In vitro acaricidal effect of tannin-rich plants against the cattle tick Rhipicephalus (Boophilus) microplus (Acari: Ixodidae)

The objectives of this study were to evaluate the in vitro acaricidal effects of lyophilized extracts of four tannin rich plants (Acacia pennatula, Piscidia piscipula, Leucaena leucocephala and Lysiloma latisiliquum) against diverse stages of Rhipicephalus (Boophilus) microplus, and to asses whether tannins were involved in the acaricidal effect using polyethylene glycol (PEG) to block tannins. Larval immersion (LIT) and adult immersion (AIT) tests were used to evaluate the acaricidal effect of each of the lyophilized extracts against larval and adult stages of R. microplus respectively. Larvae and adult ticks were exposed to increasing concentrations of each plant extract (0, 1200, 2400, 4800, 9600 and 19,200 μg ml(-1)) for 10 min. Larval mortality was recorded at 48 h post-incubation. Adult mortality was recorded daily over 14 days, at which point their reproductive efficiency was evaluated. PEG was added to the extracts to verify whether tannins were involved in the acaricidal effect. The effect on egg laying inhibition and larval mortality was analyzed using the GLM procedure in SAS. A Kruskal-Wallis test was used to assess the effect of PEG on LIT results. Calculation of the lethal concentration 50 (LC50) was performed using a probit analysis. All extracts reduced the viability of R. microplus larval stages (P<0.001), and viability was restored with the addition of PEG suggesting an important role of tannins in the acaricidal effect (P<0.001). The LC50 values of L. latisiliquum and P. piscipula plant extracts were 6.402 and 2.466 μg ml(-1). None of the tannin-rich plant extracts affected adult mortality (P>0.05). Lysiloma latisiliquum extract inhibited egg hatching of R. microplus (P<0.01). Tannin-rich plant extracts from A. pennatula, P. piscipula, L. leucocephala and L. latisiliquum showed potential acaricidal activity. Further in vivo studies are needed to confirm this finding.     

Intake and Selection of Goats Grazing Heterogeneous Vegetation: Effect of Gastrointestinal Nematodes and Condensed Tannins

Feeding behavior is a multifactorial process with pivotal relevance for the maintenance and survival of ruminants. This study evaluated the effect of gastrointestinal nematodes (GIN), condensed tannin (CT) content of plants, and their possible interaction on the intake and selection of feed resources by Criollo goats in a heterogeneous vegetation. An 8-wk field experiment was conducted in the tropical deciduous forest (TDF) during the rainy season with adult Criollo goats (35.1 ± 6.7 kg body weight [BW]). Goats were assigned to four groups (n = 6): 1) MOX: suppressive treatment with moxidectin (Cydectin 0.4 mg/kg BW subcutaneous every 4 wk), 2) INF: with natural GIN infection, 3) MOX + PEG: treated with moxidectin and dosed with a CT-neutralizing agent (50 g of polyethylene glycol [PEG] diluted in 50 mL of water, per os), and 4) INF + PEG: with natural GIN infection and PEG dosage. Intake was measured using a direct observation method in a 2.2-ha TDF plot. Plant availability was obtained from 30 exclusion quadrants (2 × 2 m). Selection was estimated with the Cheeson index using the ratio of consumed plants and their availability by grouping plant species as shrubs and herbs (with high CT [> 10%], medium CT [> 3% to < 10%], or low CT [< 3%]), vines, and grasses. Neither GIN infection nor CT neutralization influenced intake or selection of feed resources. All groups showed high selection toward grass species (P < 0.001), which may represent a strategy to reduce intake of crude protein and CT from shrubs and herbs. In conclusion, GIN infection and CT neutralization do not influence intake or selection of goats with browsing experience in the TDF. Selection toward grass suggested a trade-off between nutrient harvest optimization and the health risk of GIN infective larvae consumption. Thus, it is necessary to revalorize the TDF as a sustainable feed resource for goats.     

In vitro larval migration and kinetics of exsheathment of Haemonchus contortus larvae exposed to four tropical tanniniferous plant extracts

As for some temperate forage, some tropical tanniniferous plants (TTP) from browsing might represent an alternative to chemical anthelmintic. The anthelmintic effect of four TTP (Acacia pennatula, Lysiloma latisiliquum, Piscidia piscipula, Leucaena leucocephala) on Haemonchus contortus was measured using two in vitro assays. First, the effects of increasing concentrations of lyophilized extracts (150, 300, 600, 1200 microg/ml PBS) were tested on H. contortus larvae (L(3)) using the larval migration inhibition (LMI) test. An inhibitor of tannin, polyvinyl polypyrrolidone (PVPP), was used to verify whether tannins were responsible for the AH effect. Secondly, the effects of extracts on larval exsheathment were examined. Larvae (L(3)) were in contact with extracts (1200 microg/ml) for 3h, and then were exposed to an artificial exsheathment procedure with observations of the process at 10 min intervals. A general lineal model (GLM) test was used to determine the dose effect in the LMI test and the difference of the percentage of exsheathed larvae between the control and the treatment groups. A Kruskal Wallis test was used to determine the effect of PVPP on LMI results. The LMI test showed a dose-dependent anthelmintic effect for A. pennatula, L. latisiliquum and L. leucocephala (P<0.01), which disappeared after PVPP addition, confirming the role of tannins. No effect was found for P. piscipula on H. contortus in the LMI test. However, all four plant extracts interfered with the process of L(3) exsheathment which might be involved as a mechanism of action of tannins on H. contortus larvae. A. pennatula, L. latisiliquum and L. leucocephala could be used as an anthelmintic for the control of H. contortus after confirmation based on in vivo studies.     

Comparing the sensitivity of two in vitro assays to evaluate the anthelmintic activity of tropical tannin rich plant extracts against Haemonchus contortus

The present trial aimed at comparing the sensitivity of two in vitro methods, i.e. the larval migration inhibition assay (LMIA) and the larval exsheathment inhibition assay (LEIA), to evaluate the anthelmintic (AH) properties of tannin-rich plant extracts against Haemonchus contortus infective larvae. The two assays were applied on the same batch of H. contortus infective larvae exposed to water/acetonic extracts obtained from four tropical plants with different tannin contents: Acacia gaumeri, Brosimum alicastrum, Havardia albicans and Leucaena leucocephala. Increasing concentrations (0, 75, 150, 300, 600, 1200 μg/ml PBS) of lyophilized extracts were used in both in vitro assays. A general lineal model test was used to determine the dose-effect in the LMIA or the difference in the percentage of exsheathed larvae between the respective control and treated groups. The LMIA showed a dose-dependent AH effect for H. albicans (P<0.001) and A. gaumeri (P<0.05), but not for L. leucocephala and B. alicastrum. In contrast, the exsheathment process was significantly affected by all doses of H. albicans and A. gaumeri extracts and a significant dose-dependent effect was found for B. alicastrum and L. leucocephala. Calculation of lethal dose (LD) was possible with LEIA using B. alicastrum and L. leucocephala but not with H. albicans and A. gaumeri as the lowest tested concentration was achieving more than 50% inhibition. Calculation of LD with the LMIA results was not feasible. These results suggest that tannin-rich plant extracts are more potent inhibitors of the exsheathment of H. contortus L(3) larvae than their motility. This information underlines the difference of sensitivity between methodological procedures to evaluate the AH properties of plant extracts on the same nematode stage.     

Productive performance and urinary excretion of mimosine metabolites by hair sheep grazing in a silvopastoral system with high densities of Leucaena leucocephala

The aim of this study was to evaluate daily weight gain (DWG), total dry matter (DM) intake, rumen degradability of forage, and urinary excretion of mimosine metabolites by hair sheep in a silvopastoral system with high densities of Leucaena leucocephala. A completely randomized design was carried out with two treatments: treatment 1 (T1) silvopastoral system with leucaena at a density of 35,000 plants/ha and treatment 2 (T2), leucaena at a density of 55,000 plants/ha. Leucaena was associated with tropical grasses Panicum maximum and Cynodon nlemfluensis. Twenty-four male Pelibuey lambs of 23.2?±?3.4?kg live weight (LW) were used (12 lambs per treatment). Results showed differences (P?<?0.05) in DWG of T1 (106.41?±?11.66?g^?1 sheep^?1) with respect to that of T2 (81.33?±?11.81?g^?1 sheep). Voluntary intake was higher in lambs from T1 (83.81?±?04.07?g DM/kg LW^0.75) with respect to that from T2 (71.67?±?8.12?g DM/kg LW^0.75). There was a difference in color of urine between sheep of T1 and T2, the latter giving positive results for the presence of metabolites derived from mimosine (3-4 dihydroxypyridine and 2-3 dihydroxy pyridone). Rumen degradability of DM of L. leucocephala was higher (P?<?0.05) compared to that of P. maximum and C. nlemfluensis (72.94?±?0.40 vs. 67.06?±?1.50 and 63.25?±?1.51?%, respectively). It is concluded that grazing at high densities of L. leucocephala affects daily weight gain of hair sheep, possibly due to ingestion of high amounts of mimosine which may exert an adverse effect on voluntary intake.     

Gastrointestinal nematode infection does not affect selection of tropical foliage by goats in a cafeteria trial

It is important to determine whether gastrointestinal nematodes (GINs) affect foliage choice of goats leading to confirm the expression of a self-medication behavior. This study investigated the effect of GIN infection on tropical foliage selection by goats. During experimental stage 1 (10 days), goats had a natural mixed GIN infection, and at stage 2 (10 days), goats were treated with effective anthelmintics to maintain them free of GIN infection. During stage 1 the twelve adult goats (32?±?2.3 kg live weight [LW]) were assigned to three groups (n?=?4) according to their initial GIN infection status: HI group, with fecal egg count (FEC) between 1450 and 2150 eggs per g/feces (EPG); MI group, medium FEC (592–1167 EPG); and the NI group, free from GIN infection. Fresh foliage of four tropical plants were offered to goats ad libitum for 1 h daily: Gymnopodium floribundum (high condensed tannin [CT] content, 37–40 %), Mimosa bahamensis (medium CT content, 16–17 %), Leucaena leucocephala (low CT content, 3–5 %), and Viguiera dentata (negligible CT content, 0.6–0.9 %). Jacobs’ selection indexes (JSIs) were estimated for the experimental foliage based on dry matter (DM), CT, or crude protein (CP) intake. During both study stages, individual fecal egg counts were estimated. The JSI patterns of different plant species, based on DM, CT, or CP, were similar irrespective of infection level during stage 1 (HI, MI, and NI) or no GIN infection (stage 2). Thus, irrespective of GIN infection, goats actively selected M. bahamensis (high CT, low CP content) and V. dentata (negligible CT, high CP content) but avoided G. floribundum (high CT, low CP content) and L. leucocephala (medium CT and high CP content). Thus, natural GIN infection did not influence goats’ foliage selection.     

The effect of supplementary feeding on the resilience and resistance of browsing Criollo kids against natural gastrointestinal nematode infections during the rainy season in tropical Mexico

The objective was to determine the effect of supplementary feeding on the resilience and resistance of Criollo kids against natural gastrointestinal nematode (GIN) infections, when browsing native vegetation during the wet season in tropical Mexico. Thirty-four 2-month old Criollo kids, raised nematode free, were included at weaning in a 22-week trial. The kids were placed into four groups. Two groups of 8 kids were offered 100g/day soybean and sorghum meal (26%:74%, respectively fresh basis) (treated/supplemented (T-S) and infected/supplemented (I-S)). Two groups remained with no supplement for the duration of the trial (infected/non-supplemented (I-NS) (n = 10) and treated/non-supplemented (T-NS) (n = 8)). Kids in groups T-S and T-NS were drenched with 0.2mg of moxidectin/kg body weight orally (Cydectin, Fort Dodge) every 28 days. Groups I-S and I-NS were naturally infected with GIN. The animals browsed native vegetation (for an average of 7h/day) together with a herd of 120 naturally infected adult goats. Cumulative live weight gain (CLWG), packed cell volume (PCV), haemoglobin (Hb), total plasma protein and plasma albumin were recorded every 14 days as measurements of resilience. Resistance parameters (faecal egg counts (FEC) and peripheral eosinophil counts (PEC)) were also measured. Bulk faecal cultures were made for each group every 28 days. Every month a new pair of tracer kids assessed the infectivity of the vegetation browsed by the animals. The T-S group had the highest CLWG, PCV and Hb compared to the other three groups (P < 0.001). The I-S and T-NS group had similar mean CLWG and PCV (P > 0.05), while the I-NS group had the poorest CLWG, PCV and Hb (P < 0.001). The PEC of supplemented kids (I-S and T-S) was higher than in the I-NS and T-NS kids (P < 0.05). No effect of supplementary feeding was found in the FEC. Tracer kids and faecal cultures showed that kids suffered mixed infections with Haemonchus contortus, Trichostrongylus colubriformis and Oesophagostomum columbianum. Supplementary feeding improved resilience of browsing Criollo kids against natural GIN infections and was economically feasible. Improved resistance was also suggested by the PEC but was not confirmed in the FEC.     

Effects of four tropical tanniniferous plant extracts on the inhibition of larval migration and the exsheathment process of Trichostrongylus colubriformis infective stage

The anthelmintic (AH) effect of Acacia pennatula, Leucaena leucocephala, Lisyloma latisiliquum and Piscidia piscipula was evaluated in the infective larvae (L(3)) of Trichostrongylus colubriformis. Different concentrations of lyophilized extracts were tested using the larval migration inhibition (LMI) test. An inhibitor of tannins (the polyvinyl polypyrrolidone [PVPP]) was used to verify whether these compounds were responsible for the AH effects. Then, the effect of extracts on larval exsheathment was examined by observing the exsheathment process at 10-min intervals for 70 min. The LMI test showed a dose-dependant AH effect for A. pennatula, L. leucocephala and L. latisiliquum (P<0.01), but not for P. piscipula. The restoration of L(3) migration to values similar to those of controls after the addition of PVPP, indicates that tannins are involved in AH effects. Trichostrongylus colubriformis exsheathment was partially or totally blocked by the four plants extracts. Tropical tanniniferous plants evaluated in the current study may have potential as AH for the control of T. colubriformis if in vivo investigations indicate useful effects.     

Assessing the efficacy of Duddingtonia flagrans chlamydospores per gram of faeces to control Haemonchus contortus larvae

The aims were (a) to quantify the number of Duddingtonia flagrans chlamydospores per gram of faeces (CPG) recovered from sheep administered with different oral doses and, (b) to describe the relationship between CPG and eggs per gram of faeces (EPG) on the efficacy to reduce Haemonchus contortus infective larvae. Three doses of chlamydospores per kg BW were orally administered during seven days: (T1) non treated control group, (T2) 1 × 10⁶, (T3) 2.5 × 10⁶ and (T4) 5 × 10⁶. Three lambs, infected with H. contortus, were used per group. Faeces were obtained from the rectum of each lamb during the fungal administration period (days 0–6) and for six days after that period. Four coproculture replicates were made from each animal in days 2, 4, 6, 8 and 10. A higher chlamydospore dose produced higher CPG in faeces (p < 0.05), but a clear dose dependent effect was not found either in the larvae reduction or in the CPG:EPG ratio. When ratios were re-analyzed, independently of the treatment groups of origin, a better efficacy was obtained with a ratio from 5 to 10 CPG:EPG and a higher ratio (>10 per egg) showed a lower reduction efficacy (p < 0.05). The binomial analysis showed that for each unit of increment in CPG:EPG ratio there was a reduction of larvae number until a point (between 5 and 10 CPG:EPG) where no further reduction was detected. The surface response test indicated that the number of larvae was reduced by CPG until possible saturation. The highest CPG:EPG ratios did not necessarily improve efficacy of D. flagrans.     

Tocopherol Accumulation and Temporal Expression Analysis of VTE1 and VTE5 Gene Family in Fruit of Two Contrasting Avocado Genotypes

Plant Foods for Human Nutrition - α-tocopherol is found in high concentrations in avocado fruit mesocarp, however, its accumulation and genetic control during maturation and ripening has not...