Effects of selenium and vitamin E on performance, physiological response, and selenium balance in heat-stressed sheep.

Forty-two 7-mo-old Australian Merino wethers were used in a 50-d trial to investigate the effects of Se and vitamin E on the performance and physiological responses of heat-stressed sheep. Sheep were exposed to thermoneutral conditions (maximum = 24°C and minimum = 20°C) for 28 d followed by heat (maximum = 38°C and minimum = 28°C) for 22 d. Hot conditions were imposed between 0700 and 1800 h. Sheep were randomly allocated to diets containing 0.8 mg/kg Se (Sel-Plex), 150 mg/kg vitamin E, or 0.8 mg/kg Se and 150 mg/kg vitamin E for either the duration of the study (50 d) or from d 1 of the hot period until the end of the study. A control group that received no supplemental Se and vitamin E for the duration of the study was included. Feed intake was measured daily and sheep were weighed weekly. Blood samples were collected from all sheep before feeding on d 1, 21, and 49 for measurement of biochemical and enzymatic variables. The concentration of Se was determined in offered and refused feed, feces, urine, water, plasma, liver, and kidneys. Exposure to heat reduced ( < 0.05) DMI by 11.9%, ADG by 198 g, serum concentration of urea nitrogen and Se by 17.8%, and plasma total antioxidant status by 26.4%. During hot conditions, sheep receiving Se and vitamin E supplements for 50 d had reduced ( < 0.05) BW loss and elevated G:F compared to control sheep. Serum Se concentration and the plasma total antioxidant status were greatest in sheep receiving Se and vitamin E supplements for 50 d ( < 0.05). These results indicate that dietary supplementation with Se and vitamin E reduces the adverse effects of a high heat load. Additional studies are warranted to elucidate the mechanisms responsible for these effects.

Effect of various doses of injected selenium on performance and physiological responses of sheep to heat load.

The aim of the study was to determine the effects of various doses of injected Se on the physiological responses of sheep to heat load. Fifteen 9-mo-old Australian Merino wethers (mean BW = 27.2 ± 2.1 kg) were randomly assigned to 1 of 3 treatments: 0 (control), 0.5, and 5 mg of Se, which was administered as a subcutaneous sodium selenate injection (5 mg/mL Se) on d 1, 8, and 15 of exposure to heat stress. The animals were housed individually in an environmental chamber and exposed to high temperature from 0700 to 1800 h (maximum = 38°C; minimum = 24°C) and to thermoneutral temperature from 1800 to 0700 h (maximum = 24°C; minimum = 20°C) for 21 d. Rectal temperature (RT) and respiration rate (RR) were measured daily at 0800, 1200, and 1600 h. Feed intake was measured daily, and sheep were weighed on d 1, 8, 15, and 21. Blood samples were collected on d 1 and 21. The 5 mg Se treatment decreased RT by 0.3°C (P = 0.02) and BW loss by 4.5% (P < 0.05) and increased eosinophil count (P < 0.05). There were no differences (P > 0.05) between treatments in RR and DMI, serum concentrations of glucose, total protein, cholesterol, and NEFA or in blood hematology variables. The findings of this study have important implications for the sheep industry. Further studies are warranted to elucidate the dynamics of Se on productivity and health during hot conditions.

Physiological responses of Australian Merino wethers exposed to high heat load.

Twelve 9-mo-old Merino wethers (30.4 ± 3.2 kg of BW) were used in a crossover study to investigate the heat tolerance of Australian Merino sheep by testing their physiological responses to repeated heat loads that occurred during summer months. Wethers were randomly divided into 2 groups of 6 wethers each, housed individually in an environmental chamber, and subjected to 2 d of thermoneutral conditions (TNC) followed by either 7 d of TNC (maximum temperature of 24°C, minimum temperature of 16°C) or 7 d of hot conditions (maximum temperature of 38°C, minimum temperature of 28°C), and then 2 d of TNC. These treatments were applied in 2 replicates, with each replicate in a separate environmental chamber. Rectal temperature (RT) and respiration rate were measured daily at 0600, 0800, 1000, 1200, 1400, 1600, and 1800 h. Feed and water intakes were measured daily, and wethers were weighed on d 1 and 11. Blood samples were collected from each whether on d 2 and 6, and serum was assayed for concentrations of creatine, glucose, total protein, cholesterol, NEFA, calcium, sodium, and potassium. Exposure to a high ambient temperature resulted in an 0.8°C increase in RT (P < 0.001), an increase in respiration rate (P < 0.001) by 66 breaths/min, and a 2.7 L/d increase in water intake (P < 0.0001). Feed intake decreased by 22% (P < 0.0001), BW decreased by 5.2% (P < 0.03), and creatine concentration was reduced (P < 0.05). No differences (P > 0.05) between treatments were observed for any of the remaining serum variables. These results indicate that Australian Merino sheep were able to maintain RT within the normal range during exposure to a prolonged increase in heat and that they recovered quickly from the negative effect of heat stress within 2 d of conditions returning to TNC. It would appear that they have a high heat tolerance, and further studies are needed to examine the effects of a greater heat load to determine the temperature-humidity index thresholds for Australian Merino sheep.

In vitro effects of hydrochloric and lactic acids on bioelectric properties of equine gastric squamous mucosa.

REASONS FOR PERFORMING STUDY: Volatile fatty acids, byproducts of carbohydrate fermentation by resident bacteria, have been implicated in causing nonglandular (NG) gastric ulcers. Lactic acid (LA), also produced by stomach bacteria, may cause gastric ulcers when exposed to the equine NG mucosa. OBJECTIVES: To investigate the in vitro effects of LA on equine NG mucosa bioelectric properties, sodium transport and tissue resistance. METHODS: Gastric tissues obtained from 13 mature horses were studied in Ussing chambers. Short-circuit current (Isc) and potential difference (PD) were measured, and electrical resistance (R) and conductance (G) calculated for tissues after addition of HCl and LA (5, 10, 20 and 40 mmol/l) in normal Ringer's solution (NRS). RESULTS: Mucosa exposed to HCl or LA (5, 10 and 20 mmol/l) in NRS (pH 1.5 and to a lesser extent pH 4.0) had a significant decrease in Isc and PD. Mucosa exposed to a high concentration of LA (40 mmol/l) in NRS (LRS) at pH 1.5 showed an increased G, but this increase was not significant. Values returned to baseline after solutions were returned to pH 7.0. Histological changes were consistent with HCl-induced (pH <4.0) acid damage. CONCLUSIONS: HCl induced alteration in bioelectric properties of equine NG mucosa whereas addition of LRS did not, other than those changes seen with HCl alone.

The characterization of lactic acid producing bacteria from the rumen of dairy cattle grazing on improved pasture supplemented with wheat and barley grain.

AIMS: To identify and characterize the major lactic acid bacteria in the rumen of dairy cattle grazing improved pasture of rye grass and white clover and receiving a maize silage and grain supplement with and without virginiamycin. METHODS AND RESULTS: Eighty-five bacterial isolates were obtained from the rumen of 16 Holstein-Friesian dairy cows. The isolates were initially grouped on the basis of their Gram morphology and by restriction fragment length polymorphism analysis of the PCR amplified 16S rDNA. A more definitive analysis was undertaken by comparing the 16S rDNA sequences. Many of the isolates were closely related to other previously characterized rumen bacteria, including Streptococcus bovis, Lactobacillus vitulinus, Butyrivibrio fibrisolvens, Prevotella bryantii and Selenomonas ruminantium. The in vitro production of L- and/or D-lactate was seen with all but five of the isolates examined, many of which were also resistant to virginiamycin. CONCLUSION: Supplementation of grain with virginiamycin may reduce the risk of acidosis but does not prevent its occurrence in dairy cattle grazing improved pasture. SIGNIFICANCE AND IMPACT OF THE STUDY: This study shows that lactic acid production is caused, not only by various thoroughly researched types of bacteria, but also by others previously identified in the rumen but not further characterized.

Evaluation of a PCR detection method for Escherichia coli O157:H7/H- bovine faecal samples.

AIMS: Combinations of PCR primer sets were evaluated to establish a multiplex PCR method to specifically detect Escherichia coli O157:H7 genes in bovine faecal samples. METHODS AND RESULTS: A multiplex PCR method combining three primer sets for the E. coli O157:H7 genes rfbE, uidA and E. coli H7 fliC was developed and tested for sensitivity and specificity with pure cultures of 27 E. coli serotype O157 strains, 88 non-O157 E. coli strains, predominantly bovine in origin and five bacterial strains other than E. coli. The PCR method was very specific in the detection of E. coli O157:H7 and O157:H- strains, and the detection limit in seeded bovine faecal samples was <10 CFU g(-1) faeces, following an 18-h enrichment at 37 degrees C, and could be performed using crude DNA extracts as template. CONCLUSIONS: A new multiplex PCR method was developed to detect E. coli O157:H7 and O157:H-, and was shown to be highly specific and sensitive for these strains both in pure culture and in crude DNA extracts prepared from inoculated bovine faecal samples. SIGNIFICANCE AND IMPACT OF THE STUDY: This new multiplex PCR method is suitable for the rapid detection of E. coli O157:H7 and O157:H- genes in ruminant faecal samples.

Changes in equine hindgut bacterial populations during oligofructose-induced laminitis.

In the horse, carbohydrate overload is thought to play an integral role in the onset of laminitis by drastically altering the profile of bacterial populations in the hindgut. The objectives of this study were to develop and validate microbial ecology methods to monitor changes in bacterial populations throughout the course of experimentally induced laminitis and to identify the predominant oligofructose-utilizing organisms. Laminitis was induced in five horses by administration of oligofructose. Faecal specimens were collected at 8 h intervals from 72 h before to 72 h after the administration of oligofructose. Hindgut microbiota able to utilize oligofructose were enumerated throughout the course of the experiment using habitat-simulating medium. Isolates were collected and representatives identified by 16S rRNA gene sequencing. The majority of these isolates collected belonged to the genus Streptococcus, 91% of which were identified as being most closely related to Streptococcus infantarius ssp. coli. Furthermore, S. infantarius ssp. coli was the predominant oligofructose-utilizing organism isolated before the onset of lameness. Fluorescence in situ hybridization probes developed to specifically target the isolated Streptococcus spp. demonstrated marked population increases between 8 and 16 h post oligofructose administration. This was followed by a rapid population decline which corresponded with a sharp decline in faecal pH and subsequently lameness at 24-32 h post oligofructose administration. This research suggests that streptococci within the Streptococcus bovis/equinus complex may be involved in the series of events which precede the onset of laminitis in the horse.

Characterization of Streptococcus bovis from the rumen of the dromedary camel and Rusa deer.

AIMS: Isolation and characterization of Streptococcus bovis from the dromedary camel and Rusa deer. METHODS AND RESULTS: Bacteria were isolated from the rumen contents of four camels and two deer fed lucerne hay by culturing on the semi-selective medium MRS agar. Based on Gram morphology and RFLP analysis seven isolates, MPR1, MPR2, MPR3, MPR4, MPR5, RD09 and RD11 were selected and putatively identified as Streptococcus. The identity of these isolates was later confirmed by comparative DNA sequence analysis of the 16S rRNA gene with the homologous sequence from S. bovis strains, JB1, C14b1, NCFB2476, SbR1, SbR7 and Sb5, from cattle and sheep, and the Streptococcus equinus strain NCD01037T. The percentage similarity amongst all strains was >99%, confirming the identification of the camel isolates as S. bovis. The strains were further characterized by their ability to utilize a range of carbohydrates, the production of volatile fatty acids (VFA) and lactate and the determination of the doubling time in basal medium 10 supplemented with glucose. All the isolates produced l-lactate as a major fermentation end product, while four of five camel isolates produced VFA. The range of carbohydrates utilized by all the strains tested, including those from cattle and sheep were identical, except that all camel isolates and the deer isolate RD11 were additionally able to utilize arabinose. CONCLUSIONS: Streptococcus bovis was successfully isolated from the rumen of camels and deer, and shown by molecular and biochemical characterization to be almost identical to S. bovis isolates from cattle and sheep. SIGNIFICANCE AND IMPACT OF THE STUDY: Streptococcus bovis is considered a key lactic acid producing bacterium from the gastrointestinal tract of ruminants, and has been implicated as a causative agent of lactic acidosis. This study is the first report of the isolation and characterization of S. bovis from the dromedary camel and Rusa deer, and suggests a major contributive role of this bacterium to fermentative acidosis.

Lactobacillus ruminis is a predominant lactic acid producing bacterium in the caecum and rectum of the pig.

AIMS: To identify the predominant lactic acid producing bacteria in the small intestine, caecum and the rectum of the healthy pig. METHODS AND RESULTS: Samples obtained from the large intestine of healthy pigs post-mortem were cultured using a modified agar-MRS medium in roll tubes. Thirteen isolates were selected on the basis of their morphological characteristics and Gram stain reaction for gene sequencing. These isolates were characterized by DNA sequence analysis of 16S rDNA. Eight isolates were identified as Lactobacillus ruminis, two as Enterococcus faecium, one as Mitsuokella multiacidus and two as Escherichia coli. CONCLUSION: This is the first report of Lact. ruminis as the dominant lactic acid bacteria in the large intestine of the pig. SIGNIFICANCE AND IMPACT OF THE STUDY: The results suggest that Lact. ruminis is a dominant bacterium in the large intestine of the healthy pig. Future work should focus on the role of this bacterium in relation to the physiological function of the intestine and the health of the animal.