Effects of dietary energy and starch concentrations for newly received feedlot calves: I. Growth performance and health. (1/18)

Crossbred calves (n = 572; initial BW = 186 +/- 27 kg) purchased from northern Texas, Arkansas, and southeast Oklahoma auction markets were delivered to the Willard Sparks Beef Research Center, Stillwater, OK, and used to study the effects of dietary energy and starch concentrations on performance and health of newly received feedlot calves during a 42-d receiving period. On arrival, calves were assigned randomly to one of two dietary energy levels (0.85 or 1.07 Mcal NEg/kg DM) and one of two dietary starch levels (34 or 48% of ME from starch) in a 2 x 2 factorial arrangement of treatments. Cattle were weighed and serum samples were collected on d 0, 7, 14, 28, and 42. Individual animal records of morbidity were kept for all cases of respiratory and other disease. Nasal swabs were collected from each morbid animal and cultured for upper-respiratory pathogens. There were no energy x starch level interactions for performance or health response variables. Daily gain (1.14 kg/d) and gain efficiency (ADG:DMI = 0.179) were not affected by increasing dietary energy or starch concentrations. Calves fed low-energy diets consumed (P < 0.05) more DM. No difference (P = 0.54) was detected in morbidity for calves fed high-energy (62.4% calves treated) compared with low-energy (65.8% calves treated) diets; however, calves fed the high-starch diets had numerically (P = 0.11) greater morbidity than calves fed low-starch diets (68.8 vs. 59.4% calves treated, respectively). There were no energy or starch effects on Mannheimia haemolytica or Pasteurella multocida antibody titers; however, day effects (P < 0.02) occurred. On d 7, 14, and 28, calves had antibody titers for P. multocida that were greater (P < 0.05) than titers on d 0. In addition, calves had greater antibody titers to M. haemolytica on d 7 and 14 than on d 0. Nasal swabs revealed that calves fed the high-energy diets tended (P = 0.06) to have a lower percentage of morbid calves with P. multocida during the first antimicrobial treatment and a lower percentage of Haemophilus somnus isolates during the first (P = 0.01) and second (P = 0.06) antimicrobial treatments than calves fed the low-energy diets. Although animal performance was not influenced, the present data suggest that feeding the high-energy diet decreased the percentage of P. multocida and H. somnus pathogens in calves that received one or more antimicrobial treatments.  (+info)

Immunohistochemical study of Hemophilus somnus, Mycoplasma bovis, Mannheimia hemolytica, and bovine viral diarrhea virus in death losses due to myocarditis in feedlot cattle. (2/18)

The purpose of this study was to determine the presence of Hemophilus somnus, Mycoplasma bovis, Mannheimia hemolytica, and bovine viral diarrhea virus (BVDV) in lesional tissues of feeder calves dying with myocarditis. Tissues from the heart and lungs of 92 calves dying with myocarditis in Alberta feedlots were immunohistochemically stained for the antigens of these agents. Tissues from 44 calves dying from noninfectious causes and 35 calves dying with pneumonia were tested as controls. Hemophilus somnus was found in cardiac lesions in the majority of myocarditis cases (70/92). Mycoplasma bovis was concurrently demonstrated in the hearts of 4/92 affected calves. No bacterial pathogens were found in heart tissues from the control groups of calves. Bovine viral diarrhea virus was demonstrated in the tissues of 4/92 myocarditis cases compared with those of 13/35 calves dying from pneumonia and 0/44 calves dying from noninfectious causes. The results demonstrate that H. somnus is the principle pathogen associated with myocarditis in feedlot calves and that the presence of BVDV is more common in these calves compared with calves dying of noninfectious causes. The findings also suggest that BVDV is an important pathogen in calves dying with gross postmortem lesions of pneumonia.  (+info)

Haemophilus somnus possesses two systems for acquisition of transferrin-bound iron. (3/18)

Haemophilus somnus strain 649 was found to acquire iron from ovine, bovine, and goat transferrins (Tfs). Expression of Tf receptors, as evaluated by solid-phase binding assays, required the organisms to be grown under iron-restricted conditions in the presence of Tf. Competition binding assays revealed the presence of two distinct Tf-binding receptor systems, one specific for bovine Tf and the other capable of binding all three ruminant Tfs. Affinity isolation procedures using total membranes yielded three putative bovine Tf-binding polypeptides and one putative ovine and goat Tf-binding polypeptide. PCR amplification followed by DNA sequence analyses revealed that H. somnus strain 649 possesses genes that encode a bipartite TbpA-TbpB receptor along with a homolog of the Histophilus ovis single-component TbpA receptor. Expression of TbpB and the single-component TbpA would appear to be subject to a form of phase variation involving homopolymeric nucleotide tracts within the structural genes.  (+info)

Stimulation of P2X receptors enhances lipooligosaccharide-mediated apoptosis of endothelial cells. (4/18)

Exposure of endothelial cells to lipid A-containing molecules, such as lipopolysaccharide (LPS) or lipooligosaccharide (LOS), causes the release of purinergic compounds [e.g., adenosine 5'-triphosphate (ATP)] and can lead to apoptosis. The P2X family of purinergic receptors (e.g., P2X(7)) has been reported to modulate LPS signaling events and to participate in apoptosis. We investigated the role that P2X receptors play in the apoptosis that follows exposure of bovine endothelial cells to Haemophilus somnus LOS. Addition of P2X inhibitors, such as periodate-oxidized ATP (oATP) or pyridoxal-phosphate-6-azophenyl-2',4'-disulfonic acid tetrasodium, significantly reduced LOS-induced apoptosis. Incubation of endothelial cells with apyrase, which degrades ATP, diminished LOS-induced apoptosis of endothelial cells. Concomitant addition of P2X agonists [e.g., 2',3'-(4-benzoyl)-benzoyl ATP or ATP] to LOS-treated endothelial cells significantly enhanced caspase-3 activation. The P2X antagonist oATP significantly blocked caspase-8 but not caspase-9 activation in LOS-treated endothelial cells. Together, these data indicate that stimulation of P2X receptors enhances LOS-induced apoptosis of endothelial cells, possibly as a result of endogenous release of ATP, which results in caspase-8 activation.  (+info)

Effects of sub-minimum inhibitory concentration antibiotic levels and temperature on growth kinetics and outer membrane protein expression in Mannheimia haemolytica and Haemophilus somnus. (5/18)

The objective of this study was to determine the effects of sub-minimum inhibitory concentrations (sub-MICs) of 2 veterinary antibiotic preparations, chlortetracycline (CTC) and chlortetracycline-sulfamethazine (CTC + SMZ), on growth kinetics and outer membrane protein expression in Mannheimia haemolytica and Haemophilus somnus at normal and febrile body temperatures. Sub-minimum inhibitory concentrations of both antibiotics reduced the growth rates of M. haemolytica and H. somnus. Growth of both species was not inhibited when grown at 41 degrees C compared to 37 degrees C. There was no detectable consistent effect of antibiotic or temperature on outer membrane protein expression for either species. Our study indicates that sub-MIC levels of CTC and CTC + SMZ markedly impair growth of clinical M. haemolytica and H. somnus isolates, potentially allowing more effective host clearance during infection.  (+info)

Generation and molecular characterization of new temperature-sensitive plasmids intended for genetic engineering of Pasteurellaceae. (6/18)

Temperature-sensitive (TS) plasmids were generated through chemical mutagenesis of a derivative of the streptomycin resistance parent plasmid pD70, isolated from Mannheimia hemolytica serotype 1. Three TS plasmids which failed to replicate at or above 42 degrees C in M. hemolytica but which were fully functional below 31 degrees C were selected for further analysis. Two of the TS plasmids were shown by sequencing to possess unique single-base-pair mutations. The third TS plasmid contained a unique base pair substitution and a second mutation that had been previously identified. These mutations were clustered within a 200-bp region of the presumed plasmid origin of replication. Site-directed single-nucleotide substitutions were introduced into the wild-type pD70 origin of replication to confirm that mutations identified by sequencing had conferred thermoregulated replication. Deletion analysis on the wild-type pD70 plasmid replicon revealed that approximately 720 bp are necessary for plasmid maintenance. Replication of the TS plasmids was thermoregulated in Pasteurella multocida and Haemophilus somnus as well. To consistently transform H. somnus with TS plasmid, in vitro DNA methylation with commercially available HhaI methyltransferase was necessary to protect against the organism's restriction enzyme HsoI (recognition sequence 5'-GCGC-3') characterized herein.  (+info)

Construction of in-frame aroA deletion mutants of Mannheimia haemolytica, Pasteurella multocida, and Haemophilus somnus by using a new temperature-sensitive plasmid. (7/18)

A temperature-sensitive (TS) plasmid was generated from the endogenous streptomycin resistance plasmid of Mannheimia hemolytica and used to engineer in-frame aroA deletion mutants of Mannheimia hemolytica, Pasteurella multocida, and Haemophilus somnus. TS replacement plasmids carrying in-frame aroA deletions were constructed for each target species and introduced into host cells by electroporation. After recovery in broth, cells were spread onto plates containing antibiotic and incubated at 30 degrees C, the permissive temperature for autonomous plasmid replication. Transfer of transformants to selective plates cultured at a nonpermissive temperature for plasmid replication selected for single-crossover mutants consisting of replacement plasmids that had integrated into host chromosomes by homologous recombination. Transfer of the single-crossover mutants back to a permissive temperature without antibiotic selection drove plasmid resolution, and, depending on where plasmid excision occurred, either deletion mutants or wild-type cells were generated. The system used here represents a broadly applicable means for generating unmarked mutants of Pasteurellaceae species.  (+info)

Haemophilus somnus (Histophilus somni) in bighorn sheep. (8/18)

Respiratory disease and poor lamb recruitment have been identified as limiting factors for bighorn-sheep populations. Haemophilus somnus (recently reclassified as Histophilus somni) is associated with respiratory disease in American bison, domestic sheep, and cattle. It is also harbored in their reproductive tracts and has been associated with reproductive failure in domestic sheep and cattle. Therefore, reproductive tract and lung samples from bighorn sheep were evaluated for the presence of this organism. Organisms identified as H. somnus were isolated from 6 of 62 vaginal but none of 12 preputial swab samples. Antigen specific to H. somnus was detected by immunohistochemical study in 4 of 12 formalin-fixed lung tissue samples of bighorn sheep that died with evidence of pneumonia. Notably, H. somnus was found in alveolar debris in areas of inflammation. The 6 vaginal isolates and 2 H. somnus isolates previously cultured from pneumonic lungs of bighorn sheep were compared with 3 representative isolates from domestic sheep and 2 from cattle. The profiles of major outer membrane proteins and antigens for all of the isolates were predominantly similar, although differences that may be associated with the host-parasite relationship and virulence were detected. The DNA restriction fragment length profiles of the bighorn-sheep isolates had similarities not shared with the other isolates, suggesting distinct phylogenetic lines. All of the isolates had similar antimicrobial profiles, but the isolates from the bighorn sheep produced less pigment than those from the domestic livestock, and growth of the former was not enhanced by CO2. Wildlife biologists and diagnosticians should be aware of the potential of these organisms to cause disease in bighorn sheep and of growth characteristics that may hinder laboratory detection.  (+info)