Effect of flavan-3-ols on in vitro egg hatching, larval development and viability of infective larvae of Trichostrongylus colubriformis

The effects of flavan-3-ols (the monomer units of condensed tannins (CT)) and their galloyl derivatives on the viability of eggs, the development of first stage (L1) larvae, and the viability of the infective larvae of Trichostrongylus colubriformis were investigated under in vitro conditions. Each of the flavan-3-ol gallates showed some inhibition of egg hatching at 100 μg/ml, and 100% inhibition at 1000 μg/ml, with epigallocatechin gallate being the most effective in the egg hatch (EH) assay. In contrast, none of the flavan-3-ols were able to completely inhibit egg hatching. The flavan-3-ols and galloyl derivatives dose-dependently inhibited the development of infective larvae as assessed by the larval development (LD) assay. A larval migration inhibition (LMI) assay was used to assess the effect of flavan-3-ols and their galloyl derivatives on the motility of the infective third-stage (L3) larvae of T. colubriformis. In general, the flavan-3-ol gallates were more effective than the flavan-3-ols at immobilising the infective larvae as evidenced by their ability to inhibit more (P<0.05–0.01) larvae from passing through the LMI sieves. At 500 μg/ml, epigallocatechin gallate inhibited significantly more (P<0.1) larvae from passing through the sieves than did catechin gallate, epicatechin gallate, or gallocatechin gallate. Comparisons were made between the flavan-3-ols and their galloyl derivatives with the in vitro effects of CT extracts from several forage legumes, which have exhibited effects on parasites in vivo. The forage legumes tested at 200–500 μg/ml reduced the proportion of eggs that hatch, with comparable results to those obtained using the flavan-3-ols. The activities may be influenced by the prodelphinidin: procyanidin (PD:PC) ratios: CT extracts from Lotus pendunculatus and sainfoin have PD:PC ratios of 70:30 and 77:23, respectively, whereas the less active CT extract from Lotus corniculatus has a PD:PC ratio of 27:73. The active CT extracts from forage legumes have epigallocatechin as the dominant flavan-3-ol extender unit, and epigallocatechin is the most active flavan-3-ol in both the EH and LD assays.     

Effective degradation of tannic acid by immobilized rumen microbes of a sika deer (Cervus nippon yesoensis) in winter

In winter seasons, wild sika deer (Cervus nippon yesoensis) inhabiting the Shiretoko Peninsula of Hokkaido Island, Japan, mainly graze woody materials (bark and twigs, etc.) as their feed source. Most of the tree species that they feed upon contain a high level of hydrolysable tannins within the inner bark. Tannins generally lead to low protein digestion and nutrient loss to these herbivorous mammals due to tannization of proteins. In winter months, it is speculated that wild sika deer develop a mechanism to degrade the tannins which are contained in their feed sources, but rumen fluid obtained from sika deer in winter months did not exhibit any ability to degrade tannins in liquid culture medium. However, constant degradation of hydrolysable tannin was observed when Ca-alginate gel beads were used for microbial immobilization and culturing. The gel beads that had been impregnated with 0.6×10⁴ fold-diluted rumen fluid of sika deer in winter and pre-incubated for 24 h under anaerobic conditions supplemented with a 1.5 g/L sugar were reacted with 5 g/L tannic acid solution. Under these conditions, the immobilized rumen bacteria grown in the macrogel beads effectively hydrolyzed tannic acid to release gallic acid monomers. Major bacterial colonies emerging in the Ca-alginate gel beads were identified as Streptococcus macedonicus and this bacterium (EC-D140) was regarded as the most likely candidate as the tannin-degrading bacterium.     

Influence of ruminal bypass on the pharmacokinetics and efficacy of benzimidazole anthelmintics in sheep

Oxfendazole, fenbendazole and albendazole were each administered at 5mgkg(-1) to sheep fitted with abomasal cannulae as a single bolus intra-ruminally or infused intra-abomasally at a declining exponential rate, with half-life equivalent to the rate of rumen fluid outflow. The pharmacokinetic disposition of parent compound and metabolites in plasma and abomasal fluid was determined by high performance liquid chromatography. Compared with intra-ruminal administration, intra-abomasal infusion of fenbendazole lowered the area under the concentration-time curve of drug in both plasma and abomasal fluid; intra-abomasal infusion of albendazole substantially increased maximum drug concentration and the concentration-time curve in abomasal fluid and lowered the plasma concentration time curve of the sulphoxide metabolite; intra-abomasal infusion of oxfendazole increased maximum concentration and the concentration-time curve of drug in plasma and abomasal fluid. The greater availability in abomasal fluid of oxfendazole and albendazole when given at commercial dose rates of 5 mg kg(-1) and 3.9 mg kg(-1), respectively, by intra-abomasal infusion correlated with increased efficacy of both drugs against benzimidazole-resistant Trichostrongylus colubriformis and of albendazole against benzimidazole-resistant Haemonchus contortus over that achieved by intra-ruminal administration as a single bolus.     

Identification of first-stage dorsal-spined lungworm larvae of Tunisian barbary red deer: First report of Varestrongylus sagittatus and Elaphostrongylus cervi in Africa

Barbary red deer (Cervus elaphus barbarus) is a protected rare subspecies of red deer. The study of its Protostrongylidae fauna based only on sporadic necropsy of naturally dead animals is difficult. Therefore diagnosis of lungworms rely mainly on the identification of the first stage larvae (L1). The L1 of the different species are not readily diagnosed on morphological basis since much variation is recorded within and among dorsal-spined larvae belonging to various species. The aim of this study was to identify the dorsal-spined lungworm larvae of the Barbary red deer. A discriminant function was established, using the measurements of L1 lungworms recorded from red deer in the literature, then applied to identify 220 dorsal-spined larvae extracted from 25 Tunisian Barbary red deer fresh fecal samples. Also the ITS2 region of rDNA of four pools of larvae (n?=?25–60) were amplified, sequenced and analyzed. Using discriminant analysis of morphological traits, Elaphostrongylus cervi and Varestongylus sagittatus were identified. Molecular identification confirmed the presence of E. cervi which is the most prevalent species. This study represents the first identification of V. sagittatus and E. cervi in North Africa.     

Brugia malayi: effects of antibacterial agents on larval viability and development in vitro

Recent studies have suggested that intracellular Wolbachia bacteria are necessary for reproduction and survival of adult filarial worms. We now report results of in vitro studies of effects of antibacterial antibiotics (tetracycline, rifampicin, chloramphenicol, azithromycin, and doxycycline) on Brugia malayi infective larvae (L3) motility and molting. All of the antibiotics tested except chloramphenicol decreased L3 motility by 50% or more at 10 days, with minimal effective concentrations (MECs) of 20-100 microg/ml. Tetracyclines, rifampicin, and chloramphenicol inhibited L3 to L4 molting by 12 days in a concentration- and time-dependent manner, with MECs in the range of 1-20 microg/ml. These studies show that antibiotics active against Rickettsiaceae inhibit B. malayi L3 molting at low concentrations in vitro; higher concentrations kill the larvae. While it is possible that antibiotics directly affect filarial L3, we believe it is more likely that the effects seen are indirect effects related to bacterial killing.     

The effect of condensed tannins from Lotus pedunculatus and Lotus corniculatus on the growth of proteolytic rumen bacteria in vitro and their possible mode of action

Five strains of proteolytic rumen bacteria were treated with condensed tannins (CT) purified from Lotus pedunculatus and Lotus corniculatus to investigate their effect on the growth of these bacteria in vitro. Streptococcus bovis NCFB 2476, Eubacterium sp. C124b, Prevotella bryantii B14, Butyrivibrio fibrisolvens H17c, and Clostridium proteoclasticum B316T were tested against 200, 400, and 600 microg CT x mL(-1) extracted from L. pedunculatus and L. corniculatus. In the absence of CT, all bacterial strains showed typical growth and reached maximum optical density (OD) after 6-8 h of incubation in a plant protein medium. Growth of Eubacterium sp., P. bryantii, and B. fibrisolvens was inhibited (P < 0.01-0.001) more by the CT from L. pedunculatus than by the CT from L. corniculatus. All strains continued to grow in the presence of 200 microg x mL(-1) of the CT from L. pedunculatus, but attained significantly (P < 0.05-0.01) lower maximum OD600 values than (minus CT) controls, except for S. bovis. At 400 and 600 microg x mL(-1), the addition of CT from L. pedunculatus inhibited (P < 0.05-0.001) the growth of all bacterial strains tested compared with controls. The growth of Eubacterium sp. and P. bryantii was stimulated for the first 4-6 h of incubation (P < 0.001) by 200 microg x mL(-1) of CT from L. corniculatus, but then declined leading to a significant difference in OD values compared with the controls. At 400 microg x mL(-1), the CT from L. corniculatus reduced (P < 0.05-0.01) the growth of all strains except S. bovis, while 600 microg x mL(-1) inhibited (P < 0.01-0.001) the growth of all strains. To study the mechanism of CT action, the degradation of the large subunit (LSU) of ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco; Fraction 1 Leaf protein) was followed after bacterial cells or Rubisco were preincubated with CT extracted from L. corniculatus and L. pedunculatus. Both preincubations decreased LSU degradation, but they differed in their response to polyethylene glycol (PEG) addition. Addition of PEG to CT-Rubisco preincubations negated the effects of CT, while PEG addition to CT-bacteria preincubations did not. This implies that the CT-bacterial interaction is stronger than the CT-Rubisco interaction or the interaction is of a different type. Also, L. pedunculatus CT reduced the degradation of the LSU to a greater extent than the CT from L. corniculatus when preincubated with bacteria.     

Alternative temperate forages containing secondary compounds for improving sustainable productivity in grazing ruminants

The use of alternative temperate forages to improve the sustainable productivity of grazing ruminants, relative to grass-based pastures, is reviewed. Particular emphasis is placed upon forages containing secondary compounds for sustainable control of internal parasites, for increasing reproductive rate in sheep, reducing bloat risk in cattle and for reducing methane production as a means of lowering greenhouse gas emissions.

Of the forages reviewed, the herb chicory (Chicorium intybus) and the condensed tannin-containing legumes Lotus corniculatus L. and sulla (Hedysarum coronarium) offered the most advantages. Chicory and sulla promoted faster growth rates in young sheep and deer in the presence of internal parasites, and showed reduced methane production in other studies. L. corniculatus was not as effective as chicory and sulla in promoting growth of lambs in the presence of internal parasites. Grazing on L. corniculatus was associated with increases in reproductive rate in sheep, increases in milk production in both ewes and dairy cows and reduced methane production, effects that were mainly due to its content of condensed tannins (CT). Grazing ewes on L. corniculatus during mating and very early pregnancy may also reduce lamb mortality. However, there are no data on the effect of mating ewes, which are grazing chicory on their reproductive performance, an important omission. Risk of rumen frothy bloat in cattle grazing legumes is reduced when the forage contains 5 g CT/kg dry matter (DM) or greater. Gene transfer techniques aimed at achieving this for lucerne (Medicago sativa) have made progress, but CT concentration needs to be further increased from calculated values of 0.75–1.25 g CT/kg DM in the transformed plants. Bloat control may be achievable in genetically transformed legumes before increased amino acid absorption, as the concentration of CT required for bloat control is lower (5 versus 30–40 g/kg DM) than that required to cause increased amino acid absorption and is not affected by differences in CT structure.

Key plant characteristics for improved sustainable productivity are a high ratio of readily fermentable: structural carbohydrate and the presence of CT and certain other secondary compounds.

Taking into account both nutritional and agronomic considerations, chicory is considered one of the best emerging plants for grazing livestock, with L. corniculatus being more suitable for areas with dry summers and warm winters. Some of the agronomic limitations of L. corniculatus and sulla could be reduced by mechanical harvesting and their inclusion as a component in total mixed rations (TMR), instead of grazing.     

In vitro stress selection of nematophagous fungi for biocontrol of parasitic nematodes in ruminants

Laboratory experiments were designed to select nematophagous fungi that were able to survive in vitro conditions simulating passage through the gastro-intestinal tract of cattle. All of the tests were conducted at 39 degrees C. In a primary stress selection step in diluted rumen fluid, 21 isolates were obtained. Each of the primary stress selected isolates was tested in synthetic saliva, rumen fluid simulating the activity in the rumen, rumen fluid followed by pepsin-hydrochloric acid treatment simulating the additional effect of ruminal and abomasal activity, pepsin-hydrochloric acid solution simulating conditions in the abomasum and finally in a trypsin solution as an example of enzyme activity in the gut. The effect of the rumen fluid alone, or rumen fluid followed by pepsin-hydrochloric acid treatment, were responsible for the reduction in surviving fungal isolates. Only six of thirteen isolates belonging to the genus Arthrobotrys survived while seven of eight isolates of the genus Duddingtonia survived. Fourteen isolates were tested for their predatory capacity in a dung pat bioassay. Fungi of the genera Arthrobotrys and Duddingtonia reduced the development of Ostertagia ostertagi third stage larvae by approximately 75% and 96% respectively compared to the number of larvae that developed from fungus-free control pats.     

Hypergastrinaemia, abomasal bacterial population densities and pH in sheep infected with Ostertagia circumcincta.

Serum gastrin and pepsinogen concentrations, food intake, abomasal pH and abomasal aerotolerant and anaerobic bacterial populations were measured in sheep infected with Ostertagia circumcincta to search for links between hypergastrinaemia, food intake and changes in the abomasal environment. Abomasal pH and serum gastrin and pepsinogen concentrations were elevated in each of five sheep infected via abomasal cannulae with 150000 exsheathed larval stage three, followed 11 days later by 100000 sheathed larvae given intraruminally. Unparasitised abomasa contained aerotolerant bacterial population densities of between 10(3) and 10(6) cells ml(-1) and these did not change significantly following parasitism. In contrast, anaerobic bacterial population densities increased markedly by about 10(4)-fold following parasitism. Anaerobic numbers changed rapidly when abomasal pH increased from 2.5 to 3.5. At pH 4 and above, anaerobic bacterial numbers approached levels expected in rumen contents but parameters other than pH did not relate to bacterial numbers. Brief periods when serum gastrin was lower than expected, coinciding with raised abomasal pH, were not explicable by increased bacterial numbers. Food intake, which decreased for a variable period from around Day 5 p.i., correlated poorly with serum gastrin concentration, suggesting hypergastrinaemia is not the sole cause of anorexia in parasitised animals. The survival of substantial numbers of rumen bacteria in the abomasum at only slightly raised pH may significantly lower the bacterial protein available to the sheep.     

Condensed Tannins' Effect on White-Tailed Deer Forage Digestibility in Mississippi

Abstract: Condensed tannins (CT) can reduce digestibility of forages for white-tailed deer (Odocoileus virginianus), potentially confounding estimates of diet quality and nutritional carrying capacity. We collected 143 spring and 142 summer samples of 8 important deer forage species from 22 properties in Mississippi, USA, and tested for CT content using a modified butanol-HCl assay. Three species (partridge pea [Chamaecrista fasciculata], southern dewberry [Rubus trivialis], and roundleaf greenbrier [Smilax rotundifolia]) contained CT, ranging from 0.11% to 6.46% dry weight. Summer CT concentration was greater than in spring for 2 species. We ranked soil samples from least to most fertile using 8 chemical characteristics and found a positive correlation between fertility and CT concentration for 1 species and no correlation for 2 species. We tested effects of CT concentration on in vitro dry matter disappearance (IVDMD) and in vitro protein digestibility using samples of partridge pea and roundleaf greenbrier and rumen fluid from 3 free-ranging deer. Average IVDMD was reduced 1.9% for each 1% increase in CT concentration. In vitro protein digestibility was reduced 2.5% for each 1% increase in CT concentration. Assuming that our methods reflect the effects of CT on in vivo digestibility, maximum loss of available crude protein (CP) in our samples was 3.0 g/100 g dry-weight forage, and only 13 of the 112 CT-containing forage samples (12%) would have decreased available CP by >1 g/100 g dry-weight forage. Deer consuming equal portions of sampled forages would lose <1% of dietary CP to CT. Comparisons of foraging area quality using crude protein estimates should be unaffected by CT under reasonable restrictions of similar habitat types, soil fertility, and time. Given the ability of deer to forage selectively and the abundance of alternative forages in Mississippi, the potential for CT to substantially affect spring or summer diet quality of deer appears minimal.     

Effect of aqueous leaf extract of Irvingia gabonensis on gastrointestinal tract in rodents

Effect of the aqueous leaf extract of I. gabonensis on the gastrointestinal tract was investigated on isolated rabbit jejunum, guinea pig ileum, gastrointestinal motility, castor oil-induced diarrhoea in mice and castor oil-induced fluid accumulation in rats. The results showed that the extract exhibited a concentration-dependent relaxation of spontaneous pendular movement of isolated rabbit jejunum and guinea pig ileum, and attenuated both acetylcholine-induced contraction of rabbit jejunum and histamine-induced contraction of guinea pig ileum. The extract (100, 200 and 400 mg/kg) also caused a significant dose-dependent decrease of gastrointestinal motility in mice (40.12, 39.45 and 37.45%), intestinal fluid accumulation in rats (71.43, 81.63 and 83.27%), and remarkably protected mice against castor oil-induced diarrhoea [58.33, 75 and 91.67% (Di Carlo score)] respectively. Preliminary phytochemical screening of the aqueous leaf extract of I. gabonensis revealed the presence of saponins, tannins, phenols and phlobatanins.     

Lotus corniculatus condensed tannins decrease in vivo populations of proteolytic bacteria and affect nitrogen metabolism in the rumen of sheep

Condensed tannins in forage legumes improve the nutrition of sheep by reducing ruminal degradation of plant protein and increasing crude protein flow to the intestine. However, the effects of condensed tannins in forage legumes on rumen bacterial populations in vivo are poorly understood. The aim of this study was to investigate the specific effects of condensed tannins from Lotus corniculatus on four proteolytic rumen bacteria in sheep during and after transition from a ryegrass (Lolium perenne)-white clover (Trifolium repens) diet (i.e., low condensed tannins) to a Lotus corniculatus diet (i.e., higher condensed tannins). The bacterial populations were quantified using a competitive polymerase chain reaction. Lotus corniculatus was fed with or without ruminal infusions of polyethylene glycol (PEG), which binds to and inactivates condensed tannins, enabling the effect of condensed tannins on bacterial populations to be examined. When sheep fed on ryegrass-white clover, populations of Clostridium proteoclasticum B316T, Butyrivibrio fibrisolvens C211a, Eubacterium sp. C12b, and Streptococcus bovis B315 were 1.5 x 10(8), 1.1 x 10(6), 4.6 x 10(8), and 7.1 x 10(6) mL(-1), respectively. When the diet was changed to Lotus corniculatus, the average populations (after 8-120 h) of C. proteoclasticum, B. fibrisolvens, Eubacterium sp., and S. bovis decreased (P < 0.001) to 2.4 x 10(7), 1.1 x 10(5), 1.1 x 10(8), and 2.5 x 10(5) mL(-1), respectively. When PEG was infused into the rumen of sheep fed Lotus corniculatus, the populations of C. proteoclasticum, B. fibrisolvens, Eubacterium sp., and S. bovis were higher (P < 0.01-0.001) than in sheep fed Lotus corniculatus without the PEG infusion, with average populations (after 8-120 h) of 4.9 x 10(7), 3.8 x 10(5), 1.9 x 10(8), and 1.0 x 10(6), respectively. Sheep fed the Lotus corniculatus diet had lower rumen proteinase activity, ammonia, and soluble nitrogen (P < 0.05-0.001) than sheep that were fed Lotus corniculatus plus PEG. The Lotus corniculatus diet reduced rumen nitrogen digestibility (P < 0.05) and ammonia pool size and increased the flow of undegraded feed nitrogen to the abomasum. The nitrogen intake, rumen non-ammonia nitrogen pool size, rumen microbial non-ammonia nitrogen pool size, and abomasal microbial non-ammonia nitrogen fluxes were similar both in sheep fed only Lotus corniculatus and in sheep fed Lotus corniculatus plus PEG, but nonmicrobial non-ammonia nitrogen flux to the abomasum was higher (P < 0.01) for the sheep fed only Lotus corniculatus. Although condensed tannins in Lotus corniculatus reduced the populations of some proteolytic bacteria, total ruminal microbial protein and microbial protein outflow to the abomasum were unchanged, suggesting a species-specific effect of condensed tannins on bacteria in the rumen.     

Heterologous transmission with Dictyocaulus capreolus from roe deer (Capreolus capreolus) to cattle (Bos taurus)

Eight Swedish Red Breed cattle, about 2 months old, were experimentally infected with a Swedish isolate of Dictyocaulus viviparus (Dviv-Se) from cattle and D. capreolus from roe deer. The aims were to determine whether the roe deer lungworm is infective to cattle or if it can induce seroconversion in cattle against D. viviparus as measured with an ELISA. Four calves which were given 500 Dviv-Se infective larvae (L3) each by larval dosing for two successive days developed patent infection between days 23 and 25 post-inoculation (PI). Larval output varied among the calves and during the patent period. However, maximum recovery occurred between 28 and 56 days PI with peak shedding on day 37 PI. Shedding ceased at day 58 PI and adult worms were recovered from one calf at necropsy (day 67 PI). No immature worms were recovered from the lungs at necropsy. Seroconversion was detected on days 35-42 PI. One Dviv-Se infected calf became seronegative on day 67 PI whereas the other calves still remained seropositive during this period. Prepatency and patency periods of D. viviparus and serological findings in this study basically conform to previous studies. Each calf that was infected with 400 L3 of D. capreolus for two successive days, and about 800 L3 of the same species about 8 weeks later, did not develop to patency based on faecal and post-mortem examinations. Consequently, under the conditions of this study, D. capreolus was not infective to cattle. Two of the four calves that were infected with L3 from roe deer were challenged with L3 cultured from faeces of the Dviv-Se-infected calves. This infection did not develop to patency. Whether this was due to cross-protection as a result of the prior priming with L3 from roe deer is not clear. However, if it is so, it opens up the possibility of using D. capreolus L3 for preventing bovine dictyocauliasis.     

Effect of feeding dehydrated and ensiled tanniferous sainfoin (Onobrychis viciifolia) on nitrogen and mineral digestion and metabolism of lambs

The effects of tanniferous sainfoin on digestion and metabolism have been investigated in 12 lambs in an incomplete cross-over design (n = 6). Effects of condensed tannins (CT) were evaluated by comparing dehydrated and ensiled sainfoin treated with and without polyethylene glycol (PEG). Dehydrated and ensiled grass-clover mixtures served as controls. The lambs were fed the treatment diets, including a mineral supplement, for 21 d. During the last 7 d excreta, rumen fluid and blood were sampled. The CT of sainfoin decreased rumen fluid ammonia concentration (p < 0.001) and increased the plasma concentration mainly of essential amino acids (p < 0.001). Body retention of phosphorus, calcium and magnesium was lower with sainfoin compared to PEG-treated sainfoin (p < 0.05). Sainfoin without PEG resulted in lower digestibilities of organic matter and neutral detergent fibre than sainfoin with PEG and the grass-clover mixture (p < 0.001). Ensiling of sainfoin led to the lowest N-retention. In conclusion, the reduction in ruminal ammonia and urine-N losses by sainfoin CT did not improve N-retention.     

Effects of sainfoin (Onobrychis viciifolia) extract and monomers of condensed tannins on the association of abomasal nematode larvae with fundic explants

Tannin-rich forages offer an alternative to anthelmintic chemicals to control gastrointestinal nematodes. However, the mode of action of such bioactive plants still needs to be assessed. Previous studies have shown that extracts of tannin-rich plants interfere with the first phase of host invasion, i.e., the exsheathment of infective larvae (L3s). In the current study, we examined the hypothesis that exposure to tannins could also affect the second phase of larval establishment, i.e., the tissue association/penetration of the exsheathed L3s into the digestive mucosae. An in vitro direct challenge technique using fundic explants was applied in this study. The main parasite model was Haemonchus contortus. The objectives were to verify: (i) whether a modification of the association/penetration of L3s with the mucosae occurred after contact with sainfoin extract; (ii) whether this is a dose-dependent phenomenon; (iii) whether tannins were responsible for these effects; (iv) whether these effects were dependent on the parasite species; and (v) how the biochemical structure of tannins might influence these effects. Following 3 h contact with sainfoin extract at 1,200 μg/ml, the penetration of exsheathed L3s of H. contortus and Teladorsagia circumcincta into fundic explants was significantly reduced. Moreover, a dose–response relationship was found for H. contortus. For both nematodes, the changes were totally alleviated after addition of polyvinyl polypyrrolidone, an inhibitor of tannins, to the sainfoin extract, suggesting that tannins play a major role in the observed effects. Comparison of results obtained with different monomers of condensed tannins confirms a relationship between structure and activity, the prodelphinidin monomers and galloyl-derivatives being more effective than the procyanidin monomers. Combined with the delay or the inhibition of larval exsheathment previously shown, these effects could explain how tanniniferous plants reduce the establishment of infective larvae in small ruminants.     

Effect of Various Essential Oils Isolated from Douglas Fir Needles upon Sheep and Deer Rumen Microbial Activity

The effects of essential oils isolated from Douglas fir needles on sheep and deer rumen microbial activity were tested by use of an anaerobic manometric technique. Rumen microorganisms were obtained from a sheep which had been fed mainly on alfalfa hay and dried range grass. One deer used in this study had access to Douglas fir trees the year around, whereas the other deer had no access to Douglas fir. All of the monoterpene hydrocarbons isolated from Douglas fir needles—α-pinene, β-pinene, limonene, myrcene, camphene, Δ³-carene, and terpinolene—promoted only slightly or had no effect on deer rumen microbial activity, whereas all of them promoted activity in sheep rumen microbes, except Δ³-carene and terpinolene, which inhibited activity. Of the oxygenated monoterpenes, all monoterpene alcohols—α-terpineol, terpinen-4-ol, linalool, citronellol, and fenchyl alcohol—strongly inhibited the rumen microbial activity of both sheep and deer. Monoterpene esters (bornyl acetate) produced mild inhibition for both sheep and deer microbes, and citronellyl acetate inhibited rumen microbial activity in sheep, whereas it promoted activity in both deer. Monoterpene aldehyde (citronellal) inhibited the activity of rumen microbes from both sheep and deer having no access to Douglas fir from the Hopland Field Station, whereas they produced no effect upon the deer having access to Douglas fir from the Masonite forest. Rumen microbial activity for sheep and deer was promoted slightly with aliphatic ester (ethyl-n-caproate). There was a marked difference between sheep and deer rumen microbes as affected by addition of the various essential oils. The monoterpene hydrocarbons promoted activity more on sheep rumen microbes than on deer, and the monoterpene alcohols inhibited sheep rumen microbial activity more than that of deer. Furthermore, the deer rumen microbes from Hopland Field Station were affected more than the deer from Masonite forest.     

Bacterial Mechanisms to Overcome Inhibitory Effects of Dietary Tannins

High concentrations of tannins in fodder plants inhibit gastrointestinal bacteria and reduce ruminant performance. Increasing the proportion of tannin-resistant bacteria in the rumen protects ruminants from antinutritional effects. The reason for the protective effect is unclear, but could be elucidated if the mechanism(s) by which tannins inhibit bacteria and the mechanisms of tannin resistance were understood. A review of the literature indicates that the ability of tannins to complex with polymers and minerals is the basis of the inhibitory effect on gastrointestinal bacteria. Mechanisms by which bacteria can overcome inhibition include tannin modification/degradation, dissociation of tannin–substrate complexes, tannin inactivation by high-affinity binders, and membrane modification/repair and metal ion sequestration. Understanding the mechanism of action of tannins and the mechanism(s) bacteria use to overcome the inhibitory effects will allow better management of the rumen ecosystem to reduce the antinutritional effects of tannin-rich fodder plants and thereby improve ruminant production.     

单宁酸对淡色库蚊抗氰戊菊酯品系和敏感品系幼虫生长发育的影响

单宁酸是一种植物次生代谢物, 为探索其用于蚊幼虫防治的可能性,在室内测定了其对淡色库蚊Culex pipiens pallens抗氰戊菊酯品系和敏感品系1～4龄幼虫的毒性,并观察了其对存活幼虫生长发育的影响。结果表明,淡色库蚊敏感品系幼虫对单宁酸的敏感性比抗氰戊菊酯品系的要高,1～4龄幼虫分别高6.4、4.9、4.7和2.0倍。4个龄期幼虫中,无论是敏感品系还是抗氰戊菊酯品系,均是1龄幼虫对单宁酸的敏感性最高,3龄幼虫最低。在1 000 mg/L单宁酸持续作用下,敏感品系和抗氰戊菊酯品系各龄幼虫的存活率,均随处理时间延长而降低。与对照相比,饲养在100 mg/L～500 mg/L单宁酸溶液中的存活幼虫发育历期延长,敏感品系和抗氰戊菊酯品系发育历期分别延长了34.5～38.3 h和59.2～93.4 h。其中,125 mg/L浓度处理的敏感品系1～4龄幼虫,其发育历期与对照的差异达到了显著水平(P<0.05);抗性品系则在250 mg/L作用下也达到了差异显著水平(P<0.05)。但100～250 mg/L单宁酸处理淡色库蚊抗氰戊菊酯品系和敏感品系1龄幼虫,对其存活幼虫的化蛹率、羽化率和成虫性比均无显著影响。表明单宁酸对淡色库蚊幼虫的影响主要是延迟其生长发育,且影响程度与蚊虫对氰戊菊酯的敏感性有关。     

L3 and adult Ostertagia (Teladorsagia) circumcincta exhibit cyanide sensitive oxygen uptake

Oxygen consumption by L3 and adult Ostertagia (Teladorsagia) circumcincta was examined in vitro to determine whether oxygen can be utilised in metabolism. The oxygen concentration in the abomasal fluid of sheep infected with O. circumcincta was also measured. Rates of consumption (in nmol O2/h/1000 worms) were 13+/-1 in sheathed L3, 34+/-6 in ex-sheathed L3, and 1944+/-495 in adult worms. Constant rates of consumption were maintained until media oxygen concentration dropped to between 10 and 20 microM. Consumption was inhibited 95% by cyanide in L3 and 74% in adults. Oxygen concentration in abomasal fluid varied between 10 and 30 microM in both infected and uninfected animals. During infection, oxygen concentration decreased slightly with increased abomasal pH, though the correlation between the two was poor (r=-0.30). In conclusion, O. circumcincta can consume oxygen and oxygen concentration at the infection site is sufficient to support at least some aerobic metabolism.     

Diversity of bovine rumen methanogens In vitro in the presence of condensed tannins, as determined by sequence analysis of 16S rRNA gene library

Molecular diversity of rumen archaeal populations from bovine rumen fluid incubated with or without condensed tannins was investigated using 16S rRNA gene libraries. The predominant order of rumen archaea in the 16S rRNA gene libraries of the control and condensed tannins treatment was found to belong to a novel group of rumen archaea that is distantly related to the order Thermoplasmatales, with 59.5% (15 phylotypes) and 81.43% (21 phylotypes) of the total clones from the control and treatment clone libraries, respectively. The 16S rRNA gene library of the control was found to have higher proportions of methanogens from the orders Methanomicrobiales (32%) and Methanobacteriales (8.5%) as compared to those found in the condensed tannins treatment clone library in both orders (16.88% and 1.68% respectively). The phylotype distributed in the order Methanosarcinales was only found in the control clone library. The study indicated that condensed tannins could alter the diversity of bovine rumen methanogens.