Di-valent siRNA-mediated silencing of MSH3 blocks somatic repeat expansion in mouse models of Huntington's disease.

Huntington's disease (HD) is a severe neurodegenerative disorder caused by the expansion of the CAG trinucleotide repeat tract in the huntingtin gene. Inheritance of expanded CAG repeats is needed for HD manifestation, but further somatic expansion of the repeat tract in non-dividing cells, particularly striatal neurons, hastens disease onset. Called somatic repeat expansion, this process is mediated by the mismatch repair (MMR) pathway. Among MMR components identified as modifiers of HD onset, MutS homolog 3 (MSH3) has emerged as a potentially safe and effective target for therapeutic intervention. Here, we identify a fully chemically modified short interfering RNA (siRNA) that robustly silences Msh3 in vitro and in vivo. When synthesized in a di-valent scaffold, siRNA-mediated silencing of Msh3 effectively blocked CAG-repeat expansion in the striatum of two HD mouse models without affecting tumor-associated microsatellite instability or mRNA expression of other MMR genes. Our findings establish a promising treatment approach for patients with HD and other repeat expansion diseases.

Antibimicrobial Susceptibility of Trimethoprim-Sulfamethoxazole and 3rd-Generation Cephalosporin-Resistant Escherichia coli Isolates Enumerated Longitudinally from Feedlot Arrival to Harvest in High-Risk Beef Cattle Administered Common Metaphylactic Antimicrobials.

Multidrug resistant (MDR) Escherichia coli threaten the preservation of antimicrobials to treat infections in humans and livestock. Thus, it is important to understand where antimicrobial-resistant E. coli persist and factors that contribute to its their development. Crossbred cattle (n = 249; body weight = 244 kg ±25 kg standard deviation) were blocked by arrival date and assigned metaphylactic antimicrobial treatments of sterile saline control, tulathromycin (TUL), ceftiofur, or florfenicol at random. Trimethoprim-sulfamethoxazole (COTR) and third-generation cephalosporin (CTXR)-resistant E. coli were isolated from fecal samples on days 0, 28, 56, 112, 182, and study END (day 252 for block 1 and day 242 for block 2). Then, susceptibility testing was conducted on all confirmed isolates. MDR was detected in both COTR and CTXR E. coli isolates. In COTR isolates, the number of antimicrobials each isolate was resistant to and the minimum inhibitory concentration (MIC) for amoxicillin-clavulanic acid, ceftriaxone, and gentamicin was greatest on day 28 compared with all other days (p ≤ 0.04). Similarly, chloramphenicol MIC was greater on day 28 than on day 0 (p < 0.01). Overall, sulfisoxazole MIC was less for TUL than all other treatments (p ≤ 0.02), and trimethoprim-sulfamethoxazole MIC was greater for TUL than all other treatments (p ≤ 0.03). Finally, there was no effect of treatment, day, or treatment × day for tetracycline or meropenem MIC (p ≥ 0.07). In CTXR isolates, there was an effect of day for all antimicrobials tested except ampicillin and meropenem (p ≤ 0.06). In conclusion, administering a metaphylactic antimicrobial at feedlot arrival did influence the susceptibility of COTR and CTXR E. coli. However, MDR E. coli are widely distributed, and the MIC for most antimicrobials was not different from the initial value upon completion of the feeding period.

Longitudinal Assessment of Prevalence and Incidence of Salmonella and Escherichia coli O157 Resistance to Antimicrobials in Feedlot Cattle Sourced and Finished in Two Different Regions of the United States.

The objective was to investigate the influence of cattle origin and region of finishing on the prevalence of Salmonella, Escherichia coli O157:H7, and select antimicrobial resistance in E. coli populations. Yearling heifers (n = 190) were utilized in a 2 × 2 factorial arrangement. After determining fecal Salmonella prevalence, heifers were sorted into one of four treatments: heifers originating from South Dakota (SD) and finished in SD (SD-SD); heifers originating from SD and finished in Texas (SD-TX); heifers originating from TX and finished in SD (TX-SD); and heifers originating from TX and finished in TX (TX-TX). Fecal, pen, and water scum line samples were collected longitudinally throughout the study; hide swab and subiliac lymph node (SLN) samples were collected at study end. A treatment × time interaction was observed (p ≤ 0.01) for fecal Salmonella prevalence, with prevalence being greatest for TX-TX and TX-SD heifers before transport. From day (d) 14 through study end, prevalence was greatest for TX-TX and SD-TX heifers compared with SD-SD and TX-SD heifers. Salmonella prevalence on hides were greater (p ≤ 0.01) for heifers finished in TX compared with SD. Salmonella prevalence in SLN tended (p = 0.06) to be greater in TX-TX and SD-TX heifers compared with TX-SD and SD-SD. Fecal E. coli O157:H7 prevalence had a treatment × time interaction (p = 0.04), with SD-TX prevalence being greater than TX-SD on d 56 and SD-SD and TX-TX being intermediate. A treatment × time interaction was observed for fecal trimethoprim-sulfamethoxazole-resistant and cefotaxime-resistant E. coli O157:H7 prevalence (p ≤ 0.01). Overall, these data suggest that the region of finishing influences pathogenic bacterial shedding patterns, with the initial 14 d after feedlot arrival being critical for pathogen carriage.

Prevalence and Antimicrobial Susceptibility of Salmonella Serovars Isolated from U.S. Retail Ground Pork.

One objective of this study was to determine overall prevalence of Salmonella in ground pork from U.S. retail stores over three seasons including both case-ready and store-ground packages. Package types collected included: overwrap, chub, modified atmosphere packaging, and other (plastic or wax paper wrapped). Because package type represents different production systems and are subject to varied microbiological government regulation and testing methodologies, both USDA-FSIS and FDA Salmonella isolation protocols were performed. Another objective of the study was to determine serotypes and antimicrobial susceptibility profiles of the isolates obtained from the ground pork samples. Ground pork aliquots were subjected to real-time PCR. Recovered isolates were serotyped and minimum inhibitory concentration analysis to 15 antimicrobials was determined using microbroth dilution. Overall prevalence of Salmonella in ground pork from the 865 samples collected was 1.39%. Prevalence was not affected by package type (p = 0.29) nor grind location (case-ready vs. store-ground; p = 0.17). Season affected Salmonella prevalence (p = 0.05) with most isolates found during fall, and there was a tendency for geographic region to affect prevalence (p = 0.07). The USDA Salmonella isolation method was more effective at recovering isolates (p = 0.01) compared with the FDA methodology and yielded a kappa statistic of 0.26 as a measure of agreement. The serotypes isolated included: Infantis, 4,5,12:i:-, Brandenburg, Typhimurium var 5-, Seftenberg, and Johannesburg with only two packages containing multiple serotypes. No isolates were resistant to antibiotics commonly used to treat human Salmonella infections including extended spectrum cephalosporins or fluoroquinolones. Although the recovery of Salmonella from retail ground pork samples was rare, Salmonella Typhimurium (and its monophasic variant 4,5,12:i:-), which are among the most common serovars recovered from human infections, were recovered. Therefore, more effective strategies to further reduce or eliminate these pathogens from retail pork products are warranted.

Translocation of Orally Inoculated Salmonella Following Mild Immunosuppression in Holstein Calves and the Presence of the Salmonella in Ground Beef Samples.

The objective of this study was to determine if immunosuppression through daily dexamethasone (DEX) infusion altered Salmonella translocation from the gastrointestinal tract. Weaned Holstein steers (n = 20; body weight [BW] = 102 ± 2.7 kg) received DEX (n = 10; 0.5 mg/kg BW) or saline (control [CON]; n = 10;) for 4 days (from day -1 to 2) before oral inoculation of naldixic acid-resistant Salmonella enterica Typhimurium (SAL; 3.4 × 106 colony-forming units [CFU]/animal) on day 0. Fecal swabs were obtained daily, and blood was collected daily for hematology. At harvest (day 5), ileum, cecal fluid, lymph nodes (ileocecal, mandibular, popliteal, and subscapular), and synovial (stifle, coxofemoral, and shoulder) samples were collected for isolation of the inoculated strain of SAL. White blood cell (WBC) and neutrophil concentrations were elevated (p < 0.01) in DEX calves following each administration event. Following inoculation, 100% of DEX calves shed the experimental strain of SAL for all 5 days, 90% of CON calves shed from day 1 to 3, and 100% of CON calves shed from day 4 to 5. Greater (p < 0.01) concentrations of SAL were quantified from the cecum of DEX calves (3.86 ± 0.37 log CFU/g) compared with CON calves (1.37 ± 0.37 log CFU/g). There was no difference in SAL concentrations between DEX and CON calves in ileal tissue (p = 0.07) or ileocecal (p = 0.57), mandibular (p = 0.12), popliteal (p = 0.99), or subscapular (p = 0.83) lymph nodes. Of the stifle samples collected, 3.3% were positive for SAL, highlighting a contamination opportunity during hindquarter breakdown. While more research is needed to elucidate the interactions of immunosuppression and pathogen migration patterns, these data confirm that orally inoculated SAL can translocate from the gastrointestinal tract and be harbored in atypical locations representing a food safety risk.

Bacillus subtilis PB6 Supplementation in Weaned Holstein Steers During an Experimental Salmonella Challenge.

To evaluate the effects of a patented Bacillus subtilis probiotic, weaned Holstein steers, not shedding Salmonella (n = 40; ∼90 kg), were supplemented (CLO) or not (CON) with CLOSTAT® (13 g/hd per day; Kemin Industries, Des Moines, IA) in a starter ration for 35 d. The calves were assigned to one of four treatments in a 2 × 2 factorial design with CLO and CON calves that were orally administered Salmonella (STM) or not (NoSTM). Calves were challenged with 1.6 × 106 colony-forming unit (CFU) Salmonella Typhimurium (resistant to 50 µg/mL nalidixic acid) in 1 L of milk replacer on day 0. Blood samples were collected through jugular catheters every 6 h for 96 h, and body temperature was measured every 5 min through indwelling rectal temperature recording devices. Five calves from each treatment were harvested 48 h postchallenge, and the remaining calves were harvested 96 h postchallenge. During necropsy, tissues were collected for the isolation and quantification of the inoculated STM from various tissues. The CLOSTM group had reduced STM concentrations in the jejunum, ileum, and transverse colon 48 h after the challenge (p ≤ 0.03), but were not different 96 h postchallenge (p > 0.05). Decreased (p < 0.01) pyrexia was observed after the challenge in CLOSTM calves when compared with CONSTM calves. White blood cells and lymphocyte counts were increased (p ≤ 0.05) in CLOSTM calves after the challenge in comparison with other treatments. In calves given STM, the CLO group had greater feed intake before and after the challenge (p < 0.01) compared with the CON group. Increased serum IL-6 and IFN-γ concentrations were observed in the CONSTM group compared with other treatments. Overall, CLO reduced Salmonella presence and concentrations in gastrointestinal tissues while simultaneously reducing the severity of the challenge as indicated by blood parameters and the reduced febrile response.

Influence of prenatal transportation stress on innate immune response to an endotoxin challenge in weaned Brahman bull calves†,‡.

The objective of this study was to assess the influence of prenatal stress (PNS) on innate immune responses to an endotoxin challenge in weaned bull calves. Altered innate immune response to lipopolysaccharide (LPS), as characterized by changes in a range of variables was hypothesized in PNS bull calves. Brahman cows (n = 96; 48 stressed by transportation at five stages of gestation and 48 Controls) produced 85 calves, from which 16 uncastrated male (bull) calves from each PNS and Control treatment were selected for an LPS challenge period. Rectal temperature (RT), sickness behavior score (SBS), serum concentrations of cortisol, interferon-gamma (IFN-γ), tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6), and complete blood count (CBC) variables were assessed in response to intravenous LPS (0.25 µg/kg body weight) administration. Each reported variable increased or decreased following LPS administration. Prior to LPS, PNS bull calves exhibited increased TNF-α, IL-6, and monocyte counts, but decreased IFN-γ, eosinophils, and basophils (p < .05). Compared with Control, in response to LPS, PNS bull calves exhibited greater circulating concentrations of cortisol. PNS bull calves exhibited lower (p < .05) eosinophil and basophil counts at time 0 (time of LPS administration) but similar counts to Control bull calves 2 h after LPS. PNS bull calves exhibited a greater change from baseline for IFN-γ and monocytes in response to LPS administration. No other variables were influenced by prenatal treatment (p > .05). These findings suggest that PNS did not adversely affect basal or induced components of the innate immune response to an immunological challenge. Lay summary Our laboratory studied the influence of prenatal stress (i.e., transportation of pregnant cows) on immune function of bull calves at 8 months of age. This was accomplished by studying aspects of their innate immune response to an immunological challenge. Prenatal stress did not adversely affect basal or induced components of the innate immune response to an immunological challenge.

The digestive system of 1-week-old Jersey calves is well suited to digest, absorb, and incorporate protein and energy into tissue growth even when calves are fed a high plane of milk replacer.

The objectives of the current study were to determine the apparent digestibilities of nitrogen, organic matter, ash, and energy as well as investigate the nitrogen retention of calves fed different planes of milk replacer nutrition during the first week of life. Twelve Jersey calves were blocked by body weight at birth and randomly assigned to either a high plane of nutrition (HPN) or low plane of nutrition (LPN) treatment. The HPN calves were offered 19.2g of dry matter/kg of body weight of a 28% all-milk crude protein and 20% fat milk replacer. The LPN calves were fed 11.6g of dry matter/kg of body weight of a 20% all-milk crude protein and 20% fat milk replacer. All calves were given 3 L of pooled colostrum within 1h of birth after which they were assigned to treatments; no starter was offered during the study. Calves were given 1 d to adapt to their treatments and environment, so calves were 30 to 36 h old at the start of data collection. The study was divided into two 72-h periods. Total collection of feces occurred over each 72 h period, and total urine was collected for the last 24h of each period. Peripheral blood samples were collected at the beginning and end of each period and analyzed for plasma glucose and urea nitrogen concentrations. Data are reported as HPN vs. LPN, respectively. Fecal scores were greater for HPN calves during both periods; however, no difference was found in the dry matter percentage of feces (30.9 vs. 31.9 ± 0.06). No differences were found between treatments in either digestible or metabolizable energy efficiencies, which averaged 93.3 and 83.7%, respectively. A treatment × period interaction was found on the percentage of intake nitrogen retained, in which calves fed the HPN had a greater percentage of intake nitrogen retained during period 1 (87.9 vs. 78.4 ± 1.79%), but was not different from calves fed the LPN during period 2 (85.4 vs. 84.9 ± 1.79%). From these data therefore, we conclude that healthy neonatal calves have the capability to digest and absorb the additional nutrients with a higher plane of nutrition during the first week of postnatal life.

Survival of Escherichia coli O157:H7 transformed with either the pAK1-lux or pXEN-13 plasmids in in vitro bovine ruminal and fecal microbial fermentations.

The use of luminescent plasmids in bacteria may serve as a viable model for the real-time validation of various pre-harvest interventions on the colonization or shedding patterns of Escherichia coli O157:H7 within cattle. The objective of this study was to determine if the growth characteristics of E. coli O157:H7 in mixed ruminal and fecal microbial fluid cultures would be altered when transformed with one of the two luminescent plasmids: pAK1-lux (PAK) or pXEN-13 (XEN). Transformants harboring the luminescent plasmids were compared to the non-transformed parental strain (wild type [WT]) after incubating in mixed ruminal or fecal microbial fluid media for 6 h in triplicate (n=3). The transformants and WT exhibited similar growth rates. Within mixed ruminal microbial fluid fermentations and mixed fecal microbial fluid, all transformants grew similarly (p=0.28) through the 6-h study. The reflective light unit (RLU; photons/pixel per second) photonic emissions of each plasmid within ruminal fluid differed at 0 h (p=0.002) and 2 h (p=0.02) and within fecal fluid at 0 h (p=0.009) and 2 h (p=0.04). The RLU remained the same within rumen fluid at 4 h (p=0.22) and 6 h (p=0.80) and within fecal fluid at 4 h (p=0.06) and 6 h (p=0.29). Growth of E. coli O157:H7 transformed with the bioluminescent plasmids was not altered in comparison to the WT, suggesting that both plasmids may serve as useful models for in vivo studies.

Corrigendum: Phenotypic and genetic parameters of circadian rhythms from core body temperature profiles and their relationships with beef steers' production efficiency profiles during successive winter feeding periods.

[This corrects the article DOI: 10.3389/fgene.2023.1026601.].

Phenotypic and genetic parameters of circadian rhythms from core body temperature profiles and their relationships with beef steers' production efficiency profiles during successive winter feeding periods.

This 2-year study evaluated differences in circadian parameters obtained from measures of core body temperatures using telemetric reticulo-rumen and rectal devices during two winter feeding regimes in western Canada. The study also estimated phenotypic correlations and genetic parameters associated with circadian parameters and other production traits in each feeding regime. Each year, 80 weaned steer calves (initial age: 209 ± 11 days; BW: 264 ± 20 kg) from the same cohort were tested over two successive regimes, Fall-Winter (FW) and Winter-Spring (WS) at Lanigan, Saskatchewan, Canada. The steers received forage-based rations in both regimes where the individual feed intake was measured with automatic feeding units. During the trial, the reticulo-rumen (RTMP) and rectal (RCT) temperatures were simultaneously measured every 5 min using telemetric devices. These were used to calculate the circadian parameters (Midline Estimating Statistic Of Rhythms, amplitude, and acrophase/peak time) for both temperature measures. Growth and efficiency performance traits were also determined for all steers. Each steer was assigned into inefficient, neutral, and efficient classes based on the SD of the residual feed intake (RFI), residual gain (RG), and residual intake and gain (RIG) within each year and feeding regime. Higher (p < 0.0003) RTMP and rectal temperature MESORs were observed in the Fall-Winter compared to the Winter-Spring regime. While the two test regimes were different (p < 0.05) for the majority of the RTMP or RCT temperature parameters, they did not differ (p > 0.10) with the production efficiency profiles. The heritability estimates were higher in FW (0.78 ± 0.18 vs. 0.56 ± 0.26) than WS (0.50 ± 0.18 vs. 0.47 ± 0.22) for the rumen and rectal MESORs, respectively. There were positive genetic correlations between the two regimes for the RTMP (0.69 ± 0.21) and RCT (0.32 ± 0.59). There was a negative correlation (p < 0.001) between body temperature and ambient temperature. The high heritability estimates and genetic correlations for rumen and rectal temperature parameters demonstrate their potential as beef genetic improvement tools of economic traits associated with the parameters. However, there are limited practical implications of using only the core-body temperature as a proxy for production efficiency traits for beef steers during winter.

An updated profile of the bovine acute phase response following an intravenous lipopolysaccharide challenge.

The objective was to provide an updated profile of the bovine acute-phase response to include recent advancements in technologies and expanded hematological, cytokine, and serum chemistry variables. Beef steers (n = 32; body weight [BW] = 251 ±â€…19.5 kg) were fitted with indwelling jugular catheters 1 d before lipopolysaccharide (LPS; 0.25 µg LPS/kg BW from Escherichia coli O111:B4) administration to facilitate serial blood collection. Rectal temperature was measured using indwelling probes, and ocular temperature was measured using infrared thermal imaging. Blood samples were collected for subsequent analysis of serum chemistry, hematology, and cytokine concentrations. Pearson correlation of rectal temperature and ocular infrared temperature was 0.61 (P < 0.01) and the Spearman correlation coefficient was 0.56 (P < 0.01). Interactions of hour × method were observed for ocular and rectal measurements of body temperature in response to endotoxin exposure. Maximum observed temperature was 39.6 °C at 2.5 h for both rectal and ocular measurements. Body temperature differed by method at hours 0.5, 2.5, 4.5, 7.5, 12.5, 36.5, and 47.5 (P < 0.01), but were not different otherwise. All variables of serum chemistry and complete blood count were influenced by LPS administration, except creatinine, serum glucose, and percent basophils (P ≤ 0.02). Alanine aminotransferase and alkaline phosphatase peaked at hour 2 relative to LPS administration, returned to baseline at hour 12 and continued to decrease below the baseline value at hour 48 (P < 0.01). Total protein concentration decreased 3% in response to LPS (P = 0.01). Total white blood cell count decreased 75% after LPS administration at hour 1 (P < 0.01). Lymphocyte count recovered to baseline at hour 6; sooner than neutrophil count at hour 36. Serum cortisol concentration increased 294% relative to baseline at hour 1 followed by a sustained decrease and return to normal concentration at hour 4 (P < 0.01). Additionally, circulating cytokine concentrations changed with time in response to the LPS challenge, excluding aFGF, bFGF, IGF-1, IL-2, IL-4, MCP-1, and ANG-1 (P ≤ 0.08). Maximum observed concentration of TNF-α at hour 1 was 117% greater than the pre-challenge value (P < 0.01). Data presented herein add to existing works to understand the endocrine and immune responses of beef steers administered exogenous LPS, and incorporate recent technologies, additional biomarkers, and an expanded cytokine profile that can be used as referential data in future research.

The acute phase response is a component of innate immunity initiated by infection, inflammation, or tissue damage. Characteristics of these host responses can be emulated by administration of exogenous endotoxin and closely studied in controlled settings to understand the response to inflammatory diseases that are commonplace in livestock production. Beef steers were administered exogenous lipopolysaccharide, and responses of body temperature, serum chemistry, complete blood count, cortisol, and cytokines were quantified. A moderate correlation of body temperature measured via rectal probe or ocular infrared temperature was observed, but both methods had a similar temporal response and were sensitive to changes in body temperature. Response of serum chemistry variables highlighted the links between metabolism and the inflammatory response. The initial inflammatory response was initiated by cortisol and pro-inflammatory cytokines at hour 1 and tempered by anti-inflammatory cytokines at hours 3 and 4. Therefore, these data offer an expanded view to our understanding of the bovine acute phase response.

Removing maternal heat stress abatement during gestation modulated postnatal physiology and improved performance of Bos indicus-influenced beef offspring.

This study evaluated the growth and immune response of beef calves born from Bos indicus-influenced beef heifers provided pre- and postpartum heat abatement on pasture. On 83 ±â€…4 d prepartum (day 0), 64 Brangus crossbred beef heifers (~» B. indicus) were stratified by body weight (BW; 454 ±â€…37 kg) and body condition score (BCS; 6.3 ±â€…0.28; scale 1 to 9), and then allocated into 1 of 16 bahiagrass pastures (1 ha and 4 heifers per pasture). Treatments were randomly assigned to pastures (8 pastures per treatment) and consisted of heifers provided (SH) or not (NSH) access to artificial shade (4.5 m2 of shade area per heifer) from 83 d prepartum to 50 d postpartum (days 0 to 133). Heifers and calves were managed similarly from day 133 until the start of the breeding season (day 203). Calves were weaned on day 203 (at 119 ±â€…19 d of age), limit-fed the same drylot diet at 3.5% of BW (DM basis) days 209 to 268 (3 to 4 calves per pen; 8 pens per treatment) and vaccinated against respiratory disease pathogens on days 222 and 236. Heifer intravaginal temperatures from days 35 to 42 were lower (P ≤ 0.03) for NSH vs. SH heifers from 0000 to 0800 hours but greater (P ≤ 0.05) for NSH vs. SH heifers from 1100 to 1800 hours. Heifer intravaginal temperature from days 126 to 132 did not differ (P = 0.99) between NSH and SH heifers. Heifers assigned to NSH had greater respiration rates from days 20 to 96 (P ≤ 0.0007), greater plasma concentration of cortisol on days 35 (P = 0.07) and 55 (P = 0.02), less plasma concentration of insulin-like growth factor 1 (IGF-1) on days 35 (P = 0.10), 55, and 133 (P ≤ 0.05), and less BCS from days 55 to 203 (P ≤ 0.01) compared to SH heifers. Calves born from NSH heifers had less birth BW (P = 0.05), greater overall plasma haptoglobin concentrations (P = 0.05), greater seroconversion against bovine respiratory syncytial virus on day 222 (P = 0.02), tended to have greater ADG from days 209 to 268 (P = 0.07), and had greater BW on day 268 (P = 0.05) compared to SH offspring. Plasma concentrations of cortisol and serum titers against other respiratory disease pathogens did not differ (P ≥ 0.15) between NSH and SH offspring. Hence, removing maternal access to artificial shade: (1) increased prepartum intravaginal temperature and plasma concentrations of cortisol but reduced prepartum BCS and plasma concentrations of IGF-1 in grazing B. indicus-influenced beef heifers; and (2) increased post-weaning BW gain and had positive effects on humoral immune response of their offspring.

In dairy cattle, heat stress during late gestation negatively impacted offspring postnatal growth and immune function. The use of artificial shade is a management strategy that alleviates heat stress in environments with high temperatures and humidity. The effects of maternal access to artificial shade and its impacts on offspring performance have not been reported for Bos indicus influenced-beef cattle and were the main objective of the present study. From 83 d prepartum until 50 d postpartum, heifers had access or not to an artificial shade structure located centrally on their pastures. Overall, removing maternal access to artificial shade increased maternal internal body temperature and respiration rates, and reduced maternal body condition score from calving until the start of the breeding season and calf body weight at birth. However, calves born from heifers with no access to shade were remarkably heavier at the end of the drylot period and had lower plasma indicators of inflammatory response and positive effects to humoral immune response to vaccination.

Survival of O157:H7 and non-O157 serogroups of Escherichia coli in bovine rumen fluid and bile salts.

While Shiga toxin-producing Escherichia coli (STEC) reside asymptomatically within ruminants, particularly cattle, these strains pose a serious health risk to humans. Research related to STEC has historically focused upon O157:H7. However, with an increase in foodborne outbreaks of non-O157 origin and recent changes in testing for non-O157 by the U.S. Department of Agriculture Food Safety and Inspection Service (USDA-FSIS), there is now a critical need to understand the biological activity of non-O157 serogroups. The focus of this study was to determine whether variations exist in the ability of different serotypes of STEC to survive within bovine rumen fluid medium and bile salts. The results of this study demonstrated through viable plate count analysis that the five serotypes tested (O157:H7, O111:H8, O103:K.:H8, O145:H28, and O26:H11) were capable of growing in rumen fluid medium. However, the concentrations of the serotypes O103:K.:H8 and O26:H11 after 24 h were significantly less (p < 0.05) than that observed for the other serotypes tested. A significant decrease (p = 0.03) in the survival of O103:K.:H8 in 50 mg/mL of bovine bile salts in comparison to the other STEC strains tested was also observed. Collectively, these data suggest that non-O157 serogroups of E. coli respond differently to the environment of the bovine gastrointestinal tract. Further research is needed to elucidate how these differential physiological variations correlate with alterations in colonization success within ruminants and how they may impact human illnesses.

Sexual Dimorphic Innate Immune Response to a Viral-Bacterial Respiratory Disease Challenge in Beef Calves.

The potential for sexually dimorphic innate immune responses to respiratory disease was evaluated, where eight steers and seven heifers (280 ± 4 kg) were subjected to a viral−bacterial respiratory disease challenge utilizing bovine herpesvirus-1 (BoHV-1; intranasal; 1 × 108 PFU/nostril) and Mannheimia haemolytica (MH; intratracheal; 1.3 × 107 CFU/head) administered 72 h later. Body temperature was lesser in heifers than steers (p < 0.01). There was a sex × time interaction (p = 0.05) for white blood cells where heifers had reduced concentrations compared with steers at −72 and 0 h but greater concentrations from 36 to 60 h post-MH. Concentrations of neutrophils were lesser in heifers compared to steers from 0 to 4 h, and from 8 to 12 h (p = 0.03). Lymphocytes were greater in heifers compared to steers at 12 h and from 36 to 60 h post-MH (p < 0.01). The neutrophil−lymphocyte ratio was lesser in heifers compared to steers from 2 to 24 h and at 48 h post-MH (p < 0.01). Monocytes were greater in heifers compared to steers from 24 to 60 h post-MH (p < 0.01), while eosinophils were greater in heifers compared to steers at 48 and 60 h (p < 0.01). Serum IL-4 was lesser in heifers compared to steers at 0 h and from 2 to 72 h post-MH challenge (p = 0.02). Non-esterified fatty acid concentrations were lesser (p < 0.01) in heifers compared to steers from 2 to 4 h post-MH challenge. Urea nitrogen concentrations were greater (p < 0.01) in heifers than steers at 36 h post-MH challenge. Data from this study reveal distinct differences in the acute phase response following a respiratory disease challenge where steers produced an early response, while the response in heifers appeared to be delayed.

Stair-step strategy and immunomodulatory feed ingredient supplementation for grazing heat-stressed Bos indicus-influenced beef heifers.

On day 0 of years 1 and 2, 64 Brangus crossbred heifers per year were stratified by initial body weight (BW) and age (mean = 257 ± 20 kg and 271 ± 22 d) and allocated into 16 bahiagrass (Paspalum notatum) pastures (4 heifers per pasture per yr). Treatments were randomly allotted to pastures in a 2 × 2 factorial arrangement of treatments (4 pastures per treatment per yr). Treatments consisted of concentrate dry matter (DM) supplementation at 1.50% of BW from days 0 to 100 (CON) or concentrate DM supplementation at 1.05% of BW from days 0 to 49 and 1.95% of BW from days 50 to 100 (SST). Then, each respective supplementation strategy was added or not with immunomodulatory feed ingredients from days 0 to 100 (OMN; 4 g/45 kg of BW). Heifers were assigned to an estrus synchronization protocol from days 100 to 114. Heifers detected in estrus from days 111 to 114 were inseminated (AI) 12 h after estrus detection. Heifers not detected in estrus were timed AI on day 114. All heifers were exposed to Angus bulls from days 120 to 210 (1 bull per pasture). Effects of supplementation strategy × OMN inclusion × hour were detected (P < 0.0001) only for intravaginal temperature from days 26 to 30, which were the least (P ≤ 0.03) for SST heifers offered OMN supplementation and did not differ (P ≥ 0.17) among all remaining treatments from 0830 to 1600 hours. Effects of supplementation strategy × OMN inclusion and OMN inclusion were not detected (P ≥ 0.12) for any variable, except for the percentage of heifers detected in estrus, which was greater (P = 0.01) for heifers supplemented with vs. without OMN. Total concentrate DM offered from days 0 to 100 and heifer BW on days 0 and 56 did not differ (P ≥ 0.49) between CON and SST heifers, but SST heifers were heavier (P ≤ 0.01) on days 100 and 210 compared with CON heifers. Body surface temperature on day 25 and plasma IGF-1 concentrations on day 75 were greater (P ≤ 0.04) for SST vs. CON heifers. Percentage of pubertal heifers, heifers detected in estrus, and pregnancy to AI did not differ (P = 0.36) between SST and CON heifers but the final pregnancy percentage was greater (P = 0.04) for SST vs. CON heifers. Thus, OMN supplementation decreased the intravaginal temperature of SST heifers but failed to improve their growth and reproduction, whereas the SST strategy improved body thermoregulation, growth, and final pregnancy percentage of heat-stressed Bos indicus-influenced beef heifers compared with a constant concentrate supplementation strategy.

In Bos taurus beef heifers, altering the timing of body weight (BW) growth pattern either reduced feed costs without decreasing reproduction or enhanced reproduction without increasing feed costs. Moreover, supplementation of OmniGen-AF (OMN, a patented immunomodulatory feed ingredient) decreased internal body temperature in dairy and beef B. taurus cattle, with variable impacts on growth and reproduction. Combining both nutritional strategies for Bos indicus-beef heifers developed under heat stress conditions of tropical and subtropical environments has not been reported in the literature yet and was the main objective of the present study. For 100 d before the breeding season, heifers received either a constant supplementation amount or stair-step (SST) supplementation strategy (50 d of low followed by 50 d of high supplement amount), with or without OMN inclusion. Overall, OMN supplementation alleviated the internal body temperature of heifers but did not improve their growth and reproduction, whereas the SST strategy increased BW gain and final pregnancy percentage of B. indicus-influenced beef heifers under heat stress conditions.

Cleavage at the 586 amino acid caspase-6 site in mutant huntingtin influences caspase-6 activation in vivo.

Caspase cleavage of huntingtin (htt) and nuclear htt accumulation represent early neuropathological changes in brains of patients with Huntington's disease (HD). However, the relationship between caspase cleavage of htt and caspase activation patterns in the pathogenesis of HD remains poorly understood. The lack of a phenotype in YAC mice expressing caspase-6-resistant (C6R) mutant htt (mhtt) highlights proteolysis of htt at the 586 aa caspase-6 (casp6) site as a key mechanism in the pathology of HD. The goal of this study was to investigate how proteolysis of htt at residue 586 plays a role in the pathogenesis of HD and determine whether inhibiting casp6 cleavage of mhtt alters cell-death pathways in vivo. Here we demonstrate that activation of casp6, and not caspase-3, is observed before onset of motor abnormalities in human and murine HD brain. Active casp6 levels correlate directly with CAG size and inversely with age of onset. In contrast, in vivo expression of C6R mhtt attenuates caspase activation. Increased casp6 activity and apoptotic cell death is evident in primary striatal neurons expressing caspase-cleavable, but not C6R, mhtt after NMDA application. Pretreatment with a casp6 inhibitor rescues the apoptotic cell death observed in this paradigm. These data demonstrate that activation of casp6 is an early marker of disease in HD. Furthermore, these data provide a clear link between excitotoxic pathways and proteolysis and suggest that C6R mhtt protects against neurodegeneration by influencing the activation of neuronal cell-death and excitotoxic pathways operative in HD.

Orange peel products can reduce Salmonella populations in ruminants.

Salmonella can live undetected in the gut of food animals and be transmitted to humans. Animal diets can impact intestinal populations of foodborne pathogens, including Salmonella spp. Orange juice production results in a waste product, orange peel and orange pulp, which has a high nutritive value and is often included in cattle diets as a least-cost ration ingredient. Here we show that the inclusion of orange peel products reduced Salmonella Typhimurium populations in the gut of experimentally inoculated sheep. Sheep (n=24) were fed a cracked corn grain-based high grain diet that was supplemented with a 50%/50% (dry matter [DM], w/w) mixture of dried orange pellet and fresh orange peel to achieve a final concentration (DM, basis) of 0%, 10%, or 20% orange product (OP) for 10 days before inoculation with Salmonella Typhimurium. Sheep were experimentally inoculated with 10(10) colony forming units Salmonella Typhimurium, and fecal samples were collected every 24 h after inoculation. Sheep were humanely euthanized at 96 h after oral Salmonella inoculation. Populations of inoculated Salmonella Typhimurium were numerically reduced by OP treatment throughout the gastrointestinal tract, and this reduction only reached significant levels in the cecum (p<0.05) of sheep fed 10% OP diets. Apparent palatability issues decreased the consumption of OP in sheep fed 20% OP to intake levels below that of 10% OP (approximately 7% dry matter intake [DMI]/d feed refusal), thereby reducing the potential effects of OP feeding at this higher level. Our results demonstrate that orange peel and pellets are environmentally friendly and low-cost products that can be used as a pre-harvest intervention as part of an integrated pathogen reduction scheme.

Influence of Yeast Products on Modulating Metabolism and Immunity in Cattle and Swine.

Nutritional supplementation has been used by livestock producers for many years in order to increase animal performance, improve animal health, and reduce negative effects associated with enteric and/or respiratory pathogens. Supplements such as yeast and yeast-based products have broad applications across many livestock production systems, including poultry, aquaculture, cattle, and swine and have been shown to benefit animal production at various stages. These benefits include improvement in milk production, weight gain and feed conversion, as well as immune function. Initial research into the mode of action for these effects has focused on stimulation of the immune system by the ß-glucan fractions of yeast. However, emerging studies have revealed that some of the beneficial effects of yeast products may stem from altering metabolism, including the availability of glucose and fatty acids. These changes in metabolism, and potentially energy availability, may partially explain differences in immune function observed in yeast-supplemented livestock, as the energy demands of an activated immune system are extremely high. Thus, this paper explores the influence of yeast products on metabolism in cattle and swine, and how changes in metabolism and energy availability may contribute to improvements in immune function in supplemented animals.