R L Walker

University of California, Davis, Davis, California, United States

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Publications (33)50.28 Total impact

  • L.V. Stamm · R.L. Walker · D.H. Read
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    ABSTRACT: The etiology of bovine ulcerative mammary dermatitis (UMD) is poorly characterized. The goal of this study was to genetically analyze spirochetes present in UMD lesions. DNA prepared from UMD lesion biopsies and from spirochetes cultured from the corresponding lesion biopsies was PCR amplified using primers for the 16S rDNA-tRNA(ile) intergenic spacer region (ISR) of Treponema 16S-23S rDNA. Analysis of cloned ISR amplicons from three cultivable UMD-associated spirochetes indicated that two isolates cluster closely with cultivable papillomatous digital dermatitis (PDD)-associated and human-associated Treponema phylotypes, while the remaining isolate is unique. Analysis of ISR amplicons from UMD lesion biopsies identified additional not-yet-cultivable Treponema phylotypes. Our results revealed the presence of a genetically diverse Treponema population in an UMD lesion.
    No preview · Article · Nov 2008 · Veterinary Microbiology
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    ABSTRACT: Epizootic bovine abortion (EBA), a tick-transmitted disease of pregnant cattle grazing foothill pastures, is a major cause of reproductive failure in California and adjacent states. Affected fetuses develop a chronic disease, resulting in late-term abortion or premature calving. Despite investigations spanning 50 years, to the authors' knowledge, the etiologic agent of EBA has not yet been isolated from affected fetuses or the tick vector. The diagnosis of EBA is based on gross and microscopic lesions. Recently, documentation that the etiologic agent is susceptible to antibiotics and identification of a unique 16S deltaproteobacterial rDNA gene sequence in 90% of thymus tissues from aborted fetuses have supported the role of a bacterial infection as the cause of EBA. To determine whether bacteria could be detected in the tissues, histochemical staining and immunohistochemical procedures were used on formalin-fixed, paraffin-embedded tissues. Use of a modified Steiner silver stain revealed small numbers of intracytoplasmic bacterial rods in 37 of 42 thymic samples from EBA-affected fetuses. Improved detection was achieved by use of immunohistochemical staining with serum from EBA-affected fetuses that resulted in detection of numerous bacterial rods in the cytoplasm of histiocytic cells in the thymus from all 42 EBA-affected fetuses. Immunohistochemical examination of additional tissues from 21 field and experimental EBA cases revealed positively stained intracytoplasmic bacterial rods in many organs with inflammatory lesions. Use of the modified Steiner stain and immunohistochemical staining of tissues from negative-control fetuses failed to reveal organisms. To the authors' knowledge, this is the first report to document morphologic evidence of a bacterium associated with the lesions of EBA.
    Full-text · Article · Feb 2006 · Journal of veterinary diagnostic investigation: official publication of the American Association of Veterinary Laboratory Diagnosticians, Inc
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    ABSTRACT: An immunohistochemistry (IHC) procedure for the detection of Campylobacter fetus antigens using an avidin-biotin complex technique was performed on formalin fixed bovine and ovine fetal tissues from 26 natural cases of Campylobacter spp. abortion (four ovine and 22 bovine). The species of Campylobacter isolated included C. fetus ssp. venerealis from 13 bovine fetuses, C. fetus ssp. fetus from two ovine and one bovine fetus, Campylobacter jejuni from seven bovine fetuses, Campylobacter lari from two ovine fetuses and an unspeciated Campylobacter species in one bovine fetus. Histologic lesions identified in the aborted fetuses included placentitis, serositis, pneumonia, gastroenteritis, hepatitis and encephalitis. Campylobacter fetus antigens were identified by IHC in 13 of 13 bovine fetuses from which C. fetus ssp. venerealis was isolated and in two of two ovine fetuses from which C. fetus ssp. fetus was isolated. The IHC stains were negative in tissues from seven bovine fetuses from which C. jejuni was isolated, one bovine fetus infected with C. fetus ssp. fetus, one bovine fetus infected with the unspeciated Campylobacter and two ovine fetuses infected with C. lari. In positive cases, the IHC stain most frequently identified bacteria in the lung and gastrointestinal tract. The C. fetus IHC procedure performed on formalin fixed tissues is a practical tool for the diagnosis of natural cases of ovine and bovine abortion caused by C. fetus.
    Full-text · Article · May 2005 · Journal of Veterinary Medicine Series B
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    ABSTRACT: Tritrichomonas foetus (T. foetus) is the causative agent of bovine trichomonosis, a sexually transmitted disease leading to abortion (from 1 to 8 months gestation), infertility, and occasional pyometra. The annual losses to the U.S. beef industry are estimated to be in the hundreds of millions of dollars. Currently, the "gold standard" diagnostic test for trichomonosis in most countries is the cultivation of live organisms from reproductive secretions. The cultured organisms can then be followed by PCR assays with primers that amplify T. foetus to the exclusion of all other trichomonad species. Thus, negative results present as null data, indistinguishable from failed PCR amplification during T. foetus specific amplification. Our newly developed assay improves previously developed PCR based techniques by using diagnostic size variants from within the internal transcribed spacer 1 (ITS1) region that is between the 18S rRNA and 5.8S rRNA subunits. This new PCR assay amplifies trichomonad DNA from a variety of genera and positively identifies the causative agent in the bovine trichomonad infection. This approach eliminates false negatives found in some current assays as well as identifying the causative agent of trichomonad infection. Additionally, our assay incorporates a fluorescently labeled primer enabling high sensitivity and rapid assessment of the specific trichomonad species. Moreover, electrophoretic separation of amplified samples can be outsourced, thus eliminating the need for diagnostic laboratories to purchase expensive analysis equipment.
    Full-text · Article · Feb 2005 · Veterinary Parasitology
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    ABSTRACT: Sequence analysis of the 5.8S rRNA gene and the internal transcribed spacer regions (ITSRs) was used to compare trichomonadid protozoa (n = 39) of varying morphologies isolated from the bovine preputial cavity. A multiple sequence alignment was performed with bovine isolate sequences and other trichomonadid protozoa sequences available in GenBank. As a group, Tritrichomonasfoetus isolates (n = 7) had nearly complete homology. A similarity matrix showed low homology between the T. foetus isolates and other trichomonads recovered from cattle (<70%). Two clusters of trichomonads other than T. foetus were identified. Eighteen isolates comprised 1 group. These isolates shared >99% homology among themselves and with Pentatrichomonas hominis. The other non-T. foetus cluster (n = 14) did not exhibit a high degree of homology (<87%) with other bovine isolates or any of the trichomonad sequences available in GenBank. The sequence homology among isolates in that cluster was >99%, except for 1 isolate that varied from the others in both ITSRs (approximately 2% dissimilarity). Sequence analysis of the 5.8S rRNA gene and ITSRs was useful for comparing trichomonadid protozoa isolated from the bovine preputial cavity and demonstrated that 2 distinct types of trichomonads constituted the non-T. foetus isolates recovered from the bovine preputial cavity.
    Preview · Article · Feb 2003 · Journal of veterinary diagnostic investigation: official publication of the American Association of Veterinary Laboratory Diagnosticians, Inc
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    L V Stamm · H L Bergen · R L Walker
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    ABSTRACT: Papillomatous digital dermatitis (PDD), an emerging infectious disease of cattle, is characterized by painful, ulcerative foot lesions. The detection of high numbers of invasive spirochetes in PDD lesions suggests an important role for these organisms in the pathogenesis of PDD. PDD-associated spirochetes have phenotypic characteristics consistent with members of the genus TREPONEMA: Partial 16S ribosomal DNA (rDNA) sequence analysis of clonal isolates from California cattle showed that they comprise three phylotypes which cluster closely with human-associated Treponema spp. of the oral cavity (T. denticola and T. medium/T. vincentii) or genital area (T. phagedenis). The goal of our study was to apply 16S-23S rDNA intergenic spacer region (ISR) sequence analysis to the molecular typing of U.S. PDD-associated Treponema isolates. This methodology has potentially greater discriminatory power for differentiation of closely related bacteria than 16S rDNA analysis. We PCR amplified, cloned, and sequenced the ISRs from six California PDD-associated Treponema isolates and, for comparative purposes, one strain each of T. denticola, T. medium, T. vincentii, and T. phagedenis. Two ISRs that varied in length and composition were present in all the PDD-associated Treponema isolates and in T. denticola, T. medium, and T. phagedenis. ISR1 contained a tRNA(Ala) gene, while ISR2 contained a tRNA(Ile) gene. Only a single ISR (ISR1) was identified in T. vincentii. Comparative analyses of the ISR1 and ISR2 sequences indicated that the California PDD-associated Treponema isolates comprised three phylotypes, in agreement with the results of 16S rDNA analysis. PCR amplification of the 16S-tRNA(Ile) region of ISR2 permitted rapid phylotyping of California and Iowa PDD-associated Treponema isolates based on product length polymorphisms.
    Preview · Article · Oct 2002 · Journal of Clinical Microbiology
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    R L Walker · D H Read · D C Hayes · R W Nordhausen
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    ABSTRACT: Direct amplification and sequencing of the 16S rRNA gene and a variable region of the flagellin gene from fetal liver-associated spirochetes belonging to the Borrelia parkeri-B. turicatae tick-borne relapsing fever spirochete group with a late-term abortion in a mare are described.
    Full-text · Article · May 2002 · Journal of Clinical Microbiology
  • R L Walker · H Kinde · R J Anderson · AE Brown
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    ABSTRACT: A comparison of the VIDAS Salmonella (SLM) assay using the manufacturer's recommended sampling method or a Moore swab sampling method demonstrated that the Moore swab method detected a greater number of positive samples (83.0% vs. 67.92%). When results using a conventional culture technique were compared to the VIDAS SLM assay using Moore swab sampling, there was good agreement (97.5%). When the VIDAS SLM assay was used to test in-line milk filters and compared to results from the conventional culture method, there was also good correlation between test results (95.57%). Overall, the VIDAS SLM assay using a modified sampling method compared favorably to the conventional culture method and had the advantages of taking less time to obtain a negative or presumptive positive result, being less technically complicated and requiring less screening of non-lactose fermenting colonies from negative samples.
    No preview · Article · Aug 2001 · International Journal of Food Microbiology
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    B R Charlton · AA Bickford · R P Chin · R L Walker
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    ABSTRACT: Randomly amplified polymorphic DNA (RAPD) analysis was used to investigate the molecular epidemiology of 26 Mycoplasma gallisepticum (MG) isolates obtained from turkeys located in the central valley of California. The MG isolates were recovered from 5 different companies and 13 ranches. Each company had unique MG strains. No evidence of spread of MG between companies was detected. RAPD analysis of MG isolates within a ranch during an outbreak revealed only a single strain involved in each outbreak. RAPD analysis identified an isolate from 1 ranch with a banding pattern identical to that of the 6/85 vaccine strain, which had been used on that particular ranch. Similar RAPD banding patterns of isolates from different ranches within the same company suggested horizontal spread of MG between ranches. The use of 2 primer sets in RAPD analysis was critical to prevent misinterpretation of relationships between different isolates.
    Preview · Article · Oct 1999 · Journal of veterinary diagnostic investigation: official publication of the American Association of Veterinary Laboratory Diagnosticians, Inc
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    B R Charlton · A A Bickford · R L Walker · R Yamamoto
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    ABSTRACT: Randomly amplified polymorphic DNA (RAPD) analysis was used to differentiate 7 strains of Mycoplasma gallisepticum. Six commercially available primers or primer combinations were screened for their ability to differentiate vaccine and type strains. Although major and minor bands were produced with each primer, many of the primers were unsuitable for strain differentiation. The use of primer 6 and combined primers 3 and 4 resulted in complementary RAPD banding patterns for each M. gallisepticum strain. Eleven different isolates representing 7 different strains were segregated into 7 different patterns, corresponding to the 7 strains.
    Preview · Article · Apr 1999 · Journal of veterinary diagnostic investigation: official publication of the American Association of Veterinary Laboratory Diagnosticians, Inc
  • R L Walker · D H Read · K J Loretz · D W Hird · S L Berry
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    ABSTRACT: To determine whether a humoral response against spirochetes isolated from papillomatous digital dermatitis (PDD) lesions is elicited in dairy cattle affected with PDD. 41 cattle with PDD from 8 dairies (study population) and 30 cattle from 2 dairies free of PDD (control population). Additionally evaluated were 32 cattle from a dairy with a past history of PDD but no current disease, and 52 cattle from a dairy with high prevalence of PDD, 25 with and 27 without detectable lesions. ELISA were used to evaluate the humoral response of all cattle to representative isolates from 2 groups of spirochetes of unknown species isolated from PDD lesions. Specificity of the response was evaluated, using immune sera prepared against each of the spirochetes, and by adsorption studies of immune and field sera. The potential for confounding by an antibody response to other spirochetes associated with diseases of cattle was assessed. The antibody response (specific) to both PDD spirochete groups of cows with PDD was significantly increased, compared with that of cows from PDD-free dairies. There was no association between antibody response to PDD-associated spirochetes and antibody response to other spirochetal diseases of cattle. None of the cattle from the dairy with previous history of PDD but without current disease were classified as test positive by either PDD ELISA. There was a significant (P < 0.01) difference in classification results for both PDD ELISA for cattle with PDD from the dairy with a high herd prevalence of PDD, compared with cattle without detectable disease from the same dairy. The humoral response in cattle with PDD lesions was significantly different from that in cattle without detectable lesions, thus providing additional information regarding the potential role of spirochetes isolated from PDD lesions in the etiopathogenesis of PDD.
    No preview · Article · Aug 1997 · American Journal of Veterinary Research
  • R J Anderson · R L Walker · D W Hird · P C Blanchard
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    ABSTRACT: To identify risk factors associated with Salmonella menhaden associated disease in adult dairy cows during an outbreak in California. Case-control study. 8 case dairies that had > or = 1 adult animal that had clinical signs of salmonellosis and from which S menhaden was isolated and 22 control dairies, 16 of which were matched on the basis of herd size and county and 6 of which were matched on the basis of herd size, county, and breed (Jersey). A questionnaire was developed and reviewed with the herdsman or owner of each dairy. Primary areas of concern were herd management, disease characteristics, and feed-related information. Use of 1 particular feed mill and feeding animal fat were significant risk factors for clinical disease attributable to S menhaden infection. Feed should not be overlooked as a potential source of Salmonella organisms in dairy herds.
    No preview · Article · Feb 1997 · Journal of the American Veterinary Medical Association
  • A Rodríguez-Lainz · D W Hird · R L Walker · D H Read
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    ABSTRACT: To estimate prevalence of papillomatous digital dermatitis (PDD) in California dairies during 1993, to describe temporal and spatial patterns of PDD and response of cows to treatment, and to evaluate herd-related risk factors for PDD. Epidemiologic survey. All southern California dairies and a 50% random sample of the rest of California dairies. A questionnaire was mailed to managers of 1,429 selected dairies. Prevalence of PDD during 1993 was significantly higher for dairies in the south (75.3%) and central (68.8%) regions, compared with north coast (33.3%) and north (23.1%) regions of the state. Herd mean and median proportions of affected cows in PDD-affected herds were 11.6 (SEM = 0.9) and 5%, respectively. Most (74%) dairy managers surveyed observed PDD for the first time during 1992 or 1993. Highest PDD activity was reported as taking place in summer in the south and in fall and winter in the north coast and north regions; an obvious pattern was not determined for the central region. PDD was widespread in California dairies during 1993 and affected a high proportion of cows, especially in the south and central regions of the state.
    No preview · Article · Nov 1996 · Journal of the American Veterinary Medical Association
  • M De Rosa · R Droual · R P Chin · H L Shivaprasad · R L Walker
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    ABSTRACT: A sequence of outbreaks of respiratory disease separated by intervals of about 2 wk occurred on three turkey breeder ranches. The last two ranches affected belonged to different companies and were separated by a distance of 11 km. Mortality on the last ranch was particularly severe among certain segregated groups of turkeys that included toms, heavier birds, and birds undergoing a stressful event such as artificial insemination. On this ranch, percentages of mortality within an 18-day period were 5.2% in toms, 2.4% in hens, 7.4% in heavy toms and 5.4% in heavy hens. Turkeys from 27 to 42 wk of age were examined during the outbreaks. Gross lesions included severe lung consolidation with fibrinous exudate on the pleura and air sacs, petechiae on the epicardium, and increased cloudy fluid in the pericardial sac. Liver and spleen were moderately enlarged. Histologically, there was severe fibrinoheterophilic inflammation in the airways, pleura, and air sacs and severe perivascular interstitial edema in the lungs. Liver had acute coagulative necrosis of hepatocytes associated with occasional thrombosis at the periphery of the liver lobes. Ornithobacterium rhinotracheale was isolated from tissues of the respiratory system, such as infraorbital sinus, trachea, lung, and air sacs, but not from the liver, spleen, or bone marrow. Escherichia coli was isolated less often from lung, air sac, and trachea.
    No preview · Article · Oct 1996 · Avian Diseases
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    ABSTRACT: Salmonella enteritidis, phage type 4 (SE PT4), was isolated from five of six 27-wk-old layer chickens submitted for necropsy from a flock of 43,000. Bacteriologic and epidemiologic investigations on the ranch revealed that five of the eight flocks (n = 176,000) were infected. The prevalence of SE PT4 in randomly selected healthy birds ranged from 1.7% (in caged birds) to 50% (in free-range birds) and prevalence in culled birds (kept on dirt floor houses) ranged from 14% to 42%. The estimated overall prevalence of group D Salmonella in eggs contaminated with group D Salmonella was 2.28 per 10,000. The estimated prevalence of group D Salmonella in eggs from caged birds in three infected houses ranged from 1.5 to 4.1 per 10,000, whereas in two houses of free-range birds, prevalence was 14.9 to 19.1 per 10,000. Three of the eight flocks on the ranch remained negative for Salmonella between May 1994 and December 1995 or until removed from the ranch. Salmonella enteritidis PT4 was also isolated from 12.5% (6 of 48) of mice; 57% (four of seven) of cats; and two of two skunks tested. Environmental drag swabs and well water samples yielded multiple serotypes of Salmonella (23/180 and 5/14, respectively) but not S. enteritidis.
    No preview · Article · Jul 1996 · Avian Diseases
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    B M DeBey · P C Blanchard · R L Walker

    Full-text · Article · May 1996 · Journal of veterinary diagnostic investigation: official publication of the American Association of Veterinary Laboratory Diagnosticians, Inc
  • R.L. Walker · D.H. Read · K.J. Loretz · R.W. Nordhausen
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    ABSTRACT: Two groups of spirochetes were isolated from papillomatous digital dermatitis (PDD) lesions in dairy cattle. The two groups could be readily differentiated on the basis of morphologic and immunologic characteristics and enzymatic activity. A spirochete isolated from an interdigital dermatitis (IDD) lesion appeared morphologically and antigenically similar to spirochetes in one of the PDD groups and exhibited an identical enzyme activity pattern. The two groups of PDD spirochetes had characteristics most consistent with the genus Treponema. The PDD and IDD isolates differed morphologically from previously described bovine Treponema spp. Although spirochetes have been observed to be one of the predominant bacterial morphotypes in PDD and IDD and are found invading the stratum spinosum and dermal papillae in PDD lesions, the significance of these spirochetes in the etiopathogenesis of PDD and IDD is presently unknown.
    No preview · Article · Jan 1996 · Veterinary Microbiology
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    ABSTRACT: Isolates of Salmonella choleraesuis serotype ohio (S. ohio) recovered during an outbreak of equine neonatal salmonellosis on a Thoroughbred farm were compared with isolates of the same serotype from various animal, feed and environmental sources. Biochemical profiles, antimicrobial susceptibility patterns, phage susceptibility, plasmid profiles, restriction endonuclease analysis and ribotyping were used to compare relatedness of the strains. A total of 46 outbreak and non-outbreak associated isolates of S. ohio were studied. Differences in antimicrobial susceptibility patterns, phage susceptibility and plasmid profiles were useful for differentiating outbreak isolates from other equine isolates as well as bovine, porcine and some poultry isolates. Feed and other poultry isolates, most in geographic proximity to the outbreak, were indistinguishable from outbreak isolates by any of the methods employed. Investigative studies on the farm along with results of genotypic and phenotypic analysis of isolates suggested that contaminated feed was the most likely source of Salmonella in this outbreak.
    No preview · Article · Mar 1995 · Veterinary Microbiology
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    Full-text · Article · May 1994 · Journal of veterinary diagnostic investigation: official publication of the American Association of Veterinary Laboratory Diagnosticians, Inc
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    H Kinde · B M Daft · R L Walker · B R Charlton · R Petty
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    ABSTRACT: Between April of 1990 and March of 1992, calves on a Holstein calf ranch experienced subcutaneous decubital abscesses involving the brisket region, dorsal aspect of the carpus, and lateral aspect of the stifle joints. Fifty out of 2,500 (2%) Holstein calves between the ages of 3 and 12 weeks were affected. Needle aspirates of brisket abscesses from 8 calves and 6 live or dead calves with 1 or more decubital abscesses were submitted for examination. Two of the 6 calves in addition had bronchopneumonia. Mycoplasma bovis was isolated from all abscesses and 1 lung. Formalin fixed tissues taken from the affected areas also revealed M. bovis by immunoperoxidase staining. No evidence of joint involvement was apparent, and no mycoplasma was isolated from the joints adjacent to affected areas. Attempts to isolate mycoplasma from milk and environmental samples were unsuccessful.
    Preview · Article · May 1993 · Journal of veterinary diagnostic investigation: official publication of the American Association of Veterinary Laboratory Diagnosticians, Inc

Publication Stats

833 Citations
50.28 Total Impact Points


  • 1991-2008
    • University of California, Davis
      • • School of Veterinary Medicine
      • • California Animal Health & Food Safety Laboratory System - CAHFS
      Davis, California, United States
  • 2002
    • University of North Carolina at Chapel Hill
      • Department of Epidemiology
      North Carolina, United States