Isotrichid protozoa influence conversion of glucose to glycogen and other microbial products

The goal of this in vitro study was to determine the influence of isotrichid protozoa (IP) on the conversion of glucose (Glc) to glycogen (Glyc) and transformation of Glc into fermentation products. Treatments were ruminal inoculum mechanically processed (blended) to destroy IP (B+, verified microscopically) or not mechanically processed (B-). Accumulated microbial Glyc was measured at 3 h of fermentation with (L+; protozoa + bacteria) or without (L- predominantly protozoa) lysis of bacterial cells in the fermentation solids with 0.2 N NaOH. Two 3-h in vitro fermentations were performed using Goering-Van Soest medium in batch culture vessels supplemented with 78.75 mg of Glc/vessel in a 26.5-mL liquid volume. Rumen inoculum from 2 cannulated cows was filtered through cheesecloth, combined, and maintained under CO₂ for all procedures. At 3 h, 0.63 and 0.38 mg of Glc remained in B- and B+. Net microbial Glyc accumulation (and Glc in Glyc as % of added Glc) detected at 3 h of fermentation were 3.32 (4.69%), −1.42 (−2.01%), 6.45 (9.10%), and 3.65 (5.15%) mg for B-L-, B+L-, B-L+ and B+L+, respectively. Treatments B+ and L+ gave lower Glyc values than B- and L-, respectively. Treatment B+L- demonstrated net utilization of α-glucan contributed by inoculum with no net Glyc production. With destruction of IP, total Glyc accumulation declined by 44%, but estimated bacterial Glyc increased. Microbial accumulation of N increased 17.7% and calculated CH₄ production decreased 24.7% in B+ compared with B-, but accumulation of C in microbes, production of organic acids or C in organic acids, calculated CO₂, and carbohydrates in cell-free medium did not differ between B+ and B-. Given the short 3-h timeframe, increased N accumulation in B+ was attributed to decreased Glyc sequestration by IP rather than decreased predation on bacteria. After correction for estimates of C from AA and peptides utilized by microbes, 15% of substrate Glc C could not be accounted for in measured products in B+ or B-. Approximately 30% of substrate Glc was consumed by energetic costs associated with Glc transport and Glyc synthesis. The substantial accumulation of Glyc and changes in microbial N and Glyc accumulation related to presence of IP suggest that these factors should be considered in predicting profiles and amounts of microbial products and yield of nutrients to the cow as related to utilization of glucose. Determination of applicability of these findings to other soluble carbohydrates could be useful.     

Influence of carbohydrate source on ruminal fermentation characteristics, performance, and microbial protein synthesis in dairy cows

Eight multiparous Holstein cows (676 ± 57 kg of body weight; 121 ± 17 d-in-milk) were used in a replicated 4 × 4 Latin square design to determine the effects of 4 sources of carbohydrate on milk yield and composition, ruminal fermentation, and microbial N flow to the duodenum. Four cows in one of the Latin squares were fitted with permanent ruminal cannulae. Diets contained (DM basis) 50% forage in combinations of alfalfa hay and barley silage, and 50% concentrate. The concentrate portion of the diets contained barley, corn, wheat, or oats grain as the primary source of carbohydrate. Intake of DM ranged from 24.0 to 26.2 kg/d, and it tended to be lower in cows fed the wheat-based diet compared with those fed the barley-based diet; consequently, milk yield tended to be lower in cows fed the wheat-based diet compared with those fed the barley-based diet. Cows fed the barley- or wheat-based diets had a lower milk fat content compared with those fed the corn-based diet. Ruminal fermentation characteristics were largely unaffected by the source of dietary carbohydrate, with similar ruminal pH and volatile fatty acid and ammonia concentrations for the first 6 h after the morning feeding. Dietary treatment did not affect total tract apparent digestibility of DM, organic matter, and neutral detergent fiber; however, total tract apparent digestibility of starch in cows fed the oats-based diet was higher compared with those fed the corn-and wheat-based diets. Nitrogen that was used for productive purposes (i.e., N secreted in milk + N apparently retained by the cow) tended to be lower in cows fed the wheat-based diet compared with cows fed the barley-, corn-, or oats-based diets. Urinary purine derivative (PD) excretion was similar in cows fed the barley-, corn-, and wheat-based diets; however, purine derivative excretion was higher in cows fed the barley-based diet compared with those fed the oats-based diet. Consequently, estimated microbial N flow to the duodenum was 49 g/d higher in cows fed the barley-based diet compared with those fed the oats-based diet. Improved production performance with corn and barley diets appeared to be due to greater nutrient absorption than in cows fed oats and wheat diets, rather than improved nutrient utilization efficiency.     

Short communication: Amino acids antagonistic to the amino acids inhibitory for growth rate of mixed ruminal bacteria

Antagonism of some amino acids (AA) to the inhibitory effects of other AA (Ile, Phe, and Thr) on the growth rate of mixed ruminal bacteria was investigated. In vitro growth rate of the mixed ruminal bacteria was inhibited when the 3 inhibitory AA (1 mM each) were each added to individual control treatments in which an ammonium salt was included as a sole N source. The inhibitory effect caused by Ile was relieved by addition of Leu or Val (equimolar to Ile), and no significant inhibition was shown when both Leu and Val were added together with Ile. The growth inhibition caused by Phe was also alleviated by supplementing with Trp, and was completely negated by adding Tyr. The inhibitory effect of Thr, on the other hand, was not affected by addition of Lys or Met (which are synthesized using a common pathway with Thr), but was mitigated by supplementation with Glu, Ser, Val, Ala, or Gln. Among the antagonistic AA, Leu, Val, Trp, Tyr, and Glu were indispensable for the maximum growth rate of the ruminal bacteria under the experimental condition of supplementation of amino-N, the removal of which from a mixture of 20 protein AA caused the growth rate to decline. Removals of Ile along with Leu or Val or both, of Phe along with Trp or Tyr, and of Thr along with Glu recovered the promotion of bacterial growth rate. It was concluded that inhibitions of the bacterial growth rate caused by Ile, Phe, or Thr could be antagonized by some other AA (Leu, Val, Tyr, Trp, or Glu), and the role of these latter AA as relievers of the inhibitory effects could explain why they are indispensable for maximum growth rate of ruminal bacteria.     

Short communication: Effect of subacute ruminal acidosis on in situ fiber digestion in lactating dairy cows

Subacute ruminal acidosis (SARA) was induced by replacing 25% of the total mixed ration intake [dry matter (DM) basis] with pellets consisting of 50% wheat and 50% barley. This reduced dietary forage content (DM basis) from 39.7 to 29.8% and increased the dietary concentrate content from 60.3 to 70.2%. Induction of SARA reduced the 24- and 48-h in situ neutral detergent fiber (NDF) degradabilities of grass hay numerically from 31.5% to 24.6% (P = 0.29) and from 51.3% to 36.9% (P < 0.05), respectively. The 24- and 48-h in situ NDF degradabilities of legume hay were reduced from 35.3 to 26.3% (P < 0.05) and from 49.0 to 35.8% (P < 0.05), respectively. The 24- and 48-h in situ NDF degradabilities of corn silage were reduced from 44.0 to 37.2% (P < 0.05) and from 56.1 to 44.8% (P < 0.05), respectively. This study suggests that induction of SARA by excess feeding of wheat/barley pellets reduces the rumen digestion of NDF from grass hay, legume hay, and corn silage.     

Effect of forage:concentrate ratio on fatty acid composition of rumen bacteria isolated from ruminal and duodenal digesta

Four dairy cows were used to examine the effect of the dietary forage:concentrate ratio [35:65, 50:50, 65:35, and 80:20 on a dry matter (DM) basis] on the fatty acid composition of rumen bacteria isolated from the liquid (LAB) and solid (SAB) phase of the rumen and duodenal digesta. Rumen contents were sampled 4 h after the morning feeding. Solid and liquid phases were separated from rumen contents and duodenal bacteria from a composite duodenal sample by differential centrifugation. Total fatty acid content in bacterial DM was 1.6 to 2.8 times higher in SAB compared with LAB, and increased with dietary concentrate. In combination with published reports, the data show that bacterial fatty acid content and composition is closely related to dietary fatty acids except for C18:2n-6 and C18:3n-3. A decrease in forage:concentrate ratio increased bacterial concentration of trans-10 C18:1, and this increase was 3.4 times higher in LAB compared with SAB. Analysis of odd- and branched-chain fatty acids showed large differences between SAB and LAB, which probably reflected a difference in species composition. The variation in odd- and branched-chain fatty acids between SAB and LAB was used to estimate their relative proportions in duodenal bacteria by means of linear programming, and showed an increased proportion of SAB from 64.7 to 74.8% with increasing forage:concentrate ratio. In addition, increasing the proportion of dietary forage was closely related to the proportion of anteiso C15:0 in total odd- and branched-chain fatty acids (r_pearson = −0.771). The bacterial concentration of iso C17:0 closely reflected the bacterial growth rate as shown by the relation with cytosine:N (r_pearson = −0.729). These strong relationships suggest that odd- and branched-chain fatty acids might be used as tool to evaluate nutrient supply to rumen bacteria.     

Short communication: Characterization of enterotoxin-producing Staphylococcus aureus isolated from mastitic cows

Staphylococcus aureus is not only a common cause of bovine mastitis, but also an agent of food poisoning in humans. In an attempt to determine whether staphylococci causing bovine mastitis could also cause food poisoning, 60 isolates of presumed S. aureus were isolated in the period between March and August 2017 from 3,384 routine, composite, quarter milk samples of individual cows raised on 12 dairy farms in central Italy. Seventeen out of 60 isolates were confirmed as S. aureus after coagulase, thermonuclease, and biochemical tests. These isolates were analyzed by PCR for the presence of the nuc, sea, seb, sec, sed, and see genes. The positive isolates were nuc, 100% (17); sea, 35.29% (6); seb, 5.88% (1); sec, 5.88% (1); sed, 29.41% (5); and see, 47.06% (8). The isolates were also tested with 2 enzyme immunoassay diagnostic kits, one for the screening detection of the production of staphylococcal enterotoxins (SEA, SEB, SEC, SED, SEE) and one for the detection of specific enterotoxin produced by each isolate. Seven out of 17 (41.18%) were enterotoxin producers: 7 produced SEA (41.18%), 1 SEB (5.88%), 1 SEC (5.88%), 5 SED (29.41%), and 6 SEE (35.29%). To further characterize the isolates, they were analyzed by the Kirby Bauer test for susceptibility to 13 antimicrobials (ampicillin, ciprofloxacin, kanamycin, tetracycline, gentamicin, methicillin, nalidixic acid, erythromycin, amoxicillin/clavulanic acid, streptomycin, vancomycin, neomycin, and enrofloxacin), and we detected resistance to ampicillin (52.94%), nalidixic acid (70.59%), erythromycin (5.88%), and amoxicillin/clavulanic acid (17.65%). The isolates were sensitive to the main classes of antimicrobials used for the treatment of bovine subclinical mastitis. The presence of enterotoxin-producing isolates of S. aureus in bovine milk means that a temperature abuse or a breakdown in the thermal treatment of the milk could present a food safety risk, particularly if all enterotoxigenic isolates could potentially produce SEA in milk.     

Short communication: effectiveness of sample duplication to control error in ruminant digestion studies

Eight ruminally cannulated lactating dairy cows from a study on the effect of dietary rumen-degraded protein on production and digestion of nutrients were used to assess using sample duplication to control day-to-day variation within animals and errors associated with sampling and laboratory analyses. Two consecutive pooled omasal samples, each representing a feeding cycle, were obtained from each cow in each period. The effectiveness of sample duplication in error control was tested by comparing the variance of the difference in treatment means when taking 2 samples from each cow in each period to the variance when taking only one sample. Compared with no duplication, sample duplication improved precision by reducing variance by 50, 40, 31, 23, 23, and 9% for, respectively, rumen-undegraded protein flows, ruminal neutral detergent fiber digestibility, microbial nonammonia N flow, microbial efficiency, organic matter flow, and organic matter truly digested in the rumen. For these same variables, reductions in the standard errors of the difference between treatment means due to sample duplication represented 100, 87, 73, 59, 58, and 27% of the predicted reductions resulting from doubling the number of experimental units without sample duplication. Sample duplication can substantially reduce experimental error originating from day-to-day variation within cows, sample collection, and laboratory analyses, thus improving statistical power in ruminant digestion studies.     

Short communication: Variability in fermentation end-products and methanogen communities in different rumen sites of dairy cows

The objective of this study was to investigate differences in fermentation and methanogen communities in samples collected from 3 sites in the rumen of dairy cows. The study involved 3 ruminally cannulated nonlactating Chinese Holstein dairy cows fed a diet of 40% forage and 60% concentrate feeds. Four handfuls of whole ruminal contents were collected from the cranial sac, middle of the ventral sac, and caudodorsal blind sac of the rumen of the cows at 0, 2.5, and 6 h after the morning feeding. Concentrations of VFA, ammonia, and dissolved methane and hydrogen were analyzed. Methanogen populations and communities were analyzed targeting 16s rRNA genes. Dissolved methane concentration and pH were highest in samples from the cranial rumen. Ruminal fluid from the cranial rumen also had greater copy numbers of the Methanobrevibacter and higher Simpson indexes compared with samples from middle of the ventral rumen. In summary, cranial rumen had higher dissolved CH₄ concentration than middle and hind rumen, which might be caused by the greater population of Methanobrevibacter with higher ruminal pH.     

The effects of starch and rapidly degradable dry matter from concentrate on ruminal digestion in dairy cows fed corn silage-based diets with fixed forage proportion

This study investigated the effects of the type (starch vs. nonstarch) and rate of ruminal degradation of carbohydrates from the concentrate on digestion in dairy cows fed corn silage-based diets. Six ruminally cannulated cows were assigned to 6 treatments in a 6 × 6 Latin square design. Treatments were arranged in a 3 × 2 factorial design. Two starch levels [25 and 41% dry matter (DM) for low starch (LS) and high starch (HS) diets, respectively] were obtained by replacing starch-rich feedstuffs by nonstarch feedstuffs. These starch levels were combined with slowly, moderately, and rapidly rumen-degradable feedstuffs to obtain 3 levels of rapidly degradable carbohydrates from concentrate (18, 23, and 28% DM). These levels were estimated from the DM disappearance of concentrate after 4 h of in sacco incubation (CRDM). Wheat and corn grain were used as rapidly degradable and slowly degradable starch feedstuffs, respectively. Soybean hulls and citrus pulp were used as slowly degradable and rapidly degradable nonstarch feedstuffs, respectively. No interaction effect was found between dietary starch content and CRDM on pH range, volatile fatty acid (VFA) range, or VFA profile. Increasing CRDM led to a linear decrease in acetate-to-propionate ratio (from 2.7 to 2.1), and a linear increase in the pH and VFA ranges (from 0.86 to 1.12 pH units and from 34 to 56 mM, respectively). Feeding HS diets decreased acetate-to-propionate ratio (2.6 vs. 2.0) and increased pH range (0.89 vs. 1.04 pH units), but had no effect on VFA range. Increasing CRDM linearly decreased mean ruminal pH in LS diets but linearly increased mean ruminal pH in HS diets. Fibrolytic activity was unaffected in LS diets but decreased strongly in HS diets (from 62 to 50%). These findings suggest that pH regulation differs on a short-term and on a longer-term basis. In the short-term, increasing CRDM increased the rate of VFA production, which may have been partly buffered under LS diets due to the higher cation exchange capacity of nonstarch feedstuffs compared with starch-rich feedstuffs. In the longer term, feeding starch reduced fibrolytic activity, which may have led to lower total VFA production and higher mean pH. The results of this experiment clearly show that both fermentative characteristics of the concentrate and dietary starch content should be taken into account when formulating diets to prevent subacute ruminal acidosis and to predict VFA profile.     

Short communication: effect of carbohydrate fermentation rate on estimates of mass fermented and milk response

Current prediction equations were used to evaluate the effects of rates of fermentation of fiber or starch in individual feeds on amounts of carbohydrate fermented ruminally and milk yield responses. The small predicted increases in carbohydrate fermented and milk response associated with doubling the rates of fermentation suggest that current prediction equations are relatively insensitive to changes in rate of fermentation.     

Disappearance of cell wall monomeric components from fractions chemically isolated from alfalfa leaves and stems following in-situ ruminal digestion

Alfalfa (Medicago sativa L) leaves and stems were incubated in the rumen of an alfalfa-fed steer for 0, 12, 24 and 48 h. Residues were analysed for monosaccharides remaining in chemically isolated fractions. Pectic substances were rapidly and extensively degraded. Xyloglucans also were quite degradable, whereas glucuronoxylans were very resistant to fermentation. Cellulose disappearance was intermediate between that of xylan and xyloglucan. For all non-pectic polysaccharides, leaf material was more degradable than stem, and this difference between leaf and stem was greater for cellulose than for most hemicellulosic fractions. Acetyl groups associated with the oxalate-insoluble fractions were degraded similarly to xylans. Alkali-labile phenolic acid content was lower in stems than in leaves, but contributed only 4 g kg^?1 of leaf DM. Less than 10% of alkali-labile phenolics were associated with hemicelluloses. Structures which limit alfalfa xylan fermentation may include acetyl groups and linkages of uronic acid side chains to phenolic material.     

Short communication: Cold atmospheric plasma inactivation of Prototheca zopfii isolated from bovine milk

Mastitis is a serious bovine diseases that can be caused by Prototheca zopfii, yeast-like algae belonging to the family Chlorellaceae. The substantial economic losses and health damage associated with bovine mastitis emphasize the need to develop effective strategies aimed at control of the infection. Unfortunately, P. zopfii is highly resistant to most common antibacterial and antifungal agents, as well as to heat treatment. We report here the first attempt to use cold atmospheric plasma to inactivate this pathogen. We studied 20 strains of P. zopfii isolated from milk samples taken from cows with clinical or subclinical mastitis. The studies confirmed the high level of resistance of P. zopfii to typical antifungal agents, such as voriconazole, fluconazole, amphotericin B, caspofungin, anidulafungin, and micafungin. In contrast, each of the strains revealed high susceptibility to cold atmospheric plasma, >2-fold higher compared with a reference strain of Candida albicans. The obtained results are promising and open up a new approach in the fight against P. zopfii.     

Exclusion of dietary sodium bicarbonate from a wheat-based diet: effects on milk production and ruminal fermentation

Milk production, rumen fermentation, and whole-tract apparent nutrient digestibility in response to feeding 20% steam-rolled wheat with or without sodium bicarbonate were evaluated in 12 Holstein cows averaging 165 ± 16 DIM. Cows were fed 1 of 2 isoenergetic and isonitrogenous diets containing either 0 or 0.75% sodium bicarbonate on a DM basis for 21 d in a crossover design. Rumen fluid samples were obtained 18 times during the last 2 d of each period, and fecal samples were collected on 12 occasions from d 18 to 21 of each period. Removal of sodium bicarbonate from the diet did not affect DMI (21.0 kg/d), yields of milk (30.8 kg/d), or milk components (1.16, 1.01, and 1.40 kg/d for fat, protein, and lactose, respectively). Whole-tract apparent digestibility of DM, CP, ADF, and NDF did not differ between the 2 treatments (75.3, 76.6, 67.2, and 63.6%, respectively). The mean rumen pH was 6.24 and was not affected by excluding sodium bicarbonate from the diet. Rumen NH₃-N (12.31 mg/dL) and lactic acid (3.63 mM) concentrations were not different, whereas total volatile fatty acids concentration tended to increase when sodium bicarbonate was present in the diet (110 vs. 116 mM). However, average concentrations of the individual volatile fatty acids, as a proportion of total volatile fatty acids, were not affected by treatment. In conclusion, dairy cow diets can include up to 20% steam-rolled wheat without the need for added sodium bicarbonate as long as the diets are formulated to meet the fiber requirements of the cow.     

Short communication: Temperature sensibility of Prototheca blaschkeae strains isolated from bovine mastitic milk

Dairy cow mastitis associated with microalgae of the genus Prototheca has been reported worldwide. This alga is extremely resistant to most antimicrobials commonly used in mastitis therapy. In milk processing, different thermal treatments are generally efficient at inactivating and eliminating microorganisms. Until recently, no reports on Prototheca blaschkeae susceptibility to heat treatment have been described. Thus, considering the potential zoonotic risk that Prototheca may represent, the objective of this study was to test the susceptibility of P. blaschkeae field isolates retrieved from bovine mastitis to different temperature/time ratios that are generally used in the milk processing industry: 62°C/15 min and 30 min; 70°C/20 s, 15 min, and 30 min; 75°C/20 s; 90°C/1 s; and 100°C/1 s. The results showed a growth reduction of all isolates after the heat treatments, but only at 100°C was a total growth inhibition observed.     

湿热及酶解处理对糯米粉体外消化特性和血糖生成指数的影响

目的 研究酶解处理、湿热处理和湿热复合酶解处理对糯米粉体外消化特性和血糖生成指数(glycemic index, GI)的影响。方法 采用体外消化法测定了不同处理糯米粉的水解度和血糖生成指数值, 并通过X-射线衍射和激光共焦拉曼光谱实验测定了不同处理糯米的淀粉晶体结构。结果 3种处理的水解度和抗性淀粉(resistant starch, RS)相对含量增加, 慢消化淀粉(slowly digestible starch, SDS)相对含量降低; 其中湿热复合酶解处理条件下, RS相对含量最高, 为71.31%; 酶解处理条件下, RS相对含量最低, 为67.66%。酶解处理、湿热处理和湿热复合酶解处理的GI值分别为79.1、76.0和70.6。3种处理后糯米淀粉的晶体结构发生改变, 3种处理均使分子短程有序度与结晶度增加, 其中湿热复合酶解处理条件下, 结晶度最高, 为34.41%。 结论 湿热复合酶解处理后的糯米粉抗消化特性增强且GI值降低, 湿热复合酶解工艺可能通过影响糯米粉的体外消化率来降低GI值。     

Dry malt extract from barley partially replacing ground corn in diets of dairy cows: Nutrient digestibility,ruminal fermentation,and milk composition

Dry malt extract (DME) has been used in animal nutrition as an alternative source of rapidly fermentable carbohydrate. An experiment was conducted to evaluate the partial replacement of ground corn with DME in diets of dairy cows on apparent digestibility, ruminal fermentation, predicted rumen microbial protein supply, N excretion, serum urea-N concentration, and milk yield and composition. Twenty-eight Holstein cows (35.3 ± 5.88 kg/d milk yield and 148 ± 78 d in milk), 4 of which were rumen cannulated, were blocked according to the presence of rumen cannulas, parity, milk yield, and days in milk and enrolled into a crossover design experiment. Experimental periods lasted 21 d, of which the first 14 d were allowed for treatment adaptation and 7 d were used for data collection and sampling. Treatment sequences were composed of control (CON) or DME from barley (Liotécnica Tecnologia em Alimentos) replacing ground corn at 7.62% diet dry matter (~2 kg/d). Data were analyzed using the MIXED procedure of SAS (SAS Institute Inc.) modeling the fixed effects of treatment, period, and their interaction, in addition to the random effect of animal. Ruminal fermentation data were analyzed as repeated measures including time and its interaction with treatment in the previous model as fixed effects. Treatments did not affect nutrient intake or feed sorting. Dry malt extract increased apparent digestibility of CP. Feeding DME decreased ruminal pH and molar percentage of butyrate and increased molar percentage of acetate. No treatment effects were detected for predicted rumen microbial protein supply or N excretion. Cows fed DME had lower serum urea-N concentration than CON cows. Dry malt extract increased yields of actual milk, 3.5% fat-corrected milk, fat, and protein, and improved feed efficiency (fat-corrected milk ÷ dry matter intake). Cows fed DME had lower milk urea nitrogen content in comparison with CON cows. Dry malt extract can partially replace ground corn in the diet while improving milk yield and feed efficiency.     

The effects of forage proportion and rapidly degradable dry matter from concentrate on ruminal digestion in dairy cows fed corn silage-based diets with fixed neutral detergent fiber and starch contents

This study investigated the effects of the forage-to-concentrate (F:C) ratio and the rate of ruminal degradation of carbohydrates from the concentrate on digestion in dairy cows fed corn silage-based diets. Six cows with ruminal cannulas were assigned to 6 treatments in a 6 × 6 Latin square. Treatments were arranged in a 3 × 2 factorial design. Three proportions of neutral detergent fiber from forage [FNDF; 7.6, 13.2, and 18.9% of dry matter (DM)] were obtained by modifying F:C (20:80, 35:65, and 50:50). These F:C were combined with concentrates with either high or low content of rapidly degradable carbohydrates. The dietary content of rapidly degradable carbohydrates from the concentrate was estimated from the DM disappearance of concentrate after 4 h of in sacco incubation (CRDM). Thus, 2 proportions of CRDM were tested (20 and 30% of DM). Wheat and corn grain were used as rapidly and slowly degradable starch sources, respectively. Soybean hulls and citrus pulp were used as slowly and rapidly degradable fiber sources, respectively. Concentrate composition was adjusted to maintain dietary starch and neutral detergent fiber contents at 35.9 and 28.9% of DM, respectively. There was no effect of the interaction between F:C and CRDM on DM intake (DMI), ruminal fermentation, chewing activity, and fibrolytic activity. When F:C decreased, DMI increased, the mean ruminal pH linearly decreased, and the pH range linearly increased from 0.95 to 1.27 pH unit. At the same time, the acetate-to-propionate ratio decreased linearly. Decreasing F:C linearly decreased the average time spent chewing per kilogram of DMI from 35.2 to 19.5 min/kg of DMI and decreased ruminal liquid outflow from 11.6 to 9.2 L/kg of DMI, suggesting a decrease in the salivary flow. Increasing CRDM decreased DMI and increased the time during which pH was below 6.0 (3.1 vs. 4.8 h), the pH range (0.90 vs. 1.33), and the initial rate of pH drop. It also increased the volatile fatty acid range (35 vs. 59 mM), thus suggesting an increased rate of fermentation. It also decreased the acetate-to-propionate ratio (2.9 vs. 1.8). Increasing CRDM barely affected the average time spent chewing per kilogram of DMI and the ruminal liquid outflow. These results suggest that rumen pH is controlled by different mechanisms when F:C is decreased or when CRDM is increased. Consequently, FNDF is a good predictor of the chewing time, whereas CRDM is a good predictor of the pH range and volatile fatty acid profiles. Finally, considering both FNDF and CRDM improves the prediction of mean pH.     

Short communication: In vitro antimicrobial susceptibility of Prototheca wickerhamii and Prototheca zopfii isolated from bovine mastitis

Bovine mastitis caused by Prototheca spp. can assume high significance because of economic losses and the potential risk to public health. Studies on the susceptibility of Prototheca spp. to antimicrobials have demonstrated its high level of resistance. We report the susceptibility of bovine isolates of Prototheca wickerhamii and Prototheca zopfii to amphotericin B and nystatin, 2 antifungal agents commonly used in the control of protothecosis, and discuss the results. After subculture, minimum inhibitory concentrations of both antifungal drugs were determined using macrodilution and agar diffusion methods. The inoculum concentration was standardized by determination of colony-forming units per milliliter. Nystatin showed more efficacy than amphotericin B in inhibiting P. wickerhamii growth. In contrast, growth inhibition of P. zopfii was similar for both antifungal agents. This study demonstrates different in vitro susceptibility patterns of P. wickerhamii and P. zopfii, reinforcing the necessity for more investigation into drugs that can be used with clinical efficacy.     

Short communication: effect of exopolysaccharide isolated from "viili" on the adhesion of probiotics and pathogens to intestinal mucus

The strong ropy character of the Scandinavian fermented milk viili is conferred by the exopolysaccharides (EPS) produced by lactococcal strains. These biopolymers can be responsible for some health benefits. We have assessed the influence of the EPS fraction isolated from commercial viili on the adhesion of some probiotics and pathogens to human intestinal mucus. Concentrations of viili EPS greater than 0.1 mg/mL promoted a decrease in adherence of Bifidobacterium lactis Bb12 and Lactobacillus rhamnosus GG and this effect was dose-dependent. However, no modifications were detected on the adhesion levels of the pathogenic strains tested at a concentration of 1 mg/mL of EPS. Results obtained in the present work should be considered in the design of new probiotic products.     

Short communication: Effects of different selenium supplements on rumen fermentation and apparent nutrient and selenium digestibility of mid-lactation dairy cows

The aim of this study was to evaluate the dose-dependent effects of a hydroxy-analog of selenomethionine (HMSeBA) on rumen fermentation, apparent nutrient digestibility, and total selenium absorption in mid-lactation dairy cows, and to compare the effects with those of sodium selenite (SS). Fifty mid-lactation dairy cows with similar milk yields, days in milk, and parity were randomly assigned to 1 of 5 treatments according to a randomized complete block design. The cows were fed a basal diet containing 0.06 mg/kg dry matter (DM) of Se (control) or the same basal diet supplemented with SS, yielding 0.3 mg of Se/kg of DM (SS-0.3), or HMSeBA, yielding 0.1, 0.3, or 0.5 mg of Se/kg of DM (SO-0.1, SO-0.3, and SO-0.5, respectively), during the experimental period. The final content of Se in control, SS-0.3, SO-0.1, SO-0.3, and SO-0.5 was 0.06, 0.34, 0.15, 0.33, and 0.52 mg of Se/kg of DM. The experiment lasted for 10 wk, with a pretrial period of 2 wk. Supplementation with HMSeBA altered rumen fermentation by linearly increasing total volatile fatty acids and the molar proportions of propionate and butyrate but decreasing rumen pH, ammonia content, and the ratio of acetate to propionate. Compared with SS, HMSeBA enhanced the molar proportion of propionate in the rumen and the apparent digestibility of crude protein, neutral detergent fiber, acid detergent fiber, and selenium. We demonstrated that HMSeBA promoted rumen fermentation, apparent nutrient digestibility, and selenium absorption, implying that HMSeBA has a greater apparent absorption than SS.