In vitro acaricidal activity of neem (Azadirachta indica) seed extracts with known azadirachtin concentrations against Rhipicephalus microplus

The effect of four extracts from neem seeds (Azadirachta indica) containing 2000, 5000, 9000 and 10,000 ppm of azadirachtin A (AZA), quantified by high-performance liquid chromatography (HPLC) and diluted to 1.25%; 2.5%; 5.0%; 10.0% and 12.8% was verified by in vitro tests with engorged females and larvae of the cattle tick Rhipicephalus microplus. The results from the bioassays with the engorged females showed that the main toxic effect of the extracts was reduction of the reproductive parameters, with a sharp drop in the number of eggs laid and the hatching rate, mainly when the extracts were diluted to 10.0% and 12.8%. The product effectiveness (PE) calculations for all the solutions tested showed that the AZA solution at 10,000 ppm (N10) was the most effective. However, statistical analysis of the PE data obtained for the proportional AZA concentrations in the different diluted extracts showed significance (P<0.05) of the effects included in the model (extract dilution, principle effect (classificatory) of the assay (extract) and the interaction between the two), indicating significant variations due to the dilution, the test and the interaction between the two factors in the tests with engorged females. For solutions N2, N5, and N9, it was not possible to estimate LC(90) values in the dilution range tested. The lowest LC(50) was observed for extract N5, and although extract N10 was the only extract for which the LC(90) could be estimated within the range tested, the LC(50) was higher than for N5 and N9. These results suggest that substances other than AZA present in the extracts influenced the efficacy, especially up to a certain LC range. In the tests with larvae, no mortality was observed, indicating zero effectiveness of all the extracts tested. The results of the tests with engorged females showed that the neem extracts had acaricide activity, inhibiting egg laying and the larval hatching rate. Complementary studies are necessary to develop new methods to isolate and/or identify other substances besides AZA contained in this plant, to enable using products made from it as acaricides.     

In vivo anthelmintic activity of Dorycnium rectum and grape seed extract against Ostertagia (Teladorsagia) circumcincta and Trichostrongylus colubriformis in sheep

AIM: To assess the in vivo anthelmintic activity of condensed tannins (CT) in the forage species Dorycnium rectum and Medicago sativa, and in an extract from grape (Vitus vinifera) seeds (GSE), against two species of parasite, Teladorsagia (Ostertagia) circumcincta and Trichostrongylus colubriformis, at different stages of their life cycle, in sheep that were parasite-na?ve or previously exposed to nematodes. METHODS: In Trial 1, a factorial treatment structure was used to compare faecal nematode egg counts (FEC) and worm burdens in 40 weaned Romney lambs fed either the CT-containing forage D. rectum (12% dry matter; DM) or M. sativa (lucerne; 0.2% DM). Twenty na?ve and 20 previously-exposed lambs were drenched free of parasites then reinfected with known species and numbers of parasites, and housed in pens indoors on a diet of lucerne pellets and chaffed hay. Groups of lambs (n=5 lambs per group) were fed one of the forages over one of two time periods within the parasite's life cycle. Six to nine days after the last feeding of fresh forages, faecal samples were collected for FEC, and all lambs were slaughtered and worm counts conducted. In Trial 2, 12 Suffolk x Romney lambs were surgically implanted with an abomasal cannula and then housed indoors in metabolism crates. After infection with parasites, six lambs were infused continuously over a 14-day period with a commercially available CT GSE (96% DM, made up to 34 g/L in water); the remaining lambs were infused with water. During infusion, samples were collected for egg hatch and larval development assays. After infusion, samples were collected for FEC, and all lambs were slaughtered and worm counts conducted. RESULTS: In Trial 1, there was a significant (p<0.001) difference in burdens of O. circumcincta between na?ve lambs and those previously exposed to parasites, but no other differences were recorded. In Trial 2, lambs infused with GSE had significantly (p<0.05) fewer T. colubriformis at slaughter and significantly (p<0.001) fewer eggs hatched in the egg hatch assay (EHA) than for lambs infused with water. Overall, the differences attributable to GSE were small in magnitude, being an 11% drop in egg hatch, and an 18% drop in numbers of adult T. colubriformis after 14 days of continuous infusion. No other differences were recorded. CONCLUSION: The results indicate that the in vivo anthelmintic activity of these CT sources is, at best, modest and is unlikely to be of any practical value. Further, these data emphasise that in vitro activity is an unreliable indicator of in vivo efficacy for CT-containing forages and extracts.     

Thermotolerance of L3 Ostertagia (Teladorsagia) circumcincta and some of its enzymes

Parasitic nematodes of ruminants can be expected to experience temperatures in excess of 40 degrees C in faeces on pasture and, perhaps, in the host. L3 Ostertagia (Teladorsagia) circumcincta survived for at least 90 min at 45 degrees C in vitro in water, but the larvae were inactivated rapidly by only slightly higher temperatures. The glycolytic enzymes hexokinase and pyruvate kinase were inactivated in a similar temperature range, whereas malate dehydrogenase maintained its activity at temperatures in excess of 50 degrees C. These data imply that the loss of glycolytic activity might explain the loss of larval motility at temperatures between 45 degrees C and 50 degrees C.     

In vitro acaricidal effect of plant extract of neem seed oil (Azadirachta indica) on egg, immature, and adult stages of Hyalomma anatolicum excavatum (Ixodoidea: Ixodidae)

Effects of the plant extract of neem seed (Azadirachta indica) on eggs, immature, and adult stages of Hyalomma anatolicum excavatum was studied at concentrations of 1.6, 3.2, 6.4, and 12.8%. The extract was found to have a significant effect on the hatching rate of eggs. It significantly increased the hatching rate during the first 7 days post-treatment (DPT) giving incompletely developed and dead larvae; however, it cause hatching failure at DPT 15. Neem Azal F induced a significant increased in mortality rates of newly hatched larvae, unfed larvae, and unfed adults reaching 100% on 15th, 3rd, and 15th DPT, respectively. The mortality rates increased with the extract concentrations. Although, it had no significant effect on the moulting rates of fed nymphs, it caused malformation or deformities in 4% of adults moulted. It was concluded that the concentration of Neem Azal F which may be used for commercial control of this tick species were 1.6 and 3.2%.     

Acaricidal activity of extracts of neem (Azadirachta indica) oil against the larvae of the rabbit mite Sarcoptes scabiei var. cuniculi in vitro

The acaricidal activity of the petroleum ether extract, the chloroform extract and the acetic ether extract of neem (Azadirachta indica) oil against Sarcoptes scabiei var. cuniculi larvae was tested in vitro. A complementary log-log (CLL) model was used to analyze the data of the toxicity tests. The results showed that at all test time points, the petroleum ether extract demonstrated the highest activity against the larvae of S. scabiei var. cuniculi, while the activities of the chloroform extract and the acetic ether extract were similar. The activities of both the petroleum ether extract and the chloroform extract against the larvae showed the relation of time and concentration dependent. The median lethal concentration (LC(50)) of the petroleum ether extract (1.3muL/mL) was about three times that of the chloroform extract (4.1muL/mL) at 24h post-treatment. At the concentrations of 500.0muL/mL, the median lethal time (LT(50)) of the petroleum ether extract and the chloroform extract was 8.4 and 9.6h, respectively.     

Performance of broiler chicks fed on diets containing urea ammoniated neem (Azadirachta indica) kernel cake.

1. The performance, nutrient utilisation, blood profile, carcase composition, gross pathology and sensory evaluation of meat from broiler chicks fed from 3 to 42 d on diets containing urea ammoniated neem (Azadirachta indica, A.juss) kernel cake (NKC) as a protein supplement to replace peanut meal (PNM), were evaluated. NKC was ammoniated with urea at 15 (UANKC 1) or 25 g (UANKC 2)/kg and incorporated into the test diets to replace either half (134.5 g/kg (L-UANKC 1) and 127.5 g/kg (L-UANKC 2), respectively) or all (269, g/kg (H-UANKC 1) and 255.0 g/kg (H-UANKC 2), respectively) of the nitrogen provided by the PNM. 2. The growth, food intake and efficiency of nutrient utilisation of the birds were comparable between the reference and L-UANKC 1 and 2 diets but were depressed on the other UANKC diets. 3. The retention of dry matter (DM), crude fibre (CF), nitrogen-free extract, total carbohydrate, gross energy, acid detergent fibre, calcium and phosphorus were similar among groups, except lower DM and higher CF and phosphorus retentions were noted in chicks fed on the H-UANKC 1, H-UANKC 2 and L-UANKC 2 diets. All the chicks were in positive nitrogen balance and percentage nitrogen retention did not differ between the reference and test diets. 4. Haemoglobin, total erythrocyte count and aspartate amino transferase activity were unaffected by diet, but total leucocyte count was higher in chicks fed on the H-UANKC 1 and 2 diets and alanine amino transferase activities were lower in chicks fed on the test diets. Blood urea increased as the amount of urea in the diets increased. 5. Most of the physico-chemical carcase characteristics from birds fed on the L-UANKC 2 were comparable to those from birds fed on the reference diet. No bitter taste was noticed in cooked meat from any diet by the sensory panel. 6. Incorporation of L-UANKC 2 was economical and responses were comparable to those observed on the reference diet. 7. It was concluded that NKC detoxified with 25 g urea/kg can economically and successfully replace half the nitrogen of PNM in broiler diets thereby mitigating the chronic shortage of costly oil cakes in developing countries.     

Trichostrongylus colubriformis and Ostertagia circumcincta resistant to levamisole, morantel tartrate and thiabendazole: occurrence of field strains

Field strains of Trichostrongylus colubriformis and Ostertagia circumcincta, designated PF4 and PF5 respectively, were recovered from a farm on which the sole use of levamisole over a preceding 12 year period led to the development of anthelmintic resistance. The results of field observations and preliminary critical trials in both Merino and crossbred sheep showed that both species have varying degrees of resistance to three major anthelmintics; levamisole, morantel tartrate and thiabendazole. Mean worm count reductions for adult T colubriformis (PF4) for therapeutic doses of morantel tartrate, thiabendazole and levamisole in crossbreds were 45.7 per cent, 97.3 per cent and zero respectively, and for Merinos 80.7 per cent, 88.3 per cent and 92.0 per cent respectively. Against O circumcincta the corresponding reductions for crossbreds were 51.4 percent, 95.4 per cent and 20.3 per cent and for Merinos, 52.5 per cent, 73.1 per cent and 29.8 per cent. There was no statistically significant difference in the responses of both parasite species to either levamisole or morantel. This result suggests that resistance to the two chemically unrelated drugs may be co-inherited.     

The effect of long-term feeding of fresh and ensiled cassava (Manihot esculenta) foliage on gastrointestinal nematode infections in goats

The benefit of long-term feeding of fresh or ensiled cassava foliage on gastrointestinal parasite in goats was evaluated. Eighteen male goats (15.15 ± 2.83kg and between 4–6months) were randomly allocated into three treatments supplemented with 200g of wheat bran head⁻¹ day⁻¹. All groups were fed ad-libitum on either grass (CO), fresh cassava (CaF) or ensiled cassava foliage (CaS). At the beginning of the trial, each goat was inoculated with 3000 L3 containing ≈50% Haemonchus contortus. Individual LWt, FEC and PCV were measured at weekly intervals for 10weeks. At the termination of the experiment all goats were slaughtered for worm recovery and enumeration. The goats in CaF and CaS had similar weight gains while those in CO lost weight (p < 0.05) through the trial. FEC in CaF and CaS were lower (p < 0.05) than CO during the patency of parasite infections, but there was no difference between CaF and CaS goats. PCV of all groups decreased from above 30% to around 25% at the end of the trial. The compositions of established worm burdens were mainly H. contortus (19–40%) and Trichostrongylus colubriformis (55–76%). TWB did not differ among the groups, however, CaS significantly reduced H. contortus burdens, as compared to CaF and CO (p ≤ 0.005). Thus, ensiled cassava foliage reduced the H. contortus population while the fresh foliage only reduced worm fecundity.     

Utilization of unpeeled cassava (Manihot esculenta Crantz) root meal supplemented with or without charcoal by broiler chickens

A 42‐day feeding trial was conducted using 480‐day‐old, male Marshall broilers to study the utilization of unpeeled cassava root meal (UCRM) supplemented with or without 6 g/kg charcoal. The experimental design was laid out in a 3 × 2 factorial arrangement of treatments having three inclusion levels of UCRM (0, 100 and 200 g/kg) with or without 6 g/kg charcoal supplementation. Each treatment consisted of 80 birds replicated eight times with 10 birds per replicate. Main effect of inclusion level of UCRM and supplementation of charcoal showed reduced (p < 0.05) final live weight, weight gain, feed intake and apparent crude protein digestibility of the birds with increasing inclusion levels of UCRM. Birds fed diets supplemented with charcoal showed higher (p < 0.05) final live weight, weight gain and feed intake than birds fed diets without charcoal. Supplementation of charcoal in diet containing 100 g/kg UCRM resulted in improved (p < 0.05) weight gain when compared with birds fed similar diet but not supplemented with charcoal. Broilers fed diet containing no UCRM but supplemented with charcoal had the highest overall (p < 0.05) final live weight and weight gain, while birds fed diet containing 200 g/kg UCRM supplemented with charcoal recorded the poorest (p < 0.05) final live weight and weight gain. Serum glutamate oxaloacetate transaminase (SGOT) and serum thiocyanate concentration increased (p < 0.05) with increasing dietary inclusion levels of UCRM. Dietary supplementation of charcoal resulted in increased (p < 0.05) concentration of serum glucose and cholesterol and reduced (p < 0.05) SGOT concentration. Birds fed diets containing UCRM had high (p < 0.05) serum thiocyanate concentration irrespective of dietary supplementation or not with 6 g/kg charcoal. In conclusion, supplementation of diet containing up to 100 g/kg UCRM with 6 g/kg charcoal showed improved weight gain without any deleterious effect on serum metabolites.     

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.     

In vitro and in vivo characterization of p-amino-phenethyl-m-trifluoromethylphenyl piperazine (PAPP), a novel serotonergic agonist with anthelmintic activity against Haemonchus contortus, Teladorsagia circumcincta and Trichostrongylus colubriformis

The neurotransmitter serotonin (5-hydroxy tryptamine or 5HT) regulates key physiological processes in nematodes such as locomotion and feeding. PAPP (p-amino-phenethyl-m-trifluoromethylphenyl piperazine) is a known agonist of the 5-HT(1Hc) receptor of the barber pole worm, Haemonchus contortus. In this study, PAPP was highly active against L3-stage larvae of H. contortus and Trichostrongylus colubriformis in an in vitro larval migration assay, with EC50 values of 9.36 and 11.8 microM, respectively, that were comparable to levamisole (10.2 microM) and superior to pyrantel (55.39 microM). When administered orally or subcutaneously to nematode infected gerbils, PAPP provided >99% efficacy against H. contortus and >98% efficacy against Teladorsagia circumcincta at 100 mg/kg, comparable to levamisole at 10 mg/kg. Drug titration revealed significant activity down to 50 mg/kg against these two species. Spectrum was limited, however, with somewhat lower efficacy (83%) in T. colubriformis infected gerbils at 100 mg/kg. Oral delivery of hydrochloride, acetate and phosphate salts of PAPP to nematode infected gerbils did not result in an increase in either potency or spectrum. The finding that PAPP exhibits significant anthelmintic activity suggests that the nematode-specific serotonergic system is a viable target for future anthelmintic discovery.     

Effect of feeding cassava (Manihot esculenta Crantz) root meal on growth performance,hydrocyanide intake and haematological parameters of broiler chicks

The effect of feeding cassava root meal on growth performance, hydrocyanide intake, haematological indices and serum thiocyanate concentration of broiler chicks was investigated using 300-day-old male broilers. There were five dietary treatments arranged in a 2?×?2?+?1 factorial arrangement of two processing methods of cassava root (peeled and unpeeled) included at two levels (100 and 200 g/kg) plus a control diet (maize-based diet, containing no cassava root). Each treatment was replicated six times with ten birds per replicate. The feeding trial lasted for 28 days. Control-fed birds had the highest overall (P?P?P?P?P?P?P?P?P?P?P?P?P?P?相似文献   

The influence of a heavy infection with sensitive and resistant strains of Ostertagia circumcincta and with Trichostrongylus colubriformis on the pharmacokinetics of febantel in lambs

Plasma concentrations of febantel and its major metabolites, fenbendazole, oxfen-dazole and fenbendazole sulphone, were determined after oral administration of 7.5 mg/kg febantel in lambs before and 28 days after infection with 100 000 L3 larvae of a benzimidazole (BZ)- sensitive or BZ-resistant strain of Ostertagia circumcincta or with 75 000 L3 larvae of a BZ- sensitive Trichostrongylus colubriformis strain. The febantel concentrations were always low, and in only a few samples were higher than the limit of detection. A mean decrease in the area under the curve ( AUC ) for the three metabolites of 10.2%, 16.4% and 4.9% in lambs infected, respectively, with BZ-sensitive O. circumcincta , BZ-resistant O. circumcincta and T. colubriformis was observed. The C _max for all the metabolites was higher in the BZ-sensitive O. circumcincta group than in the naive sheep, while the T _max occurred earlier. The C _max and the T _max values for all the metabolites were lower in the BZ-resistant O. circumcincta group than in their own naive controls. In the T. colubriformis group the C_max values of the metabolites were lower and the T_max occurred much later.     

Development and spatial distribution of the free-living stages of Teladorsagia circumcincta and Trichostrongylus colubriformis on pasture: A pilot study

Abstract

AIMS: To measure the development of Teladorsagia (=Ostertagia) circumcincta and Trichostrongylus colubriformis eggs to third-stage infective larvae (L3) at different times of the year. Also, to measure the spatial distribution of L3 across herbage, soil and faeces, in order to assess whether spatial issues could be important in larval dynamics on pasture.

METHODS: Field plots were contaminated with sheep faeces containing approximately 20,000 eggs of each of T. circumcincta and T. colubriformis on five separate occasions, viz 01 December 1996 (summer), 18 March 1998 (autumn), 17 June 1998 (winter), 15 October 1998 (spring), and 23 July 1999 (winter). Replicate plots (n=10) were harvested at intervals for up to 12 months after deposition of faeces, and the number and distribution of L3 were measured. Larvae were sampled from faeces (where these remained), herbage, and three soil zones to a depth of 145 mm.

RESULTS: There were large differences between contamination dates in the percentage of eggs that developed to L3. For both species the highest percentage development was for eggs deposited in December (7.8% and 25.9% for T. circumcincta and T. colubriformis, respectively) and the lowest for June (0.4% and 0.03% T. circumcincta and T. colubriformis, respectively). Development in winter was often delayed, and this was always associated with a low yield of larvae, probably due to compounding mortalities associated with long periods of exposure to low temperatures.

The relative distribution of L3 present on herbage, in faeces or in the soil varied between sampling times. However, overall the most L3 were recovered from soil (74% and 66% for T.circumcincta and T. colubriformis, respectively, averaged over all samples), and the lowest recoveries were from the herbage.

CONCLUSIONS: Although the data are limited, the results indicated that the highest percentage of eggs developed to infective larvae in summer and only minimal development occurred in winter. The data do not support the view that substantial contamination of pastures with sheep parasites occurs over winter. Large numbers of larvae were recovered from soil, which indicates that, assuming they can subsequently migrate onto herbage, soil is a potentially important reservoir ofinfective larvae in New Zealand. Therefore, the spatial distribution of L3 on pasture may affect both the dynamics and transmission of parasite populations. Further work on both these issues is warranted.     

Feeding potential of cassava (Manihot esculenta Crantz) peels ensiled with Leucaena leucocephala and Gliricidia sepium assessed with West African dwarf goats

Cassava peels (CaPe) were ensiled in mixtures with Gliricidia sepium and Leucaena leucocephala, and the utilization of the mixed silages by West African dwarf (WAD) goats was assessed. Five silages were composed, comprising of 100 % ensiled CaPe (control), CaPe?+?G. sepium 2:1 (w/w; 2CGS), CaPe?+?G. sepium 1:1 (w/w; CGS), CaPe?+?L. leucocephala 2:1 (w/w; 2CLL) and CaPe?+?L. leucocephala 1:1 (w/w; CLL). All diets were supplemented with molasses (40 g/kg) before ensiling which lasted 3 months. Fifteen WAD goats (8.01?±?0.12 kg body weight) were fed one of the experimental diets (50 g/kg body weight) for 8 weeks. The control had the lowest hydrocyanic acid content (0.05 mg/kg DM), while others ranged from 6.2 to 81.3 mg/kg. Condensed tannin concentration ranged from 1.7 to 8.4 mg/kg DM, while mimosine levels were 11.6 and 12.4 mg/kg DM in 2CLL and CLL, respectively. After fermentation, all silages showed low pH (<4.5) and were different (P?<?0.05) in the lactic, acetic and butyric acid concentrations. Ratio of foliage supplementation influenced DM intake (P?<?0.05). Daily weight gains ranged from 17 (CLL) to 24 g/day in control. The digestibility coefficients of nutrients and fibre fractions differed (P?<?0.05) among diets. The values for packed cell volume, haemoglobin, red blood cells, neutrophils, lymphocytes and monocytes were also different (P?<?0.05) across the dietary treatments. Ensiling CaPe with foliages of G. sepium and L. leucocephala can be recommended for feeding WAD goats especially during the dry spells when there is little or no available forage for the animals.