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.     

Combined effect of apigenin and ferulic acid on frozen‐thawed boar sperm quality

The main aim of the present study was to evaluate the cryoprotective effect of apigenin (AP) and ferulic acid (FA) on boar sperm during cryopreservation. AP and FA were both demonstrated to be high‐efficiency antioxidants and had not previously been used to protect sperm from cryodamage. As boar sperm is sensitive to oxidative stress, suitable antioxidants are still needed for improving frozen‐thawed sperm quality. With this purpose, semen samples coming from five boars were used in this study. Ejaculates of five boars were mixed and split into 16 aliquots, in which different doses of AP and FA were added separately or together. The motility, the plasma membrane integrity, the mitochondrial activity, the acrosomal integrity, the antioxidase activities and the malondialdehyde concentration of the frozen‐thawed boar sperm were assessed. The results suggested that both AP and FA significantly improved the frozen‐thawed boar sperm quality in all these aspects when they were added to the freezing extender separately, while the highest improvement was recorded when the extender was supplemented with 0.1 mmol/L AP plus 0.15 mmol/L FA. These findings demonstrated that supplementation of freezing extender with both AP and FA had a combined, beneficial effect on frozen‐thawed boar sperm.     

A toxicity study of low‐dose rate half‐body irradiation and chemotherapy in dogs with lymphoma

Thirteen dogs with previously untreated multicentric lymphoma were enrolled in a prospective study investigating the effects of low‐dose rate total body irradiation (TBI) and chemotherapy. Dogs received either 6 or 8 Gy TBI in half‐body fractions, 2 weeks apart. Toxicity consisted of mild to moderate haematological and gastrointestinal (GI) signs. One dog died from treatment complications. Anorexia was noted independent of dose. Haematological toxicity was more common and more severe after 8 Gy treatment. GI toxicity was more likely postcaudal half‐body irradiation with 8 Gy. Other than leukotrichia, late effects from radiation were not observed. Results indicated that haematological and nonhaematological toxicity was dose dependent. However, the protocol was well tolerated and treatment intensification using a 2‐week inter‐radiation interval was possible in all dogs treated with 6 Gy. Preliminary survival data for these dogs were very encouraging, providing a strong rationale to analyse the efficacy of low‐dose rate irradiation (LDRI) in canine lymphoma.     

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.     

Comparison of acute inhalation toxicity of sulfuric acid by the inhalation and intratracheal instillation methods

Recently, intratracheal instillation has been focused on as a simple, low-cost alternative to the inhalation method. In this study, intratracheal instillation of sulfuric acid, a typical acidic compound, was performed to compare the acute toxicity of acidic compounds that could cause damage to the respiratory system between intratracheal instillation and inhalation. Sulfuric acid was administered to male rats at doses of 0.7, 2, 7, 20, and 60 mg/kg by dividing the total dose into four doses. General condition and body weight were examined up to 14 days after administration, and macropathological and histopathological examinations were performed. The half-lethal dose was then estimated. All animals administered 20 and 60 mg/kg sulfuric acid and one animal administered 2 mg/kg sulfuric acid died within 4 h after administration. No abnormalities were observed in other animals. At 20 and 60 mg/kg, multiple red foci or diffuse red areas were macroscopically observed in the lungs. In these lesions, histopathologically, clefts between the mucosal epithelium and basement membrane and necrosis of the alveolar epithelium were observed. Deaths in these groups may have resulted from lung injury. No notable changes were observed in other animals. Therefore, the half-lethal dose of sulfuric acid by intratracheal instillation was estimated as 7–20 mg/kg. The acute toxicity by intratracheal instillation was evaluated with two-fold sensitivity since the exposure at the half-lethal sulfuric acid concentration in inhalation studies was calculated as 43.2 mg/kg.     

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.     

Investigation of lactic acid bacterial strains for meat fermentation and the product's antioxidant and angiotensin‐I‐converting‐enzyme inhibitory activities

In the lactic acid bacteria (LAB) strains screened from our LAB collection, Lactobacillus (L.) sakei strain no. 23 and L. curvatus strain no. 28 degraded meat protein and tolerated salt and nitrite in vitro. Fermented sausages inoculated strains no. 23 and no. 28 showed not only favorable increases in viable LAB counts and reduced pH, but also the degradation of meat protein. The sausages fermented with these strains showed significantly higher antioxidant activity than those without LAB or fermented by each LAB type strain. Angiotensin‐I‐converting‐enzyme (ACE) inhibitory activity was also significantly higher in the sausages fermented with strain no. 23 than in those fermented with the type strain. Higher ACE inhibitory activity was also observed in the sausages fermented with strain no. 28, but did not differ significantly from those with the type strain. An analysis of the proteolysis and degradation products formed by each LAB in sausages suggested that those bioactivities yielded fermentation products such as peptides. Therefore, LAB starters that can adequately ferment meat, such as strains no. 23 and no. 28, should contribute to the production of bioactive compounds in meat products.     

Branched‐chain amino acid ratios in low‐protein diets regulate the free amino acid profile and the expression of hepatic fatty acid metabolism‐related genes in growing pigs

Liver metabolism is affected by nutrients. The aim of this study was to explore the effects of low‐protein diets (17% crude protein, CP) supplemented with branched‐chain amino acids (BCAAs), including leucine (Leu), isoleucine (Ile) and valine (Val), on hepatic amino acid profile and lipid metabolism in growing pigs. The ratio of Leu : Ile : Val in all groups was 1 : 0.51 : 0.63 (20% crude protein, CP), 1 : 1 : 1 (17% CP), 1 : 0.75 : 0.75 (17% CP), 1 : 0.51 : 0.63 (17% CP) and 1 : 0.25 : 0.25 (17% CP) respectively. Results revealed that compared to the positive control group (1 : 0.51 : 0.63, 20% CP), the low‐protein diets significantly augmented the concentrations of most essential amino acids and non‐essential amino acids (p < .05), with the greatest values observed in the 1 : 0.25 : 0.25 group. Moreover, relative to the control, the low‐protein diets with the Leu : Ile : Val ratio ranging from 1 : 0.75 : 0.75 to 1 : 0.25 : 0.25 markedly downregulated the mRNA abundance of acetyl‐CoA carboxylase (ACC), lipoprotein lipase (LPL) and fatty acid‐binding protein 4 (FABP‐4) (p < .05), and upregulated the mRNA expression of hormone‐sensitive lipase (HSL), peroxisome proliferator‐activated receptor‐g coactivator‐1α (PGC‐1α), uncoupling protein 3 (UCP3) and liver carnitine palmitoyltransferase 1 (L‐CPT‐1) (p < .05). Therefore, our data suggest that protein‐restricted diets supplemented with optimal BCAA ratio, that is, 1 : 0.75 : 0.75–1 : 0.25 : 0.25, induce a shift from fatty acid synthesis to fatty acid oxidation in the liver of growing pigs. These effects may be associated with increased mitochondrial biogenesis.     

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.     

Effects of addition of Aspergillus oryzae culture and 2‐hydroxyl‐4‐(methylthio) butanoic acid on milk performance and rumen fermentation of dairy cows

To investigate effects of Aspergillus oryzae culture (AOC) and 2‐hydroxy‐4‐(methylthio) butanoic acid (HMB) on milk performance and rumen fermentation of dairy cows. Sixty‐four multiparous Chinese Holstein cows were randomly allocated into four experimental diets: (i) Control diet; (ii) AOC diet: 5 g AOC/day per head; (iii) HMB diet: 25 g HMB/day; and (iv) AH diet: 5 g AOC plus 25 g HMB/day. Added HMB tended to increase the yield of milk protein (P = 0.06) and 3.5% fat‐corrected milk (P = 0.08) and milk fat content (P = 0.09). Milk fat yield (P = 0.03) and the contents of milk protein (P = 0.05) were increased by adding HMB. The cows fed on AOC diet had a tendency for higher body weight (BW) gain (P = 0.08). Addition of AOC, HMB and AH increased content of microbial protein (MCP) and total volatile fatty acids (VFA) (P < 0.01) in rumen fluid. Populations of rumen fungi, Fibrobacter succinogenes and Ruminococcus flavefaciens relative to total bacterial 16S rDNA (P ≤ 0.03) and activity of carboxymethylcellulase (CMCase) (P < 0.01) were increased with added AOC or HMB. It is inferred that added AOC or HMB can increase the contents of MCP and total VFA potentially by stimulating rumen microbe populations and CMCase activity.     

Effect of applying lactic acid bacteria and propionic acid on fermentation quality and aerobic stability of oats‐common vetch mixed silage on the Tibetan plateau

The objective of this study was to evaluate effects of lactic acid bacteria and propionic acid on the fermentation quality and aerobic stability of oats‐common vetch mixed silage by using a small‐scale fermentation system on the Tibetan plateau. (i) An inoculant (Lactobacillus plantarum) (L) or (ii) propionic acid (P) or (iii) inoculant + propionic acid (PL) were used as additives. After fermenting for 60 days, silos were opened and the aerobic stability was tested for the following 15 days. The results showed that all silages were well preserved with low pH and NH₃‐N, and high lactic acid content and V‐scores. L and PL silages showed higher (P < 0.05) lactic acid and crude protein content than the control silage. P silage inhibited lactic acid production. Under aerobic conditions, L silage had similar yeast counts as the control silage (> 10⁵ cfu/g fresh matter (FM)); however, it numerically reduced aerobic stability for 6 h. P and PL silages showed fewer yeasts (< 10⁵ cfu/g FM) (P < 0.05) and markedly improved the aerobic stability (> 360 h). The result suggested that PL is the best additive as it could not only improved fermentation quality, but also aerobic stability of oats‐common vetch mixed silage on the Tibetan plateau.     

Effects of alpha‐lipoic acid supplementation on growth performance,antioxidant capacity and biochemical parameters for ammonia‐exposed broilers

In order to estimate the effect of alpha‐lipoic acid (LA) supplementation on relieving ammonia stress of broilers, 180 22‐day‐old male broilers were assigned to three groups, six replicates in each group and 10 birds per replicate. The three groups were: (1) a control group without ammonia stress; (2) exposure to 70 ppm atmospheric ammonia (AM); (3) exposure to 70 ppm atmospheric ammonia and administration of 300 mg/kg LA (AM + LA). The experimental period was 3 weeks. Results showed that average daily weight gain was increased and feed conversion ratio was decreased in the AM + LA group, compared with the AM group (P < 0.05). Total superoxide dismutase and glutathione peroxidase activities in serum, and glutathione content in liver were higher in the AM + LA group than that in the AM group (P < 0.05); however, serum malondialdehyde content was decreased by LA addition (P < 0.05). Additionally, serum glutamic‐pyruvic transaminase, creatine kinase and lactate dehydrogenase activities were reduced and albumin level was increased by LA addition (P < 0.05). In conclusion, LA addition could relieve ammonia stress to restore broiler production performance to normal levels.     

Effect of dietary microbially produced gamma‐linolenic acid and plant extracts on enzymatic and non‐enzymatic antioxidants in various broiler chicken organs

Plant extracts and fungal fermented feed with gamma‐linolenic acid‐rich microbial oils are perspective additives for use in animal nutrition as appetite and digestion stimulants, stimulants of physiological functions, for the prevention and treatment for certain pathological conditions, and as antioxidants. The activity of antioxidant enzymes and the level of reduced glutathione were measured in the plasma and in liver, heart and kidney mitochondria after 42 days of feeding broiler chickens both regular and combination diets. These were selected based on our previous experience. The administration of agrimony and gamma‐linolenic acid resulted in a significant decrease in superoxide dismutase activity in all four bodies in contrast to plant extracts. We conclude that the decrease in activity is due to decreased production, and hence dismutation, of superoxide radicals to peroxides followed by lower activity of glutathione peroxidase, which was not seen in the case of only plant extract administration. Generally, higher glutathione reductase activity would be in response to increased demands on reduced glutathione as a cofactor for the reaction catalysed by glutathione peroxidase and the utilization of glutathione itself. However, measured levels of reduced glutathione showed no change. The results argue against any oxidative stress conditions. The application of agrimony extract appears to be suitable for the antioxidant effect against peroxidation of gamma‐linolenic acid. As the efficacy of measuring the effects of diets on the oxidative stability of meat caused by selected antioxidant enzymes is rather low, additional data from the experiment will be processed to clearly assess the influence of this combination of diets.     

Effects of maternal treatment with β‐hydroxy‐β‐metylbutyrate and 2‐oxoglutaric acid on femur development in offspring of minks of the standard dark brown type

The aim of the study was to evaluate the effect of the diet, mother type and sex of the offspring on the mechanical and geometric parameters of long bones as well as bone tissue density in minks. Primiparous and multiparous dams were supplemented with β‐hydroxy β‐methylbutyrate (a metabolite of leucine, at the daily dosage of 0.02 g/kg of body weight) and/or 2‐oxoglutaric acid (a precursor of glutamine, at the daily dosage of 0.4 g/kg of body weight) during gestation. The diet did not influence bone tissue density and the length of the humerus. An increase in the length of the femur was noted in male offspring delivered by multiparous dams. The diet resulted in an increase in the weight of the humerus in males from multiparous dams and a decrease in offspring from primiparous dams. Heavier femora were noted in male offspring delivered by both types of dams. The maximum elastic strength of the humerus was higher in the offspring delivered by multiparous than primiparous dams, irrespective of the offspring sex. The diet resulted in reduction in the ultimate strength of the femur in the male offspring delivered by primiparous dams. Only females born by multiparous dams, irrespective of the diet, showed a significant increase in the cross‐sectional area of the humerus, while a significant decline was noted in males delivered by multiparous dams and in all the offspring delivered by primiparous dams. An increase in the cross‐sectional area of the femur was noted in the offspring delivered by multiparous dams, while reduction was observed in the offspring delivered by primiparous dams. These results have shown for the first time that the presence of β‐hydroxy‐β‐methylbutyrate or 2‐oxoglutaric acid in the diet of pregnant primiparous or multiparous dams unambiguously affects the geometry and mechanical properties of offspring's long bones.     

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.     

The effect of pomegranate seed oil and grapeseed oil on cis‐9, trans‐11 CLA (rumenic acid), n‐3 and n‐6 fatty acids deposition in selected tissues of chickens

The aim of this study was to determine whether diet modification with different doses of grapeseed oil or pomegranate seed oil will improve the nutritive value of poultry meat in terms of n‐3 and n‐6 fatty acids, as well as rumenic acid (cis‐9, trans‐11 conjugated linoleic acid) content in tissues diversified in lipid composition and roles in lipid metabolism. To evaluate the influence of applied diet modification comprehensively, two chemometric methods were used. Results of cluster analysis demonstrated that pomegranate seed oil modifies fatty acids profile in the most potent way, mainly by an increase in rumenic acid content. Principal component analysis showed that regardless of type of tissue first principal component is strongly associated with type of deposited fatty acid, while second principal component enables identification of place of deposition—type of tissue. Pomegranate seed oil seems to be a valuable feed additive in chickens’ feeding.     

Improving effects of dietary rumen protected γ‐aminobutyric acid additive on apparent nutrient digestibility,growth performance and health status in heat‐stressed beef cattle

This study was aimed to investigate the effects of rumen‐protected γ‐aminobutyric acid (RP‐GABA) on apparent nutrient digestibility, growth performance and health status in heat stressed beef cattle. Fifty Jinjiang Yellow cattle were randomly assigned to 5 treatments (10 animals/treatment). Treatments 1 to 5 were basal diets affixed with 0 (control), 8, 16, 24 and 32 mg of RP‐GABA/kg of body weight (BW) respectively. The trial lasted 45 days. Apparent digestibility of crude protein (CP), crude fiber (CF) and calcium (Ca) quadratically increased with increasing RP‐GABA (p < .01), while apparent digestibility of phosphorus (P) tended to quadratically increase (p = .09). Dietary supplementation with increasing RP‐GABA linearly increased DM digestibility and average daily gain (ADG) (p < .01), whereas the feed to gain (F:G) ratio linearly decreased with increasing RP‐GABA (p < .01). The average daily feed intake (ADFI) value tended to linearly increase with RP‐GABA supplementation (p = .08). Total protein (TP), blood urea nitrogen (BUN) and malondialdehyde (MDA) levels quadratically decreased (p < .01) with increasing RP‐GABA, however albumin (ALB), glucose (GLU), superoxide dismutase (SOD), triiodothyronine (T3) and thyroxine (T4) levels quadratically increased (p ≤ .01). In conclusion, the present results indicated that dietary supplementation with RP‐GABA led to improved nutrient digestibility, growth performance and antioxidant status in heat stressed beef cattle.     

菌肥与柠檬酸互作对石灰性土壤生物学特性及草坪质量的影响

在草坪的日常管理中,肥料施用是一个必不可少的环节。在实际生产中,一般通过常规的氮、磷肥施用促进草坪草的生长,达到提高和改善草坪质量的目的,而通过施用微生物肥料和柠檬酸等辅助性肥料来改善土壤微生物环境、提高土壤养分有效性,从而促进草坪的生长方面研究较少。本文研究了柠檬酸和促生菌肥的互作处理对冷季型草坪质量及土壤生物学特性的影响,从而探讨柠檬酸和促生菌肥提高草坪草质量的机理,为草坪的科学管理提供理论依据。研究表明,1)一定量菌肥配合柠檬酸增加了石灰性土壤中微生物数量,尤其是深层土壤放线菌数量,且提高了土壤蔗糖酶和脲酶的活性。2)柠檬酸的施用,显著提高了土壤中有效磷含量。3)在一定量的氮、磷肥施用水平下,柠檬酸和促生菌肥有效提高了草坪质量。