphylococci are normal commensals of the skin and mucosa, but are also opportunist pathogens. Meticillin-resistant (MR) and multidrug-resistant (MDR) isolates are increasing in human and veterinary healthcare. Healthy humans and other animals harbour a variety of staphylococci, including MR-CoPS and MR-CoNS. The main aims of the study were to characterise the population and antimicrobial resistance profiles of staphylococci from healthy non-vet visiting and non-antimicrobial treated Labrador retrievers in the UK. Nasal and perineal samples were collected from 73 Labrador retrievers; staphylococci isolated and identified using phenotypic and biochemical methods. They were also confirmed by matrix-assisted laser desorption ionisation time-of-flight mass spectrometry (MALDI-TOF-MS), PCR of the nuc gene and PCR and sequencing of the tuf gene. Disc diffusion and minimum inhibitory concentration (MIC) susceptibility tests were determined for a range of antimicrobials. In total, 102 CoPS (S. pseudintermedius n = 91, S. aureus n = 11) and 334 CoNS isolates were detected from 99% of dogs in this study. In 52% of dogs CoNS only were detected, with both CoNS and CoPS detected in 43% dogs and CoPS only detected in 4% of dogs. Antimicrobial resistance was not common among CoPS, but at least one MDR-CoNS isolate was detected in 34% of dogs. MR-CoNS were detected from 42% of dogs but no MR-CoPS were isolated. S. epidermidis (52% of dogs) was the most common CoNS found followed by S. warneri (30%) and S. equorum (27%), with another 15 CoNS species isolated from ≤ 15% of dogs. S. pseudintermedius and S. aureus were detected in 44% and 8% of dogs respectively. MR- and MDR-CoPS were rare. However a high prevalence of MR- and MDR-CoNS were found in these dogs, even though they had no prior antimicrobial treatment or admission to veterinary premises. These findings are of concern due to the potential for opportunistic
Gómez, Paula; Lozano, Carmen; Benito, Daniel; Estepa, Vanesa; Tenorio, Carmen; Zarazaga, Myriam; Torres, Carmen
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
Full Text Available Abstract Background Surface contamination of smear cheese by Listeria spp. is of major concern for the industry. Complex smear ecosystems have been shown to harbor antilisterial potential but the microorganisms and mechanisms involved in the inhibition mostly remain unclear, and are likely related to complex interactions than to production of single antimicrobial compounds. Bacterial biodiversity and population dynamics of complex smear ecosystems exhibiting antilisterial properties in situ were investigated by Temporal temperature gradient gel electrophoresis (TTGE, a culture independent technique, for two microbial consortia isolated from commercial Raclette type cheeses inoculated with defined commercial ripening cultures (F or produced with an old-young smearing process (M. Results TTGE revealed nine bacterial species common to both F and M consortia, but consortium F exhibited a higher diversity than consortium M, with thirteen and ten species, respectively. Population dynamics were studied after application of the consortia on fresh-produced Raclette cheeses. TTGE analyses revealed a similar sequential development of the nine species common to both consortia. Beside common cheese surface bacteria (Staphylococcus equorum, Corynebacterium spp., Brevibacterium linens, Microbacterium gubbeenense, Agrococcus casei, the two consortia contained marine lactic acid bacteria (Alkalibacterium kapii, Marinilactibacillus psychrotolerans that developed early in ripening (day 14 to 20, shortly after the growth of staphylococci (day 7. A decrease of Listeria counts was observed on cheese surface inoculated at day 7 with 0.1-1 × 102 CFU cm-2, when cheeses were smeared with consortium F or M. Listeria counts went below the detection limit of the method between day 14 and 28 and no subsequent regrowth was detected over 60 to 80 ripening days. In contrast, Listeria grew to high counts (105 CFU cm-2 on cheeses smeared with a defined surface culture
Species identification, slime production and oxacillin susceptibility in coagulase-negative staphylococci isolated from nosocomial specimens Identificação de espécies, produção de "slime" e sensibilidade a oxacilina em amostras de Staphylococcus coagulase-negativo isoladas de espécimes nosocomiais
Lucía E. Alcaráz
Full Text Available Ninety-two coagulase negative staphylococci (CNS (forty-five of clinical origin and forty-seven of environmental origin, collected in a hospital in San Luis, Argentina, from March to June, 1999, were identified to species level by the ID 32 Staph and API Staph System (bioMérieux. Slime production was investigated by the quantitative and qualitative methods. Oxacillin susceptibility was determined by the disk diffusion test (1 µg, the agar dilution method (0.125 to 4 mg/ml and agar screen (6 µg/ml. The presence of mecA gene was investigated by PCR. The clinical CNS species most commonly isolated were S. epidermidis, S. haemolyticus, S. hominis and S. saprophyticus. The frequency of slime production by clinical and environmental isolates was similar (25/45 and 27/47, respectively and the results obtained by the quantitative and the qualitative methods correlated well. The mecA gene was detected in all S. epidermidis, S. haemolyticus and S. hominis isolates, which were resistant to oxacillin by the phenotypic methods. However, this gene was not present in S. klossii, S. equorum, S. xylosus and S. capitis strains. The gene was neither found in two out of the six S. saprophyticus isolates, in two out of three S. cohnii subsp. urealyticum isolates and in two out of five S. cohnii subsp. cohnii isolates, all of which resulted oxacillin resistant according to MIC. The gene was not found in oxacillin-susceptible strains either. Most of the CNS isolates (enviromental and clinical that were slime producers were found to be oxacillin resistant, which makes the early detection of these microorganisms necessary to prevent their dissemination in hospitals, particularly among immunocompromised patients.Noventa e duas amostras de Staphylococcus coagulase negativo (SCN, (45 amostras clínicas e 47 ambientais, coletadas em um hospital de San Luis, Argentina, durante o período de março a junho de 1999, foram identificadas até espécies, empregando-se os
moxidectin and pyrantel treatment. The control of the small strongyles (Ciathostoma group in horses is based on the application of anthelmintics. In our country, the development of generalized resistance to benzimidazoles is limiting the chemical alternatives available to macrocyclic lactones (ivermectin and moxidectin and to pyrantel, considered as long-acting and short-acting drugs respectively. Updated information on the activity of these drugs in the field, is critical for determining its efficacy and detecting the development of anthelmintic resistance. In these horse nematodes the period of egg reappearance (ERP after treatment is considered as an early indicator of the presence of resistance. The present study evaluated the clinical efficacy and ERP after moxidectin and pirantel treatments in adult horses naturally parasitized by small strongyles from five farms from Santa Fe and Córdoba provinces. Clinical efficacy determined at day 14 or 15 post treatment using a test of reduction in the egg count ranged from 98.9 to 98.8% for the pirantel and 99.8 to 100% for moxidectin. The ERP was at least 100 days for moxidectin and 35 days for the pirantel. These results indicate that both drugs are active for the control of these nematodes and that the populations studied (some of them resistant to benzimidazoles remain currently susceptible to moxidectin as well as to pyrantel. This last drug is of limited use in Argentina, but its inclusion in the control programs against the small strongyls could reduce the dependence and the selection pressure on the macrocyclic lactones and contribute to maintain the useful life of the same ones.
Evaluation of resistance in a selected field strain of Haemonchus contortus to ivermectin and moxidectin using the Larval Migration on Agar Test Avaliação da resistência em um isolado de campo selecionado de Haemonchus contortus à ivermectina e moxidectina usando o Teste de Migração de Larvas em Ágar
Fernanda S. Fortes
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 (pHaemonchus contortus é uma das causas mais comuns e economicamente significativas de doença em produções de pequenos ruminantes em todo o mundo, e os programas de controle de parasitas nematoides - incluindo H. contortus - baseiam-se principalmente no uso de drogas anti-helmínticas. A consequência da utilização desses compostos, como sendo a única estratégia sanitária para evitar infecções por parasitas, tem sido a redução da eficácia de todos os
Gómez, Paula; Lozano, Carmen; Benito, Daniel; Estepa, Vanesa; Tenorio, Carmen; Zarazaga, Myriam; Torres, Carmen
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
Condas, Larissa A Z; De Buck, Jeroen; Nobrega, Diego B; Carson, Domonique A; Roy, Jean-Philippe; Keefe, Greg P; DeVries, Trevor J; Middleton, John R; Dufour, Simon; Barkema, Herman W
The effect of non-aureus staphylococci (NAS) in bovine mammary health is controversial. Overall, NAS intramammary infections (IMI) increase somatic cell count (SCC), with an effect categorized as mild, mostly causing subclinical or mild to moderate clinical mastitis. However, based on recent studies, specific NAS may affect the udder more severely. Some of these apparent discrepancies could be attributed to the large number of species that compose the NAS group. The objectives of this study were to determine (1) the SCC of quarters infected by individual NAS species compared with NAS as a group, culture-negative, and major pathogen-infected quarters; (2) the distribution of NAS species isolated from quarters with low SCC (<200,000 cells/mL) and high SCC (≥200,000 cells/mL), and clinical mastitis; and (3) the prevalence of NAS species across quarters with low and high SCC. A total of 5,507 NAS isolates, 3,561 from low SCC quarters, 1,873 from high SCC quarters, and 73 from clinical mastitis cases, were obtained from the National Cohort of Dairy Farms of the Canadian Bovine Mastitis Research Network. Of quarters with low SCC, high SCC, or clinical mastitis, 7.6, 18.5, and 4.3% were NAS positive, respectively. The effect of NAS IMI on SCC was estimated using mixed-effect linear regression; prevalence of NAS IMI was estimated using Bayesian analyses. Mean SCC of NAS-positive quarters was 70,000 cells/mL, which was higher than culture-negative quarters (32,000 cells/mL) and lower than major pathogen-positive quarters (129,000 to 183,000 cells/mL). Compared with other NAS species, SCC was highest in quarters positive for Staphylococcus capitis, Staphylococcus gallinarum, Staphylococcus hyicus, Staphylococcus agnetis, or Staphylococcus simulans. In NAS-positive quarters, Staphylococcus xylosus (12.6%), Staphylococcus cohnii (3.1%), and Staphylococcus equorum (0.6%) were more frequently isolated from quarters with low SCC than other NAS species, whereas Staphylococcus