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The use of plants rich in tannins (PRT) with anthelmintic properties (AH) is an alternative for the control of gastrointestinal nematodes (NGI) in species of cattle interest. The attention in plants with these properties is mainly due to the appearance and extension of parasitic populations resistant to anthelmintics. Recently, in Yucatan, the effect of AH from plant extracts on Haemonchus contortus eggs has been reponed by the hatching inhibition test. Among the main plant species with AH properties are ethanolic and hexane extracts, mainly from the foliage of Lysiloma latisiliquum, Laguncularia racemosa, Rizophora mangle and Avicennia germinans, Theobroma cacao and Coffea arabica. However, it has recently been observed that legumes are an important source of secondary metabolites such as Leucaena leucocephala, Lysiloma latisiliquum, Acacia cornígera and Mimosa bahamensis. The evidence shows that the secondary metabolites that contain some forage legumes affect various biological processes of the gastrointestinal nematodes of ruminants, so it could be an alternative for the control of helminths in the production of small ruminants.

This study assessed the ultrastructural changes caused in adult Haemonchus contortus obtained from goats fed fodder based on polyphenol-rich plants Lysiloma latisiliquum or Onobrychis viciifolia or from goats drenched with quebracho extract, Schinopsis spp. The H. contortus were obtained from artificially infected goats used as models to investigate the anthelmintic effect of feeding or drenching with the polyphenol-rich materials. Nematode populations were exposed to polyphenol-rich plant materials by feeding host goats for 8 consecutive days (D28 to D35 post-infection) with (a) L. latisiliquum fodder at 800 g fresh basis/day, (b) O. viciifolia fodder offered ad libitum, and (c) drenched with a solution containing quebracho extract (90 g/day). Meanwhile, control H. contortus were obtained from goats fed polyphenol-free diets. The H. contortus specimens were recovered from the goats on D36 post-infection, and transmission electron microscopy (TEM) was used to identify ultrastructural changes. In vivo exposure to different polyphenol-rich plant materials caused vacuolization of the nematodes’ intestinal, muscular and hypodermal cells. These alterations represent the first evidence of cell damage caused in H. contortus when hosts were fed or drenched with polyphenol-rich materials. Ultrastructural changes affecting several types of cells could explain modifications in worm motility and nutrition, eventually affecting H. contortus reproductive success. This study contributes to our understanding of the mechanisms of action of polyphenol-rich plants against H. contortus when given as nutraceuticals to goats.

The effect of maize grain supplementation on the resilience and resistance of browsing Criollo goat kids against gastrointestinal nematodes was evaluated. Five-month-old kids (n = 42), raised worm-free, were allocated to five groups: infected + not supplemented (I-NS; n = 10), infected + maize supplement at 108 g/d (I-S108; n = 8), maize supplement at 1% of body weight (BW) (I-S1%; n = 8), maize supplement at 1.5% BW (I-S1.5%; n = 8), or infected + supplemented (maize supplement 1.5% BW) + moxidectin (0.2 mg/kg BW subcutaneously every 28 d) (T-S1.5%; n = 8). Kids browsed daily (7 h) in a tropical forest for 112 days during the rainy season. Kids were weighed weekly to adjust supplementary feeding. Hematocrit (Ht), hemoglobin (Hb), and eggs per gram of feces were determined fortnightly. On day 112, five goat kids were slaughtered per group to determine worm burdens. Kids of the I-S1.5% group showed similar body-weight change, Ht and Hb, compared to kids without gastrointestinal nematodes (T-S1.5%), as well as lower eggs per gram of feces and Trichostrongylus colubriformis worm burden compared to the I-NS group (P > 0.05). Thus, among the supplement levels tested, increasing maize supplementation at 1.5% BW of kids was the best strategy to improve their resilience and resistance against natural gastrointestinal nematode infections under the conditions of forage from the tropical forest.

Abstract Background The use of Duddingtonia flagrans as a tool for the biological control of gastrointestinal nematodes (GIN) is a promising alternative to anthelmintics. The chlamydospores of D. flagrans are orally dosed and their thick cell wall gives them the capacity to resist digestion and pass through the gastrointestinal tract (GIT). Chlamydospores reaching the faeces are able to germinate and trap nematode larvae. The efficacy of this control method is based on reducing the numbers of infective larvae leaving the faeces. Techniques have recently been developed for quantifying the numbers of chlamydospores in faeces. As the number of non-digested spores could be relevant in the design and optimization of dosing programmes for the control of GIN infective larvae, the aim of the present study was to estimate the loss of D. flagrans chlamydospores during their passage through the ruminant gastrointestinal tract using in vitro and in vivo techniques. Results After in vitro rumen digestion, chlamydospore recovery was not different from the quantity originally incubated (undigested spores) (P > 0.05). In vitro rumen+abomasum digestion caused nearly 36% loss of the chlamydospores originally incubated (P < 0.05). Germination of chlamydospores classified as viable was 24.3%. Chlamydospores classified as non-viable did not germinate. Rumen digestion resulted in more spore germination (R1 = 35.7% and R2 = 53.3%) compared to no digestion (time 0 h = 8.7%). Subsequent abomasal digestion reduced germination (R1+A = 25%) or stopped it (R2+A = 0%). In vivo apparent chlamydospore digestibility in sheep showed a loss of 89.7% of the chlamydospores (P < 0.05). Conclusions The loss of chlamydospores was evident under in vitro and in vivo conditions. Negligible amounts of spores were lost during the in vitro rumen digestion. However, in vitro rumen+abomasum digestion resulted in a chlamydospore loss of approximately 36%. In vivo passage through the sheep GIT resulted in a total loss of 89.7% of the orally administered spores.

The variability in the expression of different P-glycoprotein (P-gp) genes in parasitic nematodes of ruminants such as Haemonchus contortus (Hco-pgp) may be caused by different factors including nematode biology, geographical region and anthelmintic pressure. This study analysed the relative expression level of 10 P-gp genes in two H. contortus (Hco-pgp) field isolates from Yucatan, Mexico: 1) PARAISO (IVM-resistant) and 2) FMVZ-UADY (IVM-susceptible). These isolates were compared with a susceptible reference isolate from Puebla, Mexico, namely "CENID-SAI". In all cases H. contortus adult males were used. The Hco-pgp genes (1, 2, 3, 4, 9, 10, 11, 12, 14 and 16) were analysed for each isolate using the RT-qPCR technique. The Hco-pgp expressions were pairwise compared using the 2 -ΔΔCt method and a t-test. The PARAISO isolate showed upregulation compared to the CENID-SAI isolate for Hco-pgp 1, 3, 9, 10 and 16 (P < 0.05), and the PARAISO isolate showed upregulation vs. FMVZ-UADY isolate for Hco-pgp 2 and 9 (P < 0.05), displaying 6.58- and 5.93-fold differences (P < 0.05), respectively. In contrast, similar Hco-pgp gene expression levels were recorded for FMVZ-UADY and CENID-SAI isolates except for Hco-pgp1 (P <0.1), which presented a significant upregulation (6.08-fold). The relative expression of Hco-pgp allowed confirming the IVM-resistant status of the PARAISO isolate and the IVM-susceptible status of the FMVZ-UADY isolate when compared to the CENID-SAI reference isolate. Therefore, understanding the association between the Hco-pgp genes expression of H. contortus and its IVM resistance status could help identifying the genes that could be used as molecular markers in the diagnosis of IVM resistance. However, it is important to consider the geographic origin of the nematode isolate and the deworming history at the farm of origin., Competing Interests: Declaration of competing interest Authors have no potential conflict of interest pertaining to this submission., (Copyright © 2024 Elsevier B.V. All rights reserved.)

This study applied the in vitro rumen exsheathment test (IVRET) to evaluate the exsheathment kinetics of Haemonchus contortus infective larvae (L 3 ) incubated in ruminal liquor (RL) containing acetone:water extracts of Acacia pennatula (AP), Gymnopodium floribundum (GF), Havardia albicans (HA) or Lysiloma latisiliquum (LL). The role of polyphenols in the biological activity of the evaluated extracts was also determined. Larvae were incubated in RL either alone or added with a different plant extract (AP, GF, HA, or LL) at 1200 μg/mL. Polyethylene glycol (PEG) was added to block polyphenols in each treatment (RL+PEG, AP+PEG, GF+PEG, HA+PEG, and LL+PEG). After incubation times of 0, 1, 3, 6, 9, and 24 h, the exsheathment process was stopped to count the number of ensheathed and exsheathed L 3 . A Log-Logistic model was used to determine the L 3 exsheathment kinetics in the different RL treatments. The inflection point of the respective kinetic curves, which indicates the time to reach 50 % exsheathed L 3 (T 50 ), was the only parameter that differed when comparing the exsheathment models (99 % probability of difference). The T 50 values obtained for GF, HA, and LL treatments (T 50 = 7.11 - 7.58 h) were higher in comparison to the T 50 of RL (5.72 h) (≥ 70 % probability of difference). The L 3 incubated in RL added with GF, HA, and LL extracts delayed their exsheathment at 3 and 6 h of incubation (28.71 - 48.06 % exsheathment reduction) compared to the RL treatment. The T 50 value for AP, AP+PEG, GF+PEG, HA+PEG, and LL+PEG were similar to RL and RL+PEG (T 50 = 5.34 - 6.97 h). In conclusion, the IVRET can be used to identify plants with the potential to delay the exsheathment of H. contortus L 3 in the ruminal liquor. The acetone:water extracts of G. floribundum, H. albicans, and L. latisiliquum delayed the T 50 of H. contortus exsheathment, which was evident at 3 and 6 h of incubation in ruminal liquor. The observed exsheathment delay was attributed to the polyphenol content of the extracts., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper, (Copyright © 2024 Elsevier B.V. All rights reserved.)

This study adapted the in vitro rumen incubation (IVRI) method to evaluate the biological activity of a Gymnopodium floribundum leaves extract against the exsheathment of Haemonchus contortus infective larvae (L 3 ), and to determine the role of plant polyphenols on the biological activity. The incubation protocol followed the IVRI method, adding polyethylene glycol (PEG) as a polyphenol-blocking agent. The L 3 were incubated in ruminal liquor (RL), ruminal liquor with PEG (RL+PEG), ruminal liquor with G. floribundum extract (RLE), and ruminal liquor with G. floribundum extract and PEG (RLE+PEG). Incubation condition controls included phosphate buffered saline (PBS), PBS with PEG (PBS+PEG), incubation medium (without ruminal liquor) (IM), and incubation medium with PEG (IM+PEG). The L 3 were recovered after incubation times of 0, 1, 3, 6, 9, and 24 h (39 °C). The respective L 3 exsheathment kinetics were estimated for the different treatments (RL, RL+PEG, RLE, and RLE+PEG) using Log-Logistic models. The parameters of the different models were compared to determine the impact of the extract, with or without PEG, on the L 3 exsheathment kinetics. The exsheathment in PBS and PBS+PEG remained < 2.71% at each incubation time. The exsheathment in IM and IM+PEG reached 13.58% and 17.18% at 24 h, respectively. The exsheathment percentages for RLE were lower than those for RL at 3, 6 and 9 h of incubation. The inflection point, indicating the time required to reach 50% of the maximal exsheathment (T 50 ), was the only parameter that differed between the ruminal liquor models. The T 50 in RLE (7.106 h) was higher than the values obtained for RL (5.385 h) and RL+PEG (4.923 h) (99.99% probability of being different). Such delay resulted in a reduction of exsheathment in RLE of 62% at 3 h, 38% at 6 h, and 12% at 9 h, relative to RL values. When PEG was added with the extract (RLE+PEG), the T 50 (5.045 h) was similar to that of RL and RL+PEG. The IVRI method was adapted as an in vitro rumen exsheathment test (IVRET). The IVRET showed that H. contortus L 3 exposed to G. floribundum extract delayed their exsheathment kinetics at different time points. The exsheathment delay was attributed to the polyphenol content of the extract., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

We evaluated the relationship between individual and herd GIN infection level, nutrition, production performance and anemia parameters in a tropical farm context. Fifty-four female goats were monitored to assess their body condition score (BCS, nutritional status indicator), live weight (LW) and LW gain (LWG, both used as production level indicators), FAMACHA© and hematocrit (HT, both used as anemia indicators). Goats browsed for 4 h in a tropical forest and received balanced feed and chopped grass. The eggs per gram of feces (EPG) indicated the GIN burden, with fecal samples obtained at 7:00 (AM) and 15:00 h (PM.) from each goat at six sampling points during the study. The variables and their relationship with GIN burdens were analyzed using Kruskall-Wallis, ANOVA and Friedman tests and Spearman correlations. The fecal samples obtained in the AM and PM can be equally representative of parasitic burdens (similar and highly correlated). However, the EPG of individual goats from periods of 30 days apart can be considered independent. The BCS and LWG varied between sampling times ( p < 0.05), whereas EPG, LW and HT did not ( p > 0.05). The GIN burden was negatively correlated with HT and BCS (-0.21, p = 0.01 for each one). The individual pattern of infection demonstrates the true impact of GINs on their hosts. Additionally, feeding and nutritional status may present important variations influencing the performance of the goats more than the impact of GINs under the farm conditions of the present study. However, GIN infection contributed to the variation in goat health and productivity in this tropical farm.

Ivermectin (IVM) resistance in parasitic nematodes such as Haemonchus contortus has spurred a search for substances that help to recover its efficacy. One potential agent is the natural product curcumin (CUR). In this study, CUR was combined with polyvinylpyrrolidone (PVP) (CUR/PVP) to improve its solubility and biological applicability. This study determined the effect of CUR preincubation on the effective concentration 50% (EC 50 ) of IVM in three H. contortus isolates with different susceptibilities to IVM. The IVM EC 50 was determined for three H. contortus isolates with different IVM susceptibilities using the larval migration inhibition (LMI) test. The three isolates were (i) PARAISO (IVM resistant), (ii) FMVZ-UADY (IVM susceptible), and (iii) CENID-SAI INIFAP (reference IVM susceptible). The L 3 of each isolate were preincubated for 3 h with one of three concentrations of CUR (μg curcumin/mL): CONC-1 (3.67), CONC-2 (5.67), or CONC-3 (8.48). Corresponding controls were performed without CUR. The EC 50 of IVM was determined for each isolate after they were exposed to the different CUR concentrations. The EC 50 of IVM differed between the isolates PARAISO > FMVZ-UADY > CENID-SAI INIFAP (P < 0.05). The CUR preincubation at CONC-1 did not decrease the EC 50 of IVM for any of the three isolates, suggesting a hormetic effect. By contrast, CUR preincubation at CONC-2 or CONC-3 decreased the IVM EC 50 for the PARAISO isolate (P < 0.05) compared with the reference isolate and reduced the EC 50 of IVM for the FMVZ-UADY and CENID-SAI INIFAP isolates below the EC 50 for the CENID-SAI INIFAP isolate without CUR preincubation. In conclusion, preincubation of H. contortus L 3 with CUR reduced the EC 50 of IVM for field isolates classified as resistant and susceptible to IVM. The CUR preincubation reduced the IVM resistance factor in the different isolates tested., Competing Interests: Declaration of competing interest We declare that this is an original contribution that has not been published elsewhere in substantially the same form and it is not currently under consideration elsewhere. Furthermore, we have no financial or personal interest that could be considered a conflict of interest., (Copyright © 2023 Elsevier Inc. All rights reserved.)

This study developed and evaluated an in vitro rumen incubation (IVRI) method to describe the exsheathment kinetics of Haemonchus contortus third-stage infective larvae (L 3 ) in ruminal liquor (RL). The specific objectives were (i) to standardize the IVRI method to facilitate the contact between L 3 and RL as well as the larval recovery, and (ii) to apply the IVRI method to describe the exsheathment kinetics of H. contortus and to select the best fitting nonlinear model. Incubation devices containing H. contortus larvae were incubated according to the IVRI technique in cattle RL or PBS. The incubation conditions included RL mixed with a nitrogen-rich media, maintained at 39 °C, with pH = 7.0, vented with CO 2 and manual agitation. The larvae were recovered after 0, 1, 3, 6, 9, 12, and 24 h. The exsheathed and ensheathed larvae were counted to estimate the exsheathment (%) in RL or PBS. Exsheathment in RL was analyzed with nonlinear regression models: Exponential, Gompertz, Logistic, Log-Logistic, and Weibull. The models' fit was compared to select the one that best described the exsheathment kinetics. The exsheathment in RL reached 6.52%, 20.65%, 58.22%, 69.24%, 73.08%, and 77.20% in 1, 3, 6, 9, 12, and 24 h, respectively. Although the Gompertz, Weibull, and Logistic models were adequate to describe the observed exsheathment, the Log-Logistic model had the best fit. The IVRI method using bovine RL represents a suitable tool for the study of the in vitro exsheathment kinetics of H. contortus L 3 ., (© 2023. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

This study investigated the impact of Moringa oleifera Lam. meal (MOM) on meat nutritional properties and bone quality of slow-growing layer-type male chickens raised in semi-intensive conditions. A total of 198, 72-d-old Dominant Blue D 107 male chickens, with an average weight of 1093 ± 15.2 g, were randomly assigned to three dietary treatments supplemented with 0, 3, and 6% of MOM that corresponded to T1, T2, and T3, respectively. Each treatment, consisting of six replicated floor pens of 11 birds, had access to the outdoors for 49 days. The results showed that breast muscle ash percentage was significantly greater (P ≤ 0.05) in T2 in comparison to the T1 group. Meat dry matter, protein, and fat content were not influenced by the treatments (P > 0.05). Regardless of the treatments, oleic acid (C18:1N9C) was numerically more abundant in the breast than in the leg muscle. Alternatively, femoral and tibial lengths were shorter (P ≤ 0.05) in birds fed 3% MOM than the two other groups. Moreover, birds fed with MOM had greater tibial diameter (P ≤ 0.05) than those that were fed without MOM. In addition, bone ash content and phosphorous amount were significantly higher (P ≤ 0.05) in birds fed 6% MOM compared to those fed without MOM. The data of this study indicate that up to 6% of MOM may be added to the diet of slow-growing layer-type male chickens raised with outdoor access under tropical conditions to improve bone quality traits.

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 L 3 H. 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., Competing Interests: Conflict of interest statement The authors declare that they have no conflicts of interest., (Copyright © 2022 The Authors. Published by Elsevier B.V. All rights reserved.)

We evaluated the effect of browsing experience, nutritional quality and secondary compounds of forage resources, and the interaction between these factors on the selection and intake of goats in a cafeteria trial. Twelve juvenile Criollo goats from 7 to 9 months of age, weighing 22 ± 3 kg, were divided into two groups: (a) browser goats group ( n = 6, BG), and (b) naïve goats group ( n = 6, NG), formed according to their previous browsing experience (with and without, respectively). Animals were housed in individual pens. The cafeteria experiment lasted 21 days considering pen adaptation, foliage adaptation, and measurements, which included the selection index (SI) of experimental forage resources (Chesson's alpha) and their dry matter intake (DMI/Kg 0.75 ), using a multiple Latin square design. Furthermore, correlation and regression analyses were used to assess the relationship between the aforementioned factors. The NG did not show any selection pattern, while the BG selected Piscidia piscipula and Senegalia gaumeri ( p = 0.0002). The BG consumed smaller amounts of secondary compounds compared to NG ( p = 0.0001). In the BG, the flavonoids affected negatively their selection (R 2 = 97.51, p = 0.0001), while the DMI was affected by in vitro DM digestibility and flavonoids (R 2 = 99.85; p = 0.0001). For the NG, the crude protein and organic matter contents were associated with DMI, but none had a significant relationship with SI. The BG selected and consumed forages with suitable nutritional quality avoiding those with high content of secondary compounds such as flavonoids. Conversely, NG did not show a clear pattern for their selection or intake.

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 L 3 , 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., (© 2021. Witold Stefański Institute of Parasitology, Polish Academy of Sciences.)

Purpose: Some agroindustrial by-products could be used as nutraceutical materials for small ruminants helping with their nutrition while controlling their gastrointestinal nematodes. This study evaluated the potential in vitro nutraceutical value of pod husks and leaves of three varieties of Theobroma cacao using two Haemonchus contortus isolates with different polyphenol susceptibility., Methods: Leaves and husks from three T. cacao varieties (AZT, CAL and CEY) were evaluated for their bromatological composition, in vitro dry matter digestibility and polyphenol content. Acetone:water extracts (70:30) of each plant variety were evaluated using the egg hatch and larval exsheathment inhibition tests, using two isolates (FESC and PARAISO) of H. contortus. Effective concentrations 50% (EC 50 ) were determined for both tests. The role of polyphenols was confirmed using polyvinylpolypyrrolidone. L 3 exposed to CAL leaf extract were submitted to transmission electron microscopy., Results: Both plant materials showed a good nutritional value to complement protein-rich diets for small ruminants. Extracts inhibited exsheathment of H. contortus L 3 more effectively than the egg hatching, and the leaf extracts were more active than husk extracts in the L 3 exsheathment inhibition. The FESC isolate was more sensitive to extracts. Polyphenols blocked exsheathment inhibition of leaf extracts. Structural damage was observed in the sheath and muscles of L 3 exposed to CAL leaf extracts., Conclusion: The two T. cacao materials tested showed their potential to be used as ruminant feeds. Extracts affected H. contortus by blocking L 3 exsheathment, particularly with the leaf extracts. The in vivo nutraceutical value should be confirmed in small ruminants., (© 2021. Witold Stefański Institute of Parasitology, Polish Academy of Sciences.)

In sheep, infection with Haemonchus contortus may increase the need for energy, and this demand may vary according to the infection level. In this study, the energy intake, digestibility, and energy retention of lambs artificially infected with different levels of H. contortus were estimated. A total of 24 hair sheep lambs reared parasite-free were experimentally infected with H. contortus at one of three infection levels: non-infected (n = 6); infected with 300 infective larvae (L 3 ) of H. contortus/kg body weight (BW) (n = 9); and infected with 500 H. contortus L 3 /kg BW (n = 9). The lambs were fed for an individual weight gain of 100 g/day, and intake of organic matter (OMI) and gross energy (GEI), digestible (DEI) and metabolizable energy (MEI) were measured weekly. The digestibility of organic matter (OMD) and GE (GED) and the metabolizable energy (ME) balance adjusted to zero nitrogen balance (ME adj ) were measured for each lamb during the prepatent and patent periods of infection. From day 21 post-infection (PI), the individual eggs per gram (EPG) of feces and the total number of eggs in feces (TEF) were estimated weekly. After humane slaughter on day 42 PI, the worm burden (WB) was determined. Correlation and regression analyses were performed to estimate the relationships between the parasitological variables (L 3 , EPG, TEF and WB) and the response variables (OMI, GEI, DEI, MEI, OMD, GED, ME adj ). During the prepatent period, there were no significant relationships of L 3 with the response variables (OMI, GEI, DEI, MEI, OMD, GED, ME, ME adj ). Similarly, during the patent period, no relationship was evident between infection (EPG, TEF or WB) and OMI, GEI, DEI, GED, OMD, ME or ME adj . Thus, the gradient of H. contortus infection examined in the present study did not influence energy balance in hair sheep lambs, and infection did not impose any detectable energy cost. Further studies are needed to fully assess the impact of H. contortus infection on energy metabolism in hair sheep., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Haemonchus contortus is a parasite species that affects the health and production of grazing small ruminants in different parts of the world. Scanning electron microscopy (SEM) is an important tool for the study of parasites' morphology and taxonomy as it generates images that appear 3D and are generally easier to interpret than optical microscopy images. This study used the SEM to describe the vulval types of adult H. contortus from a Mexican isolate. A total of 14 adult H. contortus females were obtained from two artificially infected goats. Females were fixed and processed by critical point drying and observed with SEM. A collection of SEM images was obtained from these parasites and those images were used to identify the structures previously described by optical microscopy studies. Two different types of vulval structures were described in this Mexican H. contortus isolate: Type 1 (vulval flap), Type 2 (epiptygma). An unusual vulval structure was reported in a single individual. The Type 1 included vulval flaps of different sizes and spatial dispositions, as well as one or more knobs in different positions around the vulva. The Type 2 shows differences in the epiptygma. The present study suggests that the Mexican H. contortus isolate used in donor animals possess intraspecific polymorphism in vulval structures., (Copyright © 2021 Elsevier B.V. All rights reserved.)

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 (L 3 )/kg LW, and the remaining nine lambs received 500 H. contortus L 3 /kg LW. Infection doses ranged from 3870 to 9,500 L 3 . 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 L 3 doses were positively associated with HT, possibly due to platelet recruitment. On week 1 PI, the infection (L 3 ) 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., (Copyright © 2021 Elsevier B.V. All rights reserved.)

The plant kingdom can influence the productivity and health of herbivores at different levels. However, demonstrating this process in a scientific manner entails substantial endeavors from different disciplines. In the present review, we will describe the features of a native vegetation system traditionally used by small ruminants and use its particularities to build an interdisciplinary approach to evaluate the nutraceutical properties of plants. Initially, we will establish the context of the low deciduous forest (LDF), considering some botanical and nutritional aspects, as well as the presence of plant secondary compounds (PSC) and gastrointestinal nematodes (GIN). Furthermore, we will focus on coevolutionary aspects that undoubtedly shaped the plants-nutrients-PSC-GIN-herbivore relationship. In addition, the concept of nutraceutical will be discussed to provide clarity and aspects to be considered for their evaluation. Then, ethological, agronomical, nutritional, PSC, parasitological and animal species issues are deepened placing emphasis on methodological approaches. Special focus is given to condensed tannins, as they are the fourth largest group of PSCs and the most studied in livestock sciences. Validation of the nutraceutical properties of plants from native vegetation systems should be seen as a process derived from many scientific disciplines that feed into each other in a cyclic manner.

In the Mediterranean region, artichoke and broccoli are major crops with a high amount of by-products that can be used as alternative feedstuffs for ruminants, lowering feed costs and enhancing milk sustainability while reducing the environmental impact of dairy production. However, nutritional quality of milk needs to be assured under these production conditions and an optimal inclusion ratio of silages should be determined. This work aimed to evaluate the effect of three inclusion levels (25%, 40%, and 60%) of these silages (artichoke plant, AP, and broccoli by-product, BB) in goat diets on milk yield, composition, and mineral and fatty profiles. Treatments with 60% inclusion of AP and BB presented the lowest milk yield. No differences were found on the milk mineral profile. Inclusion of AP in the animals' diet improved the milk lipid profile from the point of view of human health (AI, TI) compared to BB due to a lower saturated fatty acid content (C12:0, C14:0, and C16:0) and a higher concentration of polyunsaturated fatty acids (PUFA), especially vaccenic acid (C18:1 trans11) and rumenic acid (CLA cis9, trans11), without any differences with the control treatment.

Wastes from artichoke and broccoli crops and cannery industries represent an environmental problem. A viable option to this problem is ensiling them for use as ruminants feed. The aim of this study was to characterise the ensiling process of broccoli and artichoke by-products and assess their suitability to be part of the ruminant diet, as well their minimum shelf life. Twenty-one commercial round bale silos (300 kg and 0.64 m 3 ) of each by-product were made. Samples were analysed at days 0, 7, 15, 30, 60, and 200 to determine microbial populations, fermentation metabolites, nutritional components, and phytosanitary residues. Feedstuffs showed good suitability for ensiling, and stabilisation was achieved on day 30. The variables with the greatest significant differences among sampling times were microbial populations and fermentative components. There were no important dry matter losses, and some significant differences were observed in the nutritional composition, especially in crude protein and fibrous fractions, but they were not relevant for the loss of nutritional quality of silages. The phytosanitary residues determined on day 200 were below the maximum residue limits set by European legislation. So, ensiling these by-products in commercial round bale silos is a suitable and profitable technique that allows their preservation for a long time (200 days)., Competing Interests: The authors declare no conflict of interest. The funders had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript, or in the decision to publish the results.

This study identified the optimal age of Trichostrongylus colubriformis larvae (L 3 ) under tropical conditions for the in vitro evaluation of plant extracts using the larval exsheathment inhibition test (LEIT). Two T. colubriformis isolates with different anthelmintic (AH) susceptibility status were used for this study. The L 3 of both isolates were maintained on refrigeration (4-5 °C) until use. For the LEIT, the isolates were tested every week during 16 weeks, using a stock solution of Acacia pennatula acetone:water extract at different dilutions (80-1200 μg/mL). Respective positive controls (levamisole 12.5 mM) and negative controls (PBS) were included. Effective concentrations 50 % (EC 50 ), 90 % (EC 90 ) and respective 95 % confidence intervals (95 %CI) for exsheathment inhibition were calculated at 60 min after exposure to chlorine solution. Motility of L 3 (migration percentages (M%)) was recorded weekly using the larval migration test (LMT) as an indicator of L 3 fitness over time. No correlation was found between L 3 age or M% and the extract's EC 50 or EC 90 values. However, the EC 50 values for the A. pennatula extract ranged from 80 to 200 μg/mL from weeks 2-10. Beyond week 12, larval exsheathment was irregular, with higher EC 50 and EC 90 values and wider 95 %CI. The M% decreased below 85 % on week 7 for Paraiso isolate, and on week 10 for FMVZ isolate. A linear negative relationship was observed between the age of L 3 and M% for both T. colubriformis isolates. The relationship (slope) for both isolates was similar therefore a single linear equation was estimated describing all M% data (r 2  = 0.771, df = 164, P < 0.05). Thus, when using these T. colubriformis isolates under our tropical conditions for the evaluation of AH activity of plant extracts with LEIT, the optimal age of L 3 is between weeks 2-7, when M% remained above 85 %. The latter may ensure consistent and reproducible exsheathment results for T. colubriformis. Each laboratory must identify optimal conditions to perform the LEIT., Competing Interests: Declaration of Competing Interest None., (Copyright © 2020 Elsevier B.V. All rights reserved.)

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.

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.

Two new spongian furanoditerpenes, 3β-hydroxyspongia-13(16),14-dien-2-one ( 1 ) and 19-dehydroxy-spongian diterpene 17 ( 2 ), along with five known terpenes, the spongian furanoditerpenes 9-nor-3-hydroxyspongia-3,13(16),14-trien-2-one ( 3 ), 3β,19 dihydroxyspongia-13(16),14-dien-2-one (epispongiadiol) ( 4 ) and spongian diterpene 17 ( 5 ), the furanoditerpene ambliol C ( 6 ), and the sesterterpene scalarin ( 7 ), were isolated from the methanolic extract of the sponge Spongia tubulifera , collected in the Mexican Caribbean. The planar structures of the new compounds were elucidated by 1D/2D NMR and IR spectroscopic analysis, high resolution electrospray mass spectrometry (HRESIMS), and comparison of their spectral data with those reported in the literature. Absolute configurations were determined by comparison of the experimental electronic circular dichroism (ECD) spectrum with those calculated by time-dependent density functional theory (TDDFT). Compounds 1 , 4 , and 6 displayed weak cytotoxic activity against different human tumour cell lines.

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 (L 3 ). 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.

Small ruminants browsing in tropical forests readily consume the foliage of Senegalia gaumeri. A S. gaumeri methanol:water extract was recently shown to have ovicidal activity against Haemonchus contortus eggs in vitro. In the present study, the fraction of a S. gaumeri methanol:water extract with ovicidal activity against H. contortus eggs and the metabolites potentially involved in this activity were identified. Bio-guided fractionation of the S. gaumeri methanol:water extract identified high ovicidal activity (80.29%, EC 50  = 58.9 μg/mL) in the non-polar sub-fraction P1. Gas chromatography-mass spectrometry (GC-MS) identified several fatty acids: pentacosane (18.05%), heneicosane (18.05%), triacontane (30.94%), octacosane (18.05%), and hexanedioic acid bis-(2-ethylhexyl) ester (32.72%). Purification of the polar components of sub-fraction P1 led to the identification of p-coumaric acid as a major constituent. In egg hatch tests, 400 μg/mL p-coumaric acid resulted in an ovicidal effect of 8.7%, a larvae failing eclosion effect of 2.9%, and of the emerged larvae (88.4%), many were damaged. In conclusion, the low AH activity of p-coumaric acid against H. contortus eggs indicates that it is not solely responsible for the ovicidal activity of sub-fraction P1 but might act in synergy with other compounds in this fraction. However, p-coumaric acid showed potential anthelmintic effects against the larval stage of H. contortus., (Copyright © 2019 Elsevier B.V. All rights reserved.)

This study evaluated the in vitro anthelmintic (AH) activity of cinnamic acid and six analogues against eggs and larvae of Haemonchus contortus. Stock solutions of each compound (trans-cinnamic acid, p-coumaric acid, caffeic acid, trans-ferulic acid, trans-sinapic acid, 3,4-dimethoxycinnamic acid, and chlorogenic acid) were prepared in PBS:Tween-20 (1%) for use in the egg hatch test (EHT) and larval exsheathment inhibition test (LEIT) at different concentrations (25-400 μg/mL). The respective effective concentration 50% (EC 50 ) values with 95% confidence intervals were estimated. Mixtures made of all cinnamic acid and its analogues as well as some selected individual compounds were also tested in the EHT. Only ferulic and chlorogenic acids showed AH activity in the EHT (EC 50 : 245.2 μg/mL (1.26 mM) and 520.8 μg/mL (1.47 mM), respectively) (P < 0.05). A higher EC 50 (1628.10 μg/mL) of the mixture of cinnamic acid and its analogues was required to observe activity against eggs mostly blocking the larvae hatching. The analogues' mixtures tested were less active than ferulic or chlorogenic acid alone. The activity of ferulic and chlorogenic acids against eggs was associated with larvae failing to hatch, and the two compounds exhibited antagonistic effects when evaluated together. All standards had an EC 50 lower than 0.42 mM in the LEIT. Caffeic acid had the best activity in the LEIT (EC 50 0.04 mM), followed by ferulic acid (EC 50 0.11 mM) (P < 0.05). There was no clear, definitive structure-activity relationship for these non-flavonoid polyphenols against eggs or larvae of H. contortus in vitro. This study is the first to directly evaluate cinnamic acid and its derivatives as active compounds against eggs and larvae of H. contortus., (Copyright © 2019 Elsevier B.V. All rights reserved.)

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., (Copyright © 2019 Elsevier B.V. All rights reserved.)

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 LW 0.75 and 0.0056 kJ ME/kg LW 0.75 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., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Animal habituation is key to obtain reliable data on behavioural studies but detailed procedures to achieve it are scarce. This study designed a set of actions to habituate sheep and goats to human observers. Pelibuey sheep (n = 15) and Criollo goats (n = 10) were classified as (a) avoider, flight from human interaction, or (b) follower, seek human interaction. Habituation was measured by the reduction of flight distance by avoiders, or number of followers in the presence of observers. The habituation protocol consisted of a gradually increased series of five manoeuvres, either challenge (for avoiders) or evasion (for seekers), performed first inside a pen and subsequently in a grass paddock. Habituation was considered successful when animals could be observed from a 1-m distance without flight or following the observer. In the pen, habituation took 12 and 13 days for sheep and goats, respectively. Meanwhile, in the grass paddock habituation took 10 days, for both species. The number of challenge and evasion series was negatively correlated with the flight distance in sheep and with the number of followers in goats. This protocol is simple and practical to implement and enables animal habituation for behavioural studies., (Copyright © 2018 Elsevier B.V. All rights reserved.)

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% (EC 50 ) 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 (EC 50  = 401.8 μg/mL) and L 3 (EC 50  = 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 L 3 in the selected plant species.

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.

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.

Dietary tannins can affect protein digestion and absorption, be toxic, and influence food selection by being astringent and bitter tasting. Animals that usually ingest tannins may regularly secrete tannin-binding salivary proteins (TBSPs) to counteract the negative effects of tannins or TBSPs production can be induced by a tannin-rich diet. In the wild, many primates regularly eat a diet that contains tannin-rich leaves and unripe fruit and it has been speculated that they have the physiological ability to cope with dietary tannins; however, details of their strategy remains unclear. Our research details the salivary protein composition of wild and zoo-living black howler monkeys (Alouatta pigra) feeding on natural versus manufactured low-tannin diets, and examines differences in TBSPs, mainly proline-rich proteins (PRPs), to determine whether production of these proteins is dependent on the tannin content of their food. We measured the pH, flow rate, and concentration of total protein and trichloroacetic acid soluble proteins (an index of PRPs) in saliva. Howler monkeys produced slightly alkaline saliva that may aid in the binding interaction between tannin and salivary proteins. We used gel electrophoresis to describe the salivary protein profile and this analysis along with a tannin-binding assay allowed us to detect several TBSPs in all individuals. We found no differences in the characteristics of saliva between wild and zoo-living monkeys. Our results suggest that black howler monkeys always secrete TBSPs even when fed on foods low in tannins. This strategy of constantly using this salivary anti-tannin defense enables them to obtain nutrients from plants that sometimes contain high levels of tannins and may help immediately to overcome the astringent sensation of their food allowing howler monkeys to eat tanniferous plants., (© 2018 Wiley Periodicals, Inc.)