Technique for Identifying Flavobacterium columnare Using Whole-Cell Fatty Acid Profiles

Abstract

Isolates of Flavobacterium columnare (29 from diseased fish and three American Type Culture Collection cultures [ATCC 23463, 49512, 43622]) were identified by use of biochemical characteristics prior to generating whole-cell fatty acid profiles. The microbial identification system (MIS; Microbial ID, Newark, Delaware), a gas chromatography system, was used to generate the fatty acid profiles of F. columnare. The MIS contains databases of clinically and environmentally important bacteria that are represented by over 100 genera, including Flavobacterium spp. (F. aquatile [ATCC 11947] and F. mizutaii). Flavobacterium columnare is not included in the databases because it does not grow on standard media. Fatty acid profiles of F. columnare were generated with the CLIN40 protocol established by MIS after growth of the bacteria in modified Shieh broth. The fatty acid composition of F. columnare isolates determined by the CLIN40 method consisted of 10 major fatty acids (those present at levels > 1%): 11-methyl-dodecanoic acid (13:0 iso [the term “iso” designates the methyl group at the penultimate carbon atom]), 13-methyl tetradecenoic acid isomer G (15:1 iso G), 13-methyl tetradecanoic acid (15:0 iso), 12-methyl tetradecanoic acid (15:0 anteiso [the term “anteiso” designates the methyl group at the third carbon atom from the end]), pentadecanoic acid (15:0), 14-methyl pentadecanoic acid (16:0 iso), 3-hydroxy-13-methyl tetradecanoic acid (15:0 iso 3OH), 15-methyl cis-9-hexadecenoic acid (iso 17:1 ω9c), 3-hydroxy-14-methyl pentadecanoic acid (16:0 iso 3OH), and 3-hydroxy-15-methyl hexadecanoic acid (17:0 iso 3OH) (94.8% of profile). Five fatty acids found in the highest percentages from all isolates (CLIN40 method) included 15:1 iso G (16.12%), 15:0 iso (46.54%), 15:0 iso 3OH (6.81%), iso 17:1 ω9c (7.32%), and 17:0 iso 3OH (9.42%). Fatty acid profiles were also established by means of the MIS rapid protocol (RCLN50) in which identifications can be completed in 7 min instead of 20 min. Fatty acids found in the highest percentages for the RCLN50 protocol included 15:1 iso G (15.36%), 15:0 iso (43.03%), 15:0 iso 3OH (7.43%), iso 17:1 ω9c (6.83%), and 17:0 iso 3OH (10.17%). Both methods will allow reliable identification of F. columnare.     

Characterization of Francisella tularensis Strains,Comparing Their Carbon Source Utilization

Thirteen Francisella tularensis strains were isolated from 22 seropositive brown hares (Lepus europaeus) originating from different parts of Hungary, and further two from a patas monkey (Erythrocebus patas) and vervet monkey (Chlorocebus aethiops). The isolates were identified as F. tularensis ssp. holarctica on the basis of culture, morphological and biochemical characteristics. The identification was verified by polymerase chain reaction and the sequencing of the partial 16S rRNA gene. Utilization of carbon sources of the 15 F. tularensis strains was characterized with the Biolog system. The system was able to identify the strains already after 4 h of incubation, not only after the standard 24 h. After the analysis and comparison of the metabolic profiles of our strains with the Biolog database, it was concluded that not all carbon sources indicated in the database were utilized by our isolates. The Biolog software fails to distinguish the highly virulent F. tularensis ssp. tularensis and the moderately virulent F. tularensis ssp. holarctica but the Biolog microplates can be manually read to differentiate the two subspecies based on glycerol source utilization. As all the studied strains were unable to use glycerol, they could be identified as F. tularensis ssp. holarctica. The dendrogram based on the metabolic relationship of the strains shows that the isolates are very similar to each other, which correlates with the conservative genetic character of F. tularensis ssp. holarctica.     

Influence of vitamin E on organic matter fermentation,ruminal protein and fatty acid metabolism,protozoa concentrations and transfer of fatty acids

Vitamin E (Vit. E) is discussed to influence ruminal biohydrogenation. The objective of this study was to investigate the influence of a Vit. E supplementation on rumen fermentation characteristics, ruminal microbial protein synthesis as well as ruminal organic matter fermentation. Furthermore, we aimed to investigate the influence of Vit. E supplementation on short‐chain fatty acids (SCFA) and protozoa concentrations in the rumen and, in addition, on transfer rates of middle‐chain and long‐chain fatty acids into the duodenum in lactating dairy cows. Eight rumen and duodenum fistulated German Holstein cows were assigned to either a group receiving 2,327 IU/d Vit. E (138.6 IU/kg DM DL‐α‐tocopherylacetate; n = 4) or a control group (23.1 IU/kg DM; n = 4). Neither ruminal protein synthesis nor organic matter fermentation was influenced by treatment. Vit. E did not act on the concentrations of short‐chain fatty acids and protozoa in rumen fluid. Duodenal flow of C13:0 (1.3 versus 0.2 g/d, p = 0.014) and iso‐C14:0 (1.0 versus 0.5 g/d, p = 0.050) was higher in the Vit. E group. We observed a trend for higher duodenal flows for C12:0 (1.6 versus 0.9 g/d, p = 0.095) and anteiso‐C15:0 (12.2 versus 8.9 g/d, p = 0.084). Transfer rate of C12:0 tended to be higher in the Vit. E group (125.61 versus 73.96, p = 0.082). No other transfer rates were affected by treatment. Further studies are necessary to investigate the influence of Vit. E on rumen microbiota and their fatty acid production as well as on the impact of different doses of Vit. E supplementation on variables of protein synthesis efficiency.     

Profiles of Fatty Acids in Different Bone Structures of Growing Chicks

This study was concerned with the dynamics of fatty acids in tissues involved in the growth and mineralization of the femur of Starbro chicks from 1 to 50 days of age. Four chickens that died of suffocation were obtained each day. Altogether, 400 femoral bones were studied in five age groups (I–V). Compact bone, spongy bone and articular cartilage were sampled. Lipids were extracted according to Folch and fatty acids were separated using gas chromatography. The fatty acid profile was found to change with age. Fatty acids with the highest content in bone were C_18:1, C_16:0, C_18:2 and C_18:0. The highest content of fatty acids was found in spongy bone and the lowest in articular cartilage. Several correlations were revealed between individual fatty acids. The following conclusions were drawn. (1) Fatty acid profiles in compact bone, spongy bone and articular cartilage change with age of chicks. (2) Oleic (C_18:1), palmitic (C_16:0), linoleic (C_18:2), stearic (C_18:0) and arachidonic (C_20:4) acids accounted for most of the fatty acid pool. (3) Correlations in the content of fatty acids were noted between bone structures.     

The relationships between odd‐ and branched‐chain fatty acids to ruminal fermentation parameters and bacterial populations with different dietary ratios of forage and concentrate

The objectives of this study were to investigate the effect of different dietary ratios of forage and concentrate (F:C) on ruminal odd‐ and branched‐chain fatty acids (OBCFAs) contents and to evaluate the relationships between OBCFA and ruminal fermentation parameters as well as bacterial populations tested by real‐time PCR technique. The experimental design was a 3 × 3 Latin square. Three rumen‐fistulated dry Holstein cows were fed three rations with different dietary F:C ratios (F:C; 30:70, 50:50 and 70:30). The rumen samples were collected every two hours (0600, 0800, 1000, 1200, 1400, 1600, 1800, 2000, 2200, 2400, 0200 and 0400 h) over three consecutive days in each sampling period. The results showed that rumen OBCFA profiles are significantly (p < 0.05) affected by the dietary F:C ratios. The concentrations of C11:0, C13:0, iso‐C15:0, iso‐C16:0, iso‐C17:0 and C17:0 were higher in the cows fed dietary F:C ratio of 70:30 than those fed with other two rations. However, the concentrations of anteiso‐C15:0, C15:0 and total OBCFA were on the lowest level in the high forage diet. Correlation and regression analysis showed that ruminal OBCFAs had strong relationships with ruminal fermentation parameters and bacterial populations. In particular, the iso‐fatty acids had potential power to predict butyrate and isoacids metabolized in the rumen, whereas the fatty acids with 17 carbon atoms correlated with ruminal NH₃‐N content. The OBCFA contents have different relationships with fibrolytic and starch bacteria in the rumen. C17:0 and its isomers might be used to predict populations of fibrolytic bacteria.     

The effect of dietary Chlorella vulgaris supplementation on micro‐organism community,enzyme activities and fatty acid profile in the rumen liquid of goats

Microalgae might be considered as an alternative source of fat and/or protein for ruminant's diets. However, changes in populations of ruminal micro‐organisms associated with biohydrogenation process, methane and ammonia production in response to microalgae dietary supplementation have not been well characterized. Thus, 16 cross‐bred goats were divided into two groups. Each goat of both groups was fed individually with alfalfa hay and concentrates separately. The concentrates of the control group had no microalgae while those of the treated group were supplemented with 10 g lyophilized Chlorella vulgaris/kg concentrate (chlor). On the 30th experimental day, samples of rumen fluid were collected for microbial DNA extraction, fatty acid profile and enzyme activity analyses. The results showed that the chlor diet compared with the control increased significantly the populations of Methanosphaera stadtmanae, Methanobrevibacter ruminantium and Methanogens bacteria and protozoa in the rumen of goats. A significant reduction in the cellulase activity and in the abundance of Ruminococcus albus, and a significant increase in the protease activity and in the abundance of Clostridium sticklandii in the rumen liquid of goats fed with the chlor diet, compared with the control, were found. Chlorella vulgaris supplementation promoted the formation of trans C_18:1, trans‐11 C_18:1 and monounsaturated fatty acids (MUFA), while the proportions of C_18:0 and long‐chain fatty acids (LCFA) reduced significantly in the rumen liquid of goats. This shift in ruminal biohydrogenation pathway was accompanied by a significant increase in Butyrivibrio fibrisolvens trans C_18:1‐producing bacteria. In conclusion, the supplementation of diets with microalgae needs further investigation because it enhances the populations of methane‐producing bacteria and protozoa.     

Reproductive Performance of Broiler Breeder Hens Fed n-3 Fatty Acid-rich Fish Oil

Abstract

The objective of the study was to investigate the effects of oil sources in the diet on milk yield, milk composition, and fatty acid (FA) profiles in mid-lactating dairy cows. Forty-eight Chinese Holstein dairy cows averaging 150 days in milk (DIM) at the start of the experiment (body weight = 596±19 kg; milk yield = 29.7±3.00 kg/d) were used in a completely randomized block design. The animals were assigned into four dietary treatments according to DIM and milk yield, and supplemented with no oil (control), 2% flaxseed oil (FSO), 2% soybean oil (SBO), and 2% oil from extruded soybeans (ESB). The experiment lasted nine weeks including the first week for adaptation. Milk yields, milk compositions (fat, protein, and lactose), and milk FA profiles were measured. Daily milk yield from cows fed with FSO, SBO, and ESB were higher than milk yield of the control cows (27.0, 27.0, and 26.5 vs. 25.4 kg/d). Milk fat percentage of the control cows was greater than those cows fed with oil-supplemented diets. However, increasing dietary fat content resulted with no change in fat-corrected milk yield. The FA profile of milk was changed by fat supplementation. Feeding oil reduced the proportion of both short-chain (C_8:0 to C_12:0) and medium-chain (C_14:0 to C_16:1) FAs, and increased the proportion of long-chain (≥C_18:0) FAs in milk fat. Cis-9, trans-11 conjugated linoleic acid (CLA) in milk fat was increased from 0.38% for the control to 0.79, 1.51, and 1.56% of fat for the cows supplemented with FSO, SBO, and ESB, respectively. Feeding oils rich in linoleic acid (SBO and ESB) was more effective in enhancing cis-9, trans-11 CLA in milk fat than oils containing linolenic acid (FSO). There was a linear relationship between transvaccenic acid and cis-9, trans-11 CLA content in milk. Overall, feeding the FSO, SBO, and ESB diets increased monounsaturated and polyunsaturated fatty acids and decreased the saturated fatty acid in milk fat.     

Composition and fatty acid profile of milk from cows supplemented with pressed oilseed cake

This study compared the productive and nutritional parameters of milk from crossbred lactating cows managed on Panicum maximum Jacq. cv. Tanzania and with a diet supplemented with different pressed oilseed cakes. The supplements used were as follows: peanut cake, sunflower cake and palm kernel cake for replacement of soybean meal. Sixteen cows with an average weight of 544 ± 57 kg and producing 8 ± 1.4 L of milk per day were used in this study. The animals were randomly assigned to the treatments according to a Latin square design repeated over time, with four treatments, 16 animals and four experimental periods. Supplementation of the diet with peanut cake, sunflower cake and palm kernel cake compared with soybean meal in the diet of cows did not affect the average daily production or composition of the milk. The palm kernel cake promoted an increase in lauric fatty acids (C_12:0) and palmitoleic acids (C_16:1) (5.02 and 1.65%, respectively) compared with peanut cake and sunflower cake (4.13 and 4.01%, respectively). The levels of oleic fatty acids (C_18:1) were higher for the sunflower cake and palm kernel cake supplements (26.01 and 25.01%, respectively) compared with peanut cake (23.11%). The replacement of soybean meal with sunflower cake and palm kernel cake improved the nutritional quality of the milk, with lower concentrations of saturated fatty acids and higher concentrations of unsaturated fatty acids, without compromising the production or nutritional composition of the milk. © 2015 Japanese Society of Animal Science     

A preliminary study on the effect of dietary supplementation with cod liver oil on the polyunsaturated fatty acid composition of boar semen

Eight mature Norwegian Landrace boars, of proven fertility and in routine semen production for AI, were fed individually with the same basic diet for 9 weeks. One group of 4 animals served as the control, the remaining 4 boars received a daily supplement of 75 ml cod liver oil (CLO-group). Fifteen consecutive semen samples were collected from each boar. The fatty acid composition of the semen was determined, and the content of the 15 most numerous fatty acids with a chain length longer than 12 carbon atoms was followed over time. In both groups, the proportion of 16:1n-7 decreased significantly, while 16:0 and 22:6n-3 (DHA) increased. By the end of the experiment, DHA had tended to increase and 22:5n-6 to decrease to a greater extent in the CLO-group. A significant difference between the groups was seen for onen-6 PUFA (22:4n-6), which remained unchanged in the control group but decreased in the CLO-group. No change was seen in docosapentaenoic acid (22:5n-3) and eicosapentaenoic acid (20:5n-3) was not found in any sample. These results indicate that CLO supplementation affects the fatty acid composition of boar semen. There were no significant differences in the non-return rates (4–25 days) between the two groups before, during or after the experiment.Abbreviations AA arachidonic acid - AI artificial insemination - area% each fatty acid percentage of the total area of the peaks from gas chromatographic analysis of methylated fatty acids - CLO cod liver oil - DHA docosahexaneoic acid - DPA docosapentaenoic acid - EPA eicosapentaenoic acid - LA linoleic acid - LNA linolenic acid - NR non-return rate - PUFA polyunsaturated fatty acid - T _c the temperature marking the beginning of the phase transition Fatty acid nomenclature: Example, 22:6n-3: 22=number of carbon atoms, 6=number of double bonds,n-3=position of the first double bond counted from the terminate (n or methyl end of the fatty acid chain     

Fatty acid profile and carcass characteristics in castrated and uncastrated hair lambs

The aim of this study was to investigate the effects of castration on carcass characteristics and fatty acid profile of visceral fat and meat from lambs. Eighteen six-month-old Santa Inês male lambs (18.9?±?2.4 kg of body weight (BW)) were used. Animals were assigned to two treatments according to their sexual condition: uncastrated (U) or castrated (C). During a 98-day experimental period, animals were kept on an Andropogon gayanus grass pasture area of 1 ha and supplemented with 200 g/animal/day of concentrate mixture. Water and mineral salt were available ad libitum. The lambs were weighed fortnightly, and at the last day of the trial, they were slaughtered for evaluation of carcass characteristics and fatty acids profile of perirenal fat and longissimus lumborum muscle samples. Castrated lambs showed higher BW than U during most part of the experiment (p?<?0.05). Fat deposition was higher in C lambs as evidenced by their increased carcass fat cover. Meat from U lambs showed lower content of C_16:0 and higher polyunsaturated fatty acids (PUFA) (p?<?0.05) than that from C (U, 14.3 and C, 10.5%). Conjugated linoleic acid (CLA) content was not affected by castration (p?>?0.05) (U, 0.74 and C, 0.76%). The cis-9, cis-12 C_18:2n-6 (U, 10.6 and C, 6.86%) fatty acid and the PUFA:SFA (saturated fatty acids) ratio (U, 0.36 and C, 0.25) were higher in the muscle of U lambs (p?<?0.05), indicating that the meat from U animals may provide more benefits to human health than that of C.

     

Effect of short‐chain fatty acids on triacylglycerol accumulation,lipid droplet formation and lipogenic gene expression in goat mammary epithelial cells

Short‐chain fatty acids (SCFAs) are the major energy sources for ruminants and are known to regulate various physiological functions in other species. However, their roles in ruminant milk fat metabolism are still unclear. In this study, goat mammary gland epithelial cells (GMECs) were treated with 3 mmol/L acetate, propionate or butyrate for 24 h to assess their effects on lipogenesis. Data revealed that the content of triacylglycerol (TAG) and lipid droplet formation were significantly stimulated by propionate and butyrate. The expression of FABP3, SCD1, PPARG, SREBP1, DGAT1, AGPAT6 and ADRP were upregulated by propionate and butyrate treatment. In contrast, the messenger RNA (mRNA) expression of FASN and LXRα was not affected by propionate, but reduced by butyrate. Acetate had no obvious effect on the content of TAG and lipid droplets but increased the mRNA expression of SCD1 and FABP3 in GMECs. Additionally, it was observed that propionate significantly increased the relative content of mono‐unsaturated fatty acids (C18:1 and C16:1) at the expense of decreased saturated fatty acids (C16:0 and C18:0). Butyrate and acetate had no significant effect on fatty acid composition. Overall, the results from this work help enhance our understanding of the regulatory role of SCFAs on goat mammary cell lipid metabolism.     

Hydrogenated oil decreases tissue concentrations of n‐6 polyunsaturated fatty acids and may contribute to dyschondroplasia in broilers

1. In a factorial design of dietary treatments, male Ross broilers were given diets containing soyabean oil, hydrogenated soya‐bean oil (as a source of trans‐fatty acids) or feed fat with either 0 or 300 μg of added D‐biotin/kg.

2. Growth to 28 d was not influenced by the dietary treatments.

3. Length of tibiotarsal bones was reduced (P<0.05) and severity of leg bone cartilage lesions, characteristic of dyschondroplasia, was highest (P<0.05) in broilers fed on diets containing hydrogenated soyabean oil.

4. Feeding hydrogenated soyabean oil lowered (P< 0.05) the concentrations of C₂₀:_4n6 and the ratios of C_20:4n6/C_18:2n6 in liver and growth plate cartilage.

5. Growth plate cartilage from birds affected with dyschondroplasia contained lower proportions of prostaglandin precursor fatty acids compared with normal growth plate.

6. It is speculated that an inhibition of prostaglandin biosynthesis brought about by the presence of trans‐fatty acids might contribute to the occurrence of lesions similar to dyschondroplasia.     

家养观赏地图鱼肺炎克雷伯氏菌、维氏气单胞菌与黏质沙雷氏菌的分离鉴定及耐药性分析

本试验旨在通过细菌分离鉴定,明确家养观赏地图鱼的死因,筛选敏感药物。采用常规方法分离纯化细菌后,进行细菌形态学观察,并通过小白鼠致病性试验、细菌主要生化鉴定、16SrDNA序列测定分析、药敏试验及耐药基因检测等方法对分离的细菌进行鉴定及耐药分析。分离出3株革兰氏阴性短杆菌,根据细菌形态特征及理化特性,结合16SrDNA序列测定与系统发育分析结果,判定其分别为肺炎克雷伯氏菌(Klebsiella pneumoniae)、维氏气单胞菌(Aeromonas veronii)和黏质沙雷氏菌(Serratia marcescens),其中肺炎克雷伯氏菌具有较强致病性。3株细菌均对洛美沙星、氧氟沙星、阿米卡星、卡那霉素敏感,对阿莫西林、氟苯尼考、克林霉素、甲硝唑等具有较强耐药性。结果表明,肺炎克雷伯氏菌、维氏气单胞菌、黏质沙雷氏菌的混合感染是家养观赏地图鱼的死亡原因。     

Effect of methanol on methanogenesis and fermentation in the rumen simulation technique (RUSITEC)

Introduction Major methanol sources in ruminant feeds are pectins esterified with methoxyl groups. Methanol can be released by pectin esterase activity of rumen bacteria (R exova -B enkova and M arkovic 1976). It has been detected in rumen fluid of cows in vivo and in vitro (V antcheva et al. 1970, 1972). It is, however, not likely to accumulate in the rumen fluid since it can be readily used by methylotrophic organisms. Methanogenic archaebacteria such as Methanosarcina barkeri (H utten et al. 1980; M& uuml ; ller et al. 1986) are able to use methanol as energy and carbon source. Two equations have been reported for this conversion: 4CH₄ → 3CH₄ + CO₂ + 2 H₂O (1) CH₃OH + H₂ → CH₄ + H₂O (2) Methanogenesis from methanol has been shown to occur during in vitro fermentations with rumen fluid as inoculum (C zerkawski and B reckenridge 1972; P ol and D emeyer 1988). However, acidogenic micro-organisms, such as Eubacterium limosum or Butyribacterium methylotrophicum, may also utilize methanol. In rumen fluid of sheep fed a molasses-rich diet, Eubacterium limosum was the predominant methanol-utilizing bacterium (G enthner et al. 1981). Major products of these acidogenic methylotrophs are acetate and butyrate. These different fermentation end-products, methane versus fatty acids, will affect the metabolizable energy content of feedstuffs that are rich in highly methoxylated pectins or other methyl group-containing compounds. The purpose of the present study was to investigate whether the rumen simulation technique (RUSITEC) would be a suitable model to evaluate the methanogenic or acidogenic potential of methanol during ruminal fermentation. The study focused on methane production, fermentation characteristics and methanol turnover. Fermentation traits included pH, redox potential, ammonia and volatile fatty acid production, and degradation of feed constituents.     

Communications: Bacteria in the Hemolymph of the Freshwater Prawn Macrobrachium rosenbergii

Abstract

Bacteria from the hemolymph of the freshwater prawn Macrobrachium rosenbergii were identified and quantified. Total bacterial counts ranged from 0.0 to 4.6 × 10⁵ cells/1.0 mL of hemolymph. Predominant bacteria isolated included Aeromonas spp., Bacillus sp., and Pseudomonas spp. No bacteria were found in the hemolymph of prawns without lesions. The predominant species of bacteria isolated from water samples of prawn culture ponds was a chitinoclastic Bacillus sp.     

Comparison of two live Bacillus species as feed additives for improving in vitro fermentation of cereal straws

This study was performed in a 2 × 4 factorial arrangement to explore and compare the effects of inclusion of two live Bacillus additives (B. licheniformis and B. subtilis) at four doses (0, 0.25 × 10⁷, 0.50 × 10⁷ and 0.75 × 10⁷ colony‐forming units (cfu)) on in vitro gas production kinetics, fiber degradation, methane production and ruminal fermentation characteristics of maize stover and rice straw by mixed rumen microorganisms in dairy cows. The pH, concentrations of ammonia nitrogen (NH₃‐N) and isovalerate were increased (P < 0.05), while the methane (CH₄) production, ratio of acetate to propionate, and total volatile fatty acids (TVFA) concentration were decreased (P < 0.05) by the supplementation of B. licheniformis compared with that of B. subtilis. Adding B. licheniformis and B. subtilis raised (P < 0.05) or numerically raised the maximum gas production, while decreasing (P < 0.05) or numerically lowering pH and concentrations of most volatile fatty acids. The addition of B. licheniformis increased (P < 0.05) the NH₃‐N concentration but reduced CH₄ production and ratio of acetate to propionate (P < 0.05), while the NH₃‐N concentration was decreased (P < 0.05), and the CH₄ production and ratio of acetate to propionate were increased by that of B. subtilis compared to the control. Results obtained in this research suggest that B. licheniformis would be preferred as a live Bacillus additive in comparison with B. subtilis, and its optimal dose should be 0.25 × 10⁷ cfu/500 mg substrates.     

Production of dairy cows fed whole-crop cereals or ryegrass silages supplemented with a fixed amount of concentrate

Abstract

Feed intake and milk production responses to whole-crop cereal silages and ryegrass silage (RGS) supplemented with a fixed amount of concentrate were measured in three 4×4 Latin square-designed experiment with three 17-day periods using 12 Holstein cows. Diets consisted of a fixed amount of concentrate (10 kg) and one of the following silages offered ad libitum: RGS, Mondego wheat silage (MWS), Alva wheat silage (AWS), and triticale silage (TS). Silage dry matter intake and milk yield were significantly higher for RGS. Milk composition was not affected by silage treatment. The whole-crop cereal silages tended to show higher milk concentrations of anteiso C15:0, iso C15:0, C16:1 and C18:1 and lower concentrations of C18:2 and C18:3. This study suggests better response of dairy cows to a single cut RGS than to whole-crop cereal silages harvested in an early stage of maturity.     

The effect of dietary fish oil soapstock on the performance,carcass fat and flavour of broilers 1

Three hundred and thirty six male broilers were fed on diets containing two levels (2% and 4%) of either acidulated soybean oil soapstock (SOS), acidulated fish oil soapstock (FOS), or a combination of the two. Some of the replicates were changed from FOS diets to diets containing 4% SOS at 5 or 6 1/2 weeks of age.

Dietary FOS was slightly, but significantly, inferior to SOS with regard to live performance, apparently due to an adverse effect on food intake. All birds fed on FOS were unpalatable due to a pronounced “fishy” flavour, thigh meat being more objectionable than breast meat and cooked broth more than roasted meat. About 3 to 4 weeks after transfer to SOS diets, the broilers were acceptable.

Both soapstocks contained over 65% free fatty acids, but their fatty acid composition differed, with the FOS containing more C₁₂ to C₁₆ saturated fatty acids, palmitoleic acid and the typical long‐chain polyenoic fatty acids, but much less linoleic acid. These patterns were reflected in the total dietary lipids and in the abdominal adipose tissue. After the type of dietary soapstock was altered, there was a gradual change in the composition of body fat, which, after about 3 weeks, approached the fatty acid pattern of the birds fed on a diet containing 4% SOS throughout their growing period.     

Accuracy of the n-alkane technique for intake estimates in beef cattle using different sampling procedures and feeding levels

The present study aimed to evaluate the reliability of the n-alkane technique for estimating herbage intake in beef cattle receiving two different feeding levels (low and high) and to test the effect of different faeces sampling procedures (total faeces samples or rectal grab samples at 8 h intervals) on the n-alkane faecal recoveries and hence on intake estimates. Two consecutive experiments were performed with 11 non-lactating beef cows of “Asturiana de los Valles” breed, fed with lucerne hay at two feeding levels: 1.0 or 1.7 kg DM/100 kg body weight (BW). Animals received a daily dose of paper pellets containing C₂₄, C₃₂ and C₃₆ n-alkanes as external markers with the purpose of using different n-alkane pairs of adjacent chain length (C₂₃/C₂₄, C₂₅/C₂₄, C₃₁/C₃₂, C₃₃/C₃₂, C₃₅/C₃₆) for intake calculations. A 5-day equilibrium period was sufficient for external marker concentrations and n-alkane pair ratios to reach steady state in faeces. There was no effect of different sampling times (every 8 h) on the faecal excretion of n-alkane pairs C₃₁/C₃₂ and C₃₃/C₃₂. Grab samples obtained at the time of dosing (0830 h) gave the best estimate of n-alkane pair ratios in total faeces collection. There was a general trend of increasing n-alkane faecal recoveries with increased chain length, although n-alkanes C₃₀, C₃₁ and C₃₃ showed lower faecal recoveries than expected. n-Alkane recoveries were in general lower at the low feeding level, where an increase in individual variability of natural n-alkane recoveries was observed. At both feeding levels, the n-alkane pairs C₂₃/C₂₄ and C₃₅/C₃₆ gave accurate estimates of the treatment average intake, although the natural n-alkane (C₂₃ or C₃₅) faecal recoveries showed high individual variability, which could be in part due to analytical bias caused by their extremely low concentration in the diet and faeces. However, the n-alkane pairs C₃₁/C₃₂ and C₃₃/C₃₂ gave the greatest deviations of intake estimates, due to the high discrepancy between the faecal recoveries of natural and dosed n-alkanes. These results demonstrate that in beef cattle, feeding level may have some influence on the relative faecal recoveries of the n-alkane pair used for intake calculations. This effect, together with the high individual variability of n-alkane recoveries, especially under low feeding level, may produce significant deviations of the individual intake estimates.     

Fatty acid composition of ruminal bacteria and protozoa, with emphasis on conjugated linoleic acid, vaccenic acid, and odd-chain and branched-chain fatty acids

Knowledge of the fatty acid profile of microbial lipids is of great nutritional importance to the animals and, subsequently, their products. This study was conducted to examine the fatty acid profiles of mixed rumen bacteria and protozoa. Bacterial and protozoal cells were isolated by differential centrifugation of rumen contents. The main fatty acids were palmitic (16:0) and stearic (18:0) in both the bacterial and protozoal fractions. Palmitic acid was 74% greater in the protozoal fatty acids than in the bacterial fatty acids, whereas bacteria had 2.25-times greater stearic acid (18:0) proportions compared with protozoa. The total odd-chain plus branched-chain fatty acids were 16.5% of bacterial fatty acids and 11.0% of protozoal fatty acids. The anteiso-17:0 proportions in bacterial and protozoal fatty acids were 1.4 and 2.9%, respectively. The most abundant trans-18:1 isomer, vaccenic acid (18:1 trans-11), was 6.6% of total fatty acids in protozoa and 2.0% of total fatty acids in bacteria. The cis-9, trans-11 CLA was 8.6-times greater in the protozoal fraction (1.32% of total fatty acids) than in the bacterial fraction (0.15%). These results suggest that the presence of protozoa in the rumen may increase the supply of CLA and other unsaturated fatty acids for lower gut absorption by ruminants.