Multispecies and multiple anthelmintic resistance on cattle nematodes in a farm in Argentina: the beginning of high resistance?

There are very few resistance records on cattle nematodes. South American successive records have been increasing rapidly since the year 2000. In Argentina, increasing dissatisfaction based on the exclusive use of macrocyclic lactones has prompted the use of benzimidazoles in the Pampean region. The studied farm is located in the Argentina humid Pampas and had apparently poor results after anthelmintic treatments. Evaluation of resistance was firstly based on faecal egg reduction after treatment in November 2001 and June 2002 and was complemented with worm counts obtained by necropsy in June 2002. The study reports that the reduction of faecal egg excretion after benzimidazole (reduction 31-79%) or ivermectin treatments (76-97%) in November 2001 was insufficient and these poor results were confirmed in June 2002 (benzimidazoles (65-89%) and ivermectin (47-77%)). Several methods for evaluating faecal egg count reduction were performed and yielded different results. A new method based on the estimated faecal egg counts (corrected from the initial faecal egg counts and animal type using a general linear model) was used and gave higher flexibility in the interpretation of putative resistance to anthelmintics. From necropsy results, it could be concluded that Cooperia oncophora was resistant to avermectins and benzimidazoles and that Cooperia punctata, Ostertagia ostertagi and Haemonchus placei were resistant to benzimidazoles. This case of multispecies and multidrug resistance is probably not unique and could reflect the emergence of resistances in Argentina. This emergence is probably due to the intensive use of anthelmintics, the absence of refugia, and the frequent circulation of infected cattle.     

Recent developments in the management of anthelmintic resistance in small ruminants – an Australasian perspective

Abstract

There have been a number of significant advances in recent years to the theory and practice of managing anthelmintic resistance in sheep in Australasia. The general principles of resistance management are, firstly identification and mitigation of high-risk practices, secondly using effective anthelmintics, and thirdly maintaining a refuge of unselected parasites. The first of these principles has been updated recently with the findings from a series of farm-based trials in New Zealand, in which the economic benefits of both short- and long-acting anthelmintic treatments in ewes pre-lambing were found to be inconsistent and not always positive. There have also been significant changes to the second principle, particularly given the introduction of new active families onto the market. Evidence continues to favour the use of combination products to maximise efficacy and delay the onset of treatment-failure. Many farmers have readily accepted the effectiveness of maintaining a refuge of unselected parasites; the challenge for researchers and advisers is now to improve adoption of properly designed and implemented resistance management programmes. A recently completed education programme in New Zealand has demonstrated that when this is achieved, then anthelmintic resistance can be controlled, and in many cases reduced in severity.     

Is anthelmintic resistance a concern for heartworm control? What can we learn from the human filariasis control programs?

Heartworm prophylaxis is currently largely dependent on the ability of avermectins and milbemycins to arrest the development of third and fourth stages of Dirofilaria immitis for prolonged periods, without producing adulticidal effects. Major control programs, dependent on the activity of ivermectin, are being implemented for human onchocerciasis and lymphatic filariasis. The avermectins and milbemycins act on glutamate-gated and gamma-aminobutyrate-gated chloride channel subunit proteins in nematodes. Ivermectin resistance has been widely described in trichostrongylid nematodes of ruminants. There is evidence that when ivermectin resistance occurs in nematodes, there may be selection on some, but not all of the genes that code for ligand-gated chloride channel subunit proteins as well as on some ABC-transporter genes, whose products may be involved in regulating macrocyclic lactone drug concentrations at receptors, and on some structural protein genes of amphidial neurones. Although ivermectin resistance has not been reported in filarial nematodes, there have recently been reports of suboptimal responses to ivermectin in Onchocerca volvulus. Evidence has been found of ivermectin selection on at least ABC-transporter genes and some neuronal structural protein genes in O. volvulus. To date, there is no evidence of avermectin/milbemycin resistance in D. immitis, also a filarial nematode. Chemotherapy against trichostrongylids of animals, human filariae, and D. immitis, relies on avermectins or milbemycins. However, control involves targeting different stages or processes in the nematode life cycles, different control strategies, different proportions of the nematode population in refugia, and different drug dosage rates. Consideration of the proportion of the D. immitis population normally in refugia, the life cycle stage targeted, and the anthelmintic dosages used suggest that it is unlikely that significant avermectin/milbemycin resistance will be selected in D. immitis with current treatment strategies.     

Breeding for resistance to footrot – the use of hoof lesion scoring to quantify footrot in sheep

So that genetic studies can be undertaken on footrot in sheep, it is necessary that a reliable and repeatable method to categorise the phenotype is available. This paper summarises the methods used and results obtained from 1600 hoof lesion scores of 100 mixed-age ewes independently scored twice by two trained operators. Using a 5-pont scale describing the severity of foot lesions, residual correlations were used to assess agreement between scorers and scoring occasions. Data were analysed using both zero-1 and continuous data methods. The average prevalence of any score >0 was 15%, and of scores >1 was 12%. The residual correlation between scorers for SUM_FR was 0.87 and between scoring occasions it was also 0.87, indicating high repeatability or agreement both within and between scorers. No significant differences were detected between scorers or between scoring occasions for any of the traits analysed, or different analytical methods used.     

Comparison of tick resistance of crossbred Senepol × Limousin to purebred Limousin cattle

The comparison of resistance to natural tick infestation by Rhipicephalus microplus (Canestrini, 1887) of crossbred Senepol?×?Limousin and purebred Limousin cattle was investigated. The Senepol breed, originated from St Croix Island in the Caribbean is considered as a Bos taurus breed adapted to tropical conditions. Despite its B. taurus genetic background, it is believed to have a good tick resistance, but this resistance has never been assessed previously. Tick counts under natural infestation were carried out to investigate the difference of susceptibility between crossbred Senepol?×?Limousin and purebred Limousin cattle. Mixed-effect models were used to assess the effect of the breed on the number of ticks. Results show that Senepol?×?Limousin are five times less infested by ticks than purebred Limousin. These results underline the opportunity to use Senepol cattle for crossing with susceptible B. taurus breeds in tick infested areas, to combine tick resistance with beef production abilities.     

How widespread is resistance to invermectin among gastrointestinal nematodes in sheep in The Netherlands?

In Autumn 2009, a faecal egg count reduction test (FERCT) was carried out on three sheep farms. Groups of 8-11 lambs were treated with ivermectin or moxidectin, with a 14-day interval between treatment and sampling. Ivermectin resistance was present on all three farms. Treatment with ivermectin resulted in a reduction in faecal egg numbers of 94.6%, 63%, and 59%. On two farms, 14 days after treatment pooled faecal samples yielded predominantly larvae of Hamonchus contortus (100% and 98%, respectively). On the third farm, H. contortus and (probably) Teladorsagia circumcincta were resistant to ivermectin (64% and 36% of the larvae, respectively). Treatment with moxidectin resulted in a 100% reduction in egg output in sheep on all three farms. More sensitive culture techniques failed to detect any larvae in samples taken from two farms, but a few Ostertagia-type larvae, probably of T. circumcincta, were detected in samples from the third farm. It can be concluded that gastrointestinal nematodes in sheep from these three farms were resistant to ivermectin, whereas resistance to moxidectin was not detected.     

Field studies with in‐feed medication of pigs in the Netherlands using the anthelmintic thiophanate with particular reference to efficacy against Ascaris suum

Summary

Studies on the efficacy in pigs of low level in‐feed medication with the anthelmintic thiophanate at a minimum intake of 6 mg/kg/day for fourteen days are reported. A trial was conducted to compare a group of medicated fattening pigs with a similar unmedicated group on premises known to have a high challenge of Ascaris spp. Daily growth rate was improved whilst feed conversion ratio and the liver condemnation rate were reduced in the treatment group.

Routine medication of a whole herd using this regime contributed to a great improvement of the herd production when assessed by the above criteria. User studies in various geographical areas of the Netherlands involving 1500 adult pigs and 1200 fattening pigs medicated with thiophanate in‐feed for fourteen days demonstrated that the compound eliminated the faecal worm egg output and was readily accepted and tolerated by pigs.     

Differential serotonergic mediation of aggression in roosters bred for resistance and susceptibility to Marek’s disease

1. Serotonin (5-HT) is a primary regulating neurotransmitter involved in aggressive and impulsive behaviours in mammals. Previous studies have also demonstrated that the function of the serotonergic system in regulating aggression is affected by both genetic and environmental factors. The serotonergic system may display similar functions in chickens.

2. Our objective was to investigate the aggressive and impulsive behavioural response to antagonism of the 5-HT1A and 1B receptors in cocks bred for susceptibility and resistance to Marek’s disease (i.e. strain 7₂ and 6₃, respectively).

3. Cocks from strain 7₂ exhibited increased aggressive behaviours and lower brain 5-HT concentrations compared to strain 6₃ cocks.

4. Antagonism of 5-HT1A receptors increased aggressiveness and reduced serotonin turnover in strain 7₂, but not strain 6₃ cocks. 5-HT1B receptor antagonism had no effect on aggression or serotonin turnover in either strain.

5. Levels of the serotonin metabolite 5-HIAA, but not absolute central 5-HT levels, were altered in both strains following 5-HT1B antagonism, but only in strain 7₂ cocks following 5-HT1A antagonism.

6. The data suggest that 5-HT1A and 1B regulate aggression differently in high and low aggressive strains.     

Evaluation on resistance of 4 alfalfa （Medicago sativa ） cultivars to pea aphid （Acyrthosiphon pisum ）

在大田和室内条件下,采用大量蚜虫侵染幼苗法评价4个苜蓿（Medicagosativa）品种对豌豆蚜（Acyrthosiphonpisum）的抗性,以筛选出对豌豆蚜高抗的材料。结果表明,甘农5号（HA3）在大田和室内的抗性植株百分率均显著高于其他品种,分别为50．3％和48．9％,而猎人河的抗性植株百分率在大田和室内均较低,分别为4．0％和4．3％;4个苜蓿品种在大田的抗性级别为甘农5号高抗,甘农3号、金皇后、猎人河均为感虫,室内抗性级别为甘农5号抗虫,金皇后、甘农3号低抗,猎人河感虫。     

Genomic regions and pathways associated with gastrointestinal parasites resistance in Santa Inês breed adapted to tropical climate

Background: The aim of this study was to estimate variance components and to identify genomic regions and pathways associated with resistance to gastrointestinal parasites, particularly Haemonchus contortus, in a breed of sheep adapted to tropical climate. Phenotypes evaluations were performed to verify resistance to gastrointestinal parasites, and were divided into two categories: i) farm phenotypes, assessing body condition score(BCS), degree of anemia assessed by the famacha chart(FAM), fur score(FS) and feces consistency(FC); and ii) lab phenotypes,comprising blood analyses for hematocrit(HCT), white blood cell count(WBC), red blood cell count(RBC),hemoglobin(HGB), platelets(PLT) and transformed(log10) egg per gram of feces(EPG_(log)). A total of 576 animals were genotyped with the Ovine SNP12k Bead Chip(Illumina, Inc.), that contains 12,785 bialleleic SNP markers. The variance components were estimated using a single trait model by single step genomic BLUP procedure.Results: The overall linkage disequilibrium(LD) mean between pairs of markers measured by r2 was 0.23. The overall LD mean between markers considering windows up to 10 Mb was 0.07. The mean LD between adjacent SNPs across autosomes ranged from 0.02 to 0.10. Heritability estimates were low for EPG_(log)(0.11), moderate for RBC(0.18), PLT(0.17) HCT(0.20), HGB(0.16) and WBC(0.22), and high for FAM(0.35). A total of 22, 21, 23, 20, 26, 25 and 23 windows for EPG_(log)for FAM, WBC, RBC, PLT, HCT and HGB traits were identified, respectively. Among the associated windows, 10 were shown to be common to HCT and HGB traits on OAR1, OAR2, OAR3, OAR5, OAR8 and OAR15.Conclusion: The traits indicating gastrointestinal parasites resistance presented an adequate genetic variability to respond to selection in Santa Inês breed, and it is expected a higher genetic gain for FAM trait when compared to the others. The level of LD estimated for markers separated by less than 1 Mb indicated that the Ovine SNP12k Bead Chip might be a suitable tool for identifying genomic regions associated with traits related to gastrointestinal parasite resistance. Several candidate genes related to immune system development and activation, inflammatory response,regulation of lymphocytes and leukocytes proliferation were found. These genes may help in the selection of animals with higher resistance to parasites.     

What’s happened to Staphylococcus intermedius? Taxonomic revision and emergence of multi‐drug resistance

Staphylococcus intermedius has been the predominant coagulase-positive Staphylococcus isolated from canine skin and mucosae and the most commonly reported staphylococcal pathogen in small animal practice for the last 35 years. Although microbiological tests have historically indicated variability in biochemical characteristics amongst S. intermedius isolates from animals, an acceptable level of diagnostic accuracy for clinical purposes was readily achievable with routine phenotypic testing. However, three recent developments have changed our understanding of the term "S. intermedius" and have challenged veterinary bacteriologists to ensure correct species identification of pathogenic staphylococci from small animals. First, the increasing recognition of meticillin-resistant Staphylococcus aureus in small animal practice and its human health implications demand accurate species identification. Secondly, the application of molecular techniques to analysis of staphylococcal isolates has led to a revised taxonomy and canine isolates of S. intermedius being re-named S. pseudintermedius. Thirdly, the recent, rapid emergence of meticillin- and multi-drug-resistant strains of Staphylococcus pseudintermedius (MRSP) has become a major therapeutic challenge in veterinary practice worldwide, including the UK. This article discusses the background of the recent taxonomic changes within the genus Staphylococcus and reviews the key features of MRSP and its implications for day-to-day laboratory diagnosis and small animal practice.     

Genetic analysis of multi-drug resistance and the clonal dissemination of β-lactam resistance in Salmonella Infantis isolated from broilers

An epidemiologic study was conducted to investigate the incidence and characterize the antimicrobial resistance determinants, analyzing plasmid profiles, and establishing the genetic relationship among β-lactam-resistant isolates of Salmonella Infantis from broilers in Southern Japan. A total of 120 isolates were recovered from 56 flocks belonging to 44 holdings during 2004–2006. The percentages of resistance were as follows: ampicillin (24%), cephalothin (23%), cefoxitin (0%), ceftazidime (11%), cefotaxime (11%), chloramphenicol (0%), kanamycin (7.5%), ofloxacin (20%), oxytetracycline, streptomycin and sulfamethoxazole (100%) and trimethoprim (75%). The incidence of bla_TEM-encoded β-lactam resistance in 2004–2006 was significantly higher than in 1998–2003 (P < 0.001). BlnI-digested PFGE patterns generated two related clusters implicated in the dissemination of β-lactam resistance. Two types of plasmid profiles were observed and two plasmids of ca. 50 and 180-kb size were carried by β-lactam-resistant isolates. Streptomycin resistance was conferred by aadA1 (n = 116), aadA1-aadA2 (n = 1), and aadA1-strA-strB (n = 3). Resistances to kanamycin, oxytetracycline, sulfamethoxazole and trimethoprim were conferred by aphA1 (n = 9, 100%), tetA (n = 120, 100%) sul1 (n = 120, 100%) and dfrA5 (n = 90, 100%), respectively. Two types of class 1 integrons were detected: 1.0 kb (n = 120) and, 1.0/1.5 kb (n = 3). Integrons of 1.0/1.5 kb were found in isolates with the aadA1-strA-strB gene combination. For the first time, all S. Infantis isolates showed resistance to at least three classes of antimicrobial agents; and the intestinal tract of healthy poultry was a reservoir of the extended-spectrum cephalosporin-resistant isolates of serovar Infantis.