Docosahexanoic acid diet supplementation attenuates the peripheral mononuclear cell inflammatory response to exercise following LPS activation

Exercise induces changes in circulating pro- and anti-inflammatory cytokines. The aim was to investigate the effect of docosahexaenoic acid (DHA) diet supplementation on the plasma cytokine levels and on the peripheral mononuclear (PBMCs) cells cytokine production after a training season or an acute bout of exercise. Fifteen male soccer players were randomly assigned to a placebo or an experimental group. The experimental group consumed an almond-based beverage enriched with DHA, whereas the placebo group consumed the same beverage without DHA. Three blood samples were taken: in basal conditions at the beginning of the nutritional intervention and after eight weeks of training season in basal and post-exercise conditions. The DHA content increased in erythrocytes after 8 weeks of training and supplementation. Neither diet supplementation with DHA nor training season altered the basal plasma cytokines and growth factors. Only acute exercise significantly increased plasma IL6 in experimental and placebo groups. Lipopolysaccharide (LPS) activation induced the inflammatory response in PBMCs, with a significant production rate of TNFα, IL6 and IL8 mainly after acute exercise. DHA supplementation significantly reduced the rate of TNFα and IL6 production by stimulated PBMCs. Acute exercise increased the Toll-like receptor 4 (TLR4) protein levels in PBMCs, although the increase was only statistically significant in the placebo group. In conclusion, a training season does not induce significant changes in the circulating cytokine profile in well-trained soccer players. Exercise increases the PBMCs cell capabilities to produce cytokines after TLR4 stimulation with LPS and this rate of cytokine production is attenuated by diet DHA supplementation.     

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 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.     

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.     

Activation of foal neutrophils at different ages by CpG oligodeoxynucleotides and Rhodococcus equi

Toll-like receptor 9 (TLR9) activation stimulates protective immune responses against intracellular pathogens by phagocytes, including neutrophils. This study examined TLR9-mediated neutrophil activation in neonatal foals. Unmethylated CpGs, ligands for TLR9, were used to stimulate equine neutrophils, either purified or in contact with other peripheral blood leukocytes. Rhodococcus equi was used as another stimulus in parallel. TLR9 mRNA was constitutively expressed at a similar level in purified equine neutrophils across different ages from birth to adulthood, and expression was not affected by either CpG or R. equi. Purified foal neutrophils were directly sensitive to CpG stimulation, reflected by enhanced reactive oxygen species generation following fMLP stimulation, and by expressing significantly (P < 0.05) greater mRNA of IFN-γ, IL-8, IL-12p35, and significantly (P < 0.05) decreased TNF-α mRNA. In comparison, purified foal neutrophils stimulated by R. equi showed significantly (P < 0.05) increased mRNA production of IL-6, IL-8, IL-23p19, and TNF-α. Neutrophils co-cultured with other leukocytes expressed a distinct profile of cytokine mRNA than purified neutrophils in response to CpG stimulation, whereas the profile was very similar following R. equi stimulation irrespective of neutrophil purity. When co-cultured with other leukocytes, foal neutrophils were significantly (P < 0.05) activated at birth by B-class CpGs and produced IL-6, IL-8, IL-12p40, and IL-23p19 at similar magnitudes to those at 2 months of age. In foal neutrophils at birth, R. equi significantly (P < 0.05) induced all cytokines stimulated by CpGs (except IL-12p40), as well as TNF-α. Our results indicate that foal neutrophils were sensitive to CpG or R. equi activation as early as at birth, and that B-class CpGs enhanced foal neutrophil functions in vitro.     

Effects of dietary forage-to-concentrate ratios on performance and carcass characteristics of growing fat-tailed lambs

This study was conducted to evaluate the effects of using different dietary forage-to-concentrate ratios on growth performance and carcass characteristics of eighty Chall male fat-tailed lambs, averaging 165 ± 15 (SD) days of age and body weight of 38.4 ± 4.8 (SD) kg, randomly assigned to four diets containing alfalfa hay-to-concentrate ratios (DM basis) of 70:30 (C30), 50:50 (C50), 30:70 (C70), 10:90 (C90). Metabolizable energy (ME) contents were, 9.12. 9.96, 10.67, and 11.34 MJ/kg dry matter (DM) and crude protein (CP) contents were 143, 152, 161, and 174 g/kg for the C30, C50, C70, and C90 diets, respectively. Sixteen lambs (4 lambs/treatment) were slaughtered at the end of feeding period (84 days). Dry matter intake and feed conversion ratio (FCR) (i.e., kg DM/kg gain) decreased linearly (P<0.001) as concentrate level increased in the diet. However, a linear increase (P<0.001) for ME intake and a quadratic increase for average daily gain (ADG, P<0.001) and final body weight (P<0.01) were observed with increasing dietary concentrate. Slaughter weight, eye muscle area, and weights of lean, bone, neck, shoulder, rack–loin, leg, skin, head, lung, and spleen were not affected by the experimental diets. However, as dietary concentrate increased, a linear increase for dressing percentage, feet weight (P<0.001), and backfat thickness (P<0.02), a quadratic increase for empty body weight (EBW, P<0.02), weights of hot and cold carcass (P<0.001), subcutaneous fat (P<0.02), total fat (P<0.01), brisket–flank (P<0.04), and tail fat (P<0.001), a linear decrease for weights of liver (P<0.01), and heart (P<0.05), and a quadratic decrease for lean-to-fat ratio (P<0.02) were observed. This study indicates that in Chall fat-tailed lambs the increase of dietary concentrate (up to 700 g/kg) improves growth rate, FCR and dressing percentage with negative effect on carcass lean-to-fat ratio.     

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.     

Effect of fat type and lysophosphatidylcholine addition to broiler diets on performance,apparent digestibility of fatty acids,and apparent metabolizable energy content

A completely randomized design study with a 3 × 2 factorial arrangement was conducted to evaluate the effects of three different fat sources (soybean oil, tallow, and poultry fat) with or without emulsifier supplementation on performance, coefficient of total tract apparent digestibility (CTTAD) of fatty acids, and apparent metabolizable energy (AME) content in broiler chickens. Two hundred and fifty-two one-day-old male Arbor Acres broiler chickens were randomly divided into 6 different treatments: (T1) basal diet containing soybean oil without lysophosphatidylcholine (LPC) supplementation, (T2) basal diet containing soybean oil with LPC supplementation, (T3) basal diet containing tallow without LPC supplementation, (T4) basal diet containing tallow with LPC supplementation, (T5) basal diet containing poultry fat without LPC supplementation, and (T6) basal diet containing poultry fat with LPC supplementation. Body weight gains from broiler chicks fed diets containing tallow were lower (P<0.05) than the body weight gains from chicks that were fed diets containing soybean oil or poultry fat in both the starter and grower periods. Birds fed diets containing tallow had the highest FCR (P<0.05), followed by the birds that were fed diets containing poultry fat, and soybean oil. The CTTAD of C16:0, C18:2, and C18:3n3 was greater (P<0.05) for broilers fed diets containing soybean oil than for those fed diets containing tallow or poultry fat in the starter period. The addition of LPC increased (P<0.05) body weight gain of broiler chickens in the starter period and the AME of the diets in the grower period, and tended to reduce FCR (P=0.072) in the starter period. LPC supplementation increased (P<0.05) the CTTAD of C16:0, C18:1n7 and C18:1n9 in the starter period, and of C18:2, and C18:3n3 in the grower period (P<0.05). There were no significant interactions between fat sources and the addition of LPC. These data indicated that LPC supplementation can improve body weight gain of broiler chickens in the starter period. This effect may be associated with an increase of CTTAD of FA due to LPC activity.     

Effects of conservation method on fatty acid composition of silage

This experiment was conducted to investigate effects of wilting and additives on the fatty acid (FA) composition of grass silage. The crop used was timothy (Phelum pratense L., cv. Grindstad), and the additives were Proens? (formic acid and propionic acid, 60–66 g/100 g and 25–30 g/100 g, respectively), the bacterial inoculant Siloferm® Plus (Pediococcus acidilactici and Lactobacillus plantarum) and water (control). The wilted material reached a dry matter (DM) content of 336 g/kg at the first cut and 350 g/kg at the second cut. Neither wilting nor the additives had any major effect on the FA proportions, with the only differences in the concentrations of C16:0 and C18:3. Silage treated with bacterial inoculant contained a higher proportion of C16:0 (P<0.05) than silage treated with acid, and a lower (P<0.05) concentration of C18:3 than silage treated with either of the other two additives. In the silages, there were lower (P<0.05) proportions of C16:0, C18:0, C18:1 and C18:3, and higher (P<0.05) proportions of C16:1, C18:2 and other identified FAs, than in the fresh material. A wilting process shorter than 24 h, to a DM content of 330–350 g/kg, did not have any effect on the proportions of FAs in P. pratense L., cv. Grindstad. Since the different additives and wilting strategies tested in this study did not affect the proportions of FAs in silage to a major extent, the results indicate that such a process offers a robust means to avoid losses of FAs that can occur during wilting, while retaining the positive effects of wilting, such as reduced losses of nutrients through effluents.     

Effect of feeding protected fat and proteins on milk production,composition and nutrient utilization in Murrah buffaloes (Bubalus bubalis)

Objective of this study was to investigate the effect of feeding protected fat and proteins on milk production, composition and nutrient utilization in Murrah buffaloes (Bubalus bubalis). Eighteen buffaloes were divided into two groups (9 each) on the basis of most probable production ability. Buffaloes in control group (C group; most probable production ability 2204 kg) were fed chaffed wheat straw, chopped maize fodder and concentrate mixture as per requirements. Buffaloes in supplemented group (S group; most probable production ability 2211 kg) were fed same ration as C group plus 2.5% rumen protected fat (on dry matter intake basis) and formaldehyde treated mustard and groundnut oil cake (1.2 g formaldehyde/100 g crude protein) in place of unprotected cakes. Group S buffaloes were supplemented rumen protected fat and protein 60 days pre-partum to 90 days postpartum and persistence of milk production was monitored up to 210 days of lactation. Milk yield during supplementation period (90 days) in S group was 13.11 kg/d and was 19% higher (P<0.01) than the C group (11.01 kg/d), whereas after supplement withdrawal (120 days), it was 11.04 kg/d and was 15% higher (P<0.01) than the C group (9.61 kg/d). There was no effect on total solid, protein, solid-not fat (SNF) and lactose contents in the two groups, whereas milk fat yield was increased (P<0.05) and level of milk urea nitrogen was decreased (P<0.01) in S group. Moreover, the supplement produced noticeable changes in the fatty acid profile of the milk fat, i.e., reduction in the concentration of saturated fatty acids (SFA) by 19% and an increase in that of unsaturated fatty acids (USFA) by 36%. Besides, digestibility of dry matter, crude protein, acid detergent fiber and neutral detergent fiber were not affected, whereas ether extract digestibility was higher (P<0.05) in S group. There was no effect on plasma glucose, non-esterified fatty acids, triglycerides and cholesterol concentrations between two groups, whereas blood urea nitrogen concentration was lower (P<0.01) in S group. Supplementation of protected nutrients to buffaloes increased milk production and unsaturated fatty acids content in milk fat and persistence of lactation after supplements were withdrawn.     

Coping with suboptimal water temperature: modifications in blood parameters,body composition,and postingestive-driven diet selection in Nile tilapia fed two vegetable oil blends

The world tilapia production faces seasonal variations. However, very few nutritional studies have addressed suboptimal temperature. We evaluated the effect of two temperatures (20 or 30 °C) and two vegetable oil blends (one rich in corn oil (COR) and one rich linseed oil (LIN)) on tilapia growth, body composition, and blood parameters using a 2 × 2 factorial design with the following treatments: COR-20; LIN-20; COR-30; LIN-30 (Trial 1). In addition, we also evaluated the effect of postingestive signals of dietary oils when the organoleptic properties of diets were isolated (Trial 2). In the Trial 1, 256 fish (15.36 ± 0.14 g) were placed in 16 aquariums and submitted during 30 days to the 2 × 2 factorial designs: COR-20; LIN-20; COR-30; LIN-30. The temperatures were established in two independent water recirculation systems. In the Trial 2, 96 fish (34.02 ± 0.79 g) were placed in 12 aquariums and subjected to the same experimental design of Trial 1, but to evaluate fish feeding behavior. They were allowed to select the encapsulated diets provided in different feeding halls to evaluate if diet preferences are influenced by postingestive signals. As the Trial 1 results show, diets had no significant effects on growth, dietary protein use, and body centesimal composition, but 30 °C induced the best performance and protein deposition (P < 0.05). LIN-20 showed lower very-low-density lipoprotein and cortisol, but higher high-density lipoprotein (HDL), aspartate aminotransferase (AST), alanine aminotransferase (ALT), and triglycerides (TG) than COR-20 (P < 0.05). COR-30 presented higher HDL, AST, ALT, TG, and cortisol than LIN-30. The fish fed COR showed lower C20:5n-3 (EPA) and higher n-6 than fish fed LIN (P < 0.05). The fish fed LIN had high n-3 highly unsaturated fatty acid. ∑ polyunsaturated fatty acid was higher at 30 °C. Finally, the tilapia in Trial 2 showed clear diet intake regulation and preference for LIN (P < 0.05), regardless of temperature. In short, lipid sources had no influence on tilapia performance; however, temperature affects carcass lipid deposition as well as fatty acids profile. Notably, the preference for linseed oil can suggest nutritional metabolic issues, contributing to animal behavior knowledge.     

Combined effect of probiotics and specific immunoglobulin Y directed against Escherichia coli on growth performance,diarrhea incidence,and immune system in calves

Enterotoxigenic Escherichia coli (ETEC) K99 is one of the major pathogens associated with calf diarrhea. The induction of passive immunity in animals by immunoglobulin Y and using probiotics are inexpensive alternatives to antibiotics for the prevention and treatment of a number of bacterial infections, including diarrhea. Hence, the aim of this research was to evaluate the impact of dietary probiotics and ETEC K99-specific egg yolk antibody supplements, alone and in combination with each other, on health and growth parameters, diarrhea incidence and immune stimulation in newborn Holstein calves. One hundred and twenty neonatal calves were allocated randomly into 4 dietary groups (n = 30 per group) received colostrum/milk without any additives (control group), or supplemented with egg yolk powder contained E. coli K99-specific antibody (Ab group; 1 g/day), a commercial probiotic, Hypro-calves (Pro group; 3 g/day), and their combination (Ab + Pro group), from day (d) 1 to d28 of age. Analyses of the growth parameters, feed efficiency, fecal score, and microbiota and immune function were carried out on d0, 14, 21, and 28 of the experiment. Calves in Ab or Ab + Pro group had higher (P < 0.05) average daily gain compared to control and Pro groups during 0–14d. Feed efficiency of calves in Ab and Ab + Pro groups was significantly higher than that in control group during the period of 0–14d; however, no significant differences were observed in 0–28d period. Diarrhea prevalence and fecal score in Ab + Pro group were lower than control group (P < 0.05). Calves in Ab + Pro group had the lowest number of fecal E. coli in comparison to other groups on d28 (P < 0.05). Feeding Ab + Pro supplement increased (P < 0.05) concentrations of blood IgA and serum CD4 compared to the control group. Likewise, calves in Pro group had higher CD4 levels as compared to the control calves (P < 0.05). Serum concentration of interferon-gamma in control group was lower than other groups (P < 0.05). Overall, these data suggest that feeding a combination of probiotic and specific antibody against ETEC to neonate Holstein calves enhances feed efficiency, boosts immunity, and reduces diarrhea prevalence.     

Preparation of a high-immunoglobulin product from bovine colostrum

An effective colostrum management programme is the most important factor in determining the health and survival of the neonatal calf. Commercially available colostrum replacers (CR) and colostrum supplements (CS) are an alternative to colostrum on farms that do not have an adequate, high-quality colostrum supply, or those farms that want to prevent transmission of disease from cow to calf. The present study aimed to obtain a high immunoglobulin (Ig), dried bovine colostrum product that could be used as a CR or CS for dairy calves. Dried whey was made from 6 batches of colostrum and an additional 6 batches were used to produce dried whey and curds. Dried whey had higher IgG concentration (P<0.05) and lactose (P<0.05), and less fat (P<0.05) compared to curds or the original colostrum. There was a strong linear relationship between initial colostrum IgG concentration and whey (R² = 0.86) with approximately 0.45 of the initial colostral IgG residing in curd. The high IgG and the composition of colostrum whey powder suggests it could be an effective CS product for use with dairy calves. The high fat and IgG content of the curd by-product indicate that it might be a potential weaning supplement in piglets or even a product for human consumption.     

Variation in blood leucocytes,somatic cell count,yield and composition of milk of crossbred goats

Ten multiparous crossbred goats, five each of alpine × beetal (AB) and saanen × beetal (SB) were selected from the National Dairy Research Institute goat herd immediately after parturition. These were managed as per the practices followed in the institute’s goatherd. Blood and milk samples were collected at biweekly intervals from day 14 post-kidding for 22 weeks (154 days). Somatic cell count, electrical conductivity, fat, protein and lactose contents of milk were determined using standard methods. In the blood samples total leucocytes and differential leucocytes were also determined. Somatic cell counts were high immediately after parturition on day 14 of lactation and declined gradually with advanced lactation. There were individual variations (P < 0.01) in somatic cell counts between different lactation periods. Somatic cell count of milk was negatively correlated with neutrophils only (P < 0.05) and was neither correlated with milk yield, or with fat, protein, lactose content of milk. Electrical conductivity of milk was low up to four weeks of lactation and thereafter increased as the lactation advanced. Lactose content of milk declined gradually with the advancement of lactation. Fat content of milk was stable up to the eighth week and thereafter increased with advancement of lactation while the protein content of milk did not change significantly during lactation.     

Ear fibroblasts derived from Taiwan yellow cattle are more heat resistant than those from Holstein cattle

The objective of this study was to compare the thermotolerances of ear fibroblasts derived from Holstein (H) and Taiwan yellow cattle (Y) and their apoptosis-related protein expressions with (1, 3, 6, 12, and 24 h) or without heat shock treatment. The results showed that the vaginal temperatures of Y (38.4–38.5 °C) were (P<0.05) lower than that of H (38.8 °C) during the hot season. The apoptotic rates of ear fibroblasts derived from Y (6 h: 1.1%; 12 h: 1.6%; 24 h: 2.6%) were lower (P<0.05) than those of cells derived from H (6 h: 1.8%; 12 h: 4.0%; 24 h: 6.9%), respectively, after heat shock (42 °C). The expression level of apoptosis inducing factor (AIF) in ear fibroblasts derived from H was higher (P<0.05) than those derived from Y after the heat shock treatment for 6 h and 12 h, respectively. The level of cytochrome c of ear fibroblasts derived from H was higher (P<0.05) than those derived from Y after the heat shock treatment for 1–12 h, respectively. The abundances of Caspase-3, Caspase-8 and Caspase-9 of ear fibroblasts derived from H were higher (P<0.05) than those of cells derived from Y after 12 h and 24 h of heat shock, respectively; the Bcl-2/Bax ratios of ear fibroblasts derived from H were lower (P<0.05) than those from Y-derived fibroblasts after heated for 1–24 h. The expression level of HSP-70 of Y-derived ear fibroblasts was also higher (P<0.05) than that from H after the same duration of heat shock treatments. Taken together, the thermotolerance of ear fibroblasts derived from Taiwan yellow cattle was better than that of cells derived from Holstein cattle.     

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.     

Novel l-adenosine analogs as cardioprotective agents

Two l-nucleosides, l-3′-amino-3′-deoxy-N⁶-dimethyladenosine (l-3′-ADMdA) 1, previously synthesized in our laboratory, and the novel l-3′-amino-3′-deoxy-N⁶-methyladenosine-5′-N-methyluronamide (l-3′-AM-MECA) 2 were evaluated in an ischemia/reperfusion model on Langendorff perfused mouse heart. l-3′-ADMdA 1 was found to enhance functional recovery from ischemia (32.2 ± 3.7 cm H₂O/s % rate pressure product, compared to 21.3 ± 1.4 for the control and 30.7 ± 3.4 for adenosine) and increase the time to onset of ischemic contracture (14.5 ± 0.9 min, compared to 10.5 ± 1.0 min for the control and 13.6 ± 0.6 min for adenosine) comparable to adenosine. Consistent with the functional recovery data, decreased infarction area was seen in the case of 1 (19.1 ± 8.4, compared to 40.5 ± 7.2% for the control and 11.5 ± 2.1% for adenosine). In contrast, l-3′-AM-MECA 2 did not show significant functional recovery, increased onset of contracture, nor decreased infarction area compared to control. Unlike adenosine, neither 1 nor 2 induced cardiac standstill in mouse heart.     

Coumarin-thiazole and -oxadiazole derivatives: Synthesis,bioactivity and docking studies for aldose/aldehyde reductase inhibitors

In continuation of our previous efforts directed towards the development of potent and selective inhibitors of aldose reductase (ALR2), and to control the diabetes mellitus (DM), a chronic metabolic disease, we synthesized novel coumarin-thiazole 6(a–o) and coumarin-oxadiazole 11(a–h) hybrids and screened for their inhibitory activity against aldose reductase (ALR2), for the selectivity against aldehyde reductase (ALR1). Compounds were also screened against ALR1. Among the newly designed compounds, 6c, 11d, and 11g were selective inhibitors of ALR2. Whereas, (E)-3-(2-(2-(2-bromobenzylidene)hydrazinyl)thiazol-4-yl)-2H-chromen-2-one 6c yielded the lowest IC₅₀ value of 0.16 ± 0.06 μM for ALR2. Moreover, compounds (E)-3-(2-(2-benzylidenehydrazinyl)thiazol-4-yl)-2H-chromen-2-one (6a; IC₅₀ = 2.94 ± 1.23 μM for ARL1 and 0.12 ± 0.05 μM for ARL2) and (E)-3-(2-(2-(1-(4-bromophenyl)ethylidene)hydrazinyl)thiazol-4-yl)-2H-chromen-2-one (6e; IC₅₀ = 1.71 ± 0.01 μM for ARL1 and 0.11 ± 0.001 μM for ARL2) were confirmed as dual inhibitors. Furthermore, compounds 6i, 6k, 6m, and 11b were found to be selective inhibitors for ALR1, among which (E)-3-(2-(2-((2-amino-4-chlorophenyl)(phenyl)methylene)hydrazinyl)thiazol-4-yl)-2H-chromen-2-one (6m) was most potent (IC₅₀ = 0.459 ± 0.001 μM). Docking studies performed using X-ray structures of ALR1 and ALR2 with the given synthesized inhibitors showed that coumarinyl thiazole series lacks the carboxylate function that could interact with the anionic binding site being a common ALR1/ALR2 inhibitors trait. Molecular docking study with dual inhibitor 6e also suggested plausible binding modes for the ALR1 and ALR2 enzymes. Hence, the results of this study revealed that coumarinyl thiazole and oxadiazole derivatives could act as potential ALR1/ALR2 inhibitors.     

Synthesis and biological evaluation of 3-tetrazolo steroidal analogs: Novel class of 5α-reductase inhibitors

In the present study, a series of steroidal tetrazole derivatives of androstane and pregnane have been prepared in which the tetrazole moiety was appended at C-3 and 17a-aza locations. 3-Tetrazolo-3,5-androstadien-17-one (6), 3-tetrazolo-19-nor-3,5-androstadien-17-one (10), 3-tetrazolo-3,5-pregnadien-20-one (14), 17a-substituted 3-tetrazolo-17a-aza-d-homo-3,5-androstadien-17-one (26–31) and 3-(2-acetyltetrazolo)-17a-aza-d-homo-3,5-androstadien-17-one (32) were synthesized from dehydroepiandrosterone acetate (1) through multiple synthetic steps. Some of the synthesized compounds were evaluated for their in vitro 5α-reductase (5AR) inhibitory activity by measuring the conversion of [³H] androstenedione in human embryonic kidney (HEK) cells. In vivo 5α-reductase inhibitory activity also showed a significant reduction (p <0.05) in rat prostate weight. The most potent compound 14 showed 5AR-2 inhibition with IC₅₀ being 15.6 nM as compared to clinically used drug finasteride (40 nM). There was also a significant inhibition of 5AR-1 with IC₅₀ 547 nM compared to finasteride (453 nM).