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Gilson Pereira De ,oliveira - One of the best experts on this subject based on the ideXlab platform.
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Fauna helmintológica de ovinos provenientes da microrregião de Jaboticabal, estado de São Paulo, Brasil
Univ Federal Santa Maria, 2014Co-Authors: Maciel, Willian Giquelin, Felippelli Gustavo, Zanetti Lopes, Welber Daniel, Pires Teixeira, Weslen Fabricio, Cruz, Breno Cayeiro, Santos, Thais Rabelo Dos, Buzzulini Carolina, Favero Flavia, Gomes, Lucas Costa, Gilson Pereira De ,oliveiraAbstract:This study evaluated the prevalence and counting parasitism of different species of helminths of sheep from the micro-region of Jaboticabal of Sao Paulo state. For this, 66 animals naturally infected, four to 36 months of age, raised in pasture, were selected. The results of necropsy revealed the presence of seven genera and 12 species with the following prevalence and mean count: Haemonchus contortus: 100.0% (2947.2); Trichostrongylus colubriformis: 90.9% (3048.8); Cooperia curticei: 56.0% (256.5); Oesophagostomum columbianum: 48.4% (36.0); Cooperia punctata: 30.3% (94.5); Trichostrongylus axei: 22.7% (26.5); Strongyloides papillosus: 19.6% (83.0), Haemonchus contortus (L4): 7.5% (17.2), Cooperia pectinata: 10.6% (12.9), Trichuris ovis: 10.6 % (0.6); Cooperia spatulata 4.5% (0.3); Capillaria bovis: 4.5% (0.1). The mean parasitism of helminthswas 6524.7 per animal. Haemonchus contortus (adults and L4) and Trichostrongylus colubriformis corresponded to 45.4% and 46.7% of the average worm burden totally, respectively. Based in the results obtained in this study, can be concluded that the two most abundant species of helminths and important, the micro-region of Jaboticabal are Trichostrongylus colubriformis and Haemonchus contortus, and these two species amounted to 92.1% of the distribution percentage of helminths collected from all animals. These results demonstrate the importance of conducting a counts of eggs per gram of feces (EPG) in the herds of this region when FAMACHA is used on a particular property, since this method control does not allow to diagnostic the damage/clinical signs in animals infected by T. colubriformis, because this specie does not have hematophagism habit on animals
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Fauna helmintológica de ovinos provenientes da microrregião de Jaboticabal, estado de São Paulo, Brasil
Universidade Federal de Santa Maria, 2014Co-Authors: Willian Giquelin Maciel, Gustavo Felippelli, Welber Daniel Zanetti Lopes, Weslen Fabricio Pires Teixeira, Breno Cayeiro Cruz, Thais Rabelo Dos Santos, Carolina Buzzulini, Flavia Favero, Lucas Costa Gomes, Gilson Pereira De ,oliveiraAbstract:Avaliou-se a prevalência e a contagem parasitária das diferentes espécies de helmintos de ovinos provenientes da microrregião de Jaboticabal, região Noroeste de São Paulo. Para tanto, foram utilizados 66 animais naturalmente infectados, de quatro a 36 meses de idade, criados em regime extensivo. Os resultados necroscópicos revelaram a presença de sete gêneros e 12 espécies, com a seguinte prevalência e a média de parasitismo: Haemonchus contortus: 100,0% (2947,2); Trichostrongylus colubriformis: 90,9% (3048,8); Cooperia curticei: 56,0% (256,5); Oesophagostomum columbianum: 48,4% (36,0); Cooperia punctata: 30,3% (94,5); Trichostrongylus axei: 22,7% (26,5); Strongyloides papillosusi: 19,6% (83,0); Haemonchus contortus (L4): 7,5% (17,2); Cooperia pectinatai: 10,6% (12,9); Trichuris ovis: 10,6% (0,6); Cooperia spatulata 4,5% (0,3); Capillaria bovis: 4,5% (0,1). A carga parasitária média foi de 6.524,7 helmintos por animal. Haemonchus contortus (Adultos e L4) e Trichostrongylus colubriformis corresponderam a 45,4% e 46,7% da carga parasitária média total, respectivamente. Pode-se concluir que as duas espécies de helmintos mais abundantes e importantes da microrregião de Jaboticabal/São Paulo foram Trichostrongylus colubriformis e Haemonchus contortus, sendo que essas duas espécies perfizeram 92,1% da distribuição percentual dos helmintos recolhidos de todos os animais. Tais resultados demonstram a importância em se realizar um monitoramento das contagens de ovos por grama de fezes (OPG) dos rebanhos desta região, quando o método FAMACHA for empregado em uma determinada propriedade, uma vez que este método de controle, geralmente, não permite diagnosticar os danos/sinais clínicos desencadeados nos animais pelo T. colubriformis, em função de essa espécie não possuir hábito de hematofagismo sobre os hospedeiros
Alessandro Francisco Talamini Do Amarante - One of the best experts on this subject based on the ideXlab platform.
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Helminth infections and hybridization between Haemonchus contortus and Haemonchus placei in sheep from Santana do Livramento, Brazil
Colégio Brasileiro de Parasitologia Veterinaria, 2018Co-Authors: Fabiana Alves De Almeida, M. R. V. Amarante, César Cristiano Bassetto, Ana Cláudia Alexandre De Albuquerque, Renan Zappavigna Costa Starling, Alessandro Francisco Talamini Do AmaranteAbstract:Abstract The occurrence and intensity of helminth infections were evaluated in sheep from pastures shared with cattle. In 2015 and 2016, young male sheep acquired in Santana do Livramento, Rio Grande do Sul, Brazil, were finished in integrated crop-livestock system. We selected the 12 sheep that showed the highest number of nematode eggs per gram of faeces to search for worms in the gastrointestinal tract. Haemonchus contortus and Trichostrongylus colubriformis were the major parasites. H. contortus presented mean intensities of 1,159 and 257 worms in 2015 and 2016, respectively. T. colubriformis displayed mean intensities of 4,149 and 2,427 worms in 2015 and 2016, respectively. Of the 127 male specimens of Haemonchus spp. analysed by Polymerase Chain Reaction (PCR), 125 were H. contortus, one Haemonchus placei and one hybrid. Other species detected were Cooperia punctata , Cooperia pectinata, Cooperia spatulata, Cooperia curticei, Ostertagia ostertagi, Teladorsagia circumcincta , Trichostrongylus axei, Nematodirus spathiger , and Trichuris ovis. Twenty lambs presented cysts of Taenia hydatigena in the liver and mesentery. One lamb presented Coenurus cerebralis, the larval stage of Taenia multiceps, in the brain. In conclusion, sheep from pasture shared with cattle presented a high diversity of nematode species. H. contortus and H. placei co-infection occur with consequent hybridization
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PCR primers for straightforward differentiation of Haemonchus contortus, Haemonchus placei and their hybrids.
Journal of helminthology, 2017Co-Authors: M. R. V. Amarante, Michelle C. Santos, César Cristiano Bassetto, Alessandro Francisco Talamini Do AmaranteAbstract:Haemonchus contortus and Haemonchus placei are among the major parasites of small ruminants and cattle. Although infection with these nematodes is host-specific, with H. placei predominating in cattle and H. contortus in sheep, cross-infections are observed in areas where both parasites are sympatric, and hybrid offspring can occur. Therefore, a fast and precise method is required for differentiating the parasites. Identification based on spicule morphometry is the most common technique for differentiating Haemonchus species. However, because these measurements overlap between species, morphological analysis is insufficient for differentiating between helminth species. In this work, we present a reliable, conventional polymerase chain reaction (PCR)-based method that uses two species-specific primer pairs to differentiate between H. contortus and H. placei specimens and their hybrids. Each primer pair produces one single and distinct amplification band for each species, which enables the detection of hybrid specimens. These primer pairs were validated by testing eight different populations of H. contortus, H. placei and hybrids.
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host specificity and morphometrics of female Haemonchus contortus h placei and h similis nematoda trichostrongylidae in cattle and sheep from shared pastures in sao paulo state brazil
Journal of Helminthology, 2015Co-Authors: Maria Regina Lucas Da Silva, M. R. V. Amarante, K D S Bresciani, Alessandro Francisco Talamini Do AmaranteAbstract:To better evaluate the usefulness of female Haemonchus specimens for specific identification, we undertook a detailed analysis of the morphology of a collection of worms obtained from cattle and sheep in shared pastures. Based on the results, we also more precisely evaluated the host-specificity of Haemonchus contortus, H. placei and H. similis occurring sympatrically in a farm located in the western region of Sao Paulo State, Brazil. A synlophe analysis was employed to identify the three species of Haemonchus . In cattle, the predominant species was H. similis (90.9%), followed by H. placei (9.1%). With the exception of one H. placei specimen, only H. contortus was found in sheep. The longest body length was found for H. placei specimens, followed by H. contortus and then H. similis. It was possible to distinguish H. similis females from H. contortus and H. placei on the basis of vulval structure. The synlophe analysis proved to be very useful for identification of H. contortus, H. placei and H. similis in epidemiological studies involving different species of ruminants in the same pastures. The finding that H. placei and H. similis were adapted to cattle and that H. contortus was adapted to sheep also confirmed the high host-specificity of the three nematodes species.
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differentiation of Haemonchus placei from Haemonchus contortus by pcr and by morphometrics of adult parasites and third stage larvae
Revista Brasileira De Parasitologia Veterinaria, 2014Co-Authors: Michelle C. Santos, M. R. V. Amarante, Maria Regina Lucas Da Silva, Alessandro Francisco Talamini Do AmaranteAbstract:Molecular and morphological methods were evaluated to distinguish between Haemonchus contortus and Haemonchus placei species. A total of 141 H. contortus and 89 H. placei male adult specimens collected from artificially infected lambs were identified individually by PCR analysis, using a species-specific primer pair. These PCR results were used as gold standard for Haemonchus spp. identification. Haemonchus placei presented higher mean spicule and barb lengths than H. contortus (P<0.05). However, some measurements overlapped. For this reason, a discriminate function did not allow the correct identification of 13 H. contortus and one H. placei specimen. The sheath tail length of the third stage larvae (L3), which comprises the distance between the tip of the larval tail and the end of the sheath tail, were measured. Only three of the 485 H. placei larvae (0.619%) had a sheath tail shorter than 85 µm, while only four of the 500 H. contortus larvae (0.8%) presented a sheath tail longer than 85 µm. The results indicated that 6.09% of the male adult specimens would be misclassified based on the discriminate function, while only 0.71% of infective larvae would be misclassified. Therefore, identification of L3 can be used as the first method to indicate the presence of H. placei and/or H. contortus in a population of domestic ruminants.
P J Skuce - One of the best experts on this subject based on the ideXlab platform.
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development of a loop mediated isothermal amplification lamp assay for the sensitive detection of Haemonchus contortus eggs in ovine faecal samples
Veterinary Parasitology, 2014Co-Authors: Lynsey Melville, Fiona Kenyon, Sajid Javed, Iain Mcelarney, Janina Demeler, P J SkuceAbstract:Abstract A major constraint on the effective control and management of helminth parasites in livestock is the lack of rapid and reliable diagnostic tests to identify the parasite species responsible for disease and to allow informed treatment decisions to be made. In the present study, we have developed a novel DNA-based assay for the detection of Haemonchus contortus eggs in ovine faecal samples, using loop-mediated isothermal amplification (or LAMP). LAMP allows for rapid detection of H. contortus DNA under isothermal incubation conditions. The robust nature of this assay negates the need for extensive DNA extraction, allowing amplification from relatively crude samples. Preliminary results suggest that LAMP is highly specific, and does not cross-react with DNA from other common co-infecting parasites. The Haemonchus LAMP assay is also highly sensitive, exhibiting a 10 times lower detection limit than the equivalent PCR; 10−5 ng/μl and 10−4 ng/μl DNA, respectively, allowing detection in a faecal samples containing two Haemonchus eggs per gram. Translation of this assay onto a real-time platform provided rapid results and highlighted its potential as a quantitative assay which could inform treatment decisions in the future.
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anthelmintic resistance in swedish sheep flocks based on a comparison of the results from the faecal egg count reduction test and resistant allele frequencies of the β tubulin gene
Veterinary Parasitology, 2009Co-Authors: Johan Hoglund, Katarina Gustafsson, Brittlouise Ljungstrom, Annie Engstrom, Alison Donnan, P J SkuceAbstract:A faecal egg count reduction test (FECRT) survey was conducted during the grazing season 2006 and 2007 to provide an updated indication of the prevalence of anthelmintic resistance in sheep flocks in Sweden. A total of 1330 faecal samples from 90 flocks on 45 farms, with a minimum of 20 ewes each, was collected by local sheep veterinarians. Per treatment group, approximately 15 lambs were dewormed either with oral suspensions of ivermectin (Ivomec vet.) or albendazole (Valbazen vet.). The efficacy on each farm was investigated either in 2006 or 2007 by faecal egg counts collected on the day of treatment and in a new sample from the same animals 7-10 days later. Third-stage larvae (L3) were initially identified morphologically from pooled cultures. These were then used as the source of genomic DNA template for two molecular tests. The first was a PCR-based test for specific identification of Haemonchus contortus, and the second was a Pyrosequencing assay for the analysis of benzimidazole (BZ) resistance targeting the P200 mutation in the parasite's beta-tubulin gene. Larval cultures indicated that Teladorsagia and Trichostrongylus were the predominant genera, but Haemonchus was diagnosed in 37% of the flocks. The PCR results revealed an almost 100% agreement with those farms that had previously been shown to have Haemonchus present, even when the % prevalence was low (approximately 3%). Only two (4%) of the surveyed farms showed evidence of BZ-resistant worm populations, with H. contortus being the species implicated according to post-treatment larval culture results. The Pyrosequencing assay detected BZ resistant allele frequencies of >40% in the Haemonchus-positive farms and 100% resistant alleles in the clinically most resistant farms. These preliminary results suggest that the FECRT is less sensitive than the molecular test at detecting BZ resistance. However, both tests need to be interpreted carefully, bearing in mind the relative proportions of species present and the starting egg and/or larval counts. Parasitological diagnosis of "clinical" resistance was also found against ivermectin in two flocks. However, both the pre-treatment FECs and the reductions in these were low, and only three lambs that had between 100 and 450 EPG after treatment were involved.
Ray M. Kaplan - One of the best experts on this subject based on the ideXlab platform.
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Motility in the L3 stage is a poor phenotype for detecting and measuring resistance to avermectin/milbemycin drugs in gastrointestinal nematodes of livestock
Elsevier, 2018Co-Authors: Melissa M. George, Lorraine Lopez-soberal, Bob E. Storey, Sue B. Howell, Ray M. KaplanAbstract:Motility is a commonly used in vitro phenotype for assessing anthelmintic activity of candidate compounds, and for detecting anthelmintic resistance in nematodes. Third-stage larvae (L3) of parasitic nematodes are commonly used in motility-based assays because L3 are simple to obtain and can remain viable in storage for extended periods. To improve the measurement of motility of microscopic stages of nematodes, our laboratory developed the Worminator, which quantitatively measures motility of parasites. Using the Worminator, we compared the dose-response characteristics of several avermectin/milbemycin (AM) compounds using L3 from both AM-susceptible and AM-resistant Cooperia spp. (abamectin, doramectin, eprinomectin, ivermectin, moxidectin) and Haemonchus contortus (eprinomectin, ivermectin, moxidectin). Concentrations tested with the Worminator ranged from 0.156 to 40 μM. Differences in EC50 between AM-susceptible and AM-resistant isolates of Cooperia spp. and Haemonchus contortus were small, with resistance ratios ranging from 1.00 to 1.34 for Cooperia spp., 0.99 to 1.65 for Haemonchus contortus. Larval migration inhibition assays were conducted using the same isolates and were equally ineffective for detection of resistance with resistance ratios less than 2.0. These results contrast with those of the Larval Development Assay where we obtained a resistance ratio of 16.48 using the same isolates of Haemonchus contortus. Moreover, even at the highest concentration tested (40 μM), 100% inhibition of motility was never achieved and EC50 for Worminator assays were more than 100× higher than peak plasma levels achieved in vivo following treatment. These data demonstrate that dose-response characteristics for inhibition of motility in L3 of gastrointestinal nematodes of livestock do not significantly differ for AM-susceptible and AM-resistant isolates. These data challenge the suitability of motility as a phenotype for detecting and measuring resistance to AM drugs in gastrointestinal nematodes of livestock. Keywords: Motility, Resistance, L3, Avermectin, Worminator, Livestoc
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The presence of benzimidazole resistance mutations in Haemonchus placei from US cattle.
Veterinary Parasitology, 2014Co-Authors: Umer Chaudhry, Melissa Miller, T. A. Yazwinski, Ray M. Kaplan, John S. GilleardAbstract:Haemonchus populations were collected from cattle from mid-western and eastern Southern US (four and six populations, respectively) to determine the relative prevalence of Haemonchus contortus and Haemonchus placei and the frequency of the three isotype-1 β-tubulin polymorphisms associated with benzimidazole resistance. A minimum of 32 individual adult worms were genotyped at position 24 of the rDNA ITS-2 for each population to determine species identity (296 worms in total). One population from Georgia was identified as 100% H. contortus with the remaining nine populations identified as 100% H. placei. For the H. contortus population, 29 out of 32 worms carried the P200Y (TAC) isotype-1 β-tubulin and 2 out of 32 worms carried the P167Y (TAC) benzimidazole resistance associated polymorphisms respectively. For H. placei, six out of the nine populations contained the P200Y (TAC) isotype-1 β-tubulin benzimidazole resistance associated polymorphism at low frequency (between 1.6% and 9.4%) with no resistance associated polymorphisms being identified at the P198 and P167 codons. This is the first report of the P200Y (TAC) isotype-1 β-tubulin benzimidazole resistance associated polymorphism in H. placei. The presence of this mutation in multiple independent H. placei populations indicates the risk of resistance emerging in this parasite should benzimidazoles be intensively used for parasite control in US cattle.
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modification and further evaluation of a fluorescein labeled peanut agglutinin test for identification of Haemonchus contortus eggs
Veterinary Parasitology, 2010Co-Authors: Megan E Jurasek, Ray M. Kaplan, Janell K Bishopstewart, Bobby E Storey, Michael L KentAbstract:Abstract Gastrointestinal nematodes of the family Trichostrongylidae are the most important parasites of sheep, goats and other ruminants worldwide. Of this group, Haemonchus contortus is usually the most damaging species, particularly in warmer climates or during the summer. It is therefore useful to be able to rapidly differentiate infections with this nematode from other, less pathogenic, species. However, aside from Nematodirus spp., there are only subtle differences between the egg morphology within the trichostrongyles, making it very difficult to identify eggs to the species level. It has been shown previously that peanut agglutinin specifically binds to Haemonchus eggs and not those of other trichostrongyle species. By using this lectin conjugated to fluorescein isothiocyanate, binding to Haemonchus eggs can be visualized under ultraviolet illumination. We adapted this test using eggs purified by routine sugar centrifugation methods and evaluated 26 diagnostic samples from ruminants submitted to our laboratories in Oregon and Georgia. Very good correlations were seen between this test and larval culture (adjusted R2 = 0.72132; F1,25 = 65.7; p
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prevalence of anthelmintic resistance on sheep and goat farms in the southeastern united states
Javma-journal of The American Veterinary Medical Association, 2008Co-Authors: Sue B. Howell, J M Burke, J E Miller, T H Terrill, Elide Valencia, Mimi J Williams, Lisa H Williamson, Anne M Zajac, Ray M. KaplanAbstract:Objective—To determine prevalence of anthelmintic resistance on sheep and goat farms in the southeastern United States. Design—Cross-sectional study. Animals—Sheep and goats from 46 farms in 8 southern states, Puerto Rico, and St Croix in the US Virgin Islands. Procedures—Parasite eggs were isolated from fecal samples, and susceptibility to benzimidazole, imidathiazole, and avermectin-milbemycin anthelmintics was evaluated with a commercial larval development assay. Results—Haemonchus contortus was the most common parasite on 44 of 46 farms; Trichostrongylus colubriformis was the second most commonly identified parasite. Haemonchus contortus from 45 (98%), 25 (54%), 35 (76%), and 11 (24%) farms were resistant to benzimidazole, levamisole, ivermectin, and moxidectin, respectively. Resistance to all 3 classes of anthelmintics was detected on 22 (48%) farms, and resistance to all 3 classes plus moxidectin was detected on 8 farms (17%). Conclusions and Clinical Relevance—Findings provided strong evidence that...
Pettit - One of the best experts on this subject based on the ideXlab platform.
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Evaluation of immunization with gut membrane glycoproteins of Ostertagia ostertagi against homologous challenge in calves and against Haemonchus contortus in sheep.
Parasite immunology, 2000Co-Authors: Smith, PettitAbstract:Peanut and ConA lectins were used as ligands to isolate glycoproteins from detergent extracts of adult Ostertagia ostertagi membranes. As judged by their profiles following SDS-PAGE, these fractions closely resembled the equivalents from Haemonchus contortus which are derived from the nematode intestinal cell microvillar membranes and which are highly protective when used as antigens. Groups of calves were immunized with the peanut and ConA binding fractions of Ostertagia, either as separate or pooled antigens mixed with QuilA as adjuvant. All calves, including controls immunized with adjuvant only, were challenged with a single dose of infective Ostertagia larvae and faecal egg counts were monitored for 5 weeks. In two experiments where the antigen fractions were pooled, moderate (30–50%), but statistically significant reductions in egg output were observed, but the number of worms was not diminished. No significant protection was observed in a third trial where groups of calves were immunized with peanut or ConA binding proteins given separately. Two further trials were conducted in sheep immunized with the same Ostertagia fractions but challenged with Haemonchus. Irrespective of whether they were administered separately or together, the Ostertagia antigens cross protected efficiently against Haemonchus reducing egg counts by between 81% and 97% and worm numbers by between 57% and 84%.