ORIGINAL ARTICLE: 2‐Oxoglutaric acid administration diminishes fundectomy‐induced osteopenia in pigs

Partial surgical removal of the stomach (fundectomy, FX) leads to osteopenia in animals and humans. FX adversely affects the bone. 2‐oxoglutaric acid is a precursor of glutamine and hydroxyproline – the most abundant amino acid of collagen. The aim of the study was to investigate the effects of 2‐oxoglutaric acid on FX‐evoked osteopenia in pigs. Eighteen castrated male pigs of the Pu?awska breed were used. Twelve pigs were subjected to FX and divided into two groups: FX + AKG (the AKG group; AKG at the daily dosage of 0.4 g/kg of body weight) and FX + Placebo (the FXC group; received CaCO₃ as placebo). Remaining six pigs were sham‐operated (the SHO group). The pigs were euthanized at the age of 8 months and long bones were collected. Area bone mineral density (aBMD) and bone mineral content (BMC) were measured; morphology, geometry and biomechanical properties were determined. Moreover, the serum concentrations of selected hormones and one marker of bone metabolism were determined. FX caused osteopenia in the pigs and treatment with AKG greatly reduced these effects of FX in pigs. Negative effect of fundectomy on the skeletal system leading to decreased bone mass in pigs is associated with lowered body gain and activity of the gastric–hypothalamic–pituitary axis. Better definitions of each of the local and systemic hormonal and structural components associated with fundectomy‐induced decreased bone mass that separately and together determine the whole bone properties may lead to identify opportunities for prevention.     

The effects of benzoic acid supplementation on the performance of broiler chickens

To determine the efficiency of benzoic acid, a feeding experiment was carried out with 240 one‐day old ROSS 308 cockerels. Birds were divided into three dietary treatments: I (C) – no additives, II (B1) – 0.1% inclusion of benzoic acid, III (B2) – the inclusion of 0.2% benzoic acid. The performance results were similar in birds fed the control diet and the diet with 0.1% of benzoic acid. The dietary inclusion of benzoic acid at 0.2% depressed the growth of broiler chickens (p < 0.05). The dry matter of the digesta increased in the crop and caeca after benzoic acid supplementation. The pH of the caecal contents decreased following benzoic acid supplementation and was the lowest in the B2 group. No differences were found in the pH of the crop, ileal, gizzard digesta and rectum content. Lactic acid bacteria populations were the lowest in the caeca of the B1 group (p < 0.05). Coliform bacteria decreased in the caeca contents following increased benzoic acid supplementation.     

Nutritive value and non‐toxicity of Botryosphaeria zeae‐infected wheat for weaner pigs

A novel fungal infection of wheat by Botryosphaeria zeae was identified on the Darling Downs of Queensland, Australia and called ‘white grain’ because of its bleached appearance. The only nutritional changes in the wheat grain infected with B. zeae were decreases in nitrogen and total amino acids (～5%), and slight increases in the lysine, fibre and fat content, with starch unaffected. Nutrient digestibility and potential toxicity were assessed in weaner pigs housed in metabolism crates and fed this grain over a 4‐week period, as they grew from 15 to 35 kg. Digestibility of energy and nitrogen in white grain was not different from that of normal wheat. The piglets were then bled for biochemical and haematological testing, slaughtered and the entire viscera subjected to gross pathological inspection, followed by histological examination of the liver, kidney, pancreas, spleen, heart, lung, muscle and intestine. White grain did not affect palatability of diets, pig growth rates; no abnormalities were detected in tissues, while biochemical and haematological parameters did not suggest any toxic effect. Hence, wheat with white grain appears suitable for use in pig diets, but the use of this wheat for human food should be restricted until additional longer term studies are conducted.     

Effect of intake of linoleic acid and α‐linolenic acid levels on conversion into long‐chain polyunsaturated fatty acids in backfat and in intramuscular fat of growing pigs

A study was conducted to determine the effect of two levels of linoleic acid (LA) intake at either high or low α‐linolenic acid (ALA) intake on their conversion and subsequent deposition into long‐chain (20–22 C‐atoms) polyunsaturated fatty acids (LC PUFA) in muscle and backfat in growing pigs. In a 2 × 2 factorial arrangement, 32 gilts from 8 litters were assigned to one of four dietary treatments, varying in LA and ALA intakes. Low ALA and LA intakes were 0.15 and 1.31 g/(kg BW^0.75/day), respectively, and high ALA and LA intakes were 1.48 and 2.65 g/(kg BW^0.75/day) respectively. There was a close positive relation between intake of ALA and the concentration of ALA in backfat and in intramuscular fat. Dietary ALA did not affect the concentration of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), but increased docosapentaenoic acid (DPA) in backfat. High ALA intake did not significantly affect DHA but significantly increased EPA, 20:3 n‐3 and DPA concentrations in intramuscular fat. The n‐3 LC PUFA proportion in backfat was increased from approximately 1–3%, which may be useful to enrich meat with these fatty acids. The effect of ALA intake on n‐3 LC PUFA was suppressed by LA intake. Dietary ALA suppressed the concentration of n‐6 LC PUFA in blood plasma by more than 50%. When compared at equal incremental dose, the inhibiting effect of ALA on blood arachidonic acid was stronger than the stimulating effect of LA as precursor.     

Seasonal variation in abiotic factors and ferulic acid toxicity in snail‐attractant pellets against the intermediate host snail Lymnaea acuminata

Laboratory evaluation was made to access the seasonal variations in abiotic environmental factors temperature, pH, dissolved oxygen, carbon dioxide, electrical conductivity and ferulic acid toxicity in snail‐attractant pellets (SAP) against the intermediate host snail Lymnaea acuminata in each month of the years 2010 and 2011. On the basis of a 24‐h toxicity assay, it was noted that lethal concentration values of 4.03, 3.73% and 4.45% in SAP containing starch and 4.16, 4.23% and 4.29% in SAP containing proline during the months of May, June and September, respectively, were most effective in killing the snails, while SAP containing starch/proline + ferulic acid was least effective in the month of January/February (24‐h lethal concentration value was 7.67%/7.63% in SAP). There was a significant positive correlation between lethal concentration value of ferulic acid containing SAP and levels of dissolved O₂/pH of water in corresponding months. On the contrary, a negative correlation was observed between lethal concentration value and dissolved CO₂/temperature of test water in the same months. To ascertain that such a relationship between toxicity and abiotic factors is not co‐incidental, the nervous tissue of treated (40% and 80% of 24‐h lethal concentration value) and control group of snails was assayed for the activity of acetylcholinesterase (AChE) in each of the 12 months of the same year. There was a maximum inhibition of 58.43% of AChE, in snails exposed to 80% of the 24‐h lethal concentration value of ferulic acid + starch in the month of May. This work shows conclusively that the best time to control snail population with SAP containing ferulic acid is during the months of May, June and September.     

Anti‐allergic effects of Lactobacillus crispatus KT‐11 strain on ovalbumin‐sensitized BALB/c mice

In this study, we investigated the effects of oral ingestion of Lactobacillus crispatus KT‐11 strain (KT‐11) on the immune response in an allergic rhinitis mouse model, ovalbumin (OVA)‐sensitized BALB/c mice. Sneezing activity in mice that were administered a KT‐11‐supplemented diet was significantly lower than that in mice administered a KT‐11‐free diet (control diet) at age 11 weeks. We found that serum OVA‐specific immunoglobulin E (IgE) levels and total number of interleukin (IL)‐4⁺CD4⁺ spleen cells in mice that were administered a KT‐11‐supplemented diet were significantly lower than in mice administered a control diet. The ratio of spleen interferon‐γ⁺CD4⁺/IL‐4⁺CD4⁺ cells was higher in the mice administered the KT‐11‐supplemented diet compared to that in mice administered the control or L. rhamnosus GG‐supplemented diet. In contrast, the number of CD11b⁺CD80⁺ and FcεRIα⁺CD117⁺ cells was significantly lower in mice administered the KT‐11‐supplemented diet. These results suggested that KT‐11 reduced OVA‐induced allergic symptoms in BALB/c mice via the adjustment of the T helper type 1/T helper type 2 balance, and a decrease in the number of antigen‐presenting cells and high affinity IgE receptor‐positive mast cells.     

Effects of adding liquid dl‐methionine hydroxy analogue‐free acid to drinking water on growth performance and small intestinal morphology of nursery pigs

This study was conducted to evaluate the effect of adding liquid dl ‐methionine hydroxy analogue free acid (LMA) to drinking water on growth performance, small intestinal morphology and volatile fatty acids in the caecum of nursery pigs. Twenty‐four crossbred pigs (Large White × Landrace, BW ～18 kg) were divided into three groups with four replications of two piglets each. The piglets received drinking water without (control), with 0.05 or 0.10% LMA. The results indicated that adding LMA at 0.10% to drinking water significantly increased their weight gain, average daily feed intake (p < 0.05) and tended to improve the feed conversion ratio. Adding LMA to drinking water significantly increased their water intake and significantly reduced the pH of drinking water (p < 0.01), thus total plate count (p < 0.01) and Escherichia coli in drinking water was reduced (p < 0.05), while the total number of bacteria in the caecum was not significantly affected. Liquid dl ‐methionine hydroxy analogue free acid supplementation in drinking water tended to decrease pH in the stomach, duodenum, jejunum, colon and rectum. Furthermore, adding LMA at 0.10% significantly increased villous height in the duodenum, jejunum and ileum (p < 0.05), and the villous height:crypt depth ratio in the jejunum and ileum (p < 0.01) was higher, whereas acetic acid concentration in the caecum was significantly lower than in the control group. It could be concluded that adding LMA to drinking water improved growth performance of the nursery pigs because of high water quality and high nutrient utilization caused by an improvement of small intestinal morphology (not from nutritional effect of methionine source).     

The effect of dietary long‐chain omega‐3 fatty acid supplementation on owner’s perception of behaviour and locomotion in cats with naturally occurring osteoarthritis

The aim of this randomized, double‐blinded, placebo–controlled, cross‐over designed study was to demonstrate the clinical effect, registered by a survey, of a 10‐week period of omega‐3 fatty acid supplementation of the diet (1.53 g eicosapentaenoic acid (EPA) and 0.31 g DHA, both per 1000 kcal ME, equivalent to the complete diet) of 16 cats with radiologically documented, naturally occurring osteoarthritis (OA), in comparison with a 10‐week period of supplementation with corn oil (0.00 g EPA and 0.00 g DHA, both per 1000 kcal ME). Cats on the fish oil revealed higher activity level (p = 0.07), more walking up and down the stairs (p = 0.07), less stiffness during gait (p = 0.03), more interaction with the owner (p = 0.07) and higher jumps (p = 0.03) compared to those on corn oil supplementation. In conclusion, supplementation of long‐chain omega‐3 polyunsaturated fatty acids changes the owner’s perception of some aspects of behaviour and locomotion in cats with naturally occurring OA.     

The effects of early‐age thermal manipulation and daily short‐term fasting on performance and body temperatures in broiler exposed to heat stress

This study was conducted to investigate the effects of thermal manipulation at 5 days of age and short‐term fasting during the warmest part of the day on responses to prolonged heat stress of broilers. A total of 240‐day‐old Ross 308 female broiler chicks were divided into three groups: control, thermal manipulation (chicks were exposed to 36 °C for 24 h at 5 days of age) and short‐term fasting during the warmest part of the day (10.00–17.00 h). Prolonged heat stress was induced daily from 28 to 42 days by heating until the ambient temperature reached 32–35 °C between 10.00 and 17.00 h. Both thermal manipulation and short‐term fasting resulted in a decrease in rectal temperatures and haematocrit values at 35 and 41 days of age. Thermal manipulation improved body weight, feed consumption and feed conversion. However, short‐term fasting caused a reduction in body weight and a deterioration in feed conversion. Short‐term fasting lowered the percentages of carcass, whereas thermal manipulation highered breast yield. Both thermal manipulation and short‐term fasting decreased heart mass and abdominal fat.     

Effect of dietary α‐tocopherol + ascorbic acid,selenium, and iron on oxidative stress in sub‐yearling Chinook salmon (Oncorhynchus tshawytscha Walbaum)

A three‐variable central composite design coupled with surface‐response analysis was used to examine the effects of dietary α‐tocopherol + ascorbic acid (TOCAA), selenium (Se), and iron (Fe) on indices of oxidative stress in juvenile spring Chinook salmon. Each dietary factor was tested at five levels for a total of fifteen dietary combinations (diets). Oxidative damage in liver and kidney (lipid peroxidation, protein carbonyls) and erythrocytes (erythrocyte resistance to peroxidative lysis, ERPL) was determined after feeding experimental diets for 16 (early December) and 28 (early March) weeks. Only TOCAA influenced oxidative stress in this study, with most measures of oxidative damage decreasing (liver lipid peroxidation in December and March; ERPL in December; liver protein carbonyl in March) with increasing levels of TOCAA. We also observed a TOCAA‐stimulated increase in susceptibility of erythrocytes to peroxidative lysis in March at the highest levels of TOCAA. The data suggest that under most circumstances a progressive decrease in oxidative stress occurs as dietary TOCAA increases, but higher TOCAA concentrations can stimulate oxidative damage in some situations. Higher levels of TOCAA in the diet were required in March than in December to achieve comparable levels of protection against oxidative damage, which may have been due to physiological changes associated with the parr‐smolt transformation. Erythrocytes appeared to be more sensitive to variation in dietary levels of TOCAA than liver and kidney tissues. Using the March ERPL assay results as a baseline, a TOCAA level of approximately 350–600 mg/kg diet would provide adequate protection against lipid peroxidation under most circumstances in juvenile Chinook salmon.     

Olive leaves (Olea europea L.) and α‐tocopheryl acetate as feed antioxidants for improving the oxidative stability of α‐linolenic acid‐enriched eggs

Ninety‐six brown Lohmann laying hens were equally assigned into four groups with six replicates. Hens within the control group were fed a corn–soybean‐based diet supplemented with 4% linseed oil. Two other groups were given the same diet further supplemented with 5 or 10 g ground olive leaves/kg feed, while the diet of the fourth group was further supplemented with 200 mg α‐tocopheryl acetate/kg. Supplementing diets with olive leaves had no effect on egg production, feed intake and egg traits. Eggs collected 28 days after feeding the experimental diets were analysed for lipid hydroperoxides and malondialdehyde (MDA) content, fatty acid profile, α‐tocopherol concentrations and susceptibility to iron‐induced lipid oxidation. Olive leaves were also analysed for total and individual phenolics, and total flavonoids, whereas their antioxidant capacity was determined using both the DPPH (1,1‐diphenyl‐2‐picrylhydrazyl) and ABTS (2,2‐azinobis3‐ethylbenzothiazoline‐6‐sulphonic acid) radical scavenging activity assays. Results showed that neither α‐tocopheryl acetate nor olive leaves supplementation exerted (p > 0.05) any effect on the fatty acid composition of n‐3 eggs. Supplementing the diet with 5 g olive leaves/kg had no (p > 0.05) effect on the hydroperoxide levels of n‐3 eggs, while supplementing with 10 g olive leaves/kg or 200 mg α‐tocopheryl acetate/kg, the lipid hydroperoxide levels were reduced (p ≤ 0.05) compared to control. However, although hydroperoxides were reduced, MDA, a secondary lipid oxidation product, was not affected (p > 0.05). Iron‐induced lipid oxidation increased MDA values in eggs from all groups, the increase being higher (p ≤ 0.05) in the control group and the group supplemented with 5 g olive leaves/kg. The group supplemented with 10 g olive leaves/kg presented MDA values lower (p ≤ 0.05) than the control but higher (p ≤ 0.05) than the α‐tocopheryl acetate group, which presented MDA concentrations lower (p ≤ 0.05) than all other experimental diets at all incubation time points.     

Effect of ammonium‐iron‐hexa‐cyanoferrate and of the covariates age,gender, weight,season and calendar time on radiocaesium contamination of wild boars living in the wild in Bavaria

Feed with Ammonium‐iron‐hexa‐cyanoferrate (AFCF; 1250 mg AFCF/kg) was fed between March 2009 and March 2011 to wild boars in a territory of 4.5 km² (experimental group, EXP). One hundred and forty similar territories in the same county (500 km², spruce forest, agriculture) served as control (CON). Data for comparison from all territories were available from March 2005 to March 2011. Wild boars could move between, into and from the territories. Lean skeletal muscle meat (500 g) of all wild boars that were killed by humans (hunting and traffic accidents) was investigated for gamma‐radiation from ¹³⁷Cs with a becquerel monitor with a sodium iodide scintillator crystal (range of detection 20–9999 Bq/kg). The wild boars were weighed, and gender and age were determined. For the analyses of effects, multivariable regression models were fitted with the ¹³⁷Cs concentration as response variable. There was a significant difference between the ¹³⁷Cs contamination of wild boars from CON (563 ± 932 Bq/kg meat, n = 1253) and EXP (236 ± 276 Bq/kg meat; n = 45). ¹³⁷Cs contamination decreased with increasing body weight by ?5 Bq/kg meat/kg body weight increase (p < 0.05). Females had higher Bq measurements than males (by +80 Bq/kg meat, p < 0.05). Piglets were lower than adults, but turn‐coats higher. From November to May, contamination was higher (by +500 to +600 Bq/kg meat, p < 0.05) than during the rest of the year. In 2010, contamination was higher (by +200 to + 300 Bq/kg meat, p < 0.05) in comparison with the other years under observation. When all covariates were controlled for, the effect of AFCF was highly significant. Interaction analyses showed that the intervention decreased ¹³⁷Cs contamination by ?500 Bq/kg meat during November to May and by ?200 Bq/kg meat during the rest of the year. In summary, AFCF feeding reduces ¹³⁷Cs contamination of wild boars living in the wild significantly, particularly during the season from November to May.     

Effect of excess levels of lysine and leucine in wheat‐based,amino acid‐fortified diets on the mRNA expression of two selected cationic amino acid transporters in pigs

An experiment was conducted to evaluate the effect of excess levels of Leu and Lys on the expression of b^0,+ and CAT‐1 mRNA in jejunum, liver and the muscles Longissimus dorsi (LDM) and Semitendinosus (STM). Twenty pigs with an average initial BW of 16.4 ± 1.7 kg were used in a Randomized Complete Block. Dietary treatments (T) were as follows: T1, basal diet; T2, basal plus 3.5 g l ‐Lys/kg diet; T3, basal plus 1.5 g l ‐Leu/kg diet; T4, basal plus 3.5 g l ‐Lys plus 1.5 g l ‐Leu/kg diet. Diets in T1 and T3 met 100% the requirement of Lys for pigs within the 10 to 20 kg body weight range; diets in T2 and T4 contained 35% excess of Lys. Also, diets in T1 and T2 supplied 104%, whereas diets in T3 and T4 supplied 116% the requirement of Leu. The expression of b^0,+ in jejunum was reduced (p = 0.002) because of the supplementation of l ‐Leu, but l ‐Lys supplementation had no effect (p = 0.738). In contrast, the expression of b^0,+ in STM (p = 0.012) and liver (p = 0.095) was reduced by the high level of Lys, but Leu had no effect (p > 0.100). CAT‐1 expression in STM increased by high Lys (p = 0.023) and Leu (p = 0.007) levels. In liver, the expression of CAT‐1 substantially increased (p = 0.001) because of Lys. In conclusion, excess levels of dietary Lys and Leu affect the expression of cationic amino acid transporters, and this effect varies depending on the studied tissue.     

Effects of amino acids infused into the vein on ghrelin‐induced GH,insulin and glucagon secretion in lactating cows

To investigate the effects of amino acids on ghrelin‐induced growth hormone (GH), insulin and glucagon secretion in lactating dairy cattle, six Holstein cows were randomly assigned to two infusion treatments in a cross‐over design. Mixture solution of amino acids (AMI) or saline (CON) was continuously infused into the left side jugular vein via catheter for 4 h. At 2 h after the start of infusion, synthetic bovine ghrelin was single injected into the right side jugular vein through the catheter. Ghrelin injection immediately increased plasma GH, glucose and non‐esterified fatty acids (P < 0.05) with no difference between both treatments. Additionally, plasma insulin and glucagon concentrations were increased by ghrelin injection in both treatments. The peak value of plasma insulin concentration was greater in AMI compared with CON (P < 0.05). Plasma glucagon concentration showed no difference in the peak value reached at 5 min between both treatments, and then the plasma levels in AMI compared with CON showed sustained higher values (P < 0.05). After plasma glucose concentration reached the peak, the decline was greater in AMI compared with CON (P < 0.05). These results showed that the increased plasma amino acids may enhance ghrelin action which in turn enhances insulin and glucagon secretions in lactating cows.