Criollo goats limit their grass intake in the early morning suggesting a prophylactic self-medication behaviour in a heterogeneous vegetation.

The present study compared the feeding behaviour of goats in the early morning (EM = 7:00-8:30 a.m.) and late morning (LM = 9:30-11:00 a.m.) in response to their natural gastrointestinal nematode (GIN) infection status. Twelve female adult goats (37 ± 7.7 kg live weight (LW); 5 ± 1 years) with browsing experience in the tropical deciduous forest were divided into two groups (n = 6): INF group, with natural GIN infection, and non-infected (NI) group, dewormed with moxidectin (0.4 mg/kg LW subcutaneous). Feeding behaviour (dry matter intake (DMI)) of two resource types (grasses vs. shrubs + herbs) was estimated by direct observation for 4 weeks on two grazing moments (EM vs. LM). Environmental temperature and relative humidity at pasture level were measured twice weekly. The GIN egg counts and goats' LW were measured on days 0, 14 and 28. Temperature (mean ± standard deviation) at EM (26.2 ± 1.5 °C) was lower than at LM (38.7 ± 1 °C; P < 0.01). Humidity was higher on the EM (85.1 ± 2.6%) compared to LM (60.4 ± 5.6%; P < 0.01). Irrespective of the infection status, goats consumed similar amounts of grass and shrubs + herbs during EM (P > 0.05). On the other hand, the experimental groups consumed more grass than shrubs + herbs during LM (P < 0.05). The latter suggested prophylactic behaviours strategies to (a) avoid GIN infective larvae, (b) balance the protein:energy ratio of the diet and (c) avoid saturation of detoxification pathways for the secondary compounds consumed from shrubs + herbs. The constant consumption of shrubs + herbs during the study may have reduced the GIN egg count of the INF group.

Gymnopodium floribundum fodder as a model for the in vivo evaluation of nutraceutical value against Haemonchus contortus.

This study validated a protocol to identify the nutraceutical value of Gymnopodium floribundum (GF) foliage based on its effects on sheep nutrition, production and health and its impact on adult Haemonchus contortus. Thirty animals (17.8 ± 3.44 kg BW) were distributed into five experimental groups (n = 6). Groups T1-T3 received feed including 20%, 30% and 40% GF content, respectively, and group T4 received feed with 0% GF. Groups T1-T4 were all infected with 6000 H. contortus infective larvae (L3). Group T5 included six worm-free lambs fed a diet without GF. Feed intake, dry matter digestibility (DMD) and organic matter digestibility (OMD) were measured, in addition to the faecal excretion of eggs per gram (EPG) and total faecal egg count (TFEC) of H. contortus. On day 39 post-infection, lambs were humanely slaughtered to recover and count adult parasites and assess the length and fecundity of female worms. Higher levels of GF in the diet reduced DMD and OMD (P < 0.05), with the lowest values in T3 (40% GF) (P < 0.05). Body weight gain was similar for all groups (0.18 to 0.2 kg/day; P > 0.05). EPG and TFEC were lower in T1 and T3 than in T4 (P < 0.05), and the number of adult female worms was lowest in T3 (P < 0.05). In conclusion, GF leaf meal can be considered a nutraceutical when included at 40% (T3) in the diet of lambs, based on its ability to decrease the EPG and TFEC of H. contortus by reducing the female worm burden. A 40% inclusion level had no negative impact on lamb diet intake, production and health, although a reduction of digestibility was observed.

The worm burden of tracer kids and lambs browsing heterogeneous vegetation is influenced by strata harvested and not total dry matter intake or plant life form.

This study assessed the effect of total dry matter intake (DMI), plant life form and strata harvested on the gastrointestinal nematode (GIN) worm burdens of tracer kids and lambs browsing heterogeneous vegetation during the rainy season (August-November). The rainy season was divided into 6 2-week periods (P1-P6), and environmental conditions (rainy days, rainfall, temperature and humidity) were recorded daily. Five pairs each of tracer kids and tracer lambs raised free of GIN infections were used. Every 15 days, different pairs of kids and lambs were introduced to a 2.2-ha plot and co-grazed with a flock of 30 sheep and 70 goats for a period of 3 weeks. Feeding behaviour of each pair of tracers was measured in weeks 2 and 3. The continuous bite monitoring method was used to estimate total DMI, DMI of plant life forms and DMI from plants of different strata. After each 3-week period, the tracer pair was maintained indoors for 28 days and necropsied on day 29 to recover the worm burden. The feeding behaviour of the tracers was compared between periods (P2-P6) and between kids and lambs. The differences in the worm burdens of the tracers between periods were not associated with total DMI or DMI from plant life forms. Worm burdens were highest during P5 and P6 in tracer kids and lambs (P < 0.05), suggesting a build-up of infective larvae in the tropical deciduous forest (TDF). The lower worm burdens of tracer kids compared with lambs (P < 0.05) seemed to be associated with less low- and more medium-stratum ingestion.

An in vitro approach to evaluate the nutraceutical value of plant foliage against Haemonchus contortus.

Nutraceutical plants provide nutrients for the animal as well as secondary compounds that can affect the biology and survival of gastrointestinal nematodes (GIN). Current screening of nutraceutical plants is based on in vitro evidence of anthelmintic (AH) activity against different life stages of GIN, but nutritional information is omitted or scarce. This study proposes an integral in vitro screening protocol to identify the nutraceutical value of the foliage from plant species consumed by small ruminants, using Haemonchus contortus as a biological model. The leaves from Acacia collinsii, A. pennatula, Bunchosia swartziana, Gymnopodium floribundum, Havardia albicans, Leucaena leucocephala, Lysiloma latisiliquum, Mimosa bahamensis, Piscidia piscipula, and Senegalia gaumeri were evaluated for their chemical composition and in vitro digestibility. Acetone:water extracts (70:30) from leaves of each plant were evaluated using the egg hatch assay and larval exsheathment inhibition assay. Respective effective concentrations 50% (EC50) were determined for each assay. The ten plant species showed good nutritional value for ruminants, including crude protein (> 10%), metabolizable energy (> 2.9 MJ/kg DM), and varied CT content (from 1.0 to 37.6%). The best AH activity against H. contortus eggs (EC50 = 401.8 µg/mL) and L3 (EC50 = 83.1 µg/mL) was observed for S. gaumeri extract. Although all the plant species showed in vitro nutraceutical potential, the leaves of S. gaumeri had the best values. The proposed in vitro protocol showed to be useful for the integral assessment of the nutraceutical potential of different plant species as it included the nutritional value and the AH activity against eggs and L3 in the selected plant species.

Is there a negative association between the content of condensed tannins, total phenols, and total tannins of tropical plant extracts and in vitro anthelmintic activity against Haemonchus contortus eggs?

In vitro studies using plant extracts suggest a relationship between their polyphenol contents and their anthelmintic (AH) activity against Haemonchus contortus. High polyphenol content appears to increase the efficacy of plant extracts against H. contortus as assessed by the larval exsheathment inhibition assay (LEIA) while appearing to reduce the AH efficacy measured using the egg hatch assay (EHA). In addition, some plants lack AH activity. Therefore, the present study investigated the relationship between the contents of condensed tannins (CT), total phenols (TP), and total tannins (TT) in methanol:water extracts (70:30) obtained from ten tropical plant species consumed by small ruminants as well as their AH activity against H. contortus evaluated by LEIA and EHA. Extracts of Acacia collinsii, Lysiloma latisiliquum, Havardia albicans, Senegalia gaumeri, Mimosa bahamensis, Piscidia piscipula, Acacia pennatula, Gymnopodium floribundum, Leucaena leucocephala, and Bunchosia swartziana were examined. Positive correlations were found between the effective concentration 50% (EC50) (EHA) of extracts and their CT (r = 0.6809, P < 0.05, n = 10) and TP (r = 0.9152, P < 0.05, n = 10) content, suggesting that their concentration negatively affected AH activity against eggs. Based on the LEIA, there was no significant association between the EC50 and the CT, TP, or TT of all extracts evaluated. Thus, if sheep and goats consume a complex feed mixture with high amounts of CT, TP, and TT, it might be difficult to observe an AH effect against H. contortus egg hatching. However, the AH effect upon L3 establishment might be feasible.

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.

Duddingtonia flagrans chlamydospores in nutritional pellets: effect of storage time and conditions on the trapping ability against Haemonchus contortus larvae.

The study evaluated the effect of storage time and conditions of nutritional pellets (NP) containing Duddingtonia flagrans chlamydospores on its in vitro trapping ability against Haemonchus contortus L3 larvae. The treated batch (200 NP) contained 4 × 106 chlamydospores of the FTH0-8 strain, whereas the control batch (200 NP) was produced without spores. Both NP batches were exposed to four experimental storage conditions: (T1) shelves (indoors); (T2) refrigeration (4°C); (T3) outdoors under a roof; and (T4) 100% outdoors. Each group comprised 48 NP with spores and 48 NP without spores (control). The ability of D. flagrans spores to trap H. contortus L3 larvae was evaluated for 8 weeks for each storage condition. For that purpose, six randomly selected NP with spores were compared to their respective control NP. Each NP was individually crushed. The crushed material (1 g) was placed on the surface of a 2% water agar plate with 200 H. contortus L3 larvae. Plates were sealed and were incubated at room temperature for 8 days. The whole content of every plate was transferred to a Baermann apparatus to recover the remaining larvae. There was a clear larval reduction in the NP with spores, compared to the respective control NP in the four storage conditions (P< 0.05). The mean reductions ( ± SEM) of the storage conditions were 67 ± 4.9 (T2), 77 ± 6.1 (T1), 81.5 ± 3.8 (T4) and 82.1 ± 2.5 (T3). Larval reductions were similar at all times and were not affected by storage conditions or storage time (R 20.05). The long-term shelf-life of the chlamydospores in the NP suggests that this spore dosage technology is a viable option.

Estimating Apparent Nutrient Digestibility of Diets Containing Leucaena leucocephala or Moringa oleifera Leaf Meals for Growing Rabbits by Two Methods.

This study aimed to evaluate the nutrient digestibility of growing rabbits fed diets with different levels of either Leucaena leucocephala (LLM) or Moringa oleifera (MOLM) leaf meals and also to compare total collection and TiO2 marker methods for estimating digestibility. A total of 30 California growing rabbits (1.81±0.19 kg live weight on average) were randomly distributed into five experimental groups of six rabbits each and were housed in individual cages. The groups were control, 30% LLM, 40% LLM, 30% MOLM, and 40% MOLM. All groups received pelleted diets for two weeks; diets also contained 4 g/kg titanium dioxide as dietary marker. Daily feed intake was recorded during the whole experimental period and total feces were collected daily and weighed individually during four days. The results showed that there were no difference (p>0.05) in feed, dry matter (DM), organic matter (OM), crude protein (CP), digestible energy, and crude fiber (CF) intake between the control group and the other experimental groups. The apparent digestibility values of DM, OM, CP, CF, acid detergent fiber, and gross energy were the highest for control group (p = 0.001), meanwhile MOLM diets had generally higher nutrient digestibility coefficients than LLM diets. Increasing the inclusion level of leaf meal in the diet from 30% to 40% improved the digestibility of CF from 45.02% to 51.69% for LLM and from 48.11% to 55.89% for MOLM. Similar results for apparent nutrient digestibility coefficients were obtained when either total collection or indigestible marker method was used. In conclusion, the digestibility of MOLM containing diets were better than LLM diets, furthermore TiO2 as an external marker could be used as a simple, practical and reliable method to estimate nutrients digestibility in rabbit diets.

Scanning electron microscopy of Haemonchus contortus exposed to tannin-rich plants under in vivo and in vitro conditions.

The structural changes induced in adult Haemonchus contortus after in vitro and in vivo contact with a tannin-rich (TR) plant, either tzalam (Lysiloma latisiliquum) or sainfoin (Onobrychis viciifolia), were assessed using scanning electron microscopy (SEM). All the worms used in the study were adult females. The Haemonchus adult worms were obtained from the abomasum of infected donor goats. Adult H. contortus were kept in contact with the extracts of TR plants for 24h for the in vitro assays and were compared to worms maintained in PBS (control). For the in vivo studies, the adult H. contortus parasites were obtained from artificially infected goats. All the goats were fed a tannin-free diet until D27 post-infection when infection was patent. On D28 some goats were fed fodder of one of the TR plants for seven consecutive days. Thus, their H. contortus were in contact with TR fodder within the gastrointestinal tract. The control worms were obtained from goats fed only a tannin-free diet. In the in vitro assays and in vivo studies, the SEM observations revealed structural alterations in the worms after contact with TR plants when compared to the control worms (i.e.: longitudinal and transversal folds and thicker cuticular ridges, material aggregates around the buccal capsule and/or vulva or anus). The main changes concerned the cuticle and the buccal area. The structural changes found in the worms exposed to TR plants might affect their motility and nutrition with possible consequences on their reproduction, as suggested by previous in vivo trials.

Impact of Dietary Condensed Tannins and Haemonchus contortus Infection in Growing Sheep: Effects on Nutrient Intake, Digestibility, and the Retention of Energy and Nitrogen.

PURPOSE: A controlled study evaluated the effect of condensed tannins (CT) from Gymnopodium floribundum leaf meal (GF), infection with Haemonchus contortus (I) and their interaction, on feed intake, diet digestibility and retention of N (NR) and energy (ER) in hair sheep lambs. METHODS: Thirty-six, worm-free hair sheep lambs (14.9 ± 1.56 kg body weight) were housed in metabolic cages. Eighteen animals were infected with 6000 H. contortus L3, while other 18 lambs were kept non-infected. On day 28th post-infection (PI), infected lambs were assigned to three diet groups: a diet without GF (I-NONGF), a diet with GF (I + GF) and a diet with GF + polyethylene glycol (PEG) (I + GF + PEG). Non-infected (NI) lambs were assigned to similar diet groups: NI-NONGF, NI + GF and NI + GF + PEG. The packed cell volume (% PCV), ante-mortem faecal egg counts and post-mortem worm burdens were also evaluated. RESULTS: Infection did not affect digestibility, NR and ER. Meanwhile, CT intake from the GF diet reduced the digestibility of dry matter, organic matter and crude protein, as well as NR, compared to lambs consuming the NONGF and GF + PEG diets (P < 0.05). Although, the digestible energy was similar between lambs consuming NONGF and GF + PEG diets, the ER was higher for lambs consuming the control NONGF diet. Diets did not affect the PCV, or the ante-mortem and post-mortem parasitological variables. CONCLUSION: The costs on N and energy metabolism were mainly associated with the CT content of the GF diet, but other features of the diet such as the high lignin content, seemed to affect animals consuming GF meal. Meanwhile, the H. contortus infection had a non-significant impact.

Effect of three feeding levels on the pathogenesis and establishment of Haemonchus contortus in parasite-naïve Pelibuey hair sheep lambs during their first infection.

This study evaluated the effect of three feeding levels on the pathogenesis and establishment of H. contortus upon the first infection of parasite-naïve Pelibuey hair sheep lambs. Forty-two 6-month-old hair sheep lambs (24 ± 4 kg) raised parasite free from birth were used. The lambs were assigned to 3 groups (n = 14), and each was fed a diet designed for different daily weight gain (DWG): 75 g/d (Diet 1), 125 g/d (Diet 2) and 200 g/d (Diet 3). After four weeks of diet adaptation, 10 lambs/group were infected with 450 L3H. contortus/kg BW (infected), and 4 lambs/group were kept parasite-free (NInf). DWG, hematocrit (Ht), hemoglobin (Hb), peripheral eosinophils (EOS), IgG concentration against H. contortus, and eggs per gram (EPG) of feces were measured in each lamb from day 14 before infection until day 29 postinfection (PI). On day 29 PI, the lambs were slaughtered to determine the total number of adult parasites (TAW), the length of the female worms, and the number of eggs in utero (EIU). Each group reached the expected DWG (P = 0.001), and there was no effect of infection or the diet × infection interaction. Ht was lower in infected lambs than in NInf lambs, and this difference was significant for animals on Diets 1 and 2 (P = 0.044). From day 14 PI onward, Hb was lower in the infected lambs than in the NInf lambs (P = 0.001). Furthermore, compared with NInf lambs, the infected lambs had higher EOS from day 7 PI and higher IgG from day 14 PI. Neither EOS nor IgG were affected by diet. Lambs on Diet 3 had lower EPG during patency than those fed Diets 1 or 2 (days 25 and 28 PI; P = 0.002). Furthermore, lambs fed Diet 3 had lower TAW (Diet 1 vs 3 P = 0.037; Diet 2 vs 3 P = 0.049) and EIU (P = 0.004) than lambs fed Diet 1 or 2. Lambs were resilient to infection regardless of diet. Although EOS and IgG were higher in all infected animals than in Ninf animals, EPG, TAW and EIU decreased only in lambs fed Diet 3. Thus, a diet targeting a DWG of 200 g/d can significantly limit the establishment of H. contortus in Pelibuey lambs infected for the first time.

Nitrogen retention in hair sheep lambs with a gradient of Haemonchus contortus infection.

The impact of Haemonchus contortus infections on nitrogen retention (NR) of hair sheep lambs is unknown. This study estimated the feed intake, digestibility, NR, haematocrit (HT), and their relationship with the gradient of H. contortus infection in hair sheep lambs. Twenty-four hair sheep lambs, 3-4 months old, reared parasite-free were used (live weight (LW) 16.7 ± 1.7 kg). Six lambs were kept parasite-free, nine lambs received a dose of 300 H. contortus infective larvae (L3)/kg LW, and the remaining nine lambs received 500 H. contortus L3/kg LW. Infection doses ranged from 3870 to 9,500 L3. Lambs were individually fed for an average daily gain (ADG) of 100 g. On days 1 (P < 0.01) and 7 (P < 0.001) post-infection (PI), the L3 doses were positively associated with HT, possibly due to platelet recruitment. On week 1 PI, the infection (L3) reduced the crude protein (CP) intake (g/kg metabolic LW) (P < 0.05). A reduction of 1% HT corresponded to 1200 eggs per gram of faeces (EPG) for day 21 PI (P < 0.05), 2300 EPG for day 28 PI (P < 0.001), 2400 EPG for day 35 (P < 0.001), and 1300 EPG for day 41 PI (P < 0.001). However, the HT levels were never lower than 21 %. A lower CP digestibility was associated with EPG only on week 5 PI (P < 0.05). There was no significant association between EPG, Total egg in faeces, or worm burden (WB) with intake, dry matter digestibility (DMD), ADG or NR. The NR of lambs was positive along the different weeks of study irrespective of infection level. The WB was negatively associated only with HT on day 41 PI (P < 0.001), where a 1 % HT reduction was estimated for every 257 worms hosted. In conclusion, no relationship was found between H. contortus infection and DMD, ADG or NR in hair sheep lambs. The cost of H. contortus infection resulted only from the HT reduction and depended on the gradient and stage of the infection.

Do condensed tannins from Lysiloma latisiliquum have a role in its anthelmintic activity?

We would like to comment on a recent paper by Hernandez-Bolio et al. (2017, Nat. Prod. Res. DOI: 10.1080/14786419.2017.1317774) where an extract free of condensed tannin (CT) was evaluated and concluded that Lysiloma latisiliquum CT does not have a role in its anthelmintic (AH) activity. This letter discusses the AH potential of L. latisiliquum based on previously published evidence of a partial role of CT using different AH assays and Haemonchus contortus isolates. Because the experimental design employed in the above-mentioned paper did not allow a clear-cut conclusion regarding the role of CT, this suggests that a specific experiment must be designed to confirm or reject the involvement of CT fraction(s) on L. latisiliquum AH activity.

Gastrointestinal nematode infection and feeding behaviour of goats in a heterogeneous vegetation: No evidence of therapeutic self-medication.

The aim of this study was to identify modifications in the feeding behaviour of goats browsing a tropical deciduous forest (TDF) when natural gastrointestinal nematode (GIN) infection was suppressed. Continuous bite monitoring through direct observation was implemented in 12 Criollo goats (adults, non-pregnant) foraging for 4 h per day during the rainy season. In the first Period (P1, one observation point) all goats were maintained with natural GIN infection. In the second Period (P2, three observation points), goats were equally distributed into 2 groups: i) moxidectin treated group (TG) used in a suppressive scheme; and ii) naturally infected group (IG). For each observation point, goats were monitored at three timepoints per day (80 min each), for three consecutive days, to estimate their intake of dry matter (DM), condensed tannins (CT), crude protein, metabolizable energy and digestible DM. Live weight (LW), faecal samples and blood samples were obtained every 28 days to determine LW change, faecal egg counts (FEC) and packed cell volume (PCV). During P1 and P2, the TG and IG had similar LW change and PCV. During both periods, the intake of DM, CT and all macronutrients were similar for TG and IG. The suppression of GIN infection did not modify the feeding behaviour of goats. Therefore, a therapeutic self-medicative behaviour was not identified in Criollo goats browsing a TDF.

Impact of gastrointestinal parasitism on dry matter intake and live weight gain of lambs: A meta-analysis to estimate the metabolic cost of gastrointestinal nematodes.

This study aimed to estimate the effect of gastrointestinal nematodes (GIN) on the productive performance assessed by the live-weight change (LWC) of lambs, and the metabolic cost associated with parasitism by means of a meta-analysis. Data used in the meta-analyses were obtained from twenty papers selected using the following criteria: (a) lambs with (I) and without (NI) GIN; (b) lambs fed ad libitum; (c) LWC data; (d) feed consumption data; and (e) chemical composition of diets. The effect of diet composition (crude protein [CP] and metabolizable energy [ME]) on dry matter intake (DMI), and the effect of composition and nutrient intake (DMI and CP intake [CPI]) and ME intake (MEI) on LWC was evaluated using respective regression analyses. The metabolic cost of worm burden was determined as the difference in CP and ME requirements between NI and I lambs for each adult parasite. The CP and ME cost were evaluated for lambs at two different levels: maintenance level 0 g LWC and 100 g LWC. The worm burden had an impact on the DMI and LWC of lambs (P < 0.05). The association of CP x infection level was the best predictor of DMI. The association between MEI and infection level were the best predictors of LWC. The metabolic cost of GIN increased to 0.30 mg CP/kg LW0.75 and 0.0056 kJ ME/kg LW0.75 for each adult parasite. The metabolic cost was not modified by diet quality (maintenance level 0 g LW/day or 100 g LW/day). In conclusion, GIN has a negative effect on DMI and LWC. The metabolic cost of GIN infection can be covered by supplying the additional requirements for protein and energy in the diet of infected lambs.

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.

Effect of the consumption of Lysiloma latisiliquum on the larval establishment of gastrointestinal nematodes in goats.

The consumption of tannin-rich (TR) forages has been associated with negative effects against gastrointestinal nematodes and with an improved host resilience. It has been hypothesized that tannins affect the capacity of infective larvae to establish in the mucosae of the host. In this study, we aimed at testing this hypothesis using Lysiloma latisiliquum, a tropical TR tree. The objectives were: (i) to evaluate the effect of the consumption of L. latisiliquum on the establishment of nematode third-stage larvae (L3) in goats; (ii) to define the role of tannins in these effects in vivo by using an inhibitor (polyethylene glycol, PEG); and (iii) to examine a possible indirect effect of tannins on the inflammatory response in the digestive mucosa. Eighteen Criollo goats composed three experimental groups. The control group received fresh leaves of Brosimum alicastrum, a plant with a low level of tannins. Two groups received L. latisiliquum leaves either with (L.L.+PEG) or without (L.L.) daily addition of 25 g PEG. After a 7-day adaptation period, each goat was infected with both Haemonchus contortus and Trichostrongylus colubriformis (3000 L3 per species). The goats were slaughtered 5 days after infection and worm counts and histological analyses were performed. No difference in the voluntary feed intake of foliage was observed between the 3 groups. The consumption of L. latisiliquum significantly reduced the larval establishment of both nematode species compared to the control (P<0.01). For both worm species, the effects were totally alleviated with PEG (L.L.+PEG group), suggesting a major role of tannins in the observed effects. Only minor differences in the mucosal cellular response were observed between the 3 groups. These results confirm that the consumption of TR plants reduces the establishment of nematode larvae in the host and that a direct effect is principally involved.

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.

Simpler intake estimation using direct observation in small ruminants: grouping bites by plant structure and morphology.

OBJECTIVE: To validate the estimation of dry matter intake (DMI) obtained from bite categories (BC) and weight for every plant species (method 1: M1) vs. an alternative method (method 2: M2) grouping plants based on structure and leaf morphology. A dataset containing 80,813 bites and 33 plant species obtained by M1 for sheep and goats browsing a tropical forest was used. Plant species and their respective bite weight were regrouped according to M2. BC weights within each morphological group were compared using ANOVA and post hoc Tukey's honest significant difference comparisons. DMI was estimated for sheep, goats and DMI obtained with both approaches was compared using the t-test, Pearson correlation and orthogonal regression analyses. RESULTS: Dry matter intake estimations were: M1 = 369 ± 153 vs. M2 = 425 ± 161 gDM for sheep and M1 = 567 ± 190 vs. M2 = 681 ± 203 gDM for goats. DMI estimations by M1 and M2 were similar and strongly correlated. Orthogonal regression showed both procedures yielded a similar DMI estimation (P < 0.001). M2 reduces the amount of work required to estimate DMI in heterogeneous vegetation without reducing accuracy. M2 reduced the time required and made simpler to include data from larger number of animals/replicates.

Feed resource selection of Criollo goats artificially infected with Haemonchus contortus: nutritional wisdom and prophylactic self-medication.

Previous cafeteria studies suggested that a moderate natural gastrointestinal nematode (GIN) infection did not modify the resource selection of adult Criollo goats towards tannin-rich plants compared with worm-free goats. A higher infection with Haemonchus contortus could trigger a change in the resource selection behaviour towards tannin-rich foliage. Alternatively, goats might select plant species solely to meet their nutritional requirements. A cafeteria study investigated the effect of a high artificial infection with H. contortus on the feed resource selection of goats. Adult Criollo goats (37.5±4.8 kg BW) with browsing experience were distributed in two groups: the infected group (IG) with six animals artificially infected with H. contortus (6000 L3/animal); and the non-infected group (NIG) with six animals maintained worm-free. The experiment included two 5-day periods with additional 5-day adaptation period. In the first period, animals were offered foliage of five plant species with a decreasing gradient of condensed tannins (CT) (Mimosa bahamensis, Gymnopodium floribundum, Havardia albicans, Acacia pennatula, Lysiloma latisiliqum), and three plant species with negligible CT content (Leucaena leucocephala, Piscidia piscipula and Brosimum alicastrum). In the second period the foliage of B. alicastrum was withdrawn. A grain-based concentrate feed was offered daily at 1% BW in DM basis. Dry matter and nutrient intake was determined. Foliage selection of each experimental group was determined using the Chesson selection index. The H. contortus egg count per gram of faeces (EPG) was determined for infected goats twice daily. Chesson index showed a similar pattern of foliage selection on periods 1 and 2. Mean EPG of goats in IG was 2028±259 EPG during period 1 and 1 293±198 EPG during period 2 (P>0.05). During period 1, the selection pattern was highest for B. alicastrum (tannin-free), followed by a tannin-rich plant (M. bahamensis). These two plants remained as highly selected during period 2. The Chesson index showed that both experimental groups (IG and NIG) selected the same plant species in both periods. Thus, a high H. contortus infection did not affect selection of goats fed with CT-rich plants. Apparently, goats balanced their nutrient intake with the plants selected, showing evidence of nutritional wisdom. This balance may have helped to prevent excess protein in the diet and also to maintain a low GIN infection, both considered as examples of prophylactic self-medication.