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1.  Susceptibility of goats and calves after experimental inoculation or contact exposure to a Canadian strain of Mycoplasma mycoides subsp. mycoides isolated from a goat. 
Transmissibility of Mycoplasma mycoides subsp. mycoides infection from experimentally inoculated goats to other goats and calves was studied. Eight goats and six calves were housed in an 18 m2 room. Six of the goats were inoculated endobronchially with strain D44 isolated from a natural case of polyarthritis in Ontario. These six goats died within a week of Mycoplasma septicemia. The two contact goats or the six calves never showed signs of disease and M. mycoides subsp. mycoides was not recovered from these animals. The contact goats and four calves were killed 25 days after exposure. They were all seronegative, M. mycoides subsp. mycoides was not recovered at necropsy and none had pathomorphological changes attributable to this Mycoplasma. The two remaining calves were inoculated endobronchially with 10(9) CFU of strain D44 and observed for 20 days. They never showed signs of disease and did not have significant lesions at necropsy. Both developed a significant serological response to M. mycoides subsp. mycoides, although this organism was not recovered during the experimental period or at necropsy. This study did not provide evidence for transmission of M. mycoides subsp. mycoides from endobronchially inoculated goats to contact goats or calves and endobronchially inoculated calves did not develop pneumonia. This would suggest that the infection of the goat population in Canada with this pathogen would not be a significant threat to the cattle population.
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PMCID: PMC1235981  PMID: 6365296
2.  Transmission of Bovine viral diarrhea virus 1b to susceptible and vaccinated calves by exposure to persistently infected calves 
Abstract
Bovine viral diarrhea virus (BVDV) persistently infected (PI) calves represent significant sources of infection to susceptible cattle. The objectives of this study were to determine if PI calves transmitted infection to vaccinated and unvaccinated calves, to determine if BVDV vaccine strains could be differentiated from the PI field strains by subtyping molecular techniques, and if there were different rates of recovery from peripheral blood leukocytes (PBL) versus serums for acutely infected calves. Calves PI with BVDV1b were placed in pens with nonvaccinated and vaccinated calves for 35 d. Peripheral blood leukocytes, serums, and nasal swabs were collected for viral isolation and serology. In addition, transmission of Bovine herpes virus 1 (BHV-1), Parainfluenza-3 virus (PI-3V), and Bovine respiratory syncytial virus (BRSV) was monitored during the 35 d observation period.
Bovine viral diarrhea virus subtype 1b was transmitted to both vaccinated and nonvaccinated calves, including BVDV1b seronegative and seropositive calves, after exposure to PI calves. There was evidence of transmission by viral isolation from PBL, nasal swabs, or both, and seroconversions to BVDV1b. For the unvaccinated calves, 83.2% seroconverted to BVDV1b. The high level of transmission by PI calves is illustrated by seroconversion rates of nonvaccinated calves in individual pens: 70% to 100% seroconversion to the BVDV1b. Bovine viral diarrhea virus was isolated from 45 out of 202 calves in this study. These included BVDV1b in ranch and order buyer (OB) calves, plus BVDV strains identified as vaccinal strains that were in modified live virus (MLV) vaccines given to half the OB calves 3 d prior to the study. The BVDV1b isolates in exposed calves were detected between collection days 7 and 21 after exposure to PI calves. Bovine viral diarrhea virus was recovered more frequently from PBL than serum in acutely infected calves. Bovine viral diarrhea virus was also isolated from the lungs of 2 of 7 calves that were dying with pulmonary lesions. Two of the calves dying with pneumonic lesions in the study had been BVDV1b viremic prior to death. Bovine viral diarrhea virus 1b was isolated from both calves that received the killed or MLV vaccines. There were cytopathic (CP) strains isolated from MLV vaccinated calves during the same time frame as the BVDV1b isolations. These viruses were typed by polymerase chain reaction (PCR) and genetic sequencing, and most CP were confirmed as vaccinal origin. A BVDV2 NCP strain was found in only 1 OB calf, on multiple collections, and the calf seroconverted to BVDV2. This virus was not identical to the BVDV2 CP 296 vaccine strain. The use of subtyping is required to differentiate vaccinal strains from the field strains. This study detected 2 different vaccine strains, the BVDV1b in PI calves and infected contact calves, and a heterologous BVDV2 subtype brought in as an acutely infected calf. The MLV vaccination, with BVDV1a and BVDV2 components, administered 3 d prior to exposure to PI calves did not protect 100% against BVDV1b viremias or nasal shedding. There were other agents associated with the bovine respiratory disease signs and lesions in this study including Mannheimia haemolytica, Mycoplasma spp., PI-3V, BRSV, and BHV-1.
PMCID: PMC1176294  PMID: 16187545
3.  Synovial immunoglobulin and antibody in vaccinated and nonvaccinated calves challenged with Mycoplasma bovis. 
Intravenous injection of Mycoplasma bovis produced in calves arthritis with synovial infiltration of lymphocytes, macrophages and neutrophils. Necrosis was observed focally around blood vessels. Joint spaces contained fibrinopurulent exudate. Parenterally vaccinated calves had a markedly reduced frequency of arthritic joints. Immunoglobulin classes and specific antibody in joint fluids were quantitatively less than in sera but significantly greater in arthritic than in normal joints. The possible mechanisms of induction of joint fluid antibody are discussed.
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PMCID: PMC1320127  PMID: 7272847
4.  A retrospective study of 29 cases of otitis media/interna in dairy calves 
The Canadian Veterinary Journal  2012;53(9):957-962.
Epidemiological data, clinical findings, laboratory data, medical imaging, and outcomes were reviewed in 29 dairy calves with otitis media/interna. Age at admission ranged from 1 to 24 wk. The majority of calves were referred during winter. Clinical signs included drooping ear, ptosis, head tilt, abnormal nystagmus, strabismus, dysphagia, regurgitation, stiff neck, opisthotonos, facial hyperesthesia, and purulent aural discharge. Intranasal endoscopic examination of 5 animals revealed nasopharyngeal collapse in 4. Cerebrospinal fluid (CSF) was abnormal in all of 7 cases. Mycoplasma bovis was cultured from all but 1 case with external ear or tympanic bullae samples (n = 12), and Mycoplasma arginini was cultured from the remaining ear sample. Radiographs of the tympanic bullae were performed in 24 calves, tomodensitometry (CT) in 3 calves and ultrasound in 4 calves. According to medical imaging techniques or necropsy, 69% of the cases were classified as chronic. Mean duration of treatment was 23.3 d. The rate of clinical recovery was 75%.
PMCID: PMC3418781  PMID: 23450859
5.  Capsulelike surface material of Mycoplasma dispar induced by in vitro growth in culture with bovine cells is antigenically related to similar structures expressed in vivo. 
Infection and Immunity  1991;59(9):3119-3125.
Electron microscopy has been used to show that Mycoplasma dispar produces an external capsulelike material in vivo that has an affinity for both ruthenium red and polycationic ferritin. This extracellular material is lost upon passage in culture medium but can be regained with a single passage on bovine lung fibroblast (BLF) cells. To confirm that the extracellular material associated with cell-grown mycoplasmas was the same as that observed in infected calves, rabbit antibodies were produced to purified capsulelike material isolated by protease digestion of cell-grown organisms. These antibodies bound to capsulelike material on the surface of M. dispar cells colonizing the bronchial epithelium of infected calves and to capsulelike material from cell-grown mycoplasmas. Calves infected with M. dispar produced antibodies in lung secretions that were capable of binding to the purified capsulelike material. The Fab fragments of rabbit antibodies to in vitro-produced capsulelike material could block this binding, indicating that the capsulelike material was similar in both in vivo-grown and cell-grown organisms. The carbohydrate nature of the capsular material suggested by the ruthenium red and polycationic staining characteristics was confirmed by its binding to Ricinus communis agglutinin, a galactose-specific lectin. These studies confirm that capsule material produced during infections with M. dispar share antigenic determinants with the material produced under in vitro conditions and that association with mammalian cells induces production of this material.
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PMCID: PMC258142  PMID: 1715319
6.  The effect of antimicrobial treatment and preventive strategies on bovine respiratory disease and genetic relatedness and antimicrobial resistance of Mycoplasma bovis isolates in a western Canadian feedlot 
The Canadian Veterinary Journal  2013;54(12):1146-1156.
Feedlot calves (n = 3784) were systematically randomized and allocated in a 2 × 2 factorial study to receive metaphylactic oxytetracycline (OTC) on arrival or no antimicrobial, as well as florfenicol once subcutaneously or twice intramuscularly (48 h apart) if diagnosed with bovine respiratory disease (BRD). Calves of different treatment groups were comingled and followed from placement to re-implantation (~100 days). Animals receiving OTC had a reduced risk of BRD, an increased risk of arthritis, and no significant differences in average daily gain, BRD relapse, overall mortality, or BRD mortality. There were no significant differences between treatment protocols. Deep nasal swabs (n = 233) taken at arrival (n = 122), treatment (n = 77), and swabs from lungs and joints at postmortem (n = 34) were cultured for Mycoplasma bovis from 61 animals ill or dying of chronic pneumonia and arthritis and from 61 healthy calves. There was significant variation in diversity among isolates (n = 51) between study years and different cattle. Metaphylaxis or antimicrobial treatment did not affect the diversity of isolates. Except for tilmicosin, isolates were largely susceptible to tested antimicrobials.
PMCID: PMC3831389  PMID: 24293675
7.  A Mycoplasma Isolated from Cattle with Infectious Bovine Keratoconjunctivitis 
A mycoplasma has been recovered from the eyes of calves in two naturally-occurring outbreaks of infectious bovine keratoconjunctivitis; also from a third group of calves accidentally exposed to an animal which had ocular exudates from one of the outbreaks instilled into its eyes.
The severity of the ocular lesions in infectious bovine keratoconjunctivis outbreaks may be related to a mixed infection with the mycoplasma and Moraxella bovis.
Preliminary typing studies indicate the mycoplasma is not serologically related to any known bovine mycoplasma.
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PMCID: PMC1319444  PMID: 4243033
8.  Histological features of respiratory epithelium of calves held at differing temperature and humidity. 
The effect of ambient temperature and humidity on the structure of respiratory epithelium of calves was studied. Four calves of each of three experiments were acclimatized to a nonoperational environmental chamber for six days and then exposed to constant extremes of temperatures and relative humidity of one of 30 degrees C --35%, or 27 degrees C--92%, or 5 degrees C--92% respectively in this chamber for eight days each. Five calves (3 and 2) were similarly acclimatized then exposed to 1 degrees C--40%. Nasal swabs were taken from all animals at regular intervals. Swabs of three animals yielded Mycoplasma spp. and one swab yielded the virus of infectious bovine rhinotracheitis. Detailed histological studies of respiratory epithelium of nose, trachea, major bronchus and terminal bronchioli were conducted at four sites. Goblet cells were least in calves held in hot and dry air; calves held in dry air had the least polymorphonuclear cells and the greatest prevalence of hypochromatic cell layers and vacuolation of epithelial cells. Differences between experiments were evident most for sites of trachea and major bronchus.
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PMCID: PMC1277734  PMID: 922554
9.  Immunohistochemical study of Hemophilus somnus, Mycoplasma bovis, Mannheimia hemolytica, and bovine viral diarrhea virus in death losses due to myocarditis in feedlot cattle 
The Canadian Veterinary Journal  2004;45(3):231-234.
Abstract
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.
PMCID: PMC548609  PMID: 15072195
10.  The incidence of Mycoplasma dispar, Ureaplasma and conventional Mycoplasma in the pneumonic calf lung. 
This report describes the incidence of Mycoplasma dispar, ureaplasma and conventional (large colony) mycoplasma isolated from the pneumonic lungs of groups of young calves and the identification to species level of mycoplasmas in mixed populations with the aid of the indirect fluorescent antibody test. Pneumonic lung tissue yielded one or more mycoplasma species from 88% of the 153 calves cultured. The mycoplasmas identified and percent of the calves with lungs positive for each species were: M. dispar (56%), ureaplasma (44%), Mycoplasma bovis (37%), Mycoplasma arginini (33%) and Mycoplasma bovirhinis (23%). Conventional mycoplasmas isolated from two calves (1%) could not be identified using the antisera available.
PMCID: PMC1320011  PMID: 398234
11.  The association of titers to Haemophilus somnus, and other putative pathogens, with the occurrence of bovine respiratory disease and weight gain in feedlot calves. 
Serum samples were obtained from 602 calves (from 19 groups in four feedlots: three in Ontario, and one in Alberta) upon arrival at the feedlot and 28 d later. Of these calves, 202 developed bovine respiratory disease (BRD) and 400 did not develop BRD. Based on high antibody titers noted upon arrival, we infer that most calves were exposed to Haemophilus somnus prior to arrival at the feedlot. Within a group, calves with high titers on arrival had a reduced risk of developing BRD later. Most calves did not experience titer increases after arrival; however, calves that had stable or increasing titers had a relatively low risk of contracting BRD. The calves at greatest risk of BRD were those with titers on arrival of less than 6.8 units and subsequent titer decreases of more than 1 unit. The effects of both the titer on arrival and the titer change after arrival were stable when the serologic effects of a number of viruses and Mycoplasma agents were considered. Neither antibody titer on arrival nor titer change was related to weight gain differences among calves. Calves with BRD or calves with lower weight on arrival had decreased weight gains in the first 28-day feeding period. The high titers on arrival may have protected most calves against further infection with H. somnus. However, since the calves that developed BRD had large titer increases to a number of viruses and to Pasteurella haemolytica, while having decreased antibody titers to H. somnus, we infer that the existing antibodies were "used up" in combatting the agents, including H. somnus, which may have "caused" the BRD. Calves which were able to increase their antibody levels to H. somnus tended to have a reduced risk of BRD.
PMCID: PMC1189492  PMID: 9798091
12.  The Complete Genome Sequence of Mycoplasma bovis Strain Hubei-1 
PLoS ONE  2011;6(6):e20999.
Infection by Mycoplasma bovis (M. bovis) can induce diseases, such as pneumonia and otitis media in young calves and mastitis and arthritis in older animals. Here, we report the finished and annotated genome sequence of M. bovis strain Hubei-1, a strain isolated in 2008 that caused calf pneumonia on a Chinese farm. The genome of M. bovis strain Hubei-1 contains a single circular chromosome of 953,114 bp with a 29.37% GC content. We identified 803 open reading frames (ORFs) that occupy 89.5% of the genome. While 34 ORFs were Hubei-1 specific, 662 ORFs had orthologs in the M. bovis type strain PG45 genome. Genome analysis validated lateral gene transfer between M. bovis and the Mycoplasma mycoides subspecies mycoides, while phylogenetic analysis found that the closest M. bovis neighbor is Mycoplasma agalactiae. Glycerol may be the main carbon and energy source of M. bovis, and most of the biosynthesis pathways were incomplete. We report that 47 lipoproteins, 12 extracellular proteins and 18 transmembrane proteins are phase-variable and may help M. bovis escape the immune response. Besides lipoproteins and phase-variable proteins, genomic analysis found two possible pathogenicity islands, which consist of four genes and 11 genes each, and several other virulence factors including hemolysin, lipoate protein ligase, dihydrolipoamide dehydrogenase, extracellular cysteine protease and 5′-nucleotidase.
doi:10.1371/journal.pone.0020999
PMCID: PMC3120828  PMID: 21731639
13.  Response of the respiratory tract of calves kept at controlled climatic conditions to bovine Herpesvirus 1 in aerosol. 
Seven experiments with four calves each were conducted in which the calves spent at least four days of adaptation in an environmental chamber and then were subjected to climatic stress in the form of a number of constant ambient temperature and humidity combinations. On the second day of climatic stress the calves were individually exposed to measured numbers of infectious units of bovine herpesvirus 1 (BHV1, virus of infectious bovine rhinotrachetis) in aerosol. The calves were killed seven or eight days later. Mycoplasma were found in some nasal swabs and in one lung. Certain bacteria but no Pasteurella were often isolated from the lungs. Bovine herpesvirus 1 was isolated from chamber air and from most postinoculation nasal swabs, tracheas and lungs. The number of macro- and microscopic lesions did not appear to be influenced by the climatic conditions of the experiments. The histopathological changes in epithelium at all levels of the respiratory tract were described in detail.
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PMCID: PMC1277609  PMID: 208734
14.  Naturally occurring and experimentally induced mycoplasmal arthritis of cattle. 
Journal of Clinical Microbiology  1975;2(3):165-168.
Mycoplasma agalactiae subsp. bovis strain Iowa 1136 was isolated from synovial fluids of a clinical case of arthritis in cattle on pasture in Iowa. When given to calves and cows by intra-articular or intravenous injection, it caused severe and persistent joint infections with fever, lameness, and swelling of the affected joints, plus synovitis, tendonitis, and fibrinous-purulent synovial fluids of high protein content. Intramammary administration of the organism caused severe mastitis. Calves nursing the cows developed severe mycoplasmal arthritis.
PMCID: PMC274164  PMID: 1176623
15.  Infectious bovine keratoconjunctivitis I. Experimental production. 
One or both eyes of 20 calves were inoculated one or more time with variou(s combinations of microorganism (live oor killed Moraxella bovis, infectious bovine rhinotracheitis virus, bovine adenovirus, bovine parainfluenza-3 virus and Mycoplasma bovoculi) by conjunctival instillation or direct inoculation of the conjunctivea or cornea. The eyes of all the calves received natural or artificial ultraviolet irradiation. Neither the adenovirus nor parainfluenza-3 virus became established in the eye or produced keratoconjunctivitis. Both M. bovis and infectious bovine rhinotracheitis virus became established in the bovine eye and produced disease. Subconjunctival or intracorneal inoculation of M. bovis caused a severe disease, simulating natural infectious bovine keratoconjunctivitis. Only the intracorneal inoculation of mycoplasma produced severe keratoconjunctivits. Eyes that on initial exposure to M. bovis became severly inflamed were more resistant to a second or third exposure to M. bovis, presumably by enhanced local defence mechanisms.
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PMCID: PMC1277410  PMID: 163126
16.  Development of a Recombinant Protein-Based Enzyme-Linked Immunosorbent Assay for Diagnosis of Mycoplasma bovis Infection in Cattle 
Mycoplasma bovis causes a range of diseases in cattle, including mastitis, arthritis, and pneumonia. However, accurate serological diagnosis of infection remains problematic. The studies described here aimed to identify an antigen that might be used to develop a more specific and sensitive diagnostic assay. A 226-kDa immunogenic protein was consistently detected in Western blots by antibodies in sera from calves experimentally infected with M. bovis. This protein was shown to be a membrane protein with lipase activity and was named mycoplasma immunogenic lipase A (MilA). Different regions of MilA were expressed in Escherichia coli as glutathione S-transferase (GST) fusion proteins and recombinant products from the amino-terminal end shown to have strong immunoreactivity with M. bovis-specific bovine sera. The most immunoreactive fusion protein, GST-MilA-ab, was used to develop indirect IgM and IgG enzyme-linked immunosorbent assays (ELISAs). The IgM ELISA detected M. bovis-specific IgM antibody 2 weeks after infection with 97.1% sensitivity and had a specificity of 63.3%, while the IgG ELISA detected M. bovis-specific IgG 3 weeks after infection with 92.86% sensitivity and had a specificity of 98.7%, demonstrating that the IgG ELISA has potential for use as a sensitive and specific assay for detecting infection in cattle.
doi:10.1128/CVI.00670-13
PMCID: PMC3910936  PMID: 24334686
17.  Mycoplasma alkalescens demonstrated in bronchoalveolar lavage of cattle in Denmark 
Mycoplasma alkalescens is an arginine-metabolizing mycoplasma, which has been found in association with mastitis and arthritis in cattle. Routine bacteriological examination of 17 bronchoalveolar lavage samples from calves with pneumonia in a single herd in Denmark, identified M. alkalescens in eight samples. The organism was found as a sole bacterilogical findings in five of the samples as well as in combination with Mannheimia haemolytica, Haemophilus somni and Salmonella Dublin. This is the first report of isolation of M. alkalescens in Denmark.
doi:10.1186/1751-0147-49-2
PMCID: PMC1766361  PMID: 17204146
18.  Human Diseases Associated with Mycoplasmas—With an Appendix on Simple Culture Techniques 
California Medicine  1972;116(5):1-7.
The mycoplasmas (formerly called pleuropneumonia-like organisms, or pplo) are a group of pleomorphic micro-organisms characterized by lack of cell wall and ability to form colonies on agar resembling tiny fried eggs. They have been recognized as pathogens of lower mammals since 1898. Of the more than 40 known veterinary species, many are pathogens, commonly causing pneumonia, arthritis or arteritis. Of the mycoplasmas found in man, Mycoplasma pneumoniae is the only well established human pathogen. It is responsible for a variety of respiratory syndromes, of which the most frequently recognized is cold agglutinin-positive atypical pneumonia. Hematologic, neurologic and dermatologic complications of this infection have been noted. M. hominis has been implicated as a causative factor in various febrile complications of pregnancy, such as septic abortion and amnionitis. T-strain mycoplasmas are ubiquitous in the human genitourinary tract, but attempts to link their presence to disease have thus far been unsuccessful. Mycoplasmas also have been associated with neoplastic disease and with rheumatoid arthritis. The validity of these latter findings is unclear, and additional study is needed.
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PMCID: PMC1518391  PMID: 4565394
19.  Sequence and TnphoA analysis of a Mycoplasma hyorhinis protein with membrane export function. 
Journal of Bacteriology  1991;173(6):2035-2044.
Proteins translocated across the single plasma membrane of mycoplasmas (class Mollicutes) represent important components likely to affect several interactions of these wall-less microbes with their respective hosts. However, identification and functional analysis of such proteins is hampered by the lack of mutational systems in mycoplasmas and by a perceived limitation in translating recombinant mycoplasma genes containing UGA (Trp) codons in other eubacteria. Here we directly analyze a gene encoding a Mycoplasma hyorhinis protein capable of promoting its membrane translocation. It was initially detected by screening a recombinant phage genomic library with antibody from a host with M. hyorhinis-induced arthritis and was localized by Tn5 and deletion mutations affecting expression of antigenic translational products. Sequence analysis of the isolated gene predicted a hydrophilic protein, P101, containing three UGA codons and a putative signal peptide with an uncharacteristic cluster of positively charged amino acids near its C terminus. Nevertheless, lambda::TnphoA transposon mutagenesis of an Escherichia coli plasmid bearing the p101 gene resulted in p101::TnphoA fusions expressing products that could translocate as much as 48 kDa of the P101 sequence (up to the first UGA codon) across the E. coli plasma membrane. Fusion proteins containing mature P101 sequences expressed mycoplasma epitopes and were found by cell fractionation and detergent phase partitioning to be integral membrane proteins in E. coli, suggesting a lack of signal peptide cleavage in this system. Importantly, identification of P101 by direct analysis of its export function relied neither on prior identification of the mycoplasmal product nor on complete expression of the product from the cloned mycoplasma gene.
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PMCID: PMC207738  PMID: 1848219
20.  Chronic pneumonia in calves after experimental infection with Mycoplasma bovis strain 1067: Characterization of lung pathology, persistence of variable surface protein antigens and local immune response 
Background
Mycoplasma bovis is associated with pneumonia in calves characterized by the development of chronic caseonecrotic lesions with the agent persisting within the lesion. The purposes of this study were to characterize the morphology of lung lesions, examine the presence of M. bovis variable surface protein (Vsp) antigens and study the local immune responses in calves after infection with M. bovis strain 1067.
Methods
Lung tissue samples from eight calves euthanased three weeks after experimental infection with M. bovis were examined by bacteriology and pathology. Lung lesions were evaluated by immunohistochemical (IHC) staining for wide spectrum cytokeratin and for M. bovis Vsp antigens and pMB67 antigen. IHC identification and quantitative evaluation of CD4+ and CD8+ T lymphocytes and immunoglobulin (IgG1, IgG2, IgM, IgA)-containing plasma cells was performed. Additionally, expression of major histocompatibility complex class II (MHC class II) was studied by IHC.
Results
Suppurative pneumonic lesions were found in all calves. In two calves with caseonecrotic pneumonia, necrotic foci were surrounded by epithelial cells resembling bronchial or bronchiolar epithelium. In all calves, M. bovis Vsp antigens were constantly present in the cytoplasm of macrophages and were also present extracellularly at the periphery of necrotic foci. There was a considerable increase in numbers of IgG1- and IgG2-positive plasma cells among which IgG1-containing plasma cells clearly predominated. Statistical evaluation of the numbers of CD4+ and CD8+ T cells, however, did not reveal statistically significant differences between inoculated and control calves. In M. bovis infected calves, hyperplasia of bronchus-associated lymphoid tissue (BALT) was characterized by strong MHC class II expression of lymphoid cells, but only few of the macrophages demarcating the caseonecrotic foci were positive for MHC class II.
Conclusions
The results from this study show that infection of calves with M. bovis results in various lung lesions including caseonecrotic pneumonia originating from bronchioli and bronchi. There is long-term persistence of M. bovis as demonstrated by bacteriology and immunohistochemistry for M. bovis antigens, i.e. Vsp antigens and pMB67. The persistence of the pathogen and its ability to evade the specific immune response may in part result from local downregulation of antigen presenting mechanisms and an ineffective humoral immune response with prevalence of IgG1 antibodies that, compared to IgG2 antibodies, are poor opsonins.
doi:10.1186/1751-0147-54-9
PMCID: PMC3287114  PMID: 22305416
Cattle, Mycoplasma bovis; pneumonia; immunoglobulins; CD4+ T cells; CD8+ cells; MHC class II
21.  Platelet aggregation responses and virus isolation from platelets in calves experimentally infected with type I or type II bovine viral diarrhea virus. 
Altered platelet function has been reported in calves experimentally infected with type II bovine viral diarrhea virus (BVDV). The purpose of the present study was to further evaluate the ability of BVDV isolates to alter platelet function and to examine for the presence of a virus-platelet interaction during BVDV infection. Colostrum-deprived Holstein calves were obtained immediately after birth, housed in isolation, and assigned to 1 of 4 groups (1 control and 3 treatment groups). Control calves (n = 4) were sham inoculated, while calves in the infected groups (n = 4 for each group) were inoculated by intranasal instillation with 10(7) TCID50 of either BVDV 890 (type II), BVDV 7937 (type II), or BVDV TGAN (type I). Whole blood was collected prior to inoculation (day 0) and on days 4, 6, 8, 10, and 12 after inoculation for platelet function testing by optical aggregometry by using adenosine diphosphate and platelet activating factor. The maximum percentage aggregation and the slope of the aggregation curve decreased over time in BVDV-infected calves; however, statistically significant differences (Freidman repeated measures ANOVA on ranks, P < 0.05) were only observed in calves infected with the type II BVDV isolates. Bovine viral diarrhea virus was not isolated from control calves, but was isolated from all calves infected with both type II BVDV isolates from days 4 through 12 after inoculation. In calves infected with type I BVDV, virus was isolated from 1 of 4 calves on days 4 and 12 after inoculation and from all calves on days 6 and 8 after inoculation. Altered platelet function was observed in calves infected with both type II BVDV isolates, but was not observed in calves infected with type I BVDV. Altered platelet function may be important as a difference in virulence between type I and type II BVDV infection.
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PMCID: PMC1189686  PMID: 11768131
22.  The association between serological evidence of mycoplasma infection and respiratory disease in feedlot calves. 
Calves from five Ontario feedlots were bled on arrival and approximately 28 days later. Calves treated during this interval for undifferentiated respiratory disease were classified as cases and untreated calves were classified as controls. Serum was titrated blindly for antibodies to Mycoplasma bovis and Mycoplasma dispar. Indirect hemagglutination titers of 1:20 or more were assumed to reflect recent or current exposure, whereas 1:10 or less were not. The titers to M. bovis increased in all feedlots indicating active infection. The initial titers to M. dispar were higher than the titers against M. bovis, yet they increased in all feedlots except one, suggesting widespread infection with this organism. There was an increased risk (although not statistically significant) of being treated if the titer against M. bovis rose during the period. Calves with low M. dispar titers on arrival were at increased risk of being treated and titer increases were strongly associated with treatment (statistically significant). Thus, the serological results indicate high prevalence of M. bovis and M. dispar in the feedlot calves and that calves with increasing titers in particular to M. dispar are at increased risk of being treated for respiratory disease.
PMCID: PMC1255186  PMID: 3756671
23.  Impact of respiratory disease, diarrhea, otitis and arthritis on mortality and carcass traits in white veal calves 
Background
Little is known on the effects of common calf diseases on mortality and carcass traits in the white veal industry (special-fed veal), a highly integrated production system, currently criticized for the intensive pro- and metaphylactic use of antimicrobials. The objective of the present study was to determine the impact of bovine respiratory disease (BRD), diarrhea, arthritis and otitis on the economically important parameters of mortality, hot carcass weight (HCW), carcass quality, fat cover and meat color. For this purpose, a prospective study on 3519 white veal calves, housed in 10 commercial herds, was conducted. Case definitions were based on clinical observation by the producers and written treatment records were used.
Results
Calves received oral antimicrobial group treatments in the milk during 25.2% of the production time on average. With an increasing percentage of the production cycle spent on oral antimicrobials, HCW reduced, whereas the odds for insufficient fat cover or an undesirable red meat color both decreased. Of the calves, 14.8%, 5.3%, 1.5% and 1.6% were individually diagnosed and treated for BRD, diarrhea, arthritis and otitis, respectively. Overall, 5.7% of the calves died and the mortality risk was higher in the first weeks after arrival. Calves that experienced one BRD episode showed a 8.2 kg reduction in HCW, a lower fat cover and an increased mortality risk (hazard ratio (HR) = 5.5), compared to calves which were not individually diagnosed and treated for BRD. With an increasing number of BRD episodes, these losses increased dramatically. Additionally, calves, which experienced multiple BRD episodes, were more likely to have poor carcass quality and an undesirable red meat color at slaughter. Arthritis increased the mortality risk (HR = 3.9), and reduced HCW only when associated with BRD. Otitis did only increase the mortality risk (HR = 7.0). Diarrhea severely increased the mortality risk (HR = 11.0), reduced HCW by 9.2 kg on average and decreased carcass quality.
Conclusions
Despite the massive use of group and individual treatments to alleviate the most prevalent health issues at the fattening period, the effects of BRD, diarrhea, otitis and arthritis on survival and performance are still considerable, especially in cases of chronic pneumonia with or without arthritis. Controlling calf health by effective preventive and therapeutic strategies and in particular the prevention of chronic BRD is key for the profitability of veal operations.
doi:10.1186/1746-6148-9-79
PMCID: PMC3639957  PMID: 23587206
Bovine respiratory disease; Carcass weight; Diarrhea; Economics; Mortality; Veal calves
24.  The Vlp system of Mycoplasma hyorhinis: combinatorial expression of distinct size variant lipoproteins generating high-frequency surface antigenic variation. 
Journal of Bacteriology  1991;173(15):4782-4793.
Isogenic populations of Mycoplasma hyorhinis undergo in vitro high-frequency phase variation in the expression of surface lipoproteins; these products also vary markedly in size through changes in periodic protein structure (R. Rosengarten and K.S. Wise, Science 247:315-318, 1990). In this report, we rigorously define three distinct translation products comprising the Vlp (variable lipoprotein) system of M. hyorhinis SK76 and establish parameters of Vlp structural diversity and expression that distinguish the Vlp system from previously described examples of antigenic variation. VlpA, VlpB, and VlpC are prominent amphiphilic membrane lipoproteins characterized by detergent-phase fractionation and metabolic labeling with [35S]cysteine and [3H]palmitate. VlpA is distinguished from VlpB and VlpC by its selective labeling with [35S]methionine; VlpB and VlpC are distinguished by specific epitopes defined by surface-binding monoclonal antibodies (MAbs); a third MAb defines a surface epitope shared by VlpB and VlpC (but absent from VlpA). Each Vlp displays 12 to 30 spontaneous size variant forms comprising a periodic ladder that could also be generated by partial trypsin digestion of individual Vlp size variants. Different periodic intervals within VlpB and VlpC further distinguish these two products structurally. Mycoplasma colony opacity correlates inversely with Vlp size. Each Vlp undergoes independent, oscillating high-frequency phase variation in isogenic populations and can be expressed individually or concomitantly with other Vlps in a noncoordinate manner. All seven possible combinations of these three products were observed; however, no variants were found that lacked a Vlp. High-frequency size variation of each Vlp superimposed on combinatorial diversity in Vlp expression yields greater than 10(4) possible structurally distinct Vlp mosaics, of which 104 were documented along with 24 of 42 possible transitions among the seven Vlp combinations. In addition to these features, VlpA, VlpB, and VlpC were specifically recognized by serum antibodies from swine with experimental M. hyorhinis SK76-induced arthritis, indicating expression and immunogenicity of Vlps in the natural host. The structure and variation of Vlps and their known involvement in MAb-mediated modulation of mycoplasma-infected host cell properties and mycoplasma killing are discussed in relation to the surface architecture and adaptive potential of the wall-less mycoplasmas.
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PMCID: PMC208157  PMID: 1856172
25.  Performance, survival, necropsy, and virological findings from calves persistently infected with the bovine viral diarrhea virus originating from a single Saskatchewan beef herd. 
Fifty-one calves from 652 cows and heifers that calved on a Saskatchewan ranch in 1992 were identified as persistently infected with bovine viral diarrhea virus (BVDV), based on virological and necropsy findings. Herd records suggested a further 20 calves that died between birth and weaning were probably also persistently infected. Subsequent to weaning, all surviving persistently infected calves were transferred to one pen in a 10,000 head commercial feedlot, to mimic normal management practice in western Canadian beef herds. On average, when compared with healthy, BVDV-negative herdmates, persistently infected calves were "poor doers" and had poor survivability, with only 4 persistently infected calves surviving to 1 year of age. There was no difference (P > 0.05) in survival between male and female persistently infected calves. The clinical, pathological, and virological findings from these persistently infected calves varied over time. The majority of persistently infected calves had gross pathological lesions at necropsy, consistent with mucosal disease. However, approximately 25% of the persistently infected calves had gross pneumonic lesions at necropsy, with no or only mild lesions of mucosal disease. A wide variety of other lesions were also noted in persistently infected calves at necropsy. Therefore, the possibility that BVDV-induced lesions can be misdiagnosed is very real. The results of this study indicate that persistent infection with BVDV should always be considered in calves with chronic ill thrift, chronic enteritis, or respiratory disease.
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PMCID: PMC1576666  PMID: 8993782

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