Destruction of Anoplocephala perfoliata Eggs by the Nematophagous Fungus Pochonia chlamydosporia

The in vitro effect of an isolate of the nematophagous fungus Pochonia chlamydosporia (VC1) on the eggs of Anoplocephala perfoliata was evaluated. The eggs were morphologically analyzed for their integrity using light microscopy (10× objectives), plated on 9.0-cm diameter petri dishes containing 2% WA culture medium with and without fungal isolate (control), grown for 10 days, and 10 replicates were prepared per group. In all, 1000 eggs of A perfoliata were plated on petri dishes containing 2% water agar culture medium with (VC1) and without the fungal isolate (control). After 3, 5, 7, and 10 days, approximately 100 eggs were removed from each plate and classified on the basis of the following parameters: without alteration; type 1, lytic effect without morphological damage to eggshell; type 2, lytic effect with morphological alteration of embryo and eggshell; and type 3, lytic effect with morphological alteration of embryo and eggshell, in addition to hyphal penetration and internal egg colonization and destruction. The P chlamydosporia fungus demonstrated ovicidal activity (P < .05) on the eggs of A perfoliata in the studied intervals presenting type 3 effects of 35%, 42.5%, 53.83%, and 71.17% for the intervals 3, 5, 7, and 10 days, respectively. P chlamydosporia is a potential biological control agent for the eggs of A perfoliata.     

Efficacy of a drug combination of praziquantel, pyrantel embonate, and febantel against helminth infections in dogs.

Tablets containing praziquantel, pyrantel embonate, and febantel were tested for efficacy against helminths in dogs. A single treatment with this drug combination gave 100% reductions in Toxocara canis and Taenia hydatigena in experimentally induced infections in dogs. In dogs with naturally acquired infections, treatment gave > 97 to 98% reductions in fecal egg counts attributable to Toxascaris leonina, T canis, and Uncinaria stenocephala. Efficacy against Trichuris vulpis was > 92%.     

Anthelmintic activity of pyrantel tartrate against Ascaridia galli in fowls.

At a dose of 15 mg/kg body weight, pyrantel tartrate was 18-51, 99-63, 100 and 100 per cent effective in chickens treated at 10, 20, 30 and 40 days respectively after infection with Ascaridia galli. Similarly, 25 mg/kg was 14-44, 100, 100 and 99-63 per cent effective.     

Comparative efficacy of flubendazole chewable tablets and a tablet combination of febantel, pyrantel embonate and praziquantel against Trichuris vulpis in experimentally infected dogs

Fourteen of 23 dogs developing patent Trichuris vulpis infections by 120 days p.i. with 5000 embryonated eggs were allocated into three groups. One group was treated with flubendazole 220 mg chewable tablets (Flubenol) at the recommended dose regimen once daily for 3 days. The second group was given the recommended single treatment with a tablet containing 150 mg febantel, 144 mg pyrantel embonate and 50 mg praziquantel in combination (Drontal Plus). The third group remained untreated. All dogs were necropsied for worm counts 10 or 11 days after (first) treatment. No worms were recovered from the flubendazole treated dogs resulting in a significant worm count reduction of 100%. In contrast, 2 of 5 animals treated with the combination of febantel, pyrantel embonate and praziquantel remained infected; the geometric mean worm burden was reduced by 99.4% as compared to the control group but did not differ significantly from those of the controls.     

In vitro activity of various anthelmintic compounds against Haemonchus contortus larvae

Twenty-five known anthelmintic compounds were evaluated in vitro against the highly motile exsheathed non-feeding third-stage of Haemonchus contortus larvae. Activity was based on lack of motility or death of larvae after 24 h of chemical exposure. Six compounds (avermectins, closantel, levamisole, morantel, phenylhydrazone and ticarbodine) were active at a concentration of 100 μg cm^?3 or less. The most active compounds were avermectins and levamisole. When higher in vitro concentrations were used, ten compounds (bephenium, coumaphos, dichlorovos, disophenol, hygromycin b, methyridine, parbendazole, phenothiazine, pyrantel and thiabendazole) exhibited activity. Nine compounds were found to be inactive; among these were the new benzimidazoles, i.e., albendazole, fenbendazole, mebendazole and oxibendazole. Because of the inactivity of the new benzimidazoles, this in vitro system is unsuitable as a routine screening tool. Also, the system appears to favor drugs that act quickly through percuticular entry. In an initial group of 5280 untested compounds, 254 (4.8%) exhibited in vitro activity at 100 μg cm^?3 against the non-feeding larvae stage. The exogenous and in vitro cultivation techniques required for collecting, cleaning and exsheating the larvae are described.     

Clearance times and the forensic significance of the dietary anthelmintic pyrantel tartrate in performance horses

Six performance bred (Thoroughbred or Standardbred) mares were fed the anthelmintic pyrantel tartrate as a daily supplement for a period of 21 days to assure steady state concentrations would be achieved. The forensic "clearance times" and potential for analytical interference of pyrantel tartrate were then investigated. This investigation was intend- ed to enable guidelines to be established for veterinarians and trainers to avoid a "positive" test result for pyrantel which might violate existing rules or regulations. The analysis of blood and urine samples from these horses was conducted by the University of Kentucky Equine Drug Testing Laboratory, and were performed on samples obtained on days -3, -2, -1, 7, 14, 21, +1, +2, +3 and +4. Pyrantel tartrate was readily detected by standard thin layer chromatography analysis in urine samples on days 7, 14 and 21 during the administration portion of the study, and on day + 1 of the post-adminislxation time period. Further analysis of the positive urine samples by direct probe mass spectrometry confirmed the presence of pyrantel. In our analysis of equine serum samples pyrantel tartrate was not detected. Pyrantel positive urine samples were additionally ana- lyzed using enzyme-linked immunosorbent assays designed to detect fluphenazine,butorphanol, morphine, oxymorphone, fentanyl, sufentanil and etorphine, with no cross-reactivity found. We concluded that, from a drug testing perspective, the use of pyrantel is unlikely to interfere with normal post- race drug testing.     

The anthelmintic activity of some benzimidazole derivatives against Fasciola hepatica

Micronised mebendazole has been shown to have anthelmintic activity against Fasciola hepatica in rats and sheep. Activity is higher against non-egg-producing F. hepatica in bile ducts than against the migrating larval stages. Parbendazole, cambendazole and thiabendazole have been shown to be much less active, or devoid of activity, against mature and immature F. hepatica in rats, at elevated dose rates.

Reasons for the differences, in terms of molecular structure, are briefly discussed and the pitfalls of structure/activity analyses are considered.     

Comparison of two versions of larval development test to detect anthelmintic resistance in Haemonchus contortus

Larval development (LDT) and micro-agar larval development tests (MALDT) were used to compare the reliability and sensitivity of two methods for detecting anthelmintic resistance in Haemonchus contortus. The tests were conducted using three resistant and four susceptible isolates of H. contortus. Both versions of the tests provided comparable results with regard to the characterization of benzimidazole and levamisole susceptibility but neither test was sufficiently sensitive to discrimination between an ivermectin (IVM) susceptible and an IVM resistant isolate. Each test has its own merits with the LDT having the advantage of being less time-consuming.     

In vivo anthelmintic activity of Phytolacca icosandra against Haemonchus contortus in goats

Two separate controlled and blinded studies were conducted to confirm the dose and non-interference of spinosad and milbemycin oxime (MO) administered orally in combination or alone to dogs for the treatment and control of experimentally induced flea infestations (Ctenocephalides felis) and adult hookworm infections (Ancylostoma caninum). For each study, dogs were allocated randomly based on pre-treatment adult flea and hookworm egg counts to one of four treatment groups of 10 animals each. In each study, spinosad and MO in combination, using the lower half (30-45 mg/kg spinosad; 0.5-0.75 mg/kg MO) of the US commercial dose band (30-60 mg/kg spinosad; 0.5-1.0mg/kg MO) of each active ingredient, or individually alone using the full dose range, were given orally to dogs on Day 0 using a tablet formulation. A placebo control was treated similarly. In one study, on Days -1, 5, 12, 19, 28 and 35 each dog was infested with approximately 100 unfed adult C. felis obtained from the investigator's established flea colony. All dogs were infested via the same method. Forty-eight hour post-infestation flea comb counts were conducted on Days 1, 7, 14, 21, 30 and 37 and were used to determine the knockdown and residual flea activity. In the second study, on Day -27 each of 48 dogs were experimentally inoculated with 100 third-stage infective larvae of the hookworm, A. caninum. Dogs were treated on Day 0 and necropsied on Day 7 or Day 8. All nematodes in the intestinal tract were collected on Day 7 or Day 8, identified and counted by species and stage. Post-treatment, the geometric mean live flea counts were significantly different (p-value<0.0001) between the spinosad/MO combination and the spinosad only treatment groups as compared to the vehicle control group. The flea counts in the MO only group and the control group were not statistically different. The spinosad and MO combination group and the spinosad only treatment group demonstrated significantly different knockdown (100%) and post-treatment residual flea efficacy at Day 30 was 100% for both groups as compared to the vehicle control. The presence of MO in combination with spinosad did not interfere with the flea efficacy of spinosad as compared to the spinosad only group. MO alone did not demonstrate any flea efficacy. Post-treatment, the geometric mean A. caninum worm counts were significantly different (p-value<0.0001) between the spinosad and MO combination group as compared to the vehicle control group. The worm counts in the MO only group and the combination group were not statistically different. The spinosad and MO combination group (99.8% reduction) and the MO only treatment group (99.5% reduction) both demonstrated significantly different hookworm efficacy as compared to the vehicle control group. The presence of spinosad in combination with MO did not interfere with the hookworm efficacy of MO as compared to the MO only group. Spinosad alone did not demonstrate any hookworm efficacy. In summary, flavored spinosad and MO combination tablets administered orally to dogs at the lower end (30-45 mg/kg spinosad; 0.5-0.75 mg/kg MO) of the US commercial tablet unit dose range (30-60 mg/kg spinosad; 0.5-1.0mg/kg MO) were both safe and highly efficacious delivering 100% knockdown and 30 days of residual adult flea control on experimentally infested dogs as well as >99% adult hookworm efficacy evaluated under laboratory conditions. Interference between either drugs was not demonstrated for both of these dose limiting parasites.     

Determination of the activity of pyrantel tartrate against Sarcocystis neurona in gamma-interferon gene knockout mice

Equine protozoal myeloencephalitis (EPM) is the most important protozoal disease of horses in the United States. Some horse owners and equine clinicians believe that horses which are on daily pyrantel tartrate at 2.64mg/kg for helminth prophylaxis are less likely to develop EPM. The present study examined the efficacy of pyrantel tartrate in preventing clinical disease in gamma-interferon gene knockout (BALB/c-Ifng(tm1ts)) mice. No activity was seen against sporocyst-induced Sarcocystis neurona infections in mice treated prophylacticly with 4-5mg pyrantel tartrate per mouse per day in the drinking water.     

In vitro antiviral activity of chestnut and quebracho woods extracts against avian reovirus and metapneumovirus

Field evidences have suggested that a natural extract, containing tannins, could be effective against poultry enteric viral infections. Moreover previous studies have shown that vegetable tannins can have antiviral activity against human viruses. Based on this knowledge three different Chestnut (Castanea spp.) wood extracts and one Quebracho (Schinopsis spp.) wood extract, all containing tannins and currently used in the animal feed industry, were tested for in vitro antiviral activity against avian reovirus (ARV) and avian metapneumovirus (AMPV). The MTT assay was used to evaluate the 50% cytotoxic compounds concentration (CC₅₀) on Vero cells. The antiviral properties were tested before and after the adsorption of the viruses to Vero cells. Antiviral activities were expressed as IC₅₀ (concentration required to inhibit 50% of viral cytopathic effect). CC₅₀s of tested compounds were >200 μg/ml. All compounds had an extracellular antiviral effect against both ARV and AMPV with IC₅₀ values ranging from 25 to 66 μg/ml. Quebracho extract had also evident intracellular anti-ARV activity (IC₅₀ 24 μg/ml). These preliminary results suggest that the examined vegetable extracts might be good candidates in the control of some avian virus infections. Nevertheless further in vivo experiments are required to confirm these findings.