Carine Marie-Magdeleine

In vitro effects of Cucurbita moschata seed extracts on Haemonchus contortus.

This study was carried out to evaluate the in vitro effect of Cucurbita moschata seed against the parasitic nematode of small ruminants Haemonchus contortus. Three extracts (aqueous, methanolic and dichloromethane) of C. moschata seed were tested in vitro on four developmental stages of H. contortus using egg hatch assay (EHA), larval development assay (LDA), L3 migration inhibition (LMI) assay and adult worm motility (AWM) assay. The highly significant (P<0.001) ability to stop larval development (inhibition>90% for each extract) and the negative effect of the dichloromethane and methanolic extracts on adult worm motility (inhibition of motility >59.2% after 24h of incubation) compared to the negative controls, suggest anthelmintic properties of C. moschata seed against H. contortus. The active principles responsible for the activity could be secondary metabolites such as amino acid compounds or terpenoid compounds present in the extracts.

In vitro effects of Cassava (Manihot esculenta) leaf extracts on four development stages of Haemonchus contortus.

Three extracts (aqueous, methanolic and dichloromethane) of Manihot esculenta (Cassava) leaf were tested in vitro on four development stages of Haemonchus contortus using egg hatch assay (EHA), larval development assay (LDA), L3 migration inhibition assay (LMI) and adult worm motility assay (AWM). Compared to the negative control, significant effects (P<0.0001) were observed for the methanolic extract of leaf against larval development (57.6% +/-7.6), with a dose dependent effect. These results suggest that Cassava possess anthelmintic activity against H. contortus. The active principles responsible for the activity could be the terpenoids and condensed tannin compounds present in the leaf.

Potential of tannin-rich plants for modulating ruminal microbes and ruminal fermentation in sheep

The objective of this work was to study nutritional strategies for decreasing methane production by ruminants fed tropical diets, combining in vitro and in vivo methods. The in vitro approach was used to evaluate the dose effect of condensed tannins (CT) contained in leaves of Gliricidia sepium, Leucaena leucocephala, and Manihot esculenta (39, 75, and 92 g CT/kg DM, respectively) on methane production and ruminal fermentation characteristics. Tannin-rich plants (TRP) were incubated for 24 h alone or mixed with a natural grassland hay based on Dichanthium spp. (control plant), so that proportions of TRP were 0, 0.25, 0.5, 0.75, and 1.0. Methane production, VFA concentration, and fermented OM decreased with increased proportions of TRP. Numerical differences on methane production and VFA concentration among TRP sources may be due to differences in their CT content, with greater effects for L. leucocephala and M. esculenta than for G. sepium. Independently of TRP, the response to increasing doses of CT was linear for methane production but quadratic for VFA concentration. As a result, at moderate tannin dose, methane decreased more than VFA. The in vivo trial was conducted to investigate the effect of TRP on different ruminal microbial populations. To this end, 8 rumen-cannulated sheep from 2 breeds (Texel and Blackbelly) were used in two 4 × 4 Latin square designs. Diets were fed ad libitum and were composed of the same feeds used for the in vitro trial: control plant alone or combined with pellets made from TRP leaves at 44% of the diet DM. Compared to TRP, concentration of Ruminococcus flavefaciens was greater for the control diet and concentration of Ruminococcus albus was least for the control diet. The methanogen population was greater for Texel than for Blackbelly. By contrast, TRP-containing diets did not affect protozoa or Fibrobacter succinogenes numbers. Hence, TRP showed potential for mitigating methane production by ruminants. These findings suggest that TRP fed as pellets could be used to decrease methane production.

In vitro effects of Tabernaemontana citrifolia extracts on Haemonchus contortus

Tabernaemontana citrifolia (Apocynaceae) is traditionally used as an anthelmintic preparation for ruminants in Guadeloupe (French West Indies). This study was carried out to evaluate the in vitro effect of this plant against the parasitic nematode of small ruminants Haemonchus contortus. Three extracts (aqueous, methanolic and dichloromethane) of T. citrifolia fruit, leaf and root were tested on four developmental stages of the parasite, using egg hatch assay (EHA), larval development assay (LDA), L3 migration inhibition assay (LMI), and adult worm motility assay (AWM). Compared to the negative control, significant effects were observed for the different parts of T. citrifolia but with differences depending on the parasitic stage; efficacies on the larval development of H. contortus from 88.9% to 99.8% for fruit, from 72.1% to 83.8% for root and from 33.5% to 85% for leaf with dose-dependent effect for the methanolic extract. The root gave the best result on EHA (22.7% efficacy for dichloromethane extract) and AWM (56% efficacy, with dose-dependent effect for dichloromethane extract) and the leaf on LMI (49.4% efficacy). These results suggest that T. citrifolia possess anthelmintic activity against H. contortus. The active ingredients responsible for the activity could be the alkaloid compounds present in the plant parts of the plant.

In vitro effects of Musa x paradisiaca extracts on four developmental stages of Haemonchus contortus.

This study was carried out to evaluate the in vitro effect of Musa x paradisiaca stem and leaf against the parasitic nematode of small ruminants Haemonchus contortus. Three extracts (aqueous, methanolic and/or dichloromethane) of Musa x paradisiaca stem and leaf were tested in vitro on four developmental stages of H. contortus using egg hatch assay (EHA), larval development assay (LDA), L3 migration inhibition assay (LMI) and adult worm motility assay (AWM). The highly significant (P<0.0001) ability to stop larval development (inhibition >67% for each extract) and the negative effect of the dichloromethane extract of leaf on adult worm motility (43% of inhibition of motility after 24h of incubation) compared to the negative controls, suggest anthelmintic properties of Musa x paradisiaca stem and leaf against H. contortus. The active principles responsible for the activity could be secondary metabolites such as terpenoid and flavonoid compounds present in the leaf and stem of the plant.

Pumpkin (Cucurbita moschata Duchesne ex Poir.) Seeds as an Anthelmintic Agent

Publisher Summary This chapter outlines the pharmacological actions and potential uses of pumpkin and its extracts. Pumpkin seed oil contains 9.5-13% palmitic, 6-7.93% stearic, 0.04% arachidic, 37-39% oleic, and 44% linoleic acid. The seed also contains a wide range of bioactive compounds. The aqueous extract of the seeds of C. moschata showed efficacy as an anthelmintic in humans. The use of aqueous extracts of pumpkin seeds in the treatment of puppies experimentally infected with heterophyasis gave promising results, with even better results when combining extracts of areca nut and pumpkin seeds than when giving either extract alone. The secondary metabolites suspected to be responsible for anthelmintic activity in C. moschata seed are a triterpenic compound named cucurbitacin B, a non-proteic amino acid named cucurbitin, saponins, and sterols. The non-proteic amino acid cucurbitin, which is only present in the seeds, has been focused on as the active principle responsible for anthelmintic, notably taenicidal and schistosomicidal, activity. Cucurbitin is also used as an anti-allergen for the preparation of cosmetics and pharmaceuticals, particularly dermatological, products. Despite the focus on cucurbitin as the active principle, other secondary metabolites present in the seeds of C. moschata might also be considered as anthelmintics. The non-proteic amino acid cucurbitin (3-amino-pyrrolidine-3-carboxylic acid) is suspected to be the active principle. Cucurbitin was mostly reported to have no side effects, and only a weak level of toxicity was described in dogs and humans.

Comparative response of IgA and IgG activity and hematological parameters among four main beef-cattle breeds infected with gastrointestinal nematodes in the warm humid tropic of Mexico

Abstract This study determined the immunoglobulin (A and G) activity against gastrointestinal nematodes (GIN) and also the hematological parameters in four beef-calf breeds naturally infected in a tropical region of Mexico. Thirty-six infected calves were used to determine the fecal nematode egg counts (FEC), the IgA and IgG activity in serum and saliva, the packed cell volume (PCV), the plasmatic protein (PP) and the differential leukocyte counts. These parameters were measured for a five-month period. ELISA assay was performed using adult worm crude antigen (AWCA) from Haemonchus contortus, Trichostrongylus colubriformis, Mecistocirrus digitatus and Cooperia punctata. The variables were analyzed using a fixed model according to the breed group. The FEC records (means ± standard deviation) for the different breeds were: Brangus (122±115), Charolais (391±507), Guzerat (294±326) and Brown Swiss (413±395). No statistical differences (P>0.05) were found among breeds. Cooperia and Haemonchus were the main identified genera in the coprocultures. Differences in hematological parameters were observed among breeds (P<0.01); Guzerat showed the highest PCV (42.6±4.7%). The lowest leukocyte counts were observed in the Charolais breed (9.2±2.4 × 109 cells L-1). Differences between Guzerat and Charolais were only detected in eosinophils, neutrophils and lymphocytes. Increased activity of IgA was observed against M. digitatus (21% OD) compared with the response against C. punctata in serum samples (13% OD). The lowest IgA activity in serum and saliva was found in Guzerat calves. Charolais and Brown Swiss showed the highest IgA activity in serum and the highest value in saliva was found in Brangus.

Banana meal for feeding pigs: digestive utilization, growth performance and feeding behavior.

The main objective of the present work was to determine the nutritional value and the strategies of using green banana meal (BM) in growing pigs. Two trials involving a total of 96 growing pigs were designed to study the effect of the harvest stage on the nutritional and energy values of BM (trial 1) and to evaluate the consequence of feeding gradual levels of BM on growth performance and feeding behavior in growing pigs (trial 2). In trial 1, the digestive utilization of three diets including 40% BM were compared with a control (C) soybean meal-corn diet in two batches of 12 pigs. BM was obtained from fruits harvested at 750 degrees-days (DD; early harvesting stage), 900 DD (normal harvesting stage) and 1150 DD (late harvesting stage). In trial 2, 72 Large White pigs were grouped in pens of nine animals and were given ad libitum access to one of the four dietary treatments (two pens/diet) differing from the rate of inclusion of 900 DD BM (0%, 20%, 40%, 60%). The estimated energy apparent digestibility coefficients of BM increased with the harvest stage (75.5%, 80.7% and 83.2% for BM at 750, 900 and 1150 DD, respectively). Digestible energy and metabolizable energy values were higher for BM at 1150 DD (13.56 and 13.05 MJ/kg DM, respectively) than at 900 DD (13.11 and 12.75 MJ/kg DM, respectively) or at 750 DD (12.00 and 11.75 MJ/kg DM, respectively). In trial 2, average daily gain and feed conversion ratio were not affected (P>0.05) by the rate of BM inclusion (822 g/day and 2.75 kg/kg on average, respectively). Feed intake and feeding behavior parameters were not significantly influenced by the dietary treatments except for the rate of feed ingestion with a lower value for the diet with 40% of BM (27.4 v. 32.2 g/min on average; P<0.01) when compared with the other diets. Results of this study indicate that the energy value of BM increases with the harvest stage and that BM can be incorporated up to 60% in growing finishing pig diets.

Effect of condensed tannins on methane emission and ruminal microbial populations

Enteric methane (CH4) produced by domestic ruminants represents approximately 15% of the global emissions of this potent greenhouse gas. For reducing rumen CH4 emission various compounds have been tested as feed additives. Among these compounds, tannins are considered a promising group of natural additives. A meta-analysis by Jayanegara et al. (2012) showed that condensed and hydrolysable tannins might reduce CH4 production. However, it is still unclear (1) whether tannin supplementation reduces rumen CH4 in every situation and (2) to which extent this is associated with adverse effects on digestibility and their potential toxicity to some rumen micro-organisms (Goel et al., 2005). In this experiment we investigated the effect of tanniniferous tropical plants on enteric CH4 production and on numbers of methanogens, protozoa, and total and main cellulolytic bacteria.

Multi-criteria evaluation of resources for livestock farming systems under Tropics

The aim of this study was to investigate the role of coprophagous behavior in suckling rabbit on the implantation of caecal microbiota and to try improving the health status of rabbits modifying this behavior. Three groups were compared: in FM group (n=24 litters), pups had free access to maternal hard feces, in NF group (n=28), ingestion of hard feces was prevented, and in FF group (n=28), pups had access only to hard feces excreted by foreign females (n = 5, 7 and 9 feces from 2 to 13 d, 14 to 17 d, 18 to 20 d, respectively). Pup mortality, excretion and ingestion of feces were measured daily. Bacterial composition was assessed by 454 pyrosequencing of the V3-V4 region of 16S RNA genes at 14, 35, 49 and 80 d of age. The total number of feces excreted by the does from 2 to 20 d after delivery ranged widely, but was similar among groups (16.1 ± 12.6 feces doe-1). Ingestion of feces was 3 times greater in FF than in FM group (35.6 vs 9.9, p < 0.001). From 1 to 80 d of age the FF and NF groups exhibited respectively the lowest (9.3%) and highest (22.8%) pup mortality compared to FM group (15.5 %, p = 0.03). At age 14 d the caecal bacterial community was dominated by Bacteroidetes phyla (63.3%), Bacteroidaceae family (36.0%) and Bacteriodes genus (36.0%). With increasing age, Firmicutes phyla, Lachnospiraceae and Ruminococcaceae families became the dominant taxa (92.0%, 44.0 % and 37.9 %, respectively at 80 d of age). Impairment of feces ingestion delayed this ecological succession, with greater and lower relative abundance of Bacteroidaceae and Ruminococcaceae respectively in NF than the two other groups at age 35 d (p < 0.1). In conclusion, the coprophagous behaviour of suckling rabbits is implicated in the maturation of caecal bacterial microbiota and stimulation of this natural behavior improve the health status of animals and could be used to limit the use of antibiotics.Equitherapy is widely practiced and its positive effects are quite well known. However, dynamics of communication between the horse, the user and the therapist is little described. Here, we proposed a pilot study that focused on visual attention of these three partners at two moments: before and during horseback riding (HR). The triad had been observed during the equitherapy session (around 45 min) of four boys with autism (6-9 yo). Ten-second scan samplings recorded the gaze direction of each subject. Data were converted in percentage of time. Horses had mostly gazed at physical environment both before (76.2±6.6%) and during (91.6±1.6%) HR. Human they mostly gazed was the therapist before HR (8.7±3.3%). Similarly, boys mostly gazed at physical environment throughout the session (61.7±17.1% and 46.9±15.3%, respectively). Focus on social partner showed that boys mostly gazed at horse especially during HR (8.7±4.7% and 13.2±9.6% respectively). Likewise their visual attention to humans was enhanced during HR (9.0±1.5% to 19.3±3.1%). At last, before HR, therapists mostly gazed at boy (33.8±9.4%) and horse (38.4±16.7%). During HR, their attention on boy increased slightly (36.7±10.0%) but surprisingly, their attention on horse decreased extremely (7.2±0.9%) in favor of the physical environment (26.1±11.5% to 50.3±12.7%). This pilot study on visual attention during equitherapy suggested that children with autism, whose attentional skills are impaired, seemed to be more attentive to social partners during HR. In this situation, horses seemed few attentive to humans that might be explain by either apathy or working situation. Finally, the reduction of therapist’s visual attention to the horse during HR suggests a decrease in vigilance. This was an example of routine where human pays less attention to animal, which constituted one of the most accidental situations. Thus, therapists must be greatly attentive when horses are used in such therapeutic programs where safety is indispensable.Similar farrowing progress among sows housed in crates and pens Pedersen, Janni Hales; Moustsen, V. A.; Devreese, Anne Michéle; Hansen, Christian Fink Published in: European Association for Animal Production. Annual Meeting. Book of Abstracts DOI: 10.3920/978-90-8686-782-0 Publication date: 2013 Document Version Early version, also known as pre-print Citation for published version (APA): Pedersen, J. H., Moustsen, V. A., Devreese, A. M., & Hansen, C. F. (2013). Similar farrowing progress among sows housed in crates and pens. DOI: 10.3920/978-90-8686-782-0The EAAP AFWG was constituted in 2007 with a view t o enhancing the role of animal fibre in EU27, utilizing an approach based on science and technology. Although an unrecorded and neglected product, annual production of wool from 6 2m breeding sheep (Eurostat) alone is substantial at an estimated 186,000 tonnes (FAO). R ecent outputs include organised symposia and publications defining current knowledge. In rec ognising the need for better networks of scientists and technologists, a total of 5 applicat ions for financial support has been made to the EC COST Framework since 2010. Such applications , by initial pre-proposal, are assessed in 6 categories with a maximum score of 6 for each, giving a maximum score of 36. Preproposals scoring most highly, on average, are invi ted to submit a full proposal. Evaluation has been characterised by large variation in scores of individual assessors. For one example, scores of 36, 33, 32, 32, 28, 21,16 were awarded by 7 assessors, giving mean value = 28.3; SD = 7.23; CV%= 25.5. The divergence of the median v lue of 32 from the mean (28.3) shows a skewed distribution. Removal of the two low est outliers, gives a mean = 32.2; SD= 2.86; CV%= 8.9 and median = 32 and removes the skew . The use in ranking, of such a simple average of means, is clearly unreliable. Another ex ample, with a mean score of 31.25, gave rise to an invitation to submit a full proposal. Th is was done, involving 14 EU partner, and 4 international “reciprocal agreement”, countries. Th e outcome of this application was a score of 53, and below the cut-off score of 55, for furth er progression. The consensus conclusion of evaluation was that “the expected benefits are like ly to be non-European”. This conclusion is surprising and essentially without explanation. The selection of evaluators remains a concern. 64 th EAAP Annual meeting, 25-30 August, 2013, Nantes, France Symposium on South American Camelids and other Fibre AnimalsThe role of the agouti gene in white phenotype was explored in mice. The agouti signalling protein (ASP) can inhibit the differentiation of melanoblasts through the inhibition of the alfa-MSH-induced expression of microphtalmia (MITF) and its binding to a M box regulatory element. The level of microphtalmia in the cells is reduced. To investigate the role of agouti in this phenotype we characterised the transcript structures an relative mRNA expression levels in 13 white alpaca. The reverse transcription analysis of mRNA purified from skin biopsies revealed the presence of three transcripts with different 5’untranslated regions (UTRs) and color specific expression. One of the transcripts, possibly originating from a duplication event (intra-chromosomal recombination) of the agouti gene is characterised by a 5’UTR containing 142 bp of the NCPOA6 gene sequence. Furthermore, the raltive level expression analysis of mRNA demonstrates that the agouti gene has upregulated expression in white skin, suggesting a pleiotropic effect of agouti gene in the white phenotype. Resumen : Recombinacion cromosomica del gen agouti en alpacas blancas. Fue explorado en ratones el rol del gen agouti en el fenoripo blanco. La protein senalarizadora agouti (ASP) puede inibir la diferenciacion de los melanoblastos a traves de la inhibicion de la expresion alfa-MSH inducida de macroftalmia (MITF) y su union al elemento regulador Mbox. El nivel de macroftalmia en las celulas se reduce. Para investigar el rol del agouti en este fenotipo, caracterizamos las estructuras transcriptas a una relativa expresion del mRNA en 13 alpacas blancas blancas. El analisis de transcripcion reversa del mRNA purificado a partir de las biopsias de piel, revelaron la presencia de tres transciptos con diferentes regiones no traducidas del extremo 5’ (5’UTR) y expresiones de color especificas. Uno de los transcriptos, posiblemente procedente de un evento de duplicacion (recombinacion intra-cromosomico) del gen aguti se caracteriza por un 5’ UTR que contiene 142 pb del gen NCPOA6. Por otra parte, el analisis del nivel relativo de expresion del mRNA demuestra que el gen agouti tiene sobre expresion reguladora en piel blanca, lo que sugiere un efecto pleiotropico del gen agouti en el fenotipo blanco.The agouti gene encodes agouti signalling protein (ASIP) whic h regulates pheomelanin and eumelanin synthesis in mammals. To investigate the role of agouti in coat color variation of alpaca, we characterised the agouti gene on 27 black and 12 brown alpaca. The exon-4 hosts three loss-of-function recessive mutations: g.3836C >T, g.3896G>A and g.3866_3923del57, involved in eumelanin synthesis. The deletion at th e position p.C109-Rdel19 eliminates the two beta sheets and the R-F-F- motif from the agout i functional domain, which are essential against alfa-MSH. Therefore, the deleted allele app ears to lose function. The other ANPs observed at the amino acid position 98 and 118 chan ge the conserved R to C and the R-F-Fmotif into H-F-F-. The R-F-F- motif is important fo r functioning at MCRs; the disruption of this motif may result in a non functional agouti pr otein since the alteration of residues in and around R-F-F- causes a decrease in agouti protein i nhibition of alfa-MSH binding to MCRs during signal transduction. The three mutations are randomly distributed among the black alpaca. In our sample, we observed two genotypes : g.3836C>T/g.3896G>A (10 animals) and g.3836C>T/g.3866_3923del57 (17 animals). Among the brown alpaca, 2 are homozygous for the wild allele, 12 are heterozygous for g.3896G>A mutation, carriers for black phenotype.