The laboratory maintenance of field-collected Lymnaea tomentosa for the production of Fasciola hepatica metacercariae

A culture method is described for the production of Fasciola hepatica metacercariae in Lymnaea tomentosa collected from the field habitat. Two-litre glass jars, filled with artificial spring (A.S.) water and having a substratum of soil-bentonite (S.B.) agar to provide habitat and food, maintain groups of 40 to 100 snails graded according to size. Small quantities of washed lettuce are fed as a supplement. Compressed air, pre-warmed to 25°C, is piped to each jar to maintain thermoregulation, aeration, filtration and circulation of the water. Infection is achieved by reducing the A.S. water level to 3–4 mm and exposing the snails to miracidia for 6 h. During the aquatic developmental phase of 5–6 weeks, twice-weekly changes of A.S. water and the filter minimise proliferation of protozoans and Chaetogaster spp. Accumulation of mature cercariae is achieved by sustaining the infected snails on moist S.B. agar while humidification within the jar is maintained. Alternatively, groups of five to ten snails may be held in disposable petri dishes lined with S.B. agar. Maturation of cercariae may be delayed by refrigeration at 4–5°C. Upon maximum accumulation, the cercariae are harvested by dissection of the snails and metacercariae are stored in plastic containers at 4°C.     

A comparative examination of swine sera for antibody to Aujeszky virus with the conventional and a modified virus-serum neutralization test and a modified direct complement fixation test.

Sera of pigs from élite breeding herds, of boars and sows collected at slaughter-houses, and of pigs from herds known to be infected, were examined for antibody to Aujeszky virus. The conventional and a modified virus-neutralizing antibody (VNA) test and a modified direct complement fixation (CF) test were employed. In simultaneous titrations of positive sera the modified VNA test gave titers approx. 4 log2 units above the titers obtained by the conventional test. The conventional VNA test was found insufficiently sensitive. Unspecific neutralization in the modified VNA test was infrequent in serum dilution 1/2 and rare in dilution 1/4. The GF tests on sera of slaughter sows and animals from known infected herds showed a remarkable consistency with the VNA tests. Inconsistent results were obtained with but few sera. Abt. 5 % of the sera could not be examined because of complement fixation with control antigen.     

Effect of supplementing graded concentrations of non-phytate phosphorus on performance,egg quality and bone mineral variables in White Leghorn layers

1. An experiment was conducted to determine optimal non-phytate phosphorus (NPP) concentrations for White Leghorn (WL) layers (22–72 weeks) fed diet containing 38 g Ca/kg.

2. Eight diets with graded concentrations (1.5–3.25 g/kg in increments of 0.25 g) of NPP were prepared. Each diet was fed to eight pen replicates containing 88 birds in each. Performance data was evaluated in three different phases (phase I-22–37 weeks, phase II-38–53 weeks and phase III-54–72 weeks). Optimum levels of NPP were determined by fitting a quadratic polynomial (QP) regression model.

3. Egg production (EP) was not affected (P = 0.059) by the concentration of NPP and interaction between NPP and diet phase was non-significant, indicating that the lowest concentration (1.5 g/kg diet) of NPP used in the study was adequate across the three phases. However, EP was influenced by phase (P < 0.001).

4. Optimum concentration of NPP for feed intake (FI) was estimated to be 1.5, 1.71 and 2.40 g/kg diet during phases I, II and III, respectively. FI per egg mass (EM) or feed efficiency (FE) responded quadratically with NPP and also differed significantly between phases. Optimum concentration of NPP for FE during phases I, II and III was 1.5, 2.56 and 2.32 g/kg diet, respectively.

5. Egg weight (EW), EM, shell weight and thickness were not affected by NPP concentration although all of these variables (except shell weight) were influenced by phases.

6. Breaking strength of tibia and Ca contents in tibia ash were not affected by the concentration of NPP, but bone ash and P contents in tibia ash were influenced (P < 0.001) by NPP. Predicted optimal concentrations of NPP for responses for tibia ash at 44 or 72 weeks, tibia ash P at 44 weeks and tibia ash P at 72 weeks were 1.55, 2.63 and 1.5 g/kg diet, respectively.

7. Based on the results, it was concluded that WL layers required 1.5 g, 2.63 g and 2.4 g, respectively/kg diet during phase I, II and III with the calculated daily intake of 137.3, 278.3 and 262 mg NPP/b/d.     

Infection of a canine macrophage cell line with leishmania infantum: determination of nitric oxide production and anti-leishmanial activity

We have previously shown that resistance to Leishmania infantum in dogs is associated with a Th1 type of immune response. In this study, we use a canine macrophage cell line (030-D) that can readily be infected with this protozoan parasite. Our aim is to further characterize the effector mechanisms involved in killing of Leishmania parasite in dogs. We observed that activation of 030-D cells by incubation with a supernatant derived from a Leishmania-specific T cell line containing IFN-gamma, TNF-alpha and interleukin-2 (IL-2) resulted in enhanced nitric oxide (NO) production by these cells. In addition, we observed enhanced anti-leishmanial activity of infected 030-cells after activation. Both, NO production and anti-leishmanial activity were abrogated by addition of L-N(G)-nitroargininemethyl ester (L-NAME), an analogue of L-arginine. Thus, NO play an important role in the anti-leishmanial activity of these canine macrophages. We propose the infection of the 030-D cell line as a good in vitro model to further investigate parasite-host cell interactions in dogs, a natural host of Leishmania parasites.     

Analgesic effects of butorphanol and buprenorphine in conscious African grey parrots (Psittacus erithacus erithacus and Psittacus erithacus timneh)

OBJECTIVE: To evaluate effects of butorphanol tartrate and buprenorphine hydrochloride on withdrawal threshold to a noxious stimulus in conscious African grey parrots. ANIMALS: 29 African grey parrots (Psittacus erithacus erithacus and Psittacus erithacus timneh). PROCEDURE: Birds were fitted with an electrode on the medial metatarsal region of the right leg, placed into a test box, and allowed to acclimate. An electrical stimulus (range, 0.0 to 1.46 mA) was delivered to each bird's foot through an aluminum perch. A withdrawal response was recorded when the bird lifted its foot from the perch or vigorously flinched its wings. Baseline threshold to a noxious electrical stimulus was determined. Birds then were randomly assigned to receive an i.m. injection of saline (0.9% NaCl) solution, butorphanol (1.0 mg/kg of body weight), or buprenorphine (0.1 mg/kg), and threshold values were determined again. RESULTS: Butorphanol significantly increased threshold value, but saline solution or buprenorphine did not significantly change threshold values. CONCLUSIONS AND CLINICAL RELEVANCE: Butorphanol had an analgesic effect, significantly increasing the threshold to electrical stimuli in African grey parrots. Buprenorphine at the dosage used did not change the threshold to electrical stimulus. Butorphanol provided an analgesic response in half of the birds tested. Butorphanol would be expected to provide analgesia to African grey parrots in a clinical setting.