Identification of compounds from Paris polyphylla (ChongLou) active against Dactylogyrus intermedius.

The present study was designated to ascertain the anthelmintic activity of the rhizomes of Paris polyphylla and to isolate and characterize the active constituents. The methanol extract from rhizomes of P. polyphylla showed significant anthelmintic activity against Dactylogyrus intermedius with the median effective concentration (EC50) 22.5 mg L(-1). Based on this finding, the methanol extract was fractionated by silica gel column chromatography in a bioassay-guided fractionation yielding 2 bioactive compounds, the structures of these compounds were elucidated as formosanin C and polyphyllin VII. The in vivo tests revealed that formosanin C and polyphyllin VII were significantly effective against D. intermedius with EC50 values of 0.6 and 1.2 mg L(-1), respectively. The acute toxicities (LC50) of formosanin C and polyphyllin VII for grass carp were 2.8 and 2.9 mg L(-1), respectively. The overall results provide important information for the potential application of formosanin C and polyphyllin VII in the therapy of serious infection caused by D. intermedius.

Antiparasitic efficacy of dihydrosanguinarine and dihydrochelerythrine from Macleaya microcarpa against Ichthyophthirius multifiliis in richadsin (Squaliobarbus curriculus).

Ichthyophthirius multifiliis is a holotrichous protozoan that invades the gills and skin surfaces of fish and can cause morbidity and high mortality in most species of freshwater fish worldwide. The present study was undertaken to investigate the antiparasitic activity of crude extracts and pure compounds from the leaves of Macleaya microcarpa. The chloroform extract showed a promising antiparasitic activity against I. multifiliis. Based on these finding, the chloroform extract was fractionated on silica gel column chromatography in a bioactivity-guided isolation affording two compounds showing potent activity. The structures of the two compounds were elucidated as dihydrosanguinarine and dihydrochelerythrine by hydrogen and carbon-13 nuclear magnetic resonance spectrum and electron ionization mass spectrometry. The in vivo tests revealed that dihydrosanguinarine and dihydrochelerythrine were effective against I. multifiliis with median effective concentration (EC(50)) values of 5.18 and 9.43 mg/l, respectively. The acute toxicities (LC(50)) of dihydrosanguinarine and dihydrochelerythrine for richadsin were 13.3 and 18.2mg/l, respectively. The overall results provided important information for the potential application of dihydrosanguinarine and dihydrochelerythrine in the therapy of serious infection caused by I. multifiliis.

Anthelmintic and cytotoxic activities of extracts of Markhamia obtusifolia Sprague (Bignoniaceae).

The anthelmintic activity of Markhamia obtusifolia Sprague (Bignoniaceae) leaf extracts was evaluated against the ruminant gastrointestinal nematode parasite Trichostrongylus colubriformis (Nematoda: Strongylida) using the in vitro egg hatch test. Also, the cytotoxic activity of aqueous extracts of M. obtusifolia was evaluated in cell line cytotoxicity assays. The results indicated that the effective concentration (EC(50)) for the water extract of M. obtusifolia leaves (0.46 mg/mL; Confidence Interval [CI] 0.3-0.5mg/mL) was significantly lower than the EC(50) for the acetone extract of M. obtusifolia (0.8 mg/mL; CI 0.7-1mg/mL). Aqueous extracts were twice as potent as the acetone extracts. The EC(90) (0.2mg/mL; CI 0.1-0.02) for thiabendazole (positive control) was significantly lower than the EC(90) for the water extract of M. obtusifolia (10.7 mg/mL; CI 8.3-13.7 mg/mL). In the cytotoxicity bioassay, the lethal concentration (LC(50)) for the aqueous extract of M. obtusifolia was 0.476 mg/mL, which was relatively high (low toxicity) in comparison to the highly toxic berberine (LC(50)=9.80 µg/mL). The current study showed that M. obtusifolia plant extracts possess anthelmintic activity and are relatively non-cytotoxic, thus providing support for their use in traditional veterinary practices.

Anthelminthic efficacy of aqueous extract of Acanthus montanus leaf against strongylid nematodes of small ruminants.

The anthelminthic efficacy of the crude aqueous extract of Acanthus montanus (Nees) T. Anders (Acanthaceae) against strongylid nematodes of small ruminants was investigated using the in-vitro egg hatch and larval growth inhibition assays. Faecal samples collected per rectum from sheep and goats were subjected to parasitological examination using the McMaster counting technique with a yield of 700 eggs per gram (E.P.G.) of faeces. Crude aqueous leaf extract of Acanthus montanus was extracted using cold water extraction with a yield of 13.01% w/w. Egg hatch assay revealed a 91.75% reduction in egg hatch at concentration of 25 mg/ml of the extract. The extract had a 100% inhibition, at 200mg/ml concentration which was equivalent to the activity of 3.125 mg/ml albendazole. The distilled water control however, showed a 0% inhibition. Larval growth inhibition assay results showed the extract on Day 1 at 25 mg/ml and 200mg/ml concentrations yielded 67.02% and 85.26% larval inhibition. On Day 2, 100% inhibition was obtained on all concentrations of the extract except for 25 mg/ml that yielded 88.30%. Albendazole however, had a 100% larval inhibition for all concentrations on Day 2. A 100% larval inhibition was recorded on Day 3 from the 25 mg/ml concentrations. The mean percentage larval inhibition of the extract at 200mg/ml (92.63%) was comparable to the standard anthelminthic (albendazole) at 12.50mg/ml (92.28%). These findings showed that there is a pharmacological basis for the folkloric claim of the anthelminthic effect of Acanthus montanus.

Essential veterinary education on the development of antimicrobial and antiparasitic resistance: consequences for animal health and food safety and the need for vigilance.

The role played by the veterinary profession in protecting animal and public health, along with food safety, is unique. Thus, it is important that veterinarians have the necessary knowledge to be pro-active in responding to the emergence of antimicrobial resistance. Many current veterinary educational programmes do not attempt to enhance the understanding of undergraduate students of this complex situation. As a first step, a modern veterinary education programme should provide students with a holistic view of the ecology of resistance and clearly describe how antimicrobial resistance emerges. Understanding the relationships between drug use, natural selection and antimicrobial-resistant organisms, as well as gene mobilisation and recombination, and how these contribute to the emergence of resistant organisms are important facets of a modern veterinary education.

Anthelmintic efficacy of neem (Azadirachta indica A. Juss) and the homeopathic product Fator Vermes in Morada Nova sheep.

Gastrointestinal nematodes are becoming increasingly resistant to the commercial products used to control them. The cost of routine vermifuge applications on herds and the problem of residues in animal products and the environment have prompted research on the anthelmintic activity of plant extracts. This work examines the anthelmintic action of neem and the homeopathic product Fator Vermes in sheep kept in a pasture for 18 months. Forty sheep of the Morada Nova breed were divided into four treatments and the control, according to the EPG. During the experiment, each animal received 100 g/day of shredded corn and did not receive protein supplementation. In treatment 1 (control), the animals received only shredded corn. Treatment 2 received 1.6 g/(animal day) of the homeopathic product mixed with the shredded corn, and treatments 3, 4 and 5 received, respectively, 12.5, 25.0 and 37.5 g/(animal day) of dried Azadirachta indica leaves mixed with the shredded corn. The neem was administered for alternating 15-day periods and the homeopathic product daily for 18 months. There were 39 fortnightly fecal collections made to count the EPG, and fecal cultures were performed monthly. The following genera, in percentage, were identified: Haemonchus: 65.58+/-3.27, Trichostrongylus: 15.92+/-7.38 and Oesophagostomum: 18.50+/-6.22. The treatments evaluated were not effective in controlling gastrointestinal nematodes (P>0.05), whose mean log(10) counts (EPG +1) and standard errors for treatments 1-5 were respectively 3.55+/-0.28; 3.48+/-0.31; 3.90+/-0.29; 2.78+/-0.29 and 3.48+/-0.30. A significant effect (P<0.0001) was observed of the periods of the year when the 39 collections occurred. Because of the diet deficient in raw protein, the sheep had higher average EPG counts, for all the treatments, at the end of the dry season, and the opposite occurred in the middle of the rainy season.

Direct and indirect anthelmintic effects of condensed tannins in sheep.

Anthelmintic activity of condensed tannins (CT) was evaluated both in vitro and in vivo. In vitro tests included egg hatch test and paralysis/mortality assay on adult Haemonchus contortus. In vivo anthelmintic effect was determined by faecal egg count reduction test in lambs. To this end, 18 lambs were divided into three groups (low tannin, high tannin and control). The lambs of low and high tannin groups were fed diets containing 2 and 3% CT while the control group was fed on diets without CT. In vitro trials showed a dose-dependent inhibition of nematode egg hatching; whereas, there was no effect of CT on adult H. contortus. In vivo trials indicated reduction in faecal egg counts in lambs fed diets containing CT. Feed intake and nutrient digestibility of CT-fed sheep was lower and nitrogen balance was higher as compared to control. Maximum weight gain was observed in animals fed diets containing 3% CT. The direct anthelmintic effect of CT, therefore, was evidenced by inhibited egg hatching; whereas, faecal egg counts reduction in sheep was through improved nutrient utilization.

Efficacy of a herbal product against Histomonas meleagridis after experimental infection of turkey poults.

Histomoniasis (infectious enterohepatitis, blackhead) is caused by the protozoan parasite Histomonas meleagridis (H. meleagridis). After the ban of all prophylactic and therapeutic drugs in the European Union, histomoniasis is increasingly responsible for considerable economic problems to the poultry industry. The aim of this study was to investigate the effect of a herbal product with extracts from cinnamon, garlic, lemon, and rosemary on H. meleagridis in turkey poults in vivo. For this purpose, 60 two-week-old poults were divided into three groups. Group 1 received the herbal product in the feed six days before infection and in water three days before infection, then in feed and drinking water until the end of the experiment. Groups 2 and 3 were left untreated. At week 3 of age, Groups 1 and 2 were infected intracloacally with H. meleagridis. Three weeks after infection the surviving birds were euthanized and examined for pathological lesions. Mortality was 20% in Group 1 and 50% in Group 2. There were no deaths in Group 3. DNA of histomonads was detected in all examined caeca and livers of the dead birds, but was not detected in any examined organ of the surviving birds of all groups. There was no noticeable difference in the lesion scores of the dead birds between the groups. The surviving birds of all groups did not show lesions post mortem. Since all effective prophylactic and therapeutic drugs against histomoniasis were banned in the EU, under given conditions the investigated herbal product seems to be an effective alternative for the reduction of mortality in turkeys caused by histomoniasis.

Evaluation of multiple low doses of copper oxide wire particles compared with levamisole for control of Haemonchus contortus in lambs.

High levels of anthelmintic resistance in gastrointestinal nematodes (GIN) of small ruminants have created the need for alternative approaches to parasite control. Copper oxide wire particles (COWP; 2g) have proven effective in decreasing GIN infection in lambs. However, the risk of copper toxicity has limited the usefulness of this approach. Recently, smaller doses (0.5 and 1g) have proven effective in GIN control, reducing the risk of toxicity. The objective of this study was to examine the effectiveness and risk of toxicity using multiple small doses of COWP for GIN control in lambs between weaning and market weight. Dorper crossbred ram lambs were orally administered levamisole (Levasol, 8.0mg/kg; n=8), 0.5g (n=9), or 1g COWP (n=9) at weaning (Day 0; 118+/-2 days of age; late May 2005) and again at 6-week intervals for a total of four treatments. A pooled fecal culture determined that Haemonchus contortus was the predominant gastrointestinal parasite at weaning. Lambs grazed bermudagrass pastures and were supplemented with up to 500g corn/soybean meal and free choice trace mineralized salt. Fecal egg counts (FEC), packed cell volume (PCV), and plasma aspartate aminotransferase (AST) activity were determined every 14 days and lambs weighed every 28 days. GIN infection reached a peak at Day 42 (high FEC, low PCV). COWP effectively reduced FEC on Days 0 and 42 compared with the previous week, but did not reduce FEC on Days 84 and 126 (treatment by time interaction, P<0.005). Plasma AST activity and weight gains were similar among treatment groups throughout the study period. Concentrations of copper in the liver on Day 155 were greater in COWP-treated lambs (P<0.001), but all concentrations were normal. Multiple doses of COWP were as effective as levamisole for control of H. contortus without risk of copper toxicity.

Bermudagrass management in the Southern Piedmont USA. V: Gastrointestinal parasite control in cattle.

Parasite-free pastures would improve cattle health and performance, resulting in possible economic return to producers. Our objective was to determine the effect of a single series of anthelmintic treatment of steers prior to stocking on Coastal bermudagrass pastures, during five consecutive summers, on the parasite burden in cattle. The site for this experiment had been conventionally cropped for several decades, with no exposure to cattle, and would be expected to be relatively free of nematode larvae. The experimental design was a randomized complete block (landscape features) with a split plot arrangement of treatments where main plots were pasture fertilization treatments (mineral, clover plus mineral, and broiler litter) and split plots were low and high forage mass. Anthelmintic treatment included pour-on ivermectin on day -21, albendazole on day -7, and injectable ivermectin 48 h prior to stocking of pastures, with the cattle remaining in drylot during the 48-h period prior to being placed on the experimental paddocks. All steers received only one series of treatments during any given year. Yearling Angus steers (Bos taurus) were managed in a put-and-take grazing system with three "tester" steers assigned to each paddock and "grazer" steers added or removed at 28-day intervals. From 1994 to 1998, steers grazed the paddocks for a 140-day period from mid May until early October each year. Fecal samples for worm egg counts were obtained on day 0 and at 28-day intervals, thereafter. On all sampling days after day 0, samples were obtained only from tester animals. Over the 5-year period, the mean eggs per gram of feces (epg) gradually increased from 0 (following treatment) to a mean of 2.2 (range from 0.7 to 3.0) by the end of the grazing season (the last sampling date) in October. Although the epg were not zero, they were below threshold levels that would allow development of a parasite burden in cattle. In traditional management systems, cattle graze parasite-contaminated pastures; therefore, parasites negatively impact growth and productivity throughout the entire grazing period. Periodic anthelmintic treatments simply give a temporary reprieve from those parasitic infections. Conceptually, using the current grazing system, it should be possible to maintain these pastures in a parasite-free status indefinitely; however, from a drug resistance perspective, it would be most applicable in sod-based rotation systems where cattle graze from two to five years before land is returned to row-crops. By removing the effect of parasites, cattle can grow without the physiological constraints that gastrointestinal parasites place on appetite, digestion, nutrient utilization, and general well being.

Ethno-veterinary medicine: screening of Nigerian medicinal plants for trypanocidal properties.

Trypanosoma congolense and T. brucei bloodstream form parasites were propagated axenically in suitable standard media at 34 degrees C. The effects of 33 plant extracts, fractions and pure compounds were evaluated on two clones of T. brucei and drug-sensitive and multi-drug-resistant clones of T. congolense. The cytotoxic activity of the trypanocidal extracts was also evaluated on calf aorta endothelial cells in vitro. Of the extracts tested, 22% killed T. congolense IL 1180 at a concentration of 100 microg/ml while 18% killed 90-100% of T. brucei ILTat 1.4 at the same concentration. However, 6% of the active extracts killed 93% of a dyskinetoplastid form of T. brucei IL Tat 1.1, indicating that the intact kinetoplast is a target of some of the compounds tested. Of the 12 extracts that displayed activity against drug sensitive trypanosomes, 66.7% had trypanocidal activity on a multi-drug-resistant clone, T. congolense IL 3338. The extracts of Eugenia uniflora, Acacia artaxacantha, Terminalia ivorensis, T. superba and Alchornea cordifolia had median lethal concentrations of between 13 and 69 microg/ml on both the drug-sensitive, IL 1180 and multi-drug-resistant clone, IL 3338. The median lethal doses of the active plant extracts on the calf aorta endothelial cells varied between 112 and 13750 microg/ml while the calculated selective indices ranged between 0.71 and 246.8 indicating bright prospects for the development of some of these extracts as potential trypanocidal agents.