Influence of the physiological stage of Blackbelly sheep on immunological behaviour against gastrointestinal nematodes.

The influence of the physiological stage of a Blackbelly flock against natural infection by gastrointestinal parasitic nematodes (GIN) was investigated. Data on serum and saliva IgA levels and peripheral cellular immune response were recorded in 51 sheep for 6 months. The flock was divided based on their physiological stage as follows: a) pregnant ewes, b) lactating ewes, c) non-pregnant ewes, d) pre-weaning lambs, e) growing lambs and f) replacement sheep. Additionally, sheep were classified based on the parasite infection as stabled or grazing management system as follows: g) non-infected stabled sheep, h) infected and stabled sheep and i) infected grazing sheep. Faeces and blood samples were collected every fifteen days. The percentage of packed cell volume (PCV), plasma protein, (PP) and leucocyte differential counts (LDC) were recorded. Likewise, serum and saliva IgA levels were measured by the indirect ELISA technique. The number of GIN eggs excreted per g of faeces (EPG) was also determined using the McMaster technique. Data were analysed using the SAS program to identify fixed effect of physiological stages, infection time and interactions. Results showed high EPG values in stabled lactating ewes (2764) and in growing lambs (2950-4550 EPG). Pregnant ewes had higher PCV values (27%) than lactating ewes (23.5%). Infected lambs showed low PCV values (28.5%) and high PP values (5.9 g/dL) compared with non-infected lambs (31% PCV and 5.6 g/dL PP). The two most abundant GIN species identified were Haemonchus contortus (91%) and Trichostrongylus colubriformis (81%). Pregnant and non-pregnant ewes showed 29% and <20% of IgA values, respectively. High EPG, low IgA and eosinophils counts were observed until 180 days of age in infected lambs. In conclusion, the group of infected lactating ewes and lambs showed susceptibility to GIN than the other groups, including the pregnant ewes.

A technique for the quantification of Duddingtonia flagrans chlamydospores in sheep faeces.

Previous observations showed that Duddingtonia flagrans chlamydospores were visualized in McMaster chambers containing faeces of treated sheep. This trial explored the McMaster technique as a tool to quantify chlamydospores in sheep faeces. A range of individual chlamydospore doses (from 19.5 x 10(6) to 177.5 x 10(6)) were offered orally to nine lambs for 7 consecutive days. A faecal sample (5 g) was daily obtained from the rectum of each animal (from days 1 to 13) to perform the McMaster technique using a sugar flotation fluid with 1.27 g/mL density. Each chlamydospore counted in the McMaster chamber was considered as 50 chlamydospores per g of faeces (CPG). The results confirmed that the estimated CPG was associated with the daily dose offered to the animals (r(2)=0.90; P<0.001). Furthermore, the total chlamydospore dose received by each animal was strongly associated to the total quantity of CPG obtained from the bulk faeces (TCtot) (r(2)=0.96; P<0.0001). Quantification of CPG can be used as a helpful tool to determine the number of chlamydospores reaching the faeces in orally dosed animals. This could be used to evaluate the efficacy of D. flagrans for the control of gastrointestinal nematode larvae in sheep faeces.

Phenotypic expression of parasite susceptibility to Haemonchus contortus in Pelibuey sheep.

The objective of this study was to determine if the mean faecal egg count (FEC) from a first experimental Haemonchus contortus infection could be used to classify parasite-naïve Pelibuey hair sheep as parasite-resistant high responders and parasite-susceptible low responders. Twenty 6- to 7-month-old Pelibuey male sheep raised free of gastrointestinal nematodes were challenged with 7500±1412 H. contortus L3 larvae administrated orally on day 0 of the study. Faecal samples from each lamb were obtained daily from 21 to 41days post-infection (Stage I). Lambs received a second artificial infection of 8420±1545L3 larvae on day 42, with faecal samples collected from day 65 to day 78 (Stage III). The mean FEC for each lamb in Stage I was used to classify 8 lambs with means for FEC that were more than two standard errors (SE) below the overall mean (i.e., <4764 eggs per gram of feces; epg) as high responders. The remaining 12 lambs were classified as low responders. Means for FEC in Stage I were 2449±194 epg for high responders and 14,461±1044 epg for low responders (P<0.05). High responders also had lower FEC than low responders in Stage III (actual means of 650±220 vs. 5933±1990 epg; P<0.05 following log transformation to normalize the FEC distribution). Lambs were then reclassified as high and low responders based on their mean FEC in Stage III. Fourteen lambs with means for FEC that were more than one standard error (SE) below the overall mean (i.e., below 1537 epg) were classified as high responders. The remaining six lambs were classified as low responders. Use of the Stage I responder class to predict the Stage III responder class resulted in an 83.3% sensitivity but only a 50% specificity. The positive predictive value was 41.7% and the negative predictive value was 87.5%. The poor positive predictive value was caused by 5 animals with high FEC in Stage I, but low FEC in Stage III. The first infection thus identified most high-responder lambs, but a second infection may improve accuracy by separating lambs with an intermediate level of resistance from truly susceptible lambs. This protocol now requires additional validation under more practical conditions involving natural parasite infections and larger lamb numbers.

Gastrointestinal nematode populations with multiple anthelmintic resistance in sheep farms from the hot humid tropics of Mexico.

This study evaluated the status of anthelmintic resistance against the three available classes of commercial drugs in seven sheep farms in the hot humid tropics of Mexico. Drug classes included benzimidazole (BZ), ivermectin (IVM) and levamisole (LV). Respective faecal egg count reduction tests (FECRT) were performed in each farm. Faecal samples were obtained from the rectum of >100 sheep in each farm. Adult sheep shedding >150 eggs per gram of faeces (EPG) were included. In each farm, animals were allotted to one of four groups with similar mean EPG: Control Group (untreated), BZ group (albendazole sulfoxide 5mg/kg LW), IVM group (ivermectin, 0.2mg/kg LW) and LEV group (levamisole 7.5mg/kg LW). Drugs were administered subcutaneously. A second faecal sampling was performed on the same animals of each farm 14days post-treatment. The GIN genera obtained from faecal cultures were identified for each group in different farms. Percentage faecal egg count reduction (%R) and 95% confidence intervals were estimated using the RESO© software. A questionnaire was applied to farm owners to describe anthelmintic management practices. All sheep farms had GIN populations with multiple resistance to the three anthelmintic classes tested. The %R ranged from 0 to 48% for BZ, 29 to 82% for IVM and 1 to 88% for LEV. Haemonchus spp. and Trichostongylus spp. were found in all treated groups of the study farms. Resistant Oesophagostomum spp. larvae (BZ or IVM) were found in respective farms. Treatment practices in study farms included frequent mass treatment every two months with extra treatments applied individually in the presence of clinical signs. Drug dosage used visual estimation of body weight rather than the exact weight of each animal. Quarantine anthelmintic treatment of incoming stock was used but efficacy was not confirmed.

Persistence of the efficacy of copper oxide wire particles against Haemonchus contortus in sheep.

The aim was to determine the persistent efficacy of copper oxide wire particles (COWP) against Haemonchus contortus in sheep, using the harmonization guidelines protocol. Thirty-six male lambs (2 months old) reared free of gastrointestinal nematodes were used (average body weight of 10.8±3.8kg). Before and for the duration of the study, lambs were kept in raised cages with slatted floors and were offered ad libitum a complete mixed diet. Animals were divided into six groups (n=6): one non-treated control group (G0) and five groups treated with one COWP capsule (1.7g of copper oxide; Copinox(®)). Animals in each group were treated on pre-defined dates before the artificial infection was applied: days -35 (G1), -28 (G2), -21 (G3), -14 (G4) and -7 (G5). On day 0 animals were infected with 3700 H. contortus infective larvae per animal. Animals were humanely slaughtered between days 22 and 23 post-infection. The abomasums were individually washed to obtain the contents. These organs were subjected to separate artificial digestions. Adult parasites were counted from the abomasum contents and the larvae from the digested material. Worm burden geometric means were calculated for each group. A significant worm burden reduction in either of the treated groups (G1, G2, G3, G4, and G5) compared to the control (G0) was considered as persistence of the anthelmintic effect. Copper levels were determined from individual liver samples of each animal. The geometric mean worm burden of the control group (G0) was 1959. Compared to the control, worm burdens geometric means were significantly reduced in groups G1 (1108), G4 (528) and G5 (1063) (P<0.03). Efficacies in G1, G4 and G5 were 43.4%, 73.0% and 45.7% respectively. No significant reduction was found for G2 (1342) and G3 (1430). A larger quantity of Cu was found in the livers of treated animals compared to the control group (P<0.05) except for G3 (P=0.06). A negative association between Cu liver content and worm burdens was found (r=-0.42, P<0.05). Live weight gain was similar in all groups and no clinical or post-mortem manifestations of Cu toxicity were recorded in treated animals.

Evaluating the effectiveness of a Mexican strain of Duddingtonia flagrans as a biological control agent against gastrointestinal nematodes in goat faeces.

The use of Duddingtonia flagrans in the control of goat nematodes was investigated. Initially, the time of passage of chlamydospores through the digestive tract of goats was evaluated. Two groups of seven parasite-free kids were formed. Group A received a single dose of 3.5x10(6) D. flagrans chlamydospores (FTHO-8 strain) per kg of live weight. Group B did not receive any chlamydospores. Faeces were obtained from each kid daily from day 4 prior to inoculation until day 5 post-inoculation (PI) and were placed in Petri dishes containing water agar. Gastrointestinal nematode infective larvae were added to each Petri dish and incubated at 25 degrees C for 7 days. Petri dishes were examined to detect the fungus and trapped nematodes. A second trial evaluated the effect of D. flagrans on the number of gastrointestinal nematode larvae harvested from goat faecal cultures in naturally infected goats. Two groups of seven goats were formed. The treated group received a single dose of 3.5x10(6) D. flagrans chlamydospores per kg of liveweight. The control group did not receive any chlamydospores. Faeces were obtained twice daily from each kid. Two faecal cultures were made for each kid. One was incubated for 7 days and the other for 14 days. Gastrointestinal nematode larvae were recovered from each culture and counted. Percentage of larval development reduction was determined using a ratio of larvae/eggs deposited in the control and treated groups. Duddingtonia flagrans survived the digestive process of goats, and maintained its predatory activity, being observed from 21 to 81 h PI (3 to 4 days). A reduction in the infective larvae population in the treated group compared to the non-treated group was observed in both incubation periods (7 days: 5.3-36.0%; 14 days: 0-52.8%, P>0.05). Although a single inoculation of D. flagrans can induce a reduction of infective larvae collected from faeces, a different scheme of dosing may be needed to enhance the efficacy of D. flagrans in goats.