Detection with nonradioactively labeled probes of the toxin genes of different E. coli pathotypes from swine]

We tested hemolytic E. coli from 86 pigs with edema disease or colidiarrhea. They were tested serologically and with nonradioactive digoxigenin-dUTP labelled probes for the presence of enterotoxin or Shiga-like-toxin genes. By slide-agglutination we detected 38 cases with E. coli O149:K88, 28 with E. coli O139:82B and 20 with E. coli O141. E. coli of serogroup O149:K88 isolated from diarrheic pigs, reacted with the probes for LT and STb genes. Edema disease E. coli O139:82B reacted with the SLTII probe. E. coli O141, isolated from colidiarrhea or edema disease showed a diversity of toxin gene patterns. All the E. coli O141 from diarrheic pigs reacted with the probes for LT and STap in addition to SLTII. No strains isolated from pigs with edema disease possessed any of these enterotoxin genes. Gene probe technique confirmed the serological method as useful tool for diagnosing E. coli O149:K88 and O139:82B as ETEC or VTEC, respectively. On the other hand only the demonstration of toxin genes with probes could explain the pathological findings in the pigs shedding E. coli of serogroup O141.     

Relationship between terrain use and performance of beef cows grazing foothill rangeland

A study was conducted on foothills rangeland to determine whether grazing patterns differed among cow breeds and to determine whether there was a relationship between individual grazing patterns and performance of beef cows. Hereford (HH), Tarentaise (TT), 3/4 Hereford x 1/4 Tarentaise (3H1T), 1/2 Hereford x 1/2 Tarentaise (HT), and 1/4 Hereford x 3/4 Tarentaise (1H3T) cows were observed during the summers of 1997 (n = 183) and 1998 (n = 159). Locations of individual cows were recorded two to three times per week during 1.5- to 2.5-h periods in the morning by observers on horseback. Statistical models included cow breed, age, and nursing status. Data from each year were analyzed separately. During 1997 and 1998, nonlactating cows were located at greater (P < 0.05) vertical distances from water than lactating cows. In 1998, nonlactating cows used steeper (P < 0.05) slopes than lactating cows. However, nonlactating cows did not travel as far horizontally from water (P < 0.05) as lactating cows in 1997. Younger cows (3 yr) traveled further (P < 0.05) from water both vertically and horizontally than older cows (5+ yr) in 1997, but not during 1998. Tarentaise and 1H3T cows were observed at greater (P < 0.05) vertical distances from water than HH cows during both years of the study. During 1998, TT and 1H3T cows used steeper (P < 0.05) slopes than HH cows. Using residual correlations, there were no consistent relationships between topographic aspects of individual grazing locations and cow weight, height, and body condition score. In 1997, cows with earlier calving dates and correspondingly heavier calf weaning weights used areas that had greater vertical distances to water; however, in 1998 there were no relationships (P > 0.05) of calving date and weaning weight with cow location. During both years, pregnant and nonpregnant cows used terrain similarly (P > 0.05), which suggests that cow reproductive performance was not related to terrain use. Grazing patterns in foothills rangeland varied among cow breeds. Performance of cows that used more rugged topography was similar to cows using gentler terrain.     

Novel 1,3-dioxanes from apple juice and cider

Extracts obtained by XAD solid-phase extraction of apple juice and cider were separated by liquid chromatography on silica gel. Several new 1,3-dioxanes including the known 2-methyl-4-pentyl-1,3-dioxane and 2-methyl-4-[2'(Z)-pentenyl]-1,3-dioxane, were identified in the nonpolar fractions by GC/MS analysis and confirmed by chemical synthesis. The enantioselective synthesis of the stereoisomers of the 1,3-dioxanes was performed using (R)- and (R,S)-octane-1,3-diol and (R)- and (R,S)-5(Z)-octene-1,3-diol as starting material. Comparison with the isolated products indicated that the natural products consisted of a mixture of (2S,4R) and (2R,4R) stereoisomers in the ratio of approximately 10:1, except for 1,3-dioxanes generated from acetone and 2-butanone. It is assumed that the 1, 3-dioxanes are chemically formed in the apples and cider from the natural apple ingredients (R)-octane-1,3-diol, (R)-5(Z)-octene-1, 3-diol, (3R,7R)- and (3R,7S)-octane-1,3,7-triol, and the appropriate aldehydes and ketones, which are produced either by the apples or by yeast during fermentation of the apple juice.     

The neuron-specific protein PGP 9.5 is a ubiquitin carboxyl-terminal hydrolase

A complementary DNA (cDNA) for ubiquitin carboxyl-terminal hydrolase isozyme L3 was cloned from human B cells. The cDNA encodes a protein of 230 amino acids with a molecular mass of 26.182 daltons. The human protein is very similar to the bovine homolog, with only three amino acids differing in over 100 residues compared. The amino acid sequence deduced from the cDNA was 54% identical to that of the neuron-specific protein PGP 9.5. Purification of bovine PGP 9.5 confirmed that it is also a ubiquitin carboxyl-terminal hydrolase. These results suggest that a family of such related proteins exists and that their expression is tissue-specific.     

Analysis of differential protein expression in Actinobacillus pleuropneumoniae by Surface Enhanced Laser Desorption Ionisation--ProteinChip (SELDI) technology

Actinobacillus pleuropneumoniae (APP) is the aetiological agent of porcine pleuropneumonia. An increased understanding of its molecular basis of pathogenicity and vaccine development will be facilitated by the availability of sequence data from a complete genome which, by analogy to other bacteria, is predicted to encode many proteins in the molecular mass range 3-20kDa. However, conventional techniques to study bacterial protein expression, such as SDS-PAGE and 2-dimensional electrophoresis, typically focus on the 15-200kDa range. In this study we have evaluated Surface Enhanced Laser Desorption Ionisation-ProteinChip (SELDI) technology for the analysis of protein expression, in particular those of <20kDa, of APP grown under different environmental conditions. Cytoplasmic/periplasmic and outer membrane protein fractions were obtained from the APP wildtype serotype 1 strain 4074 grown in Brain Heart Infusion (BHI) broth (+different concentrations of NAD), BHI containing pig serum or defined medium. Optimum conditions for SELDI profiles included a sample size of 1 microg and the use of sinapinic acid as the energy absorbing matrix. In the <20kDa range, the SELDI profiles obtained from wild-type bacteria grown in rich medium plus 33-66% pig serum were most similar to those grown in defined medium. The SELDI profiles of extracts of the wild-type and of an rpoE mutant were similar although there were clear differences. The results suggest that SELDI is a useful complementary approach to conventional proteomic analytical methods with APP, and presumably other bacterial pathogens, being particularly suited for analysis of proteins in the <20kDa mass range.     

Effects of barley variety fed to steers on carcass characteristics and color of meat

This study evaluated the effect of barley varieties in the diets of finishing steers on carcass composition, fat, and lean color and the fatty acid profile of subcutaneous fat. Crossbred steers (391 kg initial BW) were assigned randomly to one of five finishing diets composed primarily of corn (n = 9), Morex barley (n = 9), Steptoe barley, (n = 9), or two experimental barley varieties SM3 (n = 9) and SM5 (n = 9). Grains were cracked prior to feeding. Diets were formulated (DM basis) to be isonitrogenous (2.24% N) and isocaloric (2.01 Mcal/kg NEm and 1.35 Mcal/kg NEg). Steers were slaughtered according to industry-accepted procedures when it was visually estimated that 70% of carcasses would grade USDA Choice. After a 24-h chill at 4 degrees C, carcass quality and yield grade data were collected by trained, experienced university personnel. Objective color (L*, a*, and b*) of both the LM and subcutaneous fat were measured, and samples of subcutaneous fat were removed from the 10th- to 12th-rib region for fatty acid analysis. Diet did not affect hot carcass weight (P = 0.15), fat thickness (P = 0.58), LM area (P = 0.57), percentage of internal fat (P = 0.52), yield grade (P = 0.96), marbling (P = 0.73), or quality grade (P = 0.10). However, the LM from steers fed diets formulated with Morex and SM5 barley varieties tended to be lighter (higher L* values, P = 0.08) than the LM from steers fed the corn-based diet. Additionally, fat from steers fed corn tended to be more yellow (higher Hunter b* values, P = 0.09) than fat from steers fed barley-based diets. Although grain source had only minimal effects on the fatty acid composition of subcutaneous fat samples, pentadecanoic acid (15:0) was greater (P < 0.05) in fat from steers fed SM3 and Steptoe barley varieties than in fat from steers fed corn. Stearic acid (18:0) concentrations were higher (P < 0.05) in fat samples from steers fed corn than in those fed the experimental barley lines (SM3 and SM5). Conversely, fat samples from steers fed Steptoe and SM5 barley had greater (P < 0.05) gadoleic acid (20:1) concentrations than fat from steers fed corn or Morex variety. Although the variety/line of barley included in the finishing diet may affect LM and fat color, grain-source (barley vs. corn) had little effect on beef carcass quality and yield grades and did not greatly alter the fatty acid composition of subcutaneous fat.     

Outcomes of 35 dogs with craniomaxillofacial osteosarcoma treated with stereotactic body radiation therapy

Canine craniomaxillofacial osteosarcoma (OSA) is most commonly treated surgically; however, in cases where surgery is not feasible or non-invasive treatment is desired, stereotactic body radiation therapy (SBRT) may be elected for local tumour control. In this study, we evaluated 35 dogs treated with SBRT. Nine dogs (26%) had calvarial, seven (20%) had mandibular and 19 (54%) had maxillary OSA. Median time to first event (TFE) was 171 days, and overall median survival time (MST) was 232 days. Site-specific MSTs were 144 days for mandible, 236 days for calvarium and 232 days for maxilla (p = .49). Pulmonary metastatic disease was observed in 12/35 (34%) patients and was detected pre-SBRT in six dogs (17%) and post-SBRT in the remaining six dogs (17%). Eighteen adverse events post-SBRT were documented. Per veterinary radiation therapy oncology group criteria, five were acute (14%) and three were late (9%) grade 3 events. Neurological signs in two dogs were suspected to be early-delayed effects. Cause of death was local progression for 22/35 (63%) patients, metastasis for 9/35 (26%) patients and unknown for four. On univariate analysis, administration of chemotherapy was associated with a longer TFE (p = .0163), whereas volume of gross tumour volume was associated with a shorter TFE (p = .023). Administration of chemotherapy and five fractions versus single fraction of SBRT was associated with increased survival time (p = .0021 and .049). Based on these findings, a treatment protocol incorporating chemotherapy and five fractions of SBRT could be considered for dogs with craniomaxillofacial OSA electing SBRT with careful consideration of normal tissues in the field.     

Investigation on Opisthorchis felineus occurrence and life cycle in Italy

Opisthorchiasis is a fish borne parasitic infection caused by helminths of the genus Opisthorchis (Digenea, Opisthorchiidae), affecting humans and other fish-eating mammals. Despite Opisthorchis felineus was first described in Italy in 1884, no cases of human opisthorchiasis were reported in this country until 2004; from then on, 4 outbreaks due to this species have been recorded in Central Italy. Following the more relevant of these outbreaks, involving 34 people in August 2007, snails, fishes and fecal samples collected from the Bolsena and Bracciano lakes (Central Italy) were analyzed in order to define the cycle of O. felineus in the area and investigate its prevalence in the different hosts. Pools of 20-40 snails each (4983 specimens altogether) of the genus Bithynia were analyzed by PCR for parasite DNA detection. Eight hundred and ninety-four fish belonging to 12 species were collected from the two lakes and tested for metacercariae both by muscle compression and digestion techniques. Eighty-seven fecal samples of 5 putative definitive host species were collected very close to the two lakes and tested for parasite eggs detection by formalin-ethyl acetate concentration technique. Identification at the species level of metacercariae and eggs, respectively, from fish and stool was confirmed by PCR analysis and sequencing. O. felineus DNA was detected in 0.08% (overall minimum infection rate) of snails of the genus Bithynia from the two lakes. The tench, Tinca tinca, was the only fish found infested in both lakes (prevalence 88.5%). O. felineus eggs were found only in cat feces (prevalence 46.4%). The tench represents the only threat for the human consumption in the study area while Coregonus sp., the most economically important species for the local fishery and frequently consumed raw marinated, resulted to be not infected. The high prevalence recorded both in fish and in definitive host suggests a widespread and massive presence of the parasite in the area. Further studies are needed to better investigate the possible role of some cyprinids species as intermediate hosts, in order to check their safety for human consumption.     

Dormant Season Grazing: Effect of Supplementation Strategies on Heifer Resource Utilization and Vegetation Use

Grazing dormant forage under low-input heifer development strategies typically exposes cattle to low-quality forage. Protein supplementation while grazing dormant range can enhance heifer growth and reproductive performance. We examined resource utilization of heifers and the effects of dormant season grazing on residual vegetation characteristics under two supplementation management strategies. Approximately 100 weaned composite heifer calves were randomly selected and placed into one of two supplementation treatments in each of 2 yr, one receiving a free access 62% crude protein self-fed concentrate and the other receiving a daily hand-fed 20% crude protein cake. Grazing occurred from December (2015 and 2016) through March (2016 and 2017). Thirty transects were randomly located within each pasture for measuring vegetation quality and structure before and after grazing. Daily space use and behavior was evaluated for 21 individuals within each treatment using global positioning system (GPS) collars and resource utilization functions. Heifers supplemented with concentrated protein spent more time grazing per day than heifers supplemented with cake (6.92 ± 0.18, 6.24 ± 0.17 h). Relative use by heifers in the cake treatment was negatively related to horizontal distance from the supplement delivery site early to midwinter ($\stackrel{?}{\overline{\beta }}$ = ? 0.41 ± 0.16, ? 0.53 ± 0.17). Both treatments selected grazing locations relative to standing biomass of perennial grasses ($\stackrel{?}{\overline{\beta }}$ = 0.0005 ± 0.00004) and crude protein ($\stackrel{?}{\overline{\beta }}$ = 0.12 ± 0.007). However, resource selection was highly variable among individuals for both supplementation treatments. We found no treatment effects on pre-post grazing differences in residual cover of litter, grass, forbs and shrubs (P > 0.24). However, the time period when grazing occurred had an effect on residual vegetation conditions (P < 0.01). Our results indicate high levels of variability in grazing site selection by heifers, suggesting future research should incorporate individual animal measurements in an attempt to account for individual animal variability.