Addition Of PVPP Research Articles

Two in vitro anthelmintic assays of four Paraguayan medicinal plants for proof of concept of the role of polyphenols in their biological activities and LC-HRMS analysis

Ethnopharmacological relevanceGastrointestinal nematodes (GIN) control in small ruminants has relied on the systematic use of synthetic anthelmintics (AH), their effectiveness has been progressively decreasing due to the rise and diffusion of anthelmintic resistances. The most prevalent genera affecting small ruminants were Haemonchus spp., and Trichostrongylus spp. The investigation of new anthelmintics in plants is a highly studied option, especially when it is linked to ethnobotanical knowledge and phenolic compounds. The aims of our studyFour medicinal plants mentioned in traditional medicine were selected to evaluate their anthelmintic proprieties at different stages of the life cycle of GIN, namely: Kyllinga odorata Valh., Cassia occidentalis L., Artemisia absinthium L, and Verbena litoralis Kunth and to explore the role of polyphenols in the AH activity. Materials and methodsTo evaluate the anthelmintic activity in this study, two models of GIN species, namely Haemonchus contortus (Hc) and Trichostrongylus colubriformis (Tc) were selected and tested on two in vitro assays: 1) Larval Exsheathment Inhibition Assay (LEIA) and, 2) Egg Hatch Assay (EHA). To explore the role of tannins and polyphenols in AH activity by comparing the effects of LEIA and EHA with or without polyvinylpolypyrrolidone (PVPP) and to characterize the phytochemical composition of the most active plants using ultra-high performance liquid chromatography (UHPLC) coupled with high-resolution mass spectrometry (HRMS). ResultsC. occidentalis exhibited the highest activity on LEIA (EC50 = 250.42–41.80 μg/mL) and A. absinthium on egg hatching processes (EC50 = 121.70–137.34 μg/mL) in both species of GIN. The inhibition in the development of eggs was from 67.70% to 96.36% on H. contortus, and from 78.87% to 99.65% on T. colubriformis. At the maximal dose, Additionally, it was observed that the AH on eggs varies according to the GIN species: on H. contortus the extracts tested blocked the formation of larvae Ovicidal Effect (% higher OE) and on T. colubriformis they blocked the appearance of L1 larvae, Larvae Failing Eclosion (% higher LFE). After PVPP, a reduction in AH activity on LEIA and EHA was noted, especially with C. occidentalis (87.20–67.00% of larvae exsheathment, (p < 0.05) and 40.51–24.96% of egg hatching, (p > 0.05) of both parasite species. Nine putative features were identified using HRMS and MS/MS after addition of PVPP. ConclusionsThe present study demonstrated that C. occidentalis, A. absinthium, and K. odorata, which parts have been traditionally used as medicinal plants are a valuable source of active compounds with anthelmintic activity. The medicinal use of these plants against GIN parasites was proven by in vitro analysis. Therefore exploration of the secondary metabolites of these plant extracts and testing of isolated fractions of active compounds under in vivo experiments are planned and represent a specific challenge for alternative drug research. Regarding the PVPP, in this study we hypotheses about the standard doses it was not able to completely absorb the polyphenols of extracts of K. odorata, C. occidentalis, and A. absinthium, which would lead to more studies to evaluate the role of this product in the absorption of phenolic compounds.

Clarifying effect of different fining agents on mulberry wine

AbstractFining experiments have been conducted on mulberry wine, in which five fining agents (bentonite, gelatin, casein, egg albumin and PVPP) were investigated. The effect of these agents on polyphenol content, colour intensity, antioxidant activity and aromatic profile has been determined. In general, all treatments improved the limpidity of the mulberry wines to some extent. The wine was mostly negatively affected by gelatin and led to the greatest reduction of phenol content (16.61%), colour intensity (32.26%) and antioxidant activity. Concerning aroma, the addition of PVPP, casein and bentonite both significantly altered the aroma profile. It is noteworthy that egg albumin led to the lowest diminution total phenol value (5.74%) and improvement of antioxidant capacity and colour among the tested fining agents. Furthermore, the aroma profile remained mostly unaffected by egg albumin. As consequence, the optimal fining agent for mulberry wine was egg albumin.

In Vitro Effect of Degradation in Sacco in the Rumen on the Anthelmintic Properties of Parkia Biglobosa and Pterocarpus Erinaceus on Haemonchus Contortus

Parkia biglobosa and Pterocarpus erinaceus are traditionally used to treat various ailments including helminth infections. This study was undertaken to evaluate the effect of the rumen degradability on anthelmintic properties of the two plants and to examine the possible role of tannins and/or polyphenols on these properties. Hydro-acetone extracts of in sacco degradability residues of P. biglobosa pods and P. erinaceus leaves were screened in vitro to determine the possible anti-parasitic effects against eggs and infective larvae of Haemonchus contortus. In addition, the possible involvement of tannins and/or polyphenols was examined by comparing the levels of inhibition of larval migration obtained with the same extracts, after of not addition of PVPP. Extracts of both plants induced significant egg hatch inhibition (p &lt; 0.001). The effects were dose and incubation period dependent. Extracts of both plants have shown again remarkable larval migration inhibition compared to the PBS (p &lt; 0.05). The effect of incubation period was not significant (p &gt; 0.05). These results suggest that the plants did not lose their anthelmintic properties after rumen degradation. The use of the PVPP indicated for almost all of the extracts that tannins and/or polyphenols are largely involved in the effect. Complementary investigations are necessary to understand the metabolism of these plants in the digestive tract of the animals.

A rapid and efficient SDS-based RNA isolation protocol from different tissues of coffee.

Isolation of high-quality RNA from coffee is challenging because of high level of polysaccharides, polyphenols and other secondary metabolites. In the present study, a rapid and efficient RNA extraction protocol from different tissues of coffee was optimized. Sufficiently high quality and quantity (225.6-454.8µg/g) of RNA was obtained by using the optimized protocol. The presence of two distinct bands of 28S rRNA and 18S rRNA in agarose gel proved the intactness of the RNA samples. The average spectrophotometric values of the isolated RNA ranged from 1.96 to 2.02 (A260/280) and 1.95 to 2.14 (A260/230), indicating the high quality of RNA devoid of polyphenols, polysaccharides and protein contamination. In the optimized protocol, addition of PVPP to the extraction buffer and a brief incubation of samples at 65°C and subsequent purification with potassium acetate resulted in good-quality RNA isolation. The suitability of RNA for downstream processing was confirmed by PCR amplification with cytochrome c oxidase gene-specific primers. The amplification of a single 392bp fragment using cDNA and 1.5kb fragment using genomic DNA samples confirmed the absence of DNA contamination. The present protocol is rapid and yielded good quality and quantity of RNA suitable for functional genomics studies.

In vitro action of Mimosa caesalpinifolia ketone extract on Haemonchus contortus and Trichostrongylus colubriformis

This study aimed to evaluate the in vitro anthelmintic activity of Mimosa caesalpinifolia (sabiá) ketone extract, which is rich in condensed tannins (CT), on Haemonchus contortus and Trichostrongylus colubriformis. The leaves and stems of M. caesalpinifolia were collected, dried in the shade, and ground to a dry powder from which ketone extracts were obtained for in vitro assays. The extracts were diluted in 5% acetone and PBS to concentrations in the range 0.25-2.80 mg mL-¹ and the inhibition of egg hatchability was assayed. Feces collected from goats naturally infected with H. contortus (55%) and T. colubriformis (45%) were macerated and the helminth eggs were collected to conduct in vitro egg hatchability tests. Nematode larvae collected from the feces were used for a larval exsheathment inhibition assay of M. caesalpinifolia leaf and M. caesalpinifolia leaf and stem ketone extracts at concentrations of 0.6 and 1.2 mg mL-¹; three replicates were performed for each concentration. PBS was used as a negative control. Polyvinylpolypyrrolidone (PVPP) was added to the extracts to assess the effect of phenolic compounds on anthelmintic activity. An inhibitory effect of 80.7% and 82.3% was observed on hatchability after treatment with 1.0 and 2.0 mg mL-¹ M. caesalpinifolia leaf ketone extract, respectively. An inhibition rate lower than 75% was observed for 1.0 mg mL-¹ M. caesalpinifolia leaf and stem ketone extract. The M. caesalpinifolia leaf ketone extract larval exsheathment assay resulted in inhibition of 97.3% and 99.8% at concentrations of 0.6 and 1.2 mg mL-¹, respectively. For the leaf and stem ketone extract, the larval exsheathment inhibition was 94.3% at 1.2 mg mL-¹. The leaf and leaf and stem extracts showed no inhibitory effects on larval exsheathment after the addition of PVPP. These results showed that cuticular loss was related to the action of the phenolic compounds in the M. caesalpinifolia extracts. M. caesalpinifolia leaf ketone extract inhibited egg hatchability, but the leaf and stem extract showed lower efficiency, which indicated a greater abundance of active compounds in the leaves. The M. caesalpinifolia ketone extracts were effective in the inhibition of larval exsheathment at the highest concentration tested. Based on these results, in vivo studies should be performed to demonstrate the anthelmintic activity of this plant in goats.

In vitro assessment of the anthelmintic activity of Hedysarum carnosum Desf. at different phenological stages and from six locations in Tunisia.

Gastrointestinal nematodes are compromising productivity of grazing sheep and goats. Therefore, scientists have been looking for cost-effective alternative options. Forage legumes (Fabacea Family) contain tannins that could improve livestock performance and their health as well. The present study aimed to (i) determine the in vitro anthelmintic (AH) activity of 19 acetonic extracts of Hedysarum carnosum Desf on Haemonchus contortus by a larval exsheathment assay (LEA); (ii) test the anthelmintic activity of condensed tannins using a deactivating reagent, polyvinylpolypyrrolidone (PVPP); (iii) study the effect of location and the phenological stage on the percentage of exsheathment. The LEA was used at different concentrations (150, 300, 600, 1200 µg mL-1 of acetonic extract/mL of purified buffer solution (PBS)). The larval exsheathment is concentration, location, phenological stage dependent. All extracts, caused a delay of the percentage of exsheathment over 50% so the AH activity of H. carnosum was confirmed. After addition of PVPP, the % exsheathment was similar to the 150 µg mL-1 concentration. The biplot showed that Loc1(S), Loc4(B), Loc 5(PF), Loc 6(BM) and Loc 6(PF) were isolated from other plant extract sample. Our in vitro study showed that H. carnosum seems to be a promising alternative to AH drugs.

Anthelmintic activity of acetone–water extracts against Haemonchus contortus eggs: Interactions between tannins and other plant secondary compounds

This study aimed at (i) describing the effects of acetone–water extracts obtained from a range of different plant materials, on the hatching process of Haemonchus contortus eggs under in vitro conditions and (ii) identifying the role of tannins and other plant secondary compounds (PSC), on these AH effects by using polyvinylpolypyrrolidone (PVPP), an inhibitor of tannins and other polyphenols. An egg hatch assay (EHA) was used to determine the AH effect. Acetone–water (70:30) extracts from different foliages (Lysiloma latisiliquum, Laguncularia racemosa, Rizophora mangle, Avicennia germinans) and plant by-products (Theobroma cacao seed husk and pulp, and percolated Coffea arabica) were obtained. Fresh H. contortus eggs were incubated in PBS with increasing concentrations of each extract (0, 600, 1200, 2400 and 3600μg/ml PBS). A general linear model was used to determine the dose effect of each extract. A mild ovicidal activity was only recorded for T. cacao extracts (seed husk and pulp). The main anthelmintic (AH) effect for all the extracts, except for C. arabica, was to block the eclosion of larvated eggs. The use of PVPP at 3600μg/ml PBS showed that tannins of the L. racemosa extract were responsible for blocking eclosion of larvated eggs. Extracts of L. latisiliquum, A. germinans, T. cacao seed husk and pulp also blocked eclosion of larvated eggs but the addition of PVPP indicated that tannins were not responsible for that activity. In contrast, it suggested unfavorable interactions between polyphenols and other PSC contained in those extracts, limiting the AH effect on the egg hatching process. The present results suggest that the interactions between tannins and other PSC are complex and may reduce the AH effects against H. contortus eggs.

The horse: An emerging model for human infertility research

This study aimed at (i) describing the effects of acetone–water extracts obtained from a range of different plant materials, on the hatching process of Haemonchus contortus eggs under in vitro conditions and (ii) identifying the role of tannins and other plant secondary compounds (PSC), on these AH effects by using polyvinylpolypyrrolidone (PVPP), an inhibitor of tannins and other polyphenols. An egg hatch assay (EHA) was used to determine the AH effect. Acetone–water (70:30) extracts from different foliages (Lysiloma latisiliquum, Laguncularia racemosa, Rizophora mangle, Avicennia germinans) and plant by-products (Theobroma cacao seed husk and pulp, and percolated Coffea arabica) were obtained. Fresh H. contortus eggs were incubated in PBS with increasing concentrations of each extract (0, 600, 1200, 2400 and 3600 μg/ml PBS). A general linear model was used to determine the dose effect of each extract. A mild ovicidal activity was only recorded for T. cacao extracts (seed husk and pulp). The main anthelmintic (AH) effect for all the extracts, except for C. arabica, was to block the eclosion of larvated eggs. The use of PVPP at 3600 μg/ml PBS showed that tannins of the L. racemosa extract were responsible for blocking eclosion of larvated eggs. Extracts of L. latisiliquum, A. germinans, T. cacao seed husk and pulp also blocked eclosion of larvated eggs but the addition of PVPP indicated that tannins were not responsible for that activity. In contrast, it suggested unfavorable interactions between polyphenols and other PSC contained in those extracts, limiting the AH effect on the egg hatching process. The present results suggest that the interactions between tannins and other PSC are complex and may reduce the AH effects against H. contortus eggs.

Involvement of tannins and flavonoids in the in vitro effects of Newbouldia laevis and Zanthoxylum zanthoxyloïdes extracts on the exsheathment of third-stage infective larvae of gastrointestinal nematodes

The present study aimed at examining the possible role of tannins and flavonoids on the in vitro anthelmintic properties of the extracts of two plants from the southern area of Western Africa, i.e. Newbouldia laevis and Zanthoxylum zanthoxyloïdes. Extracts of the two plants were prepared by use of acetone/water (70/30) and their anthelmintic activity was measured by use of the larval exsheathment inhibition assay (LEIA) applied on the abomasal species, Haemonchus contortus and the intestinal species Trichostrongylus colubriformis. Three concentrations of extracts were evaluated to examine the possible dose effect. In addition, the possible involvement of tannins and flavonoids was examined by comparing the levels of inhibition of larval exsheathment obtained with the same extracts, after of not addition of PVPP which forms complexes with these compounds. The results indicate significant effects with both plants and both nematode species. In the range of concentrations examined, the results were dose-dependent for N. laevis extracts but not for Z. zanthoxyloïdes because the three doses applied provoked a similar highly significant inhibition whatever the tested dose. The use of PVPP indicated for both plant and nematode species, that tannins and flavonoids are involved partly in the effect but that some other biochemical compounds were also involved in both plants.

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.

Efficacy of polyvinylpolypyrrolidone in reducing the immunotoxicity of aflatoxin in growing broilers

1. Protective action of an enzyme-linked polyvinylpolypyrrolidone (PVPP, Mycofix ® Plus) against the immunosuppressive effect of afatoxins (AF) was evaluated by determination of peripheral blood T-lymphocyte proportions and splenic plasma cell counts. Histological changes in lymphoid organs were also investigated by light microscopy. One-d-old broiler chicks (Hybro) received 2·5 mg/kg diet AF (83·06% AFB1, 12·98% AFB2, 2·84% AFG1, 1·12% AFG2) with or without PVPP (3 g/kg diet) until 21 d of age. 2. When compared with the controls, AF treatment significantly decreased peripheral T-lymphocyte counts. AF caused a slight decrease in splenic plasma cell counts. The addition of PVPP to an AF-containing diet significantly increased T-lymphocyte counts. Splenic plasma cell counts were numerically intermediate between control and AF groups. 3. The dietary addition of PVPP to AF-free diet did not significantly alter either T-lymphocyte or splenic plasma cell counts.

Effect of Variety, Ultraviolet Light Exposure, and Enological Methods on thetrans-Resveratrol Level of Wine

Resveratrol is one of the wine components related to moderate wine consumption and reduction of serum cholesterol levels. The standard addition method was used with a high pressure liquid chromatography procedure to calculate resveratrol levels. Grape variety, UV light exposure, enzyme addition, skin contact time, and the fining agents, carbon and PVPP, affected the resveratrol levels of wines from the white varieties, Chardonnay and Chardonel, and the red varieties, Cabernet Sauvignon, Cynthiana, and Noble. UV light exposure of the grape clusters significantly increased the resveratrol level in Cynthiana and Noble wine produced in 1994, but did not yield a consistent result in the other experiments from the 1995 and 1996 wines. Enzyme addition significantly increased resveratrol levels in Chardonnay wine but not Chardonel wine. Skin contact time influenced the extraction of resveratrol from the skin in the red varieties, but the maximum extraction time was dependent on the variety of the grapes. Carbon addition decreased the resveratrol level in Noble wine but not in Cynthiana in the 1994 wines. The addition of PVPP significantly lowered resveratrol level in Chardonel, Chardonnay, Cabernet Sauvignon, Cynthiana, and Noble wine. However, low levels of added PVPP addition did not cause significant loss of resveratrol.

Allelopathic effects of white clover 1. Germination and chemical bioassay

Abstract Seed of 10 pasture species, germinated in the presence of an aqueous extract of freeze-dried white clover shoot material, showed inhibition of germination, and seedling abnormalities. A rate equivalent to 800 kg/ha of white clover dry matter (DM) almost totally inhibited the germination of white clover, alsike clover, cocksfoot, and perennial ryegrass seed. The addition of PVPP was able to remove up to 75% of observed inhibition. White clover plants grown in contrasting high country environments seldom produced levels of allelochemicals different to lowland white clover although summer-dry environments tended to promote allelochemical production. A chromatographic investigation implicated phenolics as the active allelochemicals.