Rumen enzyme profile and fermentation characteristics in sheep as affected by treatment with sodium lauryl sulfate as defaunating agent and presence of ciliate protozoa

The efficiency of sodium lauryl sulfate as a defaunating agent and effect of rumen protozoa on nutrient utilization, fermentation characteristics and enzyme profile were evaluated in adult sheep maintained on a mixed ration containing 65:35% Pala (Ziziphus numularia) leaf: concentrate. Twenty-one adult Malpura sheep divided into three equal groups (DF, RF and F) were either defaunated by oral administration of sodium lauryl sulfate at the rate of 8 g/100 kg body weight (DF), or defaunated and again refaunated (RF), or maintained faunated (F). Daily dry matter intake was similar in defaunated, refaunated and faunated sheep. However, digestibility of cell wall and cell wall contents (NDF, ADF and cellulose) were lower (P < 0.01) in defaunated than refaunated and faunated sheep. Irrespective of the presence or absence of rumen protozoa, daily intake of DCP and DE were similar in the three experimental groups. Even with similar DM, DCP and DE intake, N-retention, blood glucose level, ruminal concentration of total VFA and total-N were higher (P < 0.01), while rumen pH and NH₃-N concentration were lower (P < 0.01) in defaunated sheep. Ruminal activity of amylase, xylanase, protease and urease enzymes were not influenced by presence or absence of ciliate protozoa. However, carboxymethyl cellulase enzyme activity was lower (P < 0.01) in the rumen of defaunated sheep. The total and differential counts of rumen protozoa were similar in refaunated and faunated sheep indicating lack of residual toxic effect of sodium lauryl sulfate. It is concluded that absence of ciliate protozoa increased ruminal TVFA, total-N with lower NH₃-N concentration and fibre digestibility in sheep. Moreover, sodium lauryl sulfate was fully effective for complete removal of rumen ciliate protozoa and successfully defaunated the sheep.     

Ruminal ochratoxin A degradation—Contribution of the different microbial populations and influence of diet

The mycotoxin ochratoxin A (OTA) is degraded extensively in the rumen. In this study, the relative contribution of different rumen microbial populations (MP) and the effect of diet on degradation of OTA were evaluated in a factorial design experiment. Degradation of OTA was quantified by using the Hohenheim gas test (HGT) in vitro fermentation system. Five different HGT diets were used (concentrate:forage proportions (C:F) – 10:90, 30:70, 50:50, 70:30, 90:10), and donor animals were fed diets with the respective ratio. Diets with the highest concentrate content were supplied with and without 10 g/kg sodium bicarbonate (70:30 BC and 90:10 BC). The MP included whole rumen fluid, fungi + protozoa, bacteria + protozoa, protozoa and bacteria + fungi. Protozoa numbers were counted after 24 h and OTA and ochratoxin alpha (OTα) analysed at 0, 4, 8, 12, 24 h. Area under the curve (AUC) and half-life were calculated for the latter two. The short average OTA half-life for whole rumen fluid of 2.6 h (1.3–4.5 h) demonstrates the high OTA degradation capacity of the rumen MP (i.e., standard HGT inoculum) and corresponds well with published in vivo results. Both MP and diet affected OTA degradation. Interactions among factors occurred (P<0.001), which made it necessary to do further comparisons within factor levels. Among MP, those with bacteria (bacteria + fungi and bacteria + protozoa) had lower AUC values (P<0.001) for OTA (196–673 ng/ml h, meaning higher degradation capacity, than those without bacteria (fungi + protozoa and protozoa; 701–1206 ng/ml h). Whole rumen fluid had the lowest AUC values (146–249 ng/ml h; P<0.05). Diet had a quadratic effect (P=0.001) on protozoal numbers with minimum values for the lowest and highest C:F ratios, for bacteria + protozoa, fungi + protozoa and protozoa, but no corresponding effect was found for OTA degradation parameters. While the generally high capacity to degrade OTA was confirmed, results for the contribution of different microbial groups shed new light on ruminal OTA degradation.     

Effects of dietary whole cottonseed and crude protein level on rumen protozoal population and fermentation parameters

In this investigation in vitro and in vivo trials were performed to determine the efficacy of a cottonseed to limit protozoal population and fermentation parameters. The composition of diets given to the different treatments were as follow: (1) control (without whole cottonseed), 16% crude protein (CP), 3.2% ether extract (E.E.); (2) 20% whole cottonseed, 16% CP, 6.5% E.E.; (3) 20% whole cottonseed, 13% CP, 6.4% E.E. and (4) 20% crushed whole cottonseed, 13% CP, 6.4% E.E. DM disappearance (DMD) and fermentation characteristics of the treatments were determined by in vitro incubation studies. In the in vivo trial, ruminal fluid was taken by rumenocentesis (3 h after feeding) on days 1, 3, 5, 7, 9, 11, 14, 21 and 28 from four sheep fed about treatment diets. The pH and protozoal counts were determined in each sample, while ammonia nitrogen and volatile fatty acid (VFA) were determined in samples taken on days 7, 14, 21 and 28. The in vitro DMD after 24 h incubation decreased (p < 0.01) with the addition of cottonseed in diets 3 and 4 and DMD after 72 h incubation was highest (p < 0.01) for the control diet. The fractional rate of gas production (c) for the control and diet 2 was higher (p < 0.05) than for the diets 3 and 4. Feeding crushed whole cottonseed decreased molar proportion of propionate (p < 0.05) and increased molar proportion of butyrate (p < 0.01). Low crude protein level increased the molar proportion of propionate (p < 0.05) and decreased molar proportion of butyrate (p < 0.05) and cellolytic protozoa population (p < 0.05). Feeding cottonseed decreased (p < 0.05) the total protozoa population from approximately 500,000 to 250,000 ml⁻¹ and Holotrich and cellulolytic protozoa disappeared from the rumen of sheep and only Entodinium sp., remained. This was associated with lower concentration of ammonia nitrogen in rumen fluid of sheep fed diets 4 (p < 0.05) and 2 (p < 0.01). It was concluded that cottonseed reduced rumen fauna and ammonia nitrogen, but had no effect on ruminal VFA while the crushed whole cottonseed decreased molar proportion of isovalerate only. In vivo molar proportion of propionate and butyrate and valerate were increase and decrease, respectively, by decreasing CP percentage in treatment diets.     

Effects of mulberry leaves to replace rapeseed meal on performance of sheep feeding on ammoniated rice straw diet

This study was conducted to investigate effects of leaves of mulberry tree (Morus alba) as a protein supplement to isonitrogenously replace rapeseed meal (RSM) on performance of growing lambs offered ammoniated rice straw (ABRS) (Trial 1), and to evaluate the digestive characteristics of the ABRS supplemented with different ratios of RSM and mulberry leaves in terms of in vitro gas production (Trial 2). In Trial 1, 45 Huzhou lambs were divided into five equal groups according to their body weight and gender. Lambs in each group were kept in three pens (male, female and mixed (one male and two females)), and received one of the following dietary treatments: 100 g RSM (A), 75 g RSM plus 60 g mulberry leaves (B), 50 g RSM plus 120 g mulberry leaves (C), 25 g RSM plus 180 g mulberry leaves (D), and 240 g mulberry leaves (E). All animals were given ABRS ad libitum along with 100 g ground corn per head per day. The intake of ABRS was slightly increased with the supplementary level of mulberry leaves, and hence total intake increased with the increasing level of mulberry leaves. The growth rates were higher in diets A and E than those in other treatments (P<0.05), with little difference between diets A and E, and the slowest in C. Animals of all genders showed a similar trend, though male lambs was higher in weight gain than the female. While feed efficiency was higher in diet A, concentrate consumption per kilogram of weight gain was lower when higher level of mulberry leaves was supplemented (diets D and E). Feed cost per kilogram gain was lower in diets E and A compared to other treatments. Degradation of dry matter in the rumen of sheep were higher for mulberry leaves than for RSM, but crude protein was less degraded for mulberry leaves than for RSM. The potential GP was significantly higher in diet A than those in B, C and D (P<0.05), and higher in E than in C (P<0.05) (Trial 2), indicating a negative associate effect of mulberry leaves and RSM on digestion. It is inferred that mulberry leaves may be used as a protein supplement to ammoniated straw diets to fully substitute for RSM, but these two supplements should unlikely be supplemented together to avoid the negative associate effect.     

Studies on the in vitro cultivation of ciliate protozoa from the kangaroo forestomach

The methods used for culturing rumen protozoa were found to be unsatisfactory for growth of ciliate protozoa from the kangaroo forestomach. Based on published measurements of physical parameters in the marsupial forestomach, several modifications were incorporated into the procedure, i.e., an increase in % hydrogen in the gas phase, adjustment of initial pH of the medium to 6.9–7.0 range, feed only forage as a substrate and incubate at a lower temperature (33–36 °C). Only incubation at the lower temperature increased survival time of the kangaroo protozoa. Two species of Bitricha were still viable after 28 d in culture. Cultures had to be terminated at that time. One of the species differed considerably in size and shape from previously described species and based on 18S rRNA data, may represent a new species of Bitricha. The second species, present in low numbers was identified as Bitricha oblata. In a separate trial, Macropodinium yalanbense survived for 11 d, at which time these cultures also had to be terminated.     

Tree leaves as complete feed for goat bucks

The objective was to look for the promising tree leaves, for feeding to livestock. The preliminary screening of tree leaves (nine species) by in vitro gas production technique revealed that leaves of Azadirachta indica, Melia azedarach, Morus alba and Leucaena leucocephala could serve as promising, alternate feed resource for ruminants. Therefore, in vivo evaluation of these tree leaves (except A. indica) along with that of Toona ciliate was assessed. Each species of the fresh tree leaves, supplemented with mineral mixture and common salt, was offered ad lib as complete feed to three bucks (Beetle × Anglo Nubian × French Alpine; 6 years old of 56.7 ± 1.12 kg BW). The data were analyzed by using completely randomized design. Bucks relished all the tree leaves (except T. ciliate) as indicated by higher (P < 0.05) voluntary DM intake and digestibility of nutrients. All the animals were in positive N-balance except those fed Toona ciliate. The N-retention was maximum in animals fed M. alba followed by that in L. leucocephala and M. azedarach. However, the DCP content of M. alba, M. azedarach and L. leucocephala was statistically comparable, while ME availability was higher (P < 0.05) from leaves of M. azedarach followed by that of M. alba, clearly indicated that leaves of M. azedarach, M. alba or L. leucocephala, supplemented with mineral mixture and common salt, could serve as an excellent complete feed for small ruminants.     

Effect of dietary inclusion of Ficus infectoria leaves as a protectant of proteins on the performance of lambs

This study examined the effects of dietary inclusion of tanniferous leaves of Ficus infectoria as a protectant of proteins on the performance of lambs. Twenty-four lambs were randomly divided into four groups of six each in a completely randomized block design and fed four iso-nitrogenous supplements formulated to contain 0, 1.0, 1.5 and 2.0% condensed tannins (CT) through dried and ground leaves of F. infectoria. The diets were designated as CT-1.0, CT-1.5 and CT-2.0, respectively, and fed to lambs on a basal diet of wheat straw to meet requirements for maintenance and growth. Blood-biochemical profile was monitored in all the lambs at 0, 45, 90, 135, 180 days of feeding. The average daily growth rate and wool growth for a period of 180 days showed a significant (P < 0.05) increase by the supplementation of CT at 1.5% through F. infectoria leaves. Although addition of CT up to 1.5% in the supplement did not interfere with the nutrient intake or digestibility, a depressing effect on DM, OM and ADF digestibility at 2.0% CT level was apparent without any detrimental affect on intake. Digestible crude protein (DCP) and total digestible nutrients (TDN) values of the composite diets were comparable, except for significantly (P < 0.05) lower TDN (%) in CT-2.0 than the control. Intake of DCP, digestible organic matter and TDN was comparable irrespective of dietary treatments, though lambs under CT-2.0 had significantly (P < 0.05) lower intake as compared to their counterparts given diet CT-1.0. Feeding of CT containing diets particularly at 1.5 and 2.0% levels significantly (P < 0.05) influenced N utilization and improved its retention, however, the microbial protein synthesis as estimated by urinary excretion of purine derivatives was not affected by the dietary treatments. Haematological (haemoglobin and packed cell volume) and biochemical parameters (serum glucose, total protein, albumin, globulin, A:G ratio, alanine aminotransferase and aspartate aminotransferase were similar among the dietary treatments except for significant (P < 0.05) reduction in serum urea concentration of lambs fed 1.5 and 2.0% CT in the supplement. It may be concluded that CT from F. infectoria leaves at 1.5% in supplement could be used as a protectant of proteins for improving the performance of lambs.     

A Colpoda aspera isolate from animal faeces: In vitro cultivation and identification

A colpodean ciliate was found in the faeces of experimental rabbits. It was initially cultivated in medium mixed with 2% (w/v) rabbit faeces. Subsequently, two chemically defined media, designated CA-1 and CA-2, were found to be suitable for axenical cultivation of the ciliate. The maximum abundance of the ciliate isolate in the CA media was 1–2 × 10⁵ cells/ml. The ciliate isolate was further identified with silver impregnation and molecular analysis. Features of the left oral polykinetid, somatic dikinetids, and sliverline pattern were similar to those of Colpoda aspera as described by Foissner (1993). The 18S small subunit ribosomal RNA gene of the ciliate isolate shared 99% sequence identity with that of C. aspera, with 100% coverage, and formed a sister clade in the phylogenetic tree with the reference C. aspera isolate. In addition, the trophozoite of C. aspera could proliferate over a temperature range from 25–37 °C. When resting cysts were cultivated in CA-1 medium at 30–35 °C, 98.2% of the trophozoites were detached from the cyst wall after 7 h.     

In situ degradability of seven plant protein supplements in heifers fed high concentrate diets with different forage to concentrate ratio

Four Holstein heifers (297.5 ± 27.7 kg BW) fed high concentrate diets were used in a crossover experiment in order to characterize the rumen fermentation pattern, and to estimate by the in situ method rumen degradation kinetics of alfalfa hay and seven plant protein supplements: solvent-extracted soybean meal, solvent-extracted sunflower meal, peas (Pisum sativum L.), lupin seeds (Lupinus sp.), broadbean (Vicia faba L.), horsebean (Vicia faba L. var equina) and vetch (Vicia sativa L.), in high concentrate diets with different forage to concentrate ratio. Heifers were fitted with a ruminal cannula. The experiment was performed in two 30-day periods, 15 days of diet adaptation and 15 days of sampling. At each period, heifers were offered one of two total mixed rations (12:88 versus 30:70 forage to concentrate ratio), two heifers per diet, on ad libitum basis. After the first period, heifers switched treatments. Intake of dry matter (DM), organic matter, crude protein and neutral detergent fibre (NDF), expressed as kg/day, did not differ between treatments, but DM intake, expressed as g/kg metabolic body weight (BW), was higher in the 12:88 diet. Average rumen pH was 6.0 in both diets, and the time pH was below 5.8, which is considered as a critical threshold for fibre degradation, was the same for both treatments (10.4 ± 1.6 h). Average ammonia nitrogen and volatile fatty acid (VFA) concentrations did not differ between treatments and individual VFA proportions were typical of high concentrate diets. Average effective degradability of DM (0.62 ± 0.02) and NDF (0.25 ± 0.03) of alfalfa hay were low and no differences were detected between treatments. The same extent of NDF degradation, together with the same proportions of VFA would indicate that both diets had the same fibrolytic activity. Forage to concentrate ratio did not affect rumen nitrogen degradability of any protein supplements incubated in situ. Corrected effective degradability for small particle losses of sunflower meal (0.78) was higher than legume seeds, which were not statistically different between each other and ranged from 0.63 to 0.66. Soybean meal had the lowest degradability value (0.61). These nitrogen degradation values must be considered more valid for beef cattle formulation of high concentrate diets than data obtained with forage diets.     

Intake,digestibility and nitrogen utilization of Robinia pseudoacacia foliage fed to growing goat wethers

Black locust (BL; Robinia pseudoacacia L.), a native tree of southeastern USA known to contain substantial levels of condensed tannins (CT), was fed to 32, 4 month old (20.4 kg BW) Boer cross wether goats in two randomized complete block design trials. The objectives were to examine the effects of feeding hay diets containing several levels of fresh BL foliage on intake, digestibility, and N metabolism. First year (1999) diets were H_E (100% Eastern gamagrass [EGH; Tripsacum dactyloides L.] hay), H_EG (70% EGH and a 30% mixture of 59% ground corn [GC; Zea mays L.], 36% soybean meal [SBM; Glycine max L.], and 5% minerals), 25BL99 (75% EGH and 25% BL leaves), and 50BL99 (50% EGH and 50% BL leaves). Second year (2000) diets were H_O (100% orchardgrass [OGH; Dactylis glomerata L.] hay), H_OG (70% OGH and a 30% mixture of 63% GC, 37% SBM, and 5% minerals), 50BL00 (50% OGH and 50% BL leaves), and 75BL00 (25% OGH and 75% BL leaves). In 1999, apparent digestibilities of the diets in the order listed above were 62.4, 68.2, 58.0, and 60.6% (P = 0.001) for DM and 62.8, 72.5, 56.0, and 59.1% (P = 0.001) for crude protein (CP). Acid detergent lignin digestibilities were negative for diets 25BL99 (−56.7%) and 50BL99 (−49.3%), apparently due to the formation of insoluble tannin and lignin complexes during passage through the digestive tract. Intakes of DM were similar across diets. In 2000, apparent digestibilities of diet DM (64.4, 71.7, 64.8 and 65.4%) and CP (70.0, 76.0, 66.6, and 66.5%) did not differ. Lignin digestibilities were positive for diets 50BL00 (9.4%) and 75BL00 (29.6%) unlike those for year 1999. Overall, BL contained 10% CT and 18–34% hydrolyzable tannins. In 1999, N intake, urinary N (UN) excretion and N retained were higher for diet H_EG (P = 0.01) than diet H_E whereas fecal N excretion (FN) was similar for diets H_EG, 25BL99 and 50BL99. In 1999, FN excretion as a percentage of N intake was higher (P < 0.02) in the BL diets, although UN as a percentage of N intake did not differ among diets. In 2000, N intake and FN output were higher for BL diets compared to diets H_O (P = 0.01) and H_OG (P = 0.02). Fecal N as a percentage of N intake was lower (P = 0.01) for diet H_OG (24.0%) than for diets 50BL00 (33.4%) and 75BL00 (33.5%). Conversely, urinary N as a percentage of N intake was higher for diets H_O and H_OG compared to the BL diets (P = 0.02). Increased levels of dietary BL increased FN, suggesting that tannins formed insoluble protein complexes thus hindering digestibility.     

Effect of the supplementation of a high-concentrate diet with sunflower and fish oils on ruminal fermentation in sheep

This study was conducted to test the hypothesis that the supplementation of a high-concentrate diet with lipids, reportedly a good strategy for improving the nutritional value of ruminant-derived products, may not necessarily be associated with detrimental effects on ruminal fermentation in sheep. Four ruminally cannulated adult ewes were fed a high-concentrate diet, with no oil (Control diet), for a 14-day adaptation period. Afterwards, they were fed the same basal diet but supplemented with sunflower oil [20 g/kg fresh matter (FM)] and fish oil (10 g/kg FM) (SOFO diet) for a further 11 days, to investigate the impact of the addition of oils on the ruminal fermentation of the diet. On days 0 (Control), 3 and 10 of the experimental period rumen fluid was sampled at 0, 1.5, 3, 6 and 9 h after the morning feeding, for analysis of pH, and ammonia, lactate and total volatile fatty acid (VFA) concentrations. Alfalfa hay was incubated in situ, using the nylon bag technique, for 12 and 24 h to examine the effect of oil supplementation on ruminal disappearance of dry matter (DM), crude protein (CP) and neutral-detergent fibre (NDF). On days 0 and 11, rumen fluid was collected just before the morning feeding and used to incubate alfalfa hay and the Control and SOFO diets by means of the in vitro gas production technique. The mean concentrations of acetate (87.8 mmol/L vs. 73.7 mmol/L) and butyrate (21.2 mmol/L vs. 17.7 mmol/L) were reduced by oil supplementation (P < 0.05) and the total VFA showed a tendency (P = 0.098) to be lower with the SOFO diet (139.0 mmol/L vs. 122.1 mmol/L). However, none of the other in vivo ruminal fermentation parameters were affected by the treatment (P > 0.10). The oil supplementation affected neither in situ rumen disappearance of DM, CP and NDF of alfalfa hay, nor rates of gas production (P > 0.10). On the other hand, a little, but significant reduction in cumulative gas production was observed when the experimental diets were incubated with rumen fluid derived from animals fed the oil-rich diet (P < 0.05).Overall, the results suggest that the supplementation of high-concentrate diets with sunflower oil (20 g/kg FM) plus fish oil (10 g/kg FM) had little effect on ruminal fermentation and therefore its use to improve the nutritional value of ruminant-derived products cannot be precluded.     

Rumen ciliates of domestic cattle (Bos taurus taurus) in Kastamonu,Turkey, with the description of a new species

Species composition and distribution of ciliates were investigated in the rumen contents of 25 domestic cattle (Bos taurus taurus L.) living in Kastamonu, Turkey. Forty-seven species and 37 morphotypes representing 15 genera were identified. Of them, a new species of Ostracodinium was recognized and described as Ostracodinium anatolicum n. sp. This new species has two caudal lobes. The dorsal lobe is small and rounded and the ventral lobe is triangular shaped and bent toward the dorsal side like a thick hook. Furthermore, the anterior end of the macronucleus (1/5 of the length) is bent toward the left like a hook. The density of rumen ciliates in cattle was 96.8 ± 43.3 × 10⁴ cells mL⁻¹ and the mean number of ciliate species per host was 14.2 ± 4.4. Entodinium longinucleatum, E. nanellum, E. simulans and Isotricha prostoma were the most abundant species, each with a prevalence of 88%. Entodinium chatterjeei, E. bifidum m. monospinosum, Hsiungia triciliata, Oligoisotricha bubali, Ostracodinium dogieli, O. mammosum and O. munham are new host records for cattle from Turkey.     

Predation of Salmonella enterica serovar Typhimurium by the rumen protozoon Entodinium caudatum studied in vitro by fluorescence emission

Predation of bacteria by protozoa has important implications on rumen metabolism and bacterial populations. Protozoa can also restrict the passage of pathogenic bacteria to the host’s lower gastrointestinal tract. This work aimed to evaluate the predation by Entodinium caudatum (EC) and the intraprotozoal survival of Salmonella enterica serovar Typhimurium. EC cells from a monofaunated sheep were incubated for up to 105 min with a S. enterica strain producing a green fluorescent protein. Rumen fluid from a defaunated sheep (DEF) was used as a control. Fluorescence, as an index of predation, measured in the residual (protozoal) fraction was higher in EC than in DEF. 105 min after the beginning of the incubation it was higher than 30 min after. Intracellular survival of Salmonella within EC was assessed by means of a selective medium. Amounts of Salmonella in the residual fraction were higher in EC than in DEF only after 30 min. After 105 min, each protozoa engulfed 100 Salmonella cell per min. Intraprotozoal survival of ingested Salmonella was 0.0017. Predation of S. enterica by E. caudatum occurred and increased in proportion to time, but bacterial viability inside the protozoa was lower at 105 min. This study demonstrates that fluorescence emission combined with bacterial and protozoal cultures could be a reliable method for quantifying bacterial predation and viability in vitro.     

Influence of natural magnesium sources on the in vitro fermentation and protozoan population in the rumen fluid collected from sheep

The objective of this experiment was to determine the effect of two types of caustic calcinated magnesite (caustic magnesite (CM) and Agromag (AG)) upon the end products of in vitro fermentation (total gas, methane, total and individual fatty acids, and VFA) and protozoan population in the rumen fluid collected from sheep. Both magnesium additives (CM and AG) as natural products in the dose of 0.01 g were added to the fermentation bottles containing rumen inoculum from sheep and different substrates. Meadow hay (MH), wheat straw (WS), amorphous cellulose (AC) and barley grain (BG) were used as substrates and incubated with the buffered rumen fluid using an in vitro gas measuring technique during 72 h of incubation. The rumen protozoa, Entodinium spp., Trichostomatids and large Entodiniomorphids and the total protozoan concentration were counted after 24 h of incubation. The methane production was significantly decreased with CM or AG, respectively, by 58 or 62% (MH), by 65% (WS), by 52% (AC) and by 58% (BG). The total VFA concentration was significantly lower compared to control for CM plus MH, WS, AC, BG and AG plus WS. The total VFA concentration was significantly higher compared to control for AG plus AC. The effect of the both additives on ciliate population was not uniform and depended on the substrates used and protozoan type. Ciliate population was significantly increased in Entodinium spp. (AG plus BG) and Diploplastron affinae (CM or AG plus BG) compared to control. Tested additives significantly decreased population of Entodinium spp. (AG plus MH or AC), Dasytricha ruminantium (AG plus AC), Ophryoscolex c. tricoronatus, Eremoplastron dilobum and Polyplastron multivesiculatum (CM or AG plus BG). It can be concluded that both natural magnesium sources influenced rumen fermentation patterns and protozoan population in vitro depending on the type of the substrate used; therefore, the relative efficacy of individual tested additive cannot be determined from these experiments. In vivo experiments are required in future.     

Effect of partial replacement of concentrates with jackfruit (Artocarpus heterophyllus) leaves on growth performance of kids grazing on native pasture of Tripura,India

Fifteen Black Bengal kids of about 3 months of age and body weight ranging from 3.8 to 4.9 kg were randomly distributed into three groups of five. Kids grazed native pasture 8 h/d. The kids in group I received supplementary concentrate (maize 35%, mustard cake 32%, rice bran 30%, mineral mixture 2% and common salt 1%) at approximately 2% of BW. However, 25 and 50% of the concentrate was replaced with jackfruit leaves for groups II and III, respectively. Total dry matter intake (DMI) was significantly higher in groups II and III than for group I due to greater forage consumption. Digestibility of CP (P < 0.05) decreased and that of NDF increased (P < 0.01) with increasing level of jackfruit leaves in the diet. Digestibility of ADF (P < 0.01), hemi cellulose (P < 0.05) and cellulose (P < 0.01) was higher in groups II and III in comparison to group I. Ruminal pH and TVFA concentration were not significantly different among the groups; however, rumen ammonia-N concentration decreased (P < 0.01) with increased level of jackfruit leaves in the diet. Similarly, plasma urea nitrogen and blood glucose levels were also reduced (P < 0.05) with increasing level of jackfruit leaves in the diet Average daily gain (ADG) was 47.33, 45.11 and 35.56 g/d in groups I, II and III, respectively. ADG and DMI/kg gain were not adversely affected when the level of replacement was restricted to 25%; however, at the 50% of replacement both parameters were adversely affected (P < 0.05). From the results of this experiment, it was concluded that jackfruit leaves might replace 25% of the supplemental concentrate for growing kids grazing in native pasture of northeast India.     

In vitro methane formation and carbohydrate fermentation by rumen microbes as influenced by selected rumen ciliate species

Ciliate protozoa contribute to ruminal digestion and emission of the greenhouse gas methane. Individual species of ciliates co-cultured with mixed prokaryote populations were hypothesized to utilize carbohydrate types differently. In an in vitro batch culture experiment, 0.6 g of pure cellulose or xylan was incubated for 24 h in 40-mL cultures of Entodinium caudatum, Epidinium ecaudatum, and Eudiplodinium maggii with accompanying prokaryotes. Irrespective of ciliate species, gas formation (mL) and short-chain fatty acids (SCFA) concentrations (mmol L^?1) were higher with xylan (71; 156) than with cellulose (52; 105). Methane did not differ (7.9% of total gas). The SCFA profiles resulting from fermentation of the carbohydrates were similar before and after removing the ciliates from the mixed microbial population. However, absolute methane production (mL 24 h^?1) was lower by 50% on average after removing E. caudatum and E. maggii. Methanogen copies were less without E. maggii, but not without E. ecaudatum. Within 3 weeks part of this difference was compensated. Butyrate proportion was higher in cultures with E. maggii and E. ecaudatum than with E. caudatum and only when fermenting xylan. In conclusion, the three ciliate species partly differed in their response to carbohydrate type and in supporting methane formation.     

Effects of pelleted total mixed rations with different rumen degradable protein on milk yield and composition of Jonica dairy goat

The present study was conducted in order to examine the effects of diets with different ruminal degradable protein sources on the performance of lactating goats fed pelleted total mixed rations (PTMR). Forty multiparous Jonica goats in early-lactation were divided into two equal groups and fed for 135 days the experimental diets. Two types of PTMR were formulated to be isonitrogenous and isocaloric while providing different rumen degradable levels of crude protein: (1) high-degradable protein diet (HD) containing soybean meal, sunflower meal and urea (46% N), and (2) low-degradable protein diet (LD) including only corn gluten meal. Apparent digestibility of the two PTMR was tested using Jonica bucks placed in individual pens and results indicated significant differences (P < 0.05) for crude protein, crude fibre, neutral detergent fibre and acid detergent fibre nutrients’ utilization. In milk performance trial, no significant differences (P > 0.05) were registered in dry matter intake and milk yield. Goats fed LD diet containing corn gluten meal showed the highest milk fat, protein and casein concentration, but not significant differences (P > 0.05) were observed on other milk components and renneting properties. It was concluded that a decrease of rumen degradable protein (RDP) level in PTMR does not negatively influence nutrient utilization and milk production and composition of Jonica dairy goat.     

Ruminal large and small particle kinetics in dairy cows fed red clover and grass silages harvested at two stages of growth

Passage, comminution and digestion rates of large and small particles were estimated using a rumen evacuation technique and total faecal collection with five lactating dairy cows in a 5 × 5 Latin square experiment. Two grass and two red clover silages harvested at early and late primary growth stages and a 1:1 mixture of late harvest grass and early harvest red clover were the dietary treatments. Cows received 9.0 kg supplementary concentrate per day. Ruminal contents and faeces were divided into large (>1.25 mm) and small (1.25–0.038 mm) particles by wet sieving. Indigestible neutral detergent fibre (iNDF) was determined by 12 days ruminal in situ incubation followed by neutral detergent extraction. Plant species did not affect ruminal particle size distribution, whereas advancing forage maturity decreased the proportion of large particles for both grass and red clover silage diets. Ruminal pool size of iNDF was higher (P<0.001) with red clover compared to grass silage diets. Ruminal passage rates of iNDF and potentially digestible NDF (pdNDF) increased with decreasing particle size (P<0.01). Passage rate of iNDF for small particles was slower (P<0.01) when red clover compared to grass silage diets were fed. Particle comminution rate in the rumen was slower (P<0.001) with red clover compared to grass silage diets and it increased (P<0.01) with advancing forage maturity. The contribution of particle comminution to ruminal mean retention time of iNDF in the ruminal large particle pool was smaller (P<0.01) in red clover compared to grass silage diets and it increased (P<0.05) with the mixed silage compared to the separate silages. Passage rate of pdNDF for both large and small particles was not affected by dietary treatments. Digestion rate of pdNDF for large particles was faster (P<0.001) with red clover compared to grass silage diets. Differences in ruminal passage and digestion rates of the large and small particles, in addition to differences in the passage and digestion rates of red clover compared to grass silage diets, emphasize the need to consider particle size and forage type in metabolic models predicting feed intake and fibre digestibility in ruminants.     

Recovery of ciliate communities from an oligotrophic mountain stream after a catastrophic wind damage

We studied the recovery of ciliate taxoceonoses in a mountain stream after a catastrophic windstorm that strongly affected mountainous regions of Slovakia in 2004. To this end, we analyzed changes in the community structure of ciliate assemblages from the Zubrovica stream in the Low Tatra Mts during the time frame between 2003 and 2008 by various statistical methods, including diversity and similarity indices, hierarchical clustering, multidimensional scaling, and PCA. The climax ciliate communities were characteristic for oligotrophic running waters, i.e., they were equitable (E_H = 0.75–1.00) and harbored a comparatively low number of species (2–9) with typically low abundances (10–125 ind./ml). However, the community structure changed dramatically after the devastating windstorm. There was a significant increase of species number (10–30) and abundance (260–1480 ind./ml), concomitant with a decrease of the equitability (E_H = 0.43–0.83). These changed quantitative and qualitative community parameters wore off comparatively quickly, i.e., about six months after the catastrophic windstorm, the ciliate taxocoenoses had reached a community structure similar to that before the wind damage. The present observations and those from terrestrial habitats indicate that ciliate communities have a good capability to comparatively quickly reach a climax even after a strong disturbance.     

Surface functionalized mesoporous activated carbon for the immobilization of acidic lipase and their application to hydrolysis of waste cooked oil: Isotherm and kinetic studies

This study deals with the surface functionalization of mesoporous activated carbon, using ethylenediamine and glutaraldehyde to facilitate the strong immobilization of acidic lipase (AL) onto MAC. The AL was produced from Pseudomonas gessardii by using slaughterhouse lipid waste as the substrate. The AL immobilized on functionalized mesoporous activated carbon (ALFMAC) was applied for the hydrolysis of waste cooked oil (WCO). The optimum conditions for the immobilization of AL onto functionalized mesoporous activated carbon (FMAC) were 90 min; pH 3.5; and 35 °C; which resulted at the maximum immobilization of 5440 U/g of FMAC (3.693 mg of AL/g of FMAC or the yield 2.7% or the expressed activity 103.7% or the activity per unit area of FMAC 1.08 mg of AL/m²). The ALFMAC showed better thermal and storage stabilities than the free AL. The ALFMAC retained a 98% and a 92% initial activity at 40 °C and 50 °C, respectively, while the AL showed the thermal stability (residual activities) 65% and 38%, respectively. The storage stability of ALFMAC at 4 °C showed 100% initial activity up to 15 days from the initial day of the storage, whereas AL showed only 88% initial activity up to 15 days. The FMAC and ALFMAC were characterized by using scanning electron microscopy (SEM), Fourier transform infrared (FT-IR) spectroscopy, and X-ray diffraction (XRD) analysis. The K_m values of the ALFMAC and AL were 0.112 mM and 0.411 mM, respectively. The v_max values of the ALFMAC and AL were 1.26 mM/min and 0.53 mM/min, respectively. Immobilization of AL onto FMAC obeyed the Freundlich and Redlich–Peterson isotherm models. The non-linear models of pseudo first, and second order, intra-particle diffusion, Bangham, and Boyd plot were also performed to understand the dynamic mechanism of immobilization. ALFMAC showed a 100% hydrolysis of WCO up to 21 cycles of reuse, and 60% up to 45 cycles. The hydrolysis of WCO was confirmed by using FT-IR spectra.