Dermal immune responses against Psoroptes ovis in two cattle breeds and effects of anti-inflammatory dexamethasone treatment on the development of psoroptic mange.

Psoroptic mange is a common disease of livestock, caused by Psoroptes ovis. Compared to Holstein-Friesian (HF) cattle, the Belgian Blue (BB) cattle breed is highly susceptible to the infestation. However, the mechanism for this difference is still unclear. To determine the factors responsible for this breed susceptibility, the immune response to P. ovis was studied in experimentally infested BB and HF cattle, using clinical signs, histology, immunohistochemical profiling and gene expression analysis of skin biopsies. The mite numbers and lesion area of BB cattle were greater than in HF during the whole study period. Significant influxes of eosinophils in the epidermis and dermis were detected in comparison with the pre-infestation samples in both breeds, with significantly higher eosinophils in BB at 6 weeks post infestation (wpi). Mast cell numbers were unaffected at all stages of infestation in HF, but were significantly elevated relative to pre-infestation in BB cattle at 2 and 6 wpi. The more pronounced cutaneous eosinophilia and higher IL-4 levels at 6 wpi in BB cattle suggest that a Th2-type immune response is underlying the higher susceptibility of the BB breed. In naturally infested BB cattle, development of the psoroptic mange lesions and eosinophils and CD3+ T cell areas were severely depressed after anti-inflammatory treatment with dexamethasone. Together, these results suggest that a stronger Th2-type immune response to P. ovis causes the skin lesions in psoroptic mange in BB cattle and that local anti-inflammatory treatment could potentially be an alternative to control the pathology caused by this parasite.

Anthelmintic resistance and common worm control practices in sheep farms in Flanders, Belgium.

In contrast to many other European countries, no data were available on the presence of anthelmintic resistance in gastrointestinal nematodes in sheep in Belgium. A faecal egg count reduction test was performed in 26 sheep flocks in Flanders, Northern Belgium. Results indicated widespread resistance against benzimidazoles (albendazole, fenbendazole and mebendazole), with treatment failure on all 8 farms investigated. Haemonchus contortus and Teladorsagia circumcincta were the predominant species after treatment failure. Amino acid substitutions associated with benzimidazole resistance were detected at the codon positions 167 (8%) and 200 (92%) of the isotype-1 beta tubulin gene in H. contortus, codon positions 198 (47%) and 200 (43%) in T. circumcincta and position 200 (100%) in T. colubriformis. Resistance against macrocyclic lactones (ivermectin, doramectin and moxidectin) was recorded on 7 out of 20 flocks, mainly in H. contortus and T. circumcincta. Treatment failure was also observed for closantel (in combination with mebendazole) and for monepantel, on one farm each. Trichostrongylus spp. were implicated with resistance against monepantel. A questionnaire survey on farm management and worm control measures indicated that worm control was often not sustainable. Ewes and lambs were treated frequently (on average 2.6 and 3.2 times per year), mostly without weighing. Only few sheep farmers (9%) regularly used faecal egg counts to monitor worm infections. Despite the FECRT showing otherwise, most of the farmers perceived the efficacy of anthelmintics as very good (30%) or good (54%).

Resistance against macrocyclic lactones in Psoroptes ovis in cattle.

BACKGROUND: Psoroptic mange is an important disease in Belgian Blue cattle. Treatment failure of macrocyclic lactones against Psoroptes ovis has been reported, but clear evidence of in vivo resistance is lacking. This study assessed the efficacy of macrocyclic lactone products on 16 beef farms in Belgium and the Netherlands in vivo and in vitro. METHODS: On each farm a group of animals (n = 7-14) with psoroptic mange was treated with two subcutaneous injections of a macrocyclic lactone product with 7-10 days interval (15 farms) or a single injection with a long-acting macrocyclic lactone (1 farm). In vivo efficacy was assessed by the reduction in mite counts, clinical index (proportion of the body surface affected by lesions), the proportion of the animals with negative mite counts after the first treatment round and the number of treatment rounds needed to obtain zero mites counts in all animals. A mite population was categorized as sensitive when the mite count reduction after the first treatment round > 95% and the lower limit of the uncertainty interval > 90%. Resistance was detected when both parameters were below their threshold and suspected when one parameter was too low. In vitro knockdown and mortality were evaluated in a contact test. RESULTS: The proportion of the animals with negative mite counts after the first treatment round varied from 0 to 80%. All farms needed two or more treatments rounds to obtain zero mite counts on all animals. Clinical index only started to reduce after the second treatment round. Mite populations from three farms were categorized as sensitive, one as suspected resistant and 12 as resistant. No correlation was found between in vitro lethal dose 50 and knockdown dose 50 values and in vivo efficacy parameters. CONCLUSIONS: Unambiguous treatment failure was detected on 12 out of 16 farms, confirming the presence of macrocyclic lactone resistance on Belgian Blue beef farms. In vitro parameters could not discriminate the farms based on their in vivo sensitivity. The mean reduction in mite counts and the lower limit of the confidence interval are proposed as parameters to identify acaricide resistance.

A Bayesian evaluation of two dip-stick assays for the on-site diagnosis of infection in calves suspected of clinical giardiasis.

The objective of the present study was to evaluate two commercially available dip-stick assays for the diagnosis of Giardia infections in faecal samples from calves suspected of clinical giardiasis. The dip-stick assays provide an on-site and hence quick alternative to laboratory diagnosis. A three-test Bayesian model was used, including the test results of the Coris Giardia strip (Coris Bioconcept, Gembloux, Belgium), the Speed Giardia or BVT dip-stick (Bio Veto Test/Virbac, La Seyne sur Mer, France), and the Meridian immunofluorescence assay (IFA: Meridian Diagnostics Inc., Cincinnati, OH). In total, 421 faecal samples were examined with the three diagnostic assays between October 2008 and November 2009. Overall, the number of positive samples was markedly higher using the IFA compared to both dip-stick assays, resulting in a high sensitivity (se: 88%, with a 95% probability interval (PI) 60-99%) compared to the Coris dip-stick assay (se: 28%; PI: 16-41%) and the BVT dip-stick assay (se: 26%, PI: 16-35). The specificities of all the three assays were very high (IFA sp: 94%, PI: 90-99%; Coris sp: 92%, PI 86-98%; BVT sp: 93%, PI 88-98%). A positive diagnosis by the dip-stick assays was significantly correlated with a higher cyst excretion level, as measured by IFA. The majority (76%) of the positive samples in the present study contained less than 5000cyst per gram of faeces, even though all these animals displayed clinical symptoms of diarrhea potentially due to Giardia. The low level of cyst excretion in these samples might in part explain the poor sensitivity of both dip-stick assays. Although multiple samplings might be an option to increase the sensitivity of the dip-stick assays, the laboratory based IFA seems at current to be the best option for clinical diagnosis of Giardia in calves.

The effect of a fenbendazole treatment on cyst excretion and weight gain in calves experimentally infected with Giardia duodenalis.

A total of 28 Holstein-Friesian calves were experimentally infected with 10(5)Giardia duodenalis cysts. Eleven days later, all animals were allocated into two groups of 14 animals each, based on the average pre-treatment cyst counts. Treatment was randomly assigned to one of the two groups, and all animals in the treatment group received a daily oral dosage of 15mg fenbendazole per kg bodyweight during 3 consecutive days. The calves in the control group received a placebo (water). From 3 days after treatment onwards, cyst excretion was determined three times a week during 4 consecutive weeks. The faecal consistency and general health were recorded on a daily basis, and all animals were weighed prior to treatment and weekly thereafter. At the end of the experimental period, there was a significant (P<0.001) reduction (98%) of the cumulative cyst excretion. There were no significant differences in general health between both groups, but faecal consistency was significantly lower (P<0.002) in the control group compared to the treatment group, although none of the animals displayed overt gastro-intestinal symptoms. Prior to treatment the weight did not differ between both experimental groups. At the end of the 4-week experimental period however, the animals in the treatment group gained on average 2.86kg (=102g per day) more than the animals in the control group (P<0.031). This study demonstrates for the first time a significant difference in weight gain between fenbendazole treated and untreated calves experimentally infected with G. duodenalis, although additional data need to confirm the need for treatment in natural conditions.

Field validity and feasibility of four techniques for the detection of Trichuris in simians: a model for monitoring drug efficacy in public health?

BACKGROUND: Soil-transmitted helminths, such as Trichuris trichiura, are of major concern in public health. Current efforts to control these helminth infections involve periodic mass treatment in endemic areas. Since these large-scale interventions are likely to intensify, monitoring the drug efficacy will become indispensible. However, studies comparing detection techniques based on sensitivity, fecal egg counts (FEC), feasibility for mass diagnosis and drug efficacy estimates are scarce. METHODOLOGY/PRINCIPAL FINDINGS: In the present study, the ether-based concentration, the Parasep Solvent Free (SF), the McMaster and the FLOTAC techniques were compared based on both validity and feasibility for the detection of Trichuris eggs in 100 fecal samples of nonhuman primates. In addition, the drug efficacy estimates of quantitative techniques was examined using a statistical simulation. Trichuris eggs were found in 47% of the samples. FLOTAC was the most sensitive technique (100%), followed by the Parasep SF (83.0% [95% confidence interval (CI): 82.4-83.6%]) and the ether-based concentration technique (76.6% [95% CI: 75.8-77.3%]). McMaster was the least sensitive (61.7% [95% CI: 60.7-62.6%]) and failed to detect low FEC. The quantitative comparison revealed a positive correlation between the four techniques (Rs = 0.85-0.93; p<0.0001). However, the ether-based concentration technique and the Parasep SF detected significantly fewer eggs than both the McMaster and the FLOTAC (p<0.0083). Overall, the McMaster was the most feasible technique (3.9 min/sample for preparing, reading and cleaning of the apparatus), followed by the ether-based concentration technique (7.7 min/sample) and the FLOTAC (9.8 min/sample). Parasep SF was the least feasible (17.7 min/sample). The simulation revealed that the sensitivity is less important for monitoring drug efficacy and that both FLOTAC and McMaster were reliable estimators. CONCLUSIONS/SIGNIFICANCE: The results of this study demonstrated that McMaster is a promising technique when making use of FEC to monitor drug efficacy in Trichuris.