Effect of thermal stress on HSP70 expression in dermal fibroblast of zebu (Tharparkar) and crossbred (Karan-Fries) cattle

The present studies were conducted to investigate the difference response of dermal fibroblasts to heat stress in Tharparkar and Karan-Fries cattle. Skin is the most important environmental interface providing a protective envelope to animals. In skin, dermal fibroblasts are the most regular cell constituent of dermis that is crucial for temperature homeostasis. The study aimed to examine the reactive oxygen species (ROS) formation, cytotoxicity (%) and heat shock protein 70 (HSP70) genes expression in dermal fibroblast of Tharparkar and Karan-Fries cattle and to assess whether resistance of dermal fibroblast to heat stress is breed specific. Dermal fibroblasts from ear pinna of Tharparkar and Karan-Fries cattle were exposed at 25 °C, 37 °C, 40 °C and 44 °C for 3 h to measure the ROS, cytotoxicity (%) and HSP 70 (HSPA1A, HSPA2 and HSPA8) genes’ expression. The results showed that ROS formation at low temperature (25 °C) decreased in both breeds as compared to control (37 °C) and the differences were significant (P<0.0001). Heat stress at 40 °C did not increase ROS formation significantly in Tharparkar but increased significantly (P<0.001) in Karan-Fries cattle. The overall cytotoxicity (%) was also found to be significantly different (P<0.001) between Tharparkar and Karan-Fries cattle, and on exposure to different temperatures (P<0.001). The cytotoxicity (%) in dermal fibroblast cells of Karan-fries cows was more than Tharparkar. The expression studies indicated that all HSP70 genes (HSPA8, HSPA1A and HSPA2) were up-regulated at different temperatures in both breeds. In Tharparkar, the relative mRNA expression of HSPA8 gene was higher but HSPA1A and HSPA2 genes were low as compared to Karan-Fries cattle. At 40 and 44 °C, the relative expressions of inducible HSP 70 genes (HSPA1A and HSPA2) were higher in Karan-Fries than Tharparkar. In summary, dermal fibroblast resistance to heat shock differed between breeds. Dermal fibroblasts of Tharparkar were observed to be more heat tolerant than crossbred Karan-Fries cattle. The study concludes that zebu cattle (Tharparkar) dermal fibroblasts are more adapted to tropical climatic condition than crossbreed cattle (Karan-Fries). Differences exist in dermal fibroblasts of heat adapted and non-adapted cattle.     

Rumen microbial responses in fermentation characteristics and production of CLA and methane to linoleic acid in associated with malate or fumarate

An in vitro incubation in batch was conducted to investigate the effect of propionate precursor (malate or fumarate) on fermentation characteristics, and production of CLA and methane by rumen microbes when incubated with linoleic acid (C_18:2). Sixty milligrams of C_18:2 alone (LA), 60 mg C_18:2 with 24 mM malic acid (M-LA), or 60 mg C_18:2 with 24 mM fumaric acid (F-LA) was added to 150 ml culture solution consisting of 75 ml strained rumen fluid and 75 ml McDougall's artificial saliva. Culture solution for incubation was also made without malate, fumarate, and C_18:2 (control). Two grams of feed consisting of 1.4 g concentrate and 0.6 g ground alfalfa (DM basis) was also added to the culture solution of each treatment. An in vitro incubation in batch was made anaerobically in a shaking incubator for up to 12 h at 39 °C.The pH of the culture solution was increased (P<0.0001) in M-LA or F-LA treatments from 3 h to 12 h compared with the control and LA treatments. At 12 h incubation, the concentration of total VFA in the culture solution was higher (P<0.01) in M-LA and F-LA than in control and LA treatments. Concentration of C₃ by M-LA and F-LA was increased at 3 h (P<0.01), 6 h (P<0.01) and 12 h (P<0.01) compared with control and LA. However, no difference in C₃ concentration was observed between control and LA, or between M-LA and F-LA. Accumulated total gas produced for up to 12 h incubation was increased (P<0.01) by M-LA or F-LA compared with the control. Accumulated total methane produced for up to 12 h incubation, however, was greatly reduced (P<0.01) by all the supplements compared with control, and its production from M-LA or F-LA was smaller than the LA. The M-LA or F-LA also increased (P<0.05–<0.001) the concentrations of cis9, trans11-CLA for all incubation times and trans10, cis12-CLA at 1 h (P<0.01), 3 h (P<0.05), and 12 h (P<0.05) incubation times compared with LA.It can be concluded that malate and fumarate, as propionate precursors, act as alternative electron sinks and may compete with CH₄ generation and bio-hydrogenation of C_18:2 in the utilization of metabolic H₂. The highest CLA concentration at the early incubation stage (1 h) was accompanied by reduced propionate proportion. Linoleic acid is also considered one of the potential alternatives to suppress CH₄ generation.     

Effects of different selenium source on selenium distribution,loin quality and antioxidant status in finishing pigs

The experiment was conducted to study the effects of different selenium source on selenium distribution, loin quality and antioxidant status in finishing pigs. A total of 108 castrates (Duroc × Landrace × Yorkshire) at average body weight (BW) of 60 kg were allotted to three treatments, each of which was replicated three times with 12 pigs per replicate (four per pen). The control groups received the basal diet containing 0.045 mg Se/kg. A 0.3 mg Se/kg in forms of sodium selenite or selenomethionine was added to the basal diet for the experimental groups. The total test period was 40 d. Results showed that selenomethionine-treatment increased the Hunter a (redness) value of meat color during 45 min, 8 and 16 h measurement period (P<0.05) and decreased the drip loss of loin muscle during 8 and 16 h measurement period (P<0.05), while sodium selenite-treatment only elevated the Hunter a value of meat color during 0.75 h measurement period (P<0.05) and had no significant effects on drip loss of loin muscle tissues. Both selenomethionine and sodium selenite-treatment increased the Se content in serum, muscle, liver, pancreas and kidney tissue (P<0.05), the level was substantially higher in muscle, liver and pancreas (P<0.05) in the selenomethionine treated group. In addition, both selenomethionine and sodium selenite-treatment increased glutathione peroxidase (GSH-Px) activity (P<0.05) and decreased the malondialdehyde (MDA) content in the liver and muscle (P<0.05) when compared with control group, but the level of magnitude was higher when selenomethionine was fed. The present study suggests that compared with sodium selenite, selenomethionine is more effective in depositing Se in tissues, enhances the antioxidant status, thus decreasing the volume of drip loss and stabilizing the meat color.     

Effect of dietary β-glucan on growth performance,fecal microbial shedding and immunological responses after lipopolysaccharide challenge in weaned pigs

Two experiments (Exp.) were conducted to evaluate the effects of β-glucan inclusion in the diet on growth performance and immune function after lipopolysaccharide (LPS) challenge. In Exp. 1, a total of 40 weaned pigs (progeny of Landrace×Yorkshire sows by Duroc) with an initial body weight (BW) of 7.89 ± 0.84 kg (21 ± 2 d) of age) were used in a 28-day (d) experiment to determine the effects of dietary β-glucan on growth performance. Pigs were allotted randomly to two treatments consisting of addition of 0 or 0.1 g β-glucan/kg diet with four replicate pens per treatment and five pigs per pen. Growth performance was not affected by β-glucan supplementation throughout the experiment. However, dietary β-glucan reduced (P<0.05) the number of fecal Escherichia coli. In Exp. 2, a total of 20 weaned barrows (6.22 ± 0.25 kg of BW and 21 ± 2 d of age) individually raised in metabolic cages were used to evaluate immunological responses following LPS challenge. Pigs were fed 0 or 0.1 g β-glucan/kg diet for 42 d. At the end of the trial, half of the pigs (n = 5) from each treatment were injected intraperitoneal with E. coli LPS at a concentration of 100 μg/kg BW and the other half were injected with sterile saline solution. Treatments were arranged as a 2×2 factorial, with the main effect of LPS challenge (saline vs. LPS) and β-glucan supplementation (0 g/kg vs. 0.1 g/kg). After LPS injection, blood was taken at 0, 2, 4, 6, 8 and 12 hours (h) for the blood cell counts and blood inflammatory response. Dietary β-glucan increased (P<0.05) leukocytes counts at 4, 6 and 8 h, and blood lymphocyte concentrations at 2, 4 and 6 h and LPS challenge increased (P<0.05) counts of leukocytes at 2, 4, 6 and 8 h and blood lymphocyte at 2 and 4 h post-challenge. The rectal temperature was increased (P<0.05) at 2, 4, 6 and 8 h after LPS challenge. Dietary β-glucan reduced (P<0.05) and LPS challenge increased (P<0.05) blood plasma tumor necrosis factor-α (TNF-α) concentration at 2 and 4 h post-challenge. Dietary β-glucan increased (P<0.05) the concentration of the cluster of differentiation antigens 4 cells (CD4⁺) at 2, 4 and 6 h, and of 8 (CD8⁺) at 4 and 6 h post-challenge, respectively. The LPS challenge increased (P<0.05) CD4⁺ and CD8⁺ cell concentrations at 2, 4 and 6 h post-challenge. The CD4⁺:CD8⁺ ratio was reduced (P<0.05) by LPS challenge but was increased (P<0.05) by dietary β-glucan at 2, 4, 6 and 8 h post-challenge. In conclusion, dietary β-glucan decreased E. coli numbers but did not affect growth performance in weaned pigs and may offer benefits on immune function in weaned pigs challenged with LPS.     

Effect of pig faecal donor and of pig diet composition on in vitro fermentation of sugar beet pulp

Two experiments were undertaken to investigate the influence of (1) pig bodyweight and (2) dietary fibre content of the diet on the in vitro gas production of sugar beet pulp fibre using faecal inoculum.In the first experiment, inocula prepared from young pigs (Y; 16–50 kg), growing pigs (G; 62–93 kg) and sows (S; 216–240 kg) were compared. Sugar beet pulp, hydrolysed in vitro with pepsin and then pancreatin, was used as the fermentation substrate. The cumulated gas productions over 144 h were modelled and the kinetics parameters compared. Lag times (Y: 4.6 h; G: 6.4 h; S: 9.2 h) and half-times to asymptote (Y: 14.7 h; G: 15.9 h; S: 20.8 h) increased with pig bodyweight (P<0.001) and the fractional degradation rates of the substrate differed between the pig categories (Y: 0.110 h⁻¹; G: 0.115 h⁻¹; S: 0.100 h⁻¹; P<0.001). The final gas production was not affected (P=0.10) by the inoculum source.In the second experiment hydrolysed sugar beet pulp was fermented with four inocula prepared from pigs fed diets differing in their total and soluble dietary fibre contents, i.e. low fibre diet rich in soluble fibre (LOW-S) or in insoluble fibre (LOW-I) or high fibre diet rich in soluble fibre (HIGH-S) or in insoluble fibre (HIGH-I). The total and the soluble dietary fibres influenced the kinetics of gas production. The presence of soluble fibres decreased the lag times, whatever the total dietary fibre content (2.7 h for LOW-S versus 3.5 h for LOW-I, 4.0 h for HIGH-S versus 4.4 h for HIGH-I; P<0.001). The half-times to asymptote were higher with the low fibre diets (P<0.001) and, for similar total dietary fibre contents, they were lower when the proportion of soluble fibres increased (LOW-S: 9.9 h; LOW-I: 11.4 h; HIGH-S: 8.9 h; HIGH-I: 10.1 h; P<0.001). The fractional degradation rates of the substrate were the highest with the fibre-rich diet containing a high proportion of soluble fibres (0.158 h⁻¹; P<0.001).In conclusion, the bodyweight of the faeces donors and the dietary fibre composition of the pig diet influence the in vitro fermentation kinetics of hydrolysed sugar beet pulp, but not the final gas production.     

Effects of heat stress on some reproductive parameters of male cavie (Cavia porcellus) and mitigation strategies using guava (Psidium guajava) leaves essential oil

Climate changes, particularly the increase of temperature are among the main causes behind the decline of fertility in humans as well as animals. In this study, the effects of heat stress on some reproductive parameters of male cavies and mitigation strategies using guava leaves essential oil (GLEO) were studied. For this purpose, 40 male cavies aged 2.5–3 months and weighing between 348 and 446 g were divided into 4 groups of 10 animals each and subjected to the following temperatures: Ambient temperature (20–25 °C) for the control group, 35 °C for group 1, 45 °C for group 2 and 45 °C+100 µl GLEO/kg body weight, administered by gavage to animals for group 3. Exposure time of heat was 7 h per day for 60 days. Results reveal that the relative weights of testes, epididymis, vas deferens and seminal vesicles were hardly affected by the temperature levels considered (P>0.05). The mass and individual sperm motility was significantly lower (P<0.05) in cavies exposed to the temperature of 35 and 45 °C as compared with those which received GLEO and controls. The percentages of abnormal sperm and altered sperm DNA were higher in animals exposed to temperature of 35 and 45 °C as compared with the controls. The activity of superoxide dismutase significantly increased (P<0.05) in animals exposed to temperature of 45 °C and in those of 45 °C and orally treated with GLEO, compared with cavies exposed to temperature of 45 °C without receiving GLEO. The level of malondialdehyde was significantly increased (P<0.05) in animals exposed to temperature of 35 and 45 °C, whereas the level of nitric oxide was significantly lower (P<0.05) in exposed animals as compared with controls. It was concluded that the exposure of male cavies at 35 and 45 °C for 60 days induce heat stress that causes deterioration of sperm characteristics. These effects that can be mitigated by the administration of guava leaves essential oil.     

Thermotolerance,health profile and cellular expression of HSP90AB1 in Nguni and Boran cows raised on natural pastures under tropical conditions

Boran (n=15) and Nguni (n=15) cows were used in a study to determine the effect of breed, age and coat colour on the concentration of heat shock protein 90 (HSP90AB1), physiological rectal and skin temperature, and markers of health. The cows were exposed to summer heat stress and Boran cows had higher significant (P<0.05) skin temperature (35.1±0.42 °C) as compared to the Nguni cows (36.0±0.38 °C). Nguni cows had higher body thermal gradients than the Boran cows. Boran cows had thicker skin (P<0.05) and longer hairs (24.3±2.26 mm) than their Nguni counterparts (20.2±2.00 mm). The HSP90AB1 concentration was increased in Boran cows, although breed had no significant (P>0.05) influence. Significantly (P<0.05) high urea and total cholesterol was recorded in Boran cows. Coat colour had a significant (P<0.05) effect on the weight and rectal temperature of the study animals. Coat colour and age had no significant effect (P>0.05) on the concentration of HSP90AB1, although older cows (≥9 years) had higher concentrations (5.4±1.29 ng/ml). Age had a significant (P<0.05) effect on packed cell volume, neutrophil/lymphocyte, urea, total protein and gamma-glutamyl transferase whereas cows with ≥9 years had more concentrations than young ones. Age significantly (P<0.05) influenced hair length, skin temperature and the thermal gradients. Breed was positively correlated (P<0.001) to coat colour, age, body condition score, weight and temperature humidity index while negatively correlated to urea and total cholesterol. It was concluded that Nguni cows were more adaptable to hot environments than the Boran cows as the latter were unable to balance thermal load between their bodies and the environment.     

Evaluation of total and non-fatty ether extract in feeds and cattle feces using two analytical methods

The objective of this study was to evaluate ether extract (EE) concentrations, pigments, and wax in forages (n = 14), concentrates (n = 23), and cattle feces (n = 100) using extraction methods recommended by Association of Official Analytical Chemists (AOAC; method 920.39) and American Oil Chemist's Society (AOCS; method Am 5-04). The EE contents were compared by adjusting a linear regression model for each sample type. For the feces and forage samples, the EE contents produced by the AOCS method were greater (P<0.05) compared to those obtained using the AOAC method. No differences between methods were observed in EE content of concentrates (P>0.05). Concentration of vegetable pigments and wax were evaluated by using analysis for variance. Vegetable pigments were lower (P<0.05) in the post-extraction residues using the AOCS method, than the AOAC method, indicating greater participation of vegetable pigments in the EE. No differences were observed between the methods in wax concentration of the post-extraction residues (P>0.05). The quantification method of the EE content that is recommended by AOCS is not suggested for analyses of forage and feces of ruminants because it possibly increases the removal of non-fatty material, mostly pigments, in comparison to the method recommended by AOAC.     

Effect of inoculum and pepsin–pancreatin hydrolysis on fibre fermentation measured by the gas production technique in pigs

Two experiments were undertaken to adapt the in vitro gas production technique in syringes, used for ruminants, to fibre fermentation studies in the large intestine of pigs.In a first experiment, two inocula (faeces and large intestine content) were compared at four dilution levels in a buffer solution (0.025, 0.05, 0.1 and 0.2 g ml⁻¹) with two substrates: wheat bran and sugar–beet pulp. The accumulated gas produced over 72 h was modelled and the kinetics parameters compared. The time to half asymptote was lower for the intestinal inoculum (5.5 versus 8.0 h, P<0.02), but the 2 inocula yielded similar fractional rates of degradation (0.16 h⁻¹) and gave equal final gas production (252 ml g⁻¹ substrate). No interaction (P>0.05) was observed between inocula and substrates. The dilution of the samples in the buffer solution increased (P<0.001) the lag time (from 0.9 to 2.1 h for dilution rates ranging from 0.2 to 0.025 g ml⁻¹, respectively) and decreased (P<0.001) the rates of substrate degradation (from 0.18 to 0.13 h⁻¹).A second experiment aimed to study the effect of an in vitro pepsin–pancreatin hydrolysis of the sample prior to the gas test. Six substrates were tested: maize, wheat bran, sugar–beet pulp, lupins, peas and soybean meal. The enzymatic hydrolysis affected (P<0.001) the kinetics parameters and the ranking order of the fermented substrates. The lag times also increased for all ingredients. The rate of degradation decreased when peas, lupins, maize and wheat bran were hydrolysed (P<0.001) but it increased with soybean meal (P=0.014) and sugar–beet pulp (P<0.001). Final gas production increased with peas and soybean meal (P<0.001), remained unchanged for lupins and decreased for the other substrates (P<0.001).In conclusion, the method using faeces as a source of microbial inoculum is reliable to characterise the fermentation kinetics of ingredients in the large intestine of pigs. However, it is important to hydrolyse the substrates with pepsin and pancreatin before the gas tests.     

The interaction between lactose level and crude protein concentration on piglet post-weaning performance,nitrogen metabolism,selected faecal microbial populations and faecal volatile fatty acid concentrations

A performance study and a nitrogen balance study (2×3 factorial) were conducted to investigate the interaction between lactose level (215 and 125 g/kg) (lactofeed 70; 860 g whey permeate/kg, 140 g soya bean meal/kg, Volac International, UK) and crude protein (CP) concentration (160, 185 and 210 g/kg) on post-weaning piglet performance, nitrogen metabolism, faecal microbiology and faecal volatile fatty acid concentrations. In the performance trial, 252 piglets (7.6 kg; 33 days of age) were assigned to one of six dietary treatments following a 12-day period on a commercial creep diet (17 MJ/kg DE, 16 g lysine/kg). The experimental diets were fed for 28 days (days 12–40) and were formulated to have identical digestible energy (15 MJ/kg) and total lysine (14.5 g/kg) contents. In the N balance experiment, 24 boars (20 kg live weight) were offered the same diets as in the performance trial. Faecal samples were collected for selected microbial populations. There was an interaction (P<0.05) between lactose and CP concentration in daily gain (ADG) and daily feed intake (ADFI) (P<0.01) during the weaner period (days 12–40). At the high lactose level there was a linear increase in ADG and ADFI with increasing CP. However, at the low lactose level there was no increase in ADG or ADFI above the medium CP. Pigs offered 215 g lactose/kg had a higher dry matter (P<0.001), organic matter (P<0.001), energy (P<0.001), nitrogen (P<0.01) and neutral detergent fibre (P<0.05) coefficient of total tract apparent digestibility compared to pigs offered 125 g lactose/kg. There was an interaction between lactose and CP concentration for nitrogen intake (NI) (P<0.05), urine pH (P<0.05) and selected faecal microbial populations. At the high CP level, pigs offered diets containing 215 g lactose/kg had a higher NI and a lower urine pH than pigs offered 125 g lactose/kg (P<0.05). However, the inclusion of lactose had no significant effect on either NI or urine pH at the low or medium CP concentration. At the low lactose level there was a linear increase in faecal E. coli population and a linear decrease in faecal Lactobacilli population with increasing CP. However at high lactose levels CP concentration had no effect on either E. coli or Lactobacilli populations. Pigs offered 215 g lactose/kg had a significantly higher Bifidobacteria population compared to pigs offered 125 g lactose/kg. There was a linear decrease in Bifidobacteria population as CP increased. In conclusion, at the high lactose level there was a linear increase in ADG and ADFI with increasing CP concentrations. There was no increase in these parameters above 185 g CP/kg at the low lactose level.     

Effect of thermal exposure on physiological adaptability and seminal attributes of rams under semi-arid environment

Thermal stress in hot semi-arid environment is a major limitation of sheep production in tropical and sub-tropical climatic condition. The animals tend to maintain homeostasis through physiological adjustments in a hot environment (maximum temperature reaches up to 47.5 °C). Therefore, the present study was carried out to assess the effect of thermal exposure on physiological adaptability and seminal attributes of rams under semi-arid environment. The experiment was conducted for eight weeks involving sixteen Malpura crossbred rams (GMM: Garole X Malpura X Malpura). The rams were divided equally into two groups, designated as G1 and G2, respectively. The rams in G1 (Control) group were kept in a sheep shed under naturally prevailing environment without artificial manipulation of ambient temperature (Temperature 30.48±0.38 °C; Relative Humidity 28.59±1.15%). The rams of G2 group were exposed to different temperature at different hours of the day (38 °C at 1000–1100 h; 40 °C at 1100–1200 h; 42 °C at 12:00–1300 h; 43 °C at 1300–1400 h; 44 °C at 1400–1500 h and 42 °C at 1500–1600 h) in a climatic chamber for thermal exposure. Physiological responses, blood biochemical profile, blood endocrine profile, sexual behavior and seminal attributes were measured for both the groups. Thermal exposure significantly (P<0.05) increased the water intake; respiration rate, rectal temperature and skin temperature at afternoon in rams. Exposure of rams to thermal stress (G2) significantly (P<0.05) increased cortisol level and decreased tri-ido-thyronine level. The latency period after the first ejaculation, decreased significantly (P<0.05) in G2. The percentage of rapid motile sperm, linearity and average path velocity of sperm were also altered significantly (P<0.05) in thermal exposed rams as compared to control. However, comparable feed intake, body weight, and major blood biochemical parameters, as well as acceptable semen quality attributes of all the rams indicated that the Fec B gene introgressed Malpura cross rams adapted to the thermal exposure under semi-arid tropical climate.     

In vitro evaluation of the fermentation characteristics of the carbohydrate fractions of hulless barley and other cereals in the gastrointestinal tract of pigs

An in vitro model was used to study the fermentation characteristics of carbohydrate fractions of hulless barley (hB), in comparison to hulled barley (HB), hulled oat and oat groats (OG) in the pig intestine. For this purpose, 6 hulless barley cultivars (hB), varying in β-glucan content (36–99 g/kg DM), were compared to 3 HB cultivars, 2 oat groat samples (OG), 3 oat varieties and a reference sample of wheat. The residue of a pepsin–pancreatin hydrolysis was incubated in a buffered mineral solution inoculated with pig faeces. Gas production, proportional to the amount of fermented carbohydrates, was measured for 48 h and kinetics modelled. The fermented solution was subsequently analyzed for microbial production of short-chain fatty acids (SCFA) and ammonia. In vitro dry matter degradability varied according to ingredient (P<0.001). Higher values were observed for OG, ranging from 0.88 to 0.99 as compared to oat, hB and HB, for which degradability ranged from 0.63 to 0.73, 0.68 to 0.80 and 0.69 to 0.71, respectively. A “cereal type” effect (P<0.05) was observed on fermentation kinetics parameters. Total gas production was higher (P<0.05) with hB (224 ml/g DM incubated) than with HB and oat (188 and 55 ml/g DM incubated, respectively). No difference was observed between hB cultivars (P>0.05) for total gas production but differences (P<0.001) were found for lag time and the fractional rate of degradation. Hulless barley cultivar CDC Fibar (waxy starch) and CDC McGwire (normal starch) started to ferment sooner (lag time of 0.7 and 0.9 h, respectively) than SH99250 (high amylose starch; 1.7 h). The fractional rate of degradation was similar in both hB and OG (0.15/h on average), which was higher than that of HB (0.12/h). The production of SCFA was also higher (P<0.05) with hB (6.1 mmol/g DM incubated, on average) than with HB and oat (4.9 and 2.9 mmol/g DM incubated, respectively). Similar trends were found for SCFA production expressed per g fermented carbohydrates, with higher butyrate and lower acetate ratio. In contrast, oat fermentation generated higher (P<0.05) ammonia concentration (1.4 mmol/g DM incubated, on average) than hB (1.0 mmol/g DM incubated). In summary, hulless barleys, irrespective of cultivar type had higher in vitro fermentability and produced more SCFA and less ammonia than hulled barley and oat. Thus, hulless barleys have a better potential to be used in pig nutrition to manipulate the fermentation activity in the intestine of pigs.     

Influences of flavomycin,ropadiar, and saponin on nutrient digestibility,rumen fermentation,and methane emission from sheep

This study focused on the effects of three additives given together with a hay/concentrate-based diet on nutrient digestibility, rumen fermentation, and methane emission from sheep. The basal diet consisted of 1.29 kg mixed hay and 0.43 kg concentrate mixture based on dry matter (DM). Treatments consisted of control (no additive), flavomycin⁴⁰ (250 mg/d), ropadiar from an oregano extract (250 mg/d), and saponin in the form of a yucca schidigera extract (170 mg/d). Results indicated that intake and digestibility were unaffected by treatments (P>0.05). The NH₃-N concentration of rumen liquor was lower (P<0.05) for additive treatments versus the control treatment. Higher concentrations of volatile fatty acid (VFA) were observed in the saponin (75.8 mmol/L) and ropadiar (73.1 mmol/L) treatments. The proportion of individual fatty acid of rumen liquor was unchanged, whereas lower ratio of acetate to propionate in the saponin treatment was observed (P<0.05). The average methane production expressed on digested organic matter (OM) and neutral detergent fiber (aNDFom) basis were decreased by approximately 3.3 and 12.0 g/kg, respectively in saponin, and 4.2 and 11.9 g/kg in ropadiar treatment compared to the control. Methane production was positively correlated with the concentrations of NH₃-N, and negatively correlated with total VFA and the proportion of propionate of rumen liquor (P<0.05). The study found that saponin and ropadiar could have the potential to reduce rumen methanogenesis in sheep.     

Effects of type of ionophore and carrier on in vitro ruminal dry matter disappearance,gas production,and fermentation end products of a concentrate substrate

Effects of ionophore type and carrier on in vitro ruminal digestion and fermentation patterns of a concentrate substrate were evaluated at various incubation times. Treatments were: control (no ionophore); lasalocid sodium commercial premix (Bov); lasalocid sodium mycelium cake (LasBio); laidlomycin sodium salt (LaidNa); laidlomycin propionate commercial premix (LaidPro); monensin sodium salt (Mon); and monensin sodium commercial premix (Rum). The Bov, LasBio, Mon, and Rum treatments supplied 4 μg of ionophore/mL of culture volume, whereas the LaidNa and LaidPro treatments supplied 1.33 μg of ionophore/mL. Total gas and methane production did not differ among treatments at any of the incubation times (P>0.09). Similarly, in vitro dry matter disappearance (IVDMD) was not affected by treatment (P>0.28) at 6, 18, and 24 h of incubation; however, IVDMD (P=0.03) was greater for ionophores than for the control at 12 h of incubation. Molar proportions of acetate (P<0.01), acetate:propionate (P<0.01), and total volatile fatty acid (VFA) concentrations (P<0.01) were decreased and propionate was increased (P<0.001) for the average of all ionophore-containing substrates compared with the control. Total VFA were decreased by Bov, LaidNa, and Rum, contrasted with their specific counterparts (LasBio, LaidPro, and Mon, respectively; P<0.05). Differences were detected among ionophore types for acetate (lasalocid vs. laidlomycin; P<0.05), propionate (lasalocid vs. monensin; P<0.05), and butyrate (monensin vs. lasalocid or laidlomycin; P<0.05). Capture of metabolic hydrogen in end products of fermentation was greater for ionophore-containing treatments (P<0.01) than for the control. These data suggest limited unique effects of ionophore type or carrier on IVDMD, total gas production, and methane; however, VFA proportions varied among ionophore types and carriers, which deserves further study.     

Effect of moist heat treatment on in-vitro degradability and ruminal escape protein and amino acids of mustard meal

A study was conducted to determine the effects of moist heat treatment (127°C, 117 kPa steam pressure) for 10 min on protein fractions and in-vitro crude protein (CP) degradability of mustard meal. Rumen undegraded protein (RUP) and amino acid disappearance of unheated, and heated, mustard meal were measured following 12 h of rumen incubation using two ruminally fistulated cows. Intestinal availability of RUP was estimated using an enzymatic (pepsin–pancreatin) procedure. Heat treatment reduced (p<0.05) protein solubility and increased (p<0.05) neutral detergent insoluble CP without affecting acid detergent insoluble CP of mustard meal. Relative to the control, heated mustard meal had a lower (p<0.05) effective in-vitro CP degradability (445.2 vs. 746.8 g kg⁻¹ of CP) and a higher (p<0.05) ruminal escape CP (615.1 vs. 120.2 g kg⁻¹ of CP) value. Amino acid composition was not affected by heat treatment except for the concentration of arginine and lysine which was lower (p<0.05) in heated than in unheated mustard meal. Disappearance of all amino acids following 12 h of rumen incubation was lower (p<0.05) in unheated than in heated mustard meal. Heat treatment increased (p<0.05) the amount of protein available for digestion in the small intestine from 75.7 to 518.1 g kg⁻¹ of CP. It was concluded that moist heating of mustard meal for 10 min will reduce ruminal CP and amino acid degradability without compromising the intestinal availability of ruminal undegraded protein.     

Effect of peppermint (Mentha piperita) oil on in vitro methanogenesis and fermentation of feed with buffalo rumen liquor

The effect of inclusion of peppermint (Mentha piperita) oil (at 0, 0.33, 1.0 and 2.0 μl/ml of incubation medium) on gas and methane production, fermentation of feed and microbial profile was studied in in vitro gas production test, using 200 mg of wheat straw and concentrate mixture in equal proportion as substrate in a 100 ml graduated syringe. The buffalo rumen liquor was used as inoculum and the observations were recorded at 24 h of incubation. Methane emission was reduced (P<0.001) by 19.9%, 46.0% and 75.6% at 0.33, 1.0 and 2.0 μl levels, respectively. The concentration (mM/100 ml) of total volatile fatty acids was reduced (P<0.01) by inclusion of peppermint oil at higher levels (1.0 and 2.0 μl) whereas at 0.33 μl level there was no effect. The proportion of acetate increased (P<0.05) and that of propionate decreased (P<0.001) at 1.0 and 2.0 μl levels of peppermint oil. There was a fall (P<0.001) in carboxymethylcellulase and xylanase activities and the inhibition increased with the increasing level of peppermint oil which resulted in a dose dependent decrease (P<0.05) in in vitro true digestibility of feed. At 0.33 μl level of peppermint oil, the population density of total bacteria was similar to that of control but fungi, Ruminococcus flavefaciens and methanogens increased by 4-, 6- and 2-folds, respectively, as determined with real-time PCR. At 1.0 and 2.0 μl levels the population density of total bacteria, fungi, Fibrobacter succinogens and methanogens decreased drastically and fell below the control values. The numbers of holotrichs and spirotrichs were reduced (P<0.001) by increasing dose of peppermint oil. The higher doses of peppermint oil were toxic for the rumen microbes but the lower levels could be further explored in in vivo experiments as rumen modifier to reduce methanogenesis.     

Effects of fibre level and dietary mannanoligosaccharides on digestibility,caecal volatile fatty acids and performances of growing rabbits

The current experiment with 3 trials aimed to study the effect of two levels of dietary fibre – high fibre (HF; 323 g aNDFom/kg) and low fibre (LF; 248 g aNDFom/kg) – and the effect of mannanoligosaccharides (MOS) addition (1 g/kg) to the LF diet (LFM) on the performances and health status of growing rabbits, digestibility and caecal fermentative characteristics. In the growth trial 132 rabbits of both sexes were used (11 cages with 4 rabbits per treatment) from weaning (32 days of age) to slaughter (67 days of age). Rabbits fed HF diet showed a significantly higher weight gain and live weight at 67 days than rabbits fed LF diet (2032 g vs. 1935 g) (P<0.05). Feed and digestible energy intake increased with dietary fibre level (P<0.05). During the growing period rabbits fed HF diet had a feed intake 26% higher than those fed LF diet. Feed efficiency ratio was worse in HF animals (0.334 vs. 0.385; P<0.05). Addition of MOS to LF diet did not affect growth performance parameters (P>0.05). Mortality and morbidity rate were not affected by treatments. In the digestibility 24 rabbits from 46 to 51 days of age trial were used. The HF diet resulted in a significant (P<0.05) decrease in digestibility of dry matter, organic matter and protein while the aNDFom digestibility was not significantly different between diets (P>0.05). Supplementation with MOS had no effects on digestibility (P>0.05). In the 3rd trial the caecal traits were measured in 30 rabbits with 46 days of age that received the experimental diets in the previous 14 days. Caecal production of total volatile fatty acids (VFA), acetate and propionate were significantly higher (P>0.05) on rabbits fed HF diets than on rabbits fed LF diets. The total VFA concentration increased 64% (from 5.01 to 8.20 mmol/100 ml) and acetate increased 73% (from 3.73 to 6.44 mmol/100 ml). Butyrate production was not different between diets (P>0.05). Fibre level did not affect proportions of VFA and caecal contents and caecal weights. Addition of MOS to LF diet did not affect any caecal trait (P>0.05). It was concluded that the reduction of dietary fibre level increases feed digestibility but worsens rabbit growth performances. Supplementation of low fibre diet with 1 g MOS/kg is not enough to reduce its negative effects on growth performances.     

Gene expression of apoptosis-related genes,stress protein and antioxidant enzymes in hemocytes of white shrimp Litopenaeus vannamei under nitrite stress

Apoptotic cell ratio and mRNA expression of caspase-3, cathepsin B (CTSB), heat shock protein 70 (HSP70), manganese superoxide dismutase (MnSOD), catalase (CAT), glutathione peroxidase (GPx) and thioredoxin (TRx) in hemocytes of white shrimp Litopenaeus vannamei exposed to nitrite-N (20 mg/L) was investigated at different stress time (0, 4, 8, 12, 24, 48 and 72 h). The apoptotic cell ratio and mRNA expression level of CTSB were significantly increased in shrimp exposed to nitrite-N for 48 and 72 h. Caspase-3 mRNA expression level significantly increased by 766.50% and 1811.16% for 24 and 48 h exposure, respectively. HSP70 expression level significantly increased at 8 and 72 h exposure. MnSOD mRNA expression in hemocytes up-regulated at 8 and 48 h, while CAT mRNA expression level increased at 24 and 48 h. GPx expression showed a trend that increased first and then decreased. Significant increases of GPx expression were observed at 8 and 12 h exposure. Expression level of TRx reached its highest level after 48 h exposure. These results suggest that nitrite exposure induces expression of apoptosis-related genes in hemocytes, and subsequently caused hemocyte apoptosis. Meanwhile, expression levels of HSP70 and antioxidant enzymes up-regulated to protect the hemocyte against nitrite stress.