Phylogenetic placement within Lecanoromycetes of lichenicolous fungi associated with Cladonia and some other genera.

Though most of the lichenicolous fungi belong to the Ascomycetes, their phylogenetic placement based on molecular data is lacking for numerous species. In this study the phylogenetic placement of 19 species of lichenicolous fungi was determined using four loci (LSU rDNA, SSU rDNA, ITS rDNA and mtSSU). The phylogenetic analyses revealed that the studied lichenicolous fungi are widespread across the phylogeny of Lecanoromycetes. One species is placed in Acarosporales, Sarcogyne sphaerospora; five species in Dactylosporaceae, Dactylospora ahtii, D. deminuta, D. glaucoides, D. parasitica and Dactylospora sp.; four species belong to Lecanorales, Lichenosticta alcicorniaria, Epicladonia simplex, E. stenospora and Scutula epiblastematica. The genus Epicladonia is polyphyletic and the type E. sandstedei belongs to Leotiomycetes. Phaeopyxis punctum and Bachmanniomyces uncialicola form a well supported clade in the Ostropomycetidae. Epigloea soleiformis is related to Arthrorhaphis and Anzina. Four species are placed in Ostropales, Corticifraga peltigerae, Cryptodiscus epicladonia, C. galaninae and C. cladoniicola comb. nov. (= Lettauia cladoniicola). Three new species are described, Dactylospora ahtii, Cryptodiscus epicladonia and C. galaninae.

Sharpening the species boundaries in the Cladonia mediterranea complex (Cladoniaceae, Ascomycota).

The complex Cladonia mediterranea belongs to the section Impexae and is formed by C. azorica, C. macaronesica and C.mediterranea. These species are basically distributed in the Mediterranean and Macaronesian Regions. In the present work the limits between the species of this complex are re-examined. To this end, the morphological characters were studied along with the secondary metabolites and the DNA sequences from three loci (ITS rDNA, IGS rDNA and rpb2). The morphological data were studied by principal component analysis (PCA), while the DNA sequences were analyzed using several approaches available to delimit species: genealogical concordance phylogenetic species recognition, species tree (BEAST* and spedeSTEM) and cohesion species recognition. In addition, the genealogical sorting index was used in order to assess the monophyly of the species. The different procedures used in our study turned out to be highly congruent with respect to the limits they establish, but these limits are not the ones separating the prior species. Either the morphological analysis or the different approaches to species delimitation indicate that C. mediterranea is a different species from C. macaronesica, while C. azorica and C. macaronesica, which are reduced to synonyms of C. portentosa, constitute a separate lineage.

Effect of reduction of milking frequency and supplementation of vitamin E and selenium above requirements on milk yield and composition in Assaf ewes.

The aim of this research was to study the effect of milking frequency and supplementation with a vitamin-mineral complex above requirements on intake, body weight (BW), and milk yield and composition in high-yielding Assaf ewes. Sixteen lactating Assaf ewes were used in this study, separated into 4 groups of 4 ewes each. Animals in 2 of the groups (control groups) did not receive any extra vitamin-mineral supplement, whereas animals in the other 2 groups (supplement groups) received daily an oral dose of 1g of vitamin E (1,000 IU, DL-α-tocopherol acetate) and 0.4 mg of selenium (sodium selenite anhydrous). The experiment consisted of 2 consecutive periods of 3 wk (twice-daily milking in both mammary glands) and 8 wk (once-daily milking in one mammary gland and twice-daily milking in the other gland). Intake, BW, and milk composition were controlled weekly, and milk production was recorded 3 times a week. Administration of the vitamin-mineral supplement had no effect on dry matter intake, BW, or milk production and composition. The reduction of milking from twice to once a day caused a decrease in milk production and lactose concentration and a significant increase in protein concentration, total solids, and somatic cell count, without affecting the fat content. Administration of a vitamin E and Se supplement at the doses used in the present study does not seem to exert, in the short term, a noticeable effect on the mammary gland when milking frequency is reduced.

Animal performance and fatty acid composition of lambs fed with different vegetable oils.

Twenty-seven lambs were used to investigate the effects of the inclusion of 4% hydrogenated palm oil (HPO) or sunflower oil (SFO) in the concentrate on animal performance, carcass and meat quality and fat characteristics and fatty acid composition. Animals (16.2±0.27kg initial weight) were fed concentrate (Control, HPO or SFO) and barley straw ad libitum and slaughtered at 25kg. SFO lambs tended to eat less concentrate than HPO animals (P<0.10). Neither HPO nor SFO affected any of the carcass characteristics studied, meat pH and meat and fat colour (P>0.05). SFO decreased proportions of C16:0, C18:1 cis-11 and C18:3 (P<0.05) and increased C18:1 trans (P<0.001) and C18:2/C18:3 ratio (P<0.05). Atherogenicity index was lower (P<0.05) when SFO was included in the concentrate. HPO did not affected and SFO improved fatty acid composition of fattening lambs without affecting animal performance.

Carcass and meat quality of Assaf milk fed lambs: Effect of rearing system and sex.

The effect of sex and rearing system on growth and carcass and meat characteristics of milk fed Assaf lambs was studied. Thirty-six lambs, 18 males and 18 females were used. Twelve lambs remained with their mothers throughout the experiment (NR). Within 24-36h of birth, the rest were housed individually and fed twice a day ad libitum (AAR) or at 70% of ad libitum consumption (RAR) with reconstituted cow's milk. Sex did not affect animal performance, yet females showed higher carcass and non-carcass fat deposits. NR lambs showed greater BWG than AAR fed lambs, and AAR, higher than the RAR. Differences between naturally and artificially reared lambs in CCW and killing out percentage were not significant. Empty digestive tract and mesenteric fat weights were greater for RAR than NR lambs, with the AAR lambs demonstrating intermediate values; conversely, omental fat was greater in NR lambs. Carcass ether extract content was greater for NR lambs, possibly due to the greater growth. Use of ad libitum cow's milk substitute in suckling lambs twice a day resulted in less body weight gain but similar killing out percentages compared to naturally raised lambs. A 70% restricted supply increased the days in suckling and reduced carcass fatness and compactness. Except for water loss, which was less in NR than artificially fed lambs, no differences were found in meat characteristics.

Effects of the inclusion of sodium bicarbonate and sugar beet pulp in the concentrate for fattening lambs on acid-base status and meat characteristics.

Thirty-six young Merino lambs (15.3kg live weight) were allocated into four equal groups to study the effects of the inclusion of sodium bicarbonate ('Bic', 0 vs. 2%) and sugar beet pulp ('SBP', 0 vs. 12%) in the concentrate on meat characteristics. Lambs were fed barley straw and concentrate ad libitum. When they reached 25kg a blood sample was taken and the animal slaughtered. After 24h meat characteristics (pH, colour, water holding capacity, Warner-Bratzler shear force, chemical and fatty acid - 'FA' - composition) were measured on M. longissimus thoracis et lumborum. SBP replacing 24% of the barley in the concentrate caused an increase in shear force (P<0.05) and saturated FA (P<0.05), decreasing meat unsaturated FA (P<0.05). SB increased blood base excess (P<0.05) and meat yellowness (P<0.05) and decreased meat pH (P<0.05), without altering any other meat characteristics.

Effects of sunflower soap stocks on light lamb meat quality.

Thirty-two lambs were used to study the effect of sunflower soap stocks (SS), a by-product from the vegetable oil refinery industry, on meat chemical composition, fatty acid profile, volatile compounds, and consumer acceptability. Lambs were finished (average length of fattening period 35 ± 7.3 d, 26.8 ± 0.09 kg final BW) on a pelleted total mixed ration (TMR) with no SS (00SS) or including 15, 30 or 60 g SS/kg (15SS, 30SS, and 60SS, respectively). Sunflower soap stocks decreased the percentage of SFA, increased the proportion of -MUFA ( < 0.05), and modified the levels of several odor-active lipid-derived volatile compounds ( 0.05). Consumers were able to distinguish between control and 15SS meat samples in a triangular test ( < 0.05), but a well-defined preference for meat of any of these treatments was not exhibited ( > 0.05). Atherogenicity and saturation indexes decreased by 31% and 27%, respectively, in SS groups compared to control (linear 0.05). However, certain volatile compounds (benzene and toluene) and 10-18:1 fatty acid, known potential hazards for human health, were increased in meat from lambs fed TMR with SS. For this reason, only inclusion rates up to 15 g SS/kg TMR seem to sustain a satisfactory balance between beneficial and detrimental effects on lamb meat composition and quality.

Total mixed ration pellets for light fattening lambs: effects on animal health.

Fifty male Merino lambs (6 to 8 weeks, 14.1 kg; n=10 per group) were used to study the effect of feeding system: barley straw in long form and concentrate pellets in separate troughs (Control), ad libitum alfalfa supplemented with concentrate in separate troughs (Alfalfa) or including various levels of ground barley straw in concentrate pellets (B05, B15 and B25 for 50, 150 and 250 g barley straw/kg), on rumen characteristics, acid-base status, blood cell counts and lymphocyte stimulation. Alfalfa lambs had the heaviest digestive tract contents, highest rumen pH values, lowest volatile fatty acid concentration, highest papillae counts and best mucosa colour and the greatest blood pCO2 values, lowest sodium and chloride and highest potassium concentrations (P<0.05). Including ground barley straw in the concentrate pellet or providing straw in long form separately from the concentrate reduces rumen pH and darkens ruminal mucosa as compared with alfalfa-fed lambs, thus affecting acid-base status.

Effects of linseed and quercetin added to the diet of fattening lambs on the fatty acid profile and lipid antioxidant status of meat samples.

Thirty-two Merino lambs fed barley straw and a concentrate formulated either with palm oil (CTRL group) or with linseed (+LS group), both alone or supplemented with quercetin (+QCT group or +LS+QCT group) were used to assess the effects of these dietary supplements on meat quality attributes. After being slaughtered, the longissimus thoracis muscles were used to study the fatty acid (FA) profile in detail, whilst longissimus lumborum slices were stored under refrigerated conditions to determine the lipid stability. Linseed increased the content of highly unsaturated n-3 long-chain fatty acid (20:5n-3; 22:5n-3; 22:6n-3). Interestingly, a significant increment of rumenic acid content (9c,11t-18:2) was observed when this seed was administered together with dietary quercetin. Moreover, the feeding of quercetin resulted in a reduction in the proportion of saturated FA and a decrease in lipid peroxidation of meat when the lambs were fed linseed. In conclusion, from both a nutritional and a commercial (shelf-life) point of view, it may be useful to include a source of quercetin when lambs are fed linseed diets.

The effect of quercetin dietary supplementation on meat oxidation processes and texture of fattening lambs.

Thirty two lambs were fed a total mixed ration (TMR) formulated either with palm oil (CTRL; 34 g palm oil kg(-1) TMR) or whole flaxseed (+FS, 85 g flaxseed kg(-1) TMR) alone or enriched with quercetin (+QCT, 34 g palm oil plus 2 g quercetin kg(-1) TMR; +FS+QCT, 85 g flaxseed plus 2 g quercetin kg(-1) TMR). Dietary flaxseed did not affect, in a significant manner, the lipid peroxidation of meat samples. Quercetin treatment reduced oxysterol content (P<0.05) after 7 days of refrigerated storage of fresh meat, but did not affect significantly (P>0.05) the level of lipid-derived volatiles in the headspace of the light-exposed stored cooked meat. Sensory evaluation showed flaxseed as being responsible for a negative effect on meat flavour, probably associated with a modification of the fatty acid profile whereas, unexpectedly, quercetin seemed to worsen meat tenderisation.

Effects of dietary inclusion of sunflower soap stocks on nutrient digestibility, growth performance, and ruminal and blood metabolites of light fattening lambs.

Forty-four merino lambs (6 to 8 wk old; BW 15.6 ± 0.21 kg) were used to study the effect of adding different proportions of sunflower soap stock (SS) to pelleted total mixed ration (TMR) for fattening lambs on feed intake, animal growth and dressing percentage, ruminal fluid characteristics, and blood acid-base parameters. Lambs were assigned to 1 of 4 experimental groups (11 lambs per group), each randomly assigned to 1 dietary treatment: 00SS (0 g SS/kg TMR pellet), 15SS (15 g SS/kg TMR pellet), 30SS (30 g SS/kg TMR pellet), and 60SS (60 g SS/kg TMR pellet). Lambs were individually fed the corresponding diet ad libitum. On d 19 to 23, total feces were collected and sampled from 4 lambs per group. When lambs reached 27 kg BW, they were slaughtered. Dry matter intake, ADG, and length of fattening period were not affected by the dietary treatment (P > 0.10). Animals in the 30SS group tended to show the best G:F values (P < 0.10). Dressing percentage tended to linearly decrease as SS increased (P < 0.10). Animals in the 60SS groups showed the lowest DM and fiber digestibility values (P < 0.05). Total VFA contents in ruminal fluid were not affected (P > 0.10), but the propionate proportion linearly increased (P < 0.05), and the acetate to propionate ratio tended to decrease with SS supplementation. Increasing dietary content of SS linearly decreased the cold carcass weight (P < 0.05) but did not affect other measured carcass characteristics. Ruminal fluid pH tended to decrease (P < 0.10) and mucosa color decreased as SS increased in the TMR (P < 0.05). Blood pH and Na concentration increased (P < 0.05), whereas the anion gap, CO2 pressure, and K concentration linearly decreased (P < 0.05) as SS increased. Including more than 30 g SS/kg TMR in the diet for fattening lambs reduces DM and fiber digestibility without affecting feed intake and ADG. The acidotic rumen conditions that induced a darkening of rumen mucosa were counteracted by blood acid-base parameters. The optimum level of inclusion seems to be 30 g SS/kg TMR.

Effect of sunflower oil supplementation and milking frequency reduction on sheep milk production and composition.

The aim of this study was to investigate the effects of milking frequency reduction and dietary lipid supplementation on intake, BW, and milk yield and composition in high yielding dairy ewes. Ten lactating Assaf ewes were allocated into 2 experimental groups (n=5). Ewes were fed alfalfa hay ad libitum and 34 g·kg(-1) of BW of a concentrate feed with either 0 (Control group) or 43 g of sunflower oil·kg(-1) of DM (SO group). The experiment lasted 63 d and consisted of 3 periods. During Period 1 (from d 1 to 21), ewes were milked twice a day. During Period 2 (from d 22 to 49), ewes were unilaterally milked, so that each gland of each ewe was milked either once or twice daily. During Period 3 (from d 50 to the end of the experiment), both udder halves were again milked twice daily. Intake, BW, and milk composition were controlled weekly and milk production from each half udder was recorded twice a week. Total DM intake, BW, and milk yield in Period 1 were not significantly (P>0.10) affected by dietary treatments. Milk yield tended to be increased in the ewes fed the SO diet in periods 2 (P=0.093) and 3 (P=0.067). Oil supplementation (SO diet) significantly (P<0.05) decreased milk protein and total solids concentrations in the 3 experimental periods and fat content in Period 3, and tended (P=0.077) to decline fat content in Period 2. Lactose content and somatic cell count (SCC) were unaffected (P>0.10) by dietary lipid supplementation in any of the experimental periods. There were no significant (P>0.10) differences between half udders in milk yield and composition in Period 1, and in SCC in any of the experimental periods. Fat and total solids contents were unaffected (P>0.10) by reducing milking frequency. Nevertheless, milk protein content was increased (P<0.001) when glands were milked only once daily whereas milk yield and lactose content were decreased (P=0.001). The interaction between gland and diet was significant for lactose in Period 2, suggesting a greater effect of milking frequency reduction on tight junctions in Control ewes. In fact, the ratio between glands for milk yield was significantly (P<0.05) greater in SO (0.82) than in Control (0.72) ewes. In Period 3, this ratio increased but it was still lower in Control ewes (0.92 vs. 0.78, P<0.05). Thus, milking frequency reduction and SO supplementation seem to have counteracting effects on milk production and composition. Our results suggest that SO-supplemented ewes have a better capacity of adaptation to changes in milking frequency, probably due to processes induced in the mammary gland.

Quercetin dietary supplementation of fattening lambs at 0.2% rate reduces discolouration and microbial growth in meat during refrigerated storage.

Thirty-two Merino lambs fed barley straw and a concentrate formulated either with palm oil (CTRL group) plus quercetin (QCT group) or flaxseed (FS group) plus quercetin (FS-QCT group) were used to assess the effects of this flavonoid on meat quality attributes. The animals were slaughtered after being fed for at least 5 weeks with the experimental diets. Chemical composition of longissimus thoracis (LT) muscle was not different among treatments. The longissimus lumborum (LL) samples of QCT and FS-QCT groups revealed lower discolouration (hue angle) when compared to the CTRL and FS lambs (P<0.05), whereas extract release volume (ERV) and microbiological data jointly suggest that flaxseed and quercetin may reduce the growth of microbial populations responsible for meat spoilage in quadriceps femoris (QF).

Electrolyzed water as novel technology to improve hygiene of drinking water for dairy ewes.

Tap water alone (TW) or treated with 3% of slightly acidic electrolyzed water (SAEW) were used in this experiment to study its effect on water quality, blood biochemical parameters and milk yield and composition. Each type of water was supplied to one group of 10 milking ewes for 25 days. Weekly water samples from troughs were taken. On days 1, 12 and 25, milk yield was measured, and milk and blood samples were taken. SAEW reduced (P < 0.05) bacterial counts (aerobic mesophilic, total coliform and streptococcus). Blood gases, biochemical parameters and milk yield and its composition were not affected (P > 0.05). SAEW can be used at 3% rate as a powerful and economic agent for sanitizing drinking water for dairy ewes with no effects on animal performance.

The liver antioxidant status of fattening lambs is improved by naringin dietary supplementation at 0.15% rates but not meat quality.

Twenty Assaf lambs fed barley straw plus a concentrate alone (CONTROL group) or enriched with naringin (1.5 g/kg DM, NARINGIN group) were used to assess the effect of this polyphenolic compound on meat quality attributes. Serum samples were collected for 7 weeks, then the animals were slaughtered and the livers and longissimus thoracis et lumborum muscles extracted for analysis. Triacylglycerol levels in the serum samples tended to show (P = 0.087) lower average values for the NARINGIN group when compared with the CONTROL, but no differences were observed when the meat was analysed for the intramuscular fat content. Lower thiobarbituric acid-reactive substances procedure (TBARS) values (P < 0.001) in the liver of the NARINGIN group were detected, probably as a consequence of naringenin accumulation in this organ. No significant differences were observed in the meat samples concerning TBARS or colour evolution during refrigerated storage, as not enough naringenin would have reached the muscle. Independent of naringin administration, the low levels of the most atherogenic oxysterols must be highlighted as the most important quality score in the lamb meat samples studied.

Carnosic acid dietary supplementation at 0.12% rates slows down meat discoloration in gluteus medius of fattening lambs.

Thirty-two Merino lambs fed barley straw and a concentrate alone (CONTROL) or enriched with vitamin E (VITE006) or carnosic acid (CARN006; CARN012) were used to assess the effect of these antioxidant compounds on meat quality attributes. The animals were slaughtered after being fed for at least 5weeks with the experimental diets. The longissimus lumborum samples of VITE006, CARN006 and CARN012 groups showed higher values (P<0.001) of L* (lightness) through the complete storage period under modified atmosphere when compared to the CONTROL group. Moreover, the VITE006 and CARN012 samples revealed lower discoloration when compared to the CONTROL group, these differences being more apparent in a less color stable muscle such as gluteus medius (P<0.05 for hue after 14days of refrigerated storage). Meat sensory traits were not significantly affected by carnosic acid and microbiological analyses were not conclusive at the doses administered.

Feeding vegetable oils to lactating ewes modifies the fatty acid profile of suckling lambs.

The objective of this study was to evaluate the effects of vegetable oil supplementation of ewe diets on the performance and fatty acid (FA) composition of their suckling lambs. Forty-eight pregnant Churra ewes (mean BW 64.3±0.92 kg) with their 72 newborn lambs (prolificacy=1.5) were assigned to one of four experimental diets, supplemented with 3% of hydrogenated palm (PALM), olive (OLI), soya (SOY) or linseed (LIN) oil. Lambs were nourished exclusively by suckling from their respective mothers. Ewes were milked once daily, and milk samples were taken once a week. When lambs reached 11 kg, they were slaughtered and samples were taken from musculus longissimus dorsi (intramuscular fat) and subcutaneous fat tissue. No changes were observed in milk yield, proximal composition or lamb performance (P>0.10). Milk and lamb subcutaneous and intramuscular fat samples from the PALM diet had the highest saturated fatty acid concentration, whereas those of the OLI, SOY and LIN diets had the lowest (P<0.05). The greatest monounsaturated fatty acid concentration was observed in milk from ewes fed OLI, and the least in milk and in lamb subcutaneous and intramuscular fat samples from LIN and PALM diets. Milk and lamb fat from ewes fed PALM displayed the highest 16:0 proportion and the lowest 18:0 (P<0.05). There were higher concentrations of cis-9 18:1 in OLI samples (P<0.05), more 18:2n-6 in SOY lambs and milk fat (P<0.001) and the highest levels of 18:3n-3 and 20:5n-3 in LIN samples (P<0.01). Milk and lamb subcutaneous and intramuscular samples from SOY and LIN diets contained the most cis-9, trans-11 conjugated linoleic acid, whereas PALM samples had the least (P<0.01). Sheep diet supplementation with different oils, constituting up to 3% of their diets, resulted in changes in the FA composition of milk and the subcutaneous and intramuscular fat of suckling lambs, but did not affect either milk production or lamb performance.

Naringin and vitamin E influence the oxidative stability and lipid profile of plasma in lambs fed fish oil.

Thirty two Merino lambs (15 weeks old) fed barley straw and fish oil enriched concentrate were used to assess the effect of vitamin E (6 g kg(-1) DM) and naringin (1.5-3 g kg(-1) DM) on plasma lipid peroxidation (TBARS), total antioxidant status (TAS), immune response, plasma cholesterol, and triglycerides. After 21 days feeding the experimental diets, lambs were subjected to a 4 h transportation stress period and then held 4 more hours without feed. TBARS values before stress were lower for animals consuming vitamine E and naringin when compared to control lambs (P<0.05). However, after stress all groups presented similar levels of TBARS. TAS decreased (P<0.05) in all groups in response to stress with values recovering (P<0.05) to pre-stress values following 4 h of rest. A rise (P<0.05) in serum concentrations of triacylglycerol following 21 d of fish oil supplementation was dampened in lambs consuming vitamin E or naringin. Both pre-stress TBARS and triacylglycerol-reducing effects of naringin added to fish oil enriched concentrate for fattening lambs are reported.

Vinasse added to the concentrate for fattening lambs: intake, animal performance, and carcass and meat characteristics.

Twenty-four Merino lambs (mean BW 15.4 ± 0.13 kg, 6 to 7 wk old) were used to study the effects of the addition of 0 (control), 100 (V10), and 200 (V20) g of vinasse per kilgram of concentrate on intake, animal performance, biochemical blood profile, and carcass and meat characteristics. Lambs were assigned to 1 of 3 experimental diets and fed barley straw and the corresponding concentrate ad libitum. When the animals reached 25 kg of BW, a sample of blood was taken and the lambs were slaughtered. Feed intake, growth rate, biochemical blood profile, and carcass and meat characteristics were assessed. Lambs that received the concentrates with vinasse showed a reduced concentrate intake (linear contrast, P = 0.029) and ADG (linear contrast, P = 0.004) and an increased length of fattening period (linear contrast, P = 0.002) as well as feed:gain ratio (linear contrast P = 0.011). Vinasse enhanced ruminal pH (orthogonal contrast control vs. V10 + V20; P = 0.007). Plasma glucose concentrations declined in lambs fed vinasse (linear contrast, P = 0.003), whereas plasma urea concentration increased in animals fed vinasse (linear contrast, P = 0.036). The plasma concentrations of creatinine, triglycerides, and lactate and the enzyme profile studied (alkaline phosphate, alanine transaminase, glutamate oxal-acetate transaminase, γ-glutamyl transpeptidase, and lactate dehydrogenase) were not modified in response to vinasse inclusion. Lambs in the vinasse groups had less Na(+) and nitrate and greater K(+) and nitrite plasma concentrations (linear contrasts, P < 0.05). None of the carcass characteristics studied was affected by vinasse (P > 0.10). Meat chemical composition and characteristics were unaffected (P > 0.10), but shear force was greater for lambs that received vinasse (orthogonal contrast, control vs. V10 + V20, P = 0.007). The addition of 100 or 200 g vinasse/kg of concentrate for fattening lambs reduced feed intake and growth rate and increased the feed:gain ratio and meat toughness without affecting any other carcass and meat characteristics.