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Veterinary Microbiology Apr 2016Although treponemes are consistently identified in tissue from bovine digital dermatitis (DD) lesions, the definitive etiology of this debilitating polymicrobial disease...
Although treponemes are consistently identified in tissue from bovine digital dermatitis (DD) lesions, the definitive etiology of this debilitating polymicrobial disease is still unresolved. To study the microbiomes of 27 DD-infected and 10 healthy interdigital skin samples, we used a combination of different molecular methods. Deep sequencing of the 16S rRNA gene variable regions V1-V2 showed that Treponema, Mycoplasma, Fusobacterium and Porphyromonas were the genera best differentiating the DD samples from the controls. Additional deep sequencing analysis of the most abundant genus, Treponema, targeting another variable region of the 16S rRNA gene, V3-V4, identified 15 different phylotypes, among which Treponema phagedenis-like and Treponema refringens-like species were the most abundant. Although the presence of Treponema spp., Fusobacterium necrophorum and Porphyromonas levii was confirmed by fluorescence in situ hybridization (FISH), the results for Mycoplasma spp. were inconclusive. Extensive treponemal epidermal infiltration, constituting more than 90% of the total bacterial population, was observed in 24 of the 27 DD samples. F. necrophorum and P. levii were superficially located in the epidermal lesions and were present in only a subset of samples. RT-qPCR analysis showed that treponemes were also actively expressing a panel of virulence factors at the site of infection. Our results further support the hypothesis that species belonging to the genus Treponema are major pathogens of DD and also provide sufficient clues to motivate additional research into the role of M. fermentans, F. necrophorum and P. levii in the etiology of DD.
Topics: Animals; Biodiversity; Cattle; Cattle Diseases; Digital Dermatitis; Gram-Negative Bacteria; Gram-Negative Bacterial Infections; Microbiota; RNA, Ribosomal, 16S; Skin; Virulence Factors
PubMed: 27016768
DOI: 10.1016/j.vetmic.2016.03.003 -
Journal of Clinical Microbiology Mar 2015A yet-undescribed bacterial species, tentatively named "Porphyromonas katsikii," was isolated from individuals of a small goat herd with pyogranulomatous pneumonia...
A yet-undescribed bacterial species, tentatively named "Porphyromonas katsikii," was isolated from individuals of a small goat herd with pyogranulomatous pneumonia during an outbreak of acute respiratory disease. The isolated bacteria grew in the form of black-pigmented colonies after 14 days of incubation under anaerobic conditions at 37°C on a tryptic soy blood agar medium. The bacteria were identified as a yet-undescribed Porphyromonas species by determination of the nucleotide sequence of the rrs 16S rRNA gene, and this species was tentatively named Porphyromonas katsikii. PCR amplification with specific primers for this yet-undescribed species revealed the presence of P. katsikii in the lung tissue of all affected animals, while no PCR signals were evidenced from the lungs of healthy goats or from goats with pasteurellosis caused by Mannheimia haemolytica. These data indicate P. katsikii as the causative agent of acute respiratory distress. P. katsikii is phylogenetically related to Porphyromonas somerae and Porphyromonas levii, which cause pathologies in humans and animals, respectively. P. katsikii was not detected by PCR from samples of the gingival pockets or of the faces of healthy goats.
Topics: Anaerobiosis; Animals; Bacteriological Techniques; Cluster Analysis; DNA, Bacterial; DNA, Ribosomal; Disease Outbreaks; Goat Diseases; Goats; Lung; Molecular Sequence Data; Phylogeny; Pigments, Biological; Pneumonia, Bacterial; Porphyromonas; RNA, Ribosomal, 16S; Sequence Analysis, DNA; Temperature
PubMed: 25540395
DOI: 10.1128/JCM.02682-14 -
Theriogenology Apr 2012Bovine necrotic vulvovaginitis (BNVV) is a syndrome unique to Israel characterized by necrotic lesion in the caudal vagina mainly in first calf heifers after calving,...
Bovine necrotic vulvovaginitis (BNVV) is a syndrome unique to Israel characterized by necrotic lesion in the caudal vagina mainly in first calf heifers after calving, associated with Porphyromonas levii. The objectives of this study were to analyze the impact of BNVV on reproductive performance, milk production and survival in the heard of first calf dairy heifers in affected farms, and to verify if the effects of BNVV are severity-dependent. For assessment of the severity level a scale of 4 degrees was formed, and cows were scored 4 to 6 d after calving. Data were obtained from two dairy farms during 2006-07, consisting of 603 lactations. The incidence and the severity of BNVV declined between 2006 and 2007, and severe BNVV tended to be more prevalent in the summer. The odds to conceive in the first artificial insemination of BNVV cow tended to be lower than healthy cows (OR = 0.676, P = 0.052). Cows with BNVV had longer empty period (145.8 d vs. 135.1 d of healthy cows, P = 0.031), but only severe BNVV had a negative effect on the odds of the cow to be empty at 150 d in milk (DIM) (OR = 2.05, P = 0.052). Severe BNVV also affected the mean survival time to conception (155.9 d vs. 142.3 d, P = 0.042). All BNVV severity degrees had a negative effect on milk production. The effect on milk production was not limited only to the beginning of the lactation, cows with BNVV produced 338.1 kg milk less than healthy cows (P = 0.016) in 305 d corrected lactation. The effect on milk production was not severity depended. No effect on survival time in the herd was demonstrated.
Topics: Animals; Cattle; Cattle Diseases; Female; Insemination, Artificial; Longevity; Milk; Pregnancy; Pregnancy Rate; Reproduction; Vulvovaginitis
PubMed: 22225696
DOI: 10.1016/j.theriogenology.2011.10.024 -
Microbiology and Immunology Jun 2010Bovine digital epidermitis involves different pathologies, including PDD, interdigital dermatitis, and foot rot. Bacteriological and molecular biological studies suggest...
Bovine digital epidermitis involves different pathologies, including PDD, interdigital dermatitis, and foot rot. Bacteriological and molecular biological studies suggest that these are multimicrobial infections. During our study on the isolation of treponemes from biopsies of PDD, colonies producing black pigment were isolated frequently from the primary cultures, suggesting that Porphyromonas species were present. Moreover, 16S rRNA genes of Fusobacterium necrophorum and Porphyromonas levii-like species were detected in the lesions. We therefore determined whether an immunological response could be elicited by a P. levii-like organism isolated from a PDD lesion, as well as two subspecies of F. necrophorum in the sera from cows with and without PDD. A total of 151 serum samples were collected from 85 cows with PDD lesions and 33 cows without lesions on 12 PDD-positive farms and from 33 cows on two PDD-free farms. ELISA data showed that IgG antibody levels against antigens of P. levii-like species and F. necrophorum subsp. necrophorum were significantly higher in cows on PDD-positive farms than in cows on PDD-free farms, regardless of the presence of PDD lesions in the cows on the PDD-positive farms. However, F. necrophorum subsp. funduliforme was present at low levels in both groups. The ELISA results were confirmed by western blot analysis. Furthermore, antigens of these bacteria were detected in PDD-biopsy sections examined by immunohistochemical staining. F. necrophorum subsp. necrophorum and P. levii-like species may be involved in the pathogenesis of PDD.
Topics: Animals; Antibodies, Bacterial; Antigens, Bacterial; Cattle; Cattle Diseases; Dermatitis; Enzyme-Linked Immunosorbent Assay; Female; Foot Dermatoses; Fusobacterium necrophorum; Immunohistochemistry; Porphyromonas; Rabbits
PubMed: 20536732
DOI: 10.1111/j.1348-0421.2010.00220.x -
Veterinary Therapeutics : Research in... 2009Laboratories use pigmentation, antibiotic susceptibility, and biochemical tests to identify anaerobic organisms that play a role in bovine interdigital necrobacillosis...
Laboratories use pigmentation, antibiotic susceptibility, and biochemical tests to identify anaerobic organisms that play a role in bovine interdigital necrobacillosis (bovine foot rot). In this study, 16S rRNA gene sequencing was used to identify strains to the species level that were originally classified as Prevotella or Porphyromonas spp by conventional phenotype assessment methods. Of 264 qualified strains from ceftiofur clinical trials, 241 isolates were definitively identified by 16S rRNA sequencing as Porphyromonas levii. Similarly, of 275 qualified strains from tulathromycin clinical trials, 156 isolates were definitively identified by 16S rRNA sequencing as P. levii. The predominance of P. levii in this study supports the role of this organism as an associative agent of bovine foot rot and may have implications for routine laboratory diagnosis.
Topics: Animals; Bacteroidaceae Infections; Cattle; Cattle Diseases; Foot Diseases; Porphyromonas; RNA, Ribosomal, 16S
PubMed: 20425726
DOI: No ID Found -
The Veterinary Record
Topics: Animals; Bacteroidaceae Infections; Cattle; Cattle Diseases; Disease Outbreaks; Female; Israel; Necrosis; Porphyromonas; Pregnancy; Risk Factors; Vaginal Smears; Vulvovaginitis
PubMed: 19103617
DOI: No ID Found -
Theriogenology Jul 2007A semiquantitative evaluation of potential bacterial pathogens was correlated to the severity of lesions during an outbreak of bovine necrotic vulvovaginitis (BNVV) on...
A semiquantitative evaluation of potential bacterial pathogens was correlated to the severity of lesions during an outbreak of bovine necrotic vulvovaginitis (BNVV) on an Israeli dairy herd. Bacteriologic examination of 287 vaginal swabs from 104 post-calving heifers showed a highly significant correlation between Porphyromonas levii colony forming unit numbers and the clinical scores of the lesions, when assessed by an ordinal regression statistical model. No such correlation was found for the other bacteria included in the study. Nineteen samples taken for virological examinations resulted negative for bovine herpes viruses 1, 2, 4 and 5. Thus the results of this study substantiate the essential role of P. levii in the etiology of BNVV and indicate that BHV4 is not required as a predisposing factor to the syndrome.
Topics: Animals; Cattle; Cattle Diseases; Disease Outbreaks; Female; Herpesvirus 1, Bovine; Herpesvirus 2, Bovine; Herpesvirus 4, Bovine; Herpesvirus 5, Bovine; Necrosis; Porphyromonas; Vulvovaginitis
PubMed: 17553557
DOI: 10.1016/j.theriogenology.2007.05.040 -
Veterinary Journal (London, England :... May 2008Bovine necrotic vulvovaginitis (BNVV) is characterized by the development of a necrotic vulvovaginal lesion, almost exclusively in post-parturient first-lactation cows,...
Bovine necrotic vulvovaginitis (BNVV) is characterized by the development of a necrotic vulvovaginal lesion, almost exclusively in post-parturient first-lactation cows, associated with Porphyromonas levii. The scope of this survey was to evaluate the impact of BNVV on herd productivity as a means to rationally evaluate the resources that should be allocated in dealing with the syndrome. During an outbreak of BNVV in a dairy herd, following the introduction of a large number of cows from another farm, the impact of the animals' origin (local or transferred) and BNVV (positive or negative) upon involuntary culling rate, milk yield and days between pregnancies were assessed. The results indicated that the number of days between pregnancies was significantly higher in first-lactation cows with BNVV but was not influenced by the other independent variables. None of the other variables included in this survey had any effect on the involuntary culling rate and milk yield.
Topics: Animals; Bacteroidaceae Infections; Cattle; Cattle Diseases; Dairying; Disease Outbreaks; Female; Israel; Lactation; Milk; Porphyromonas; Postpartum Period; Pregnancy; Vulvovaginitis
PubMed: 17433735
DOI: 10.1016/j.tvjl.2007.02.020 -
American Journal of Veterinary Research Nov 2006To investigate the effects of short-chain fatty acids (SCFAs) and pH on neutrophil oxidative burst, phagocytosis, and morphology after exposure to acetate, propionate,... (Comparative Study)
Comparative Study
OBJECTIVE
To investigate the effects of short-chain fatty acids (SCFAs) and pH on neutrophil oxidative burst, phagocytosis, and morphology after exposure to acetate, propionate, butyrate, or succinate at pH 5.5 and 6.7.
SAMPLE POPULATION
Neutrophils isolated from bovine blood samples and Porphyromonas levii, Prevotella spp, and Bacteroides fragilis isolated from lesions of cattle with acute interdigital phlegmon (foot rot).
PROCEDURES
Bacteria were cultured in strictly anaerobic conditions. Bacterial SCFA production was measured with high-performance liquid chromatography. Neutrophils were isolated, stimulated with phorbol 12-myristate 13-acetate (PMA) or opsonized zymosan (OZ), and incubated with dihydroethidium or dichlorofluorescein diacetate to measure production of O(2)and H(2)O(2), respectively. Phagocytosis was assessed after exposure to serum-opsonized bacteria. Cellular morphology was assessed with differential staining.
RESULTS
All bacteria produced at least 3 of the 4 SCFAs. Production of both O(2) and H(2)O(2) was markedly curtailed in PMA-stimulated neutrophils exposed to SCFA at pH 5.5, compared with production at pH 6.7. Succinate caused a significant dose-dependent decrease in O(2) production at pH 6.7 in OZ-stimulated neutrophils. Monoprotic SCFAs elicited a significant increase in H(2)O(2) production in OZ-stimulated neutrophils at pH 6.7 but a significant decrease at pH 5.5. Monoprotic SCFAs significantly increased phagocytosis at pH 6.7 but decreased phagocytic activity at pH 5.5. Cellular necrosis was observed in cells exposed to SCFAs at pH 5.5.
CONCLUSIONS AND CLINICAL RELEVANCE
Establishment and persistence of anaerobic bacteria in cattle with foot rot infection may result in part from neutrophil dysfunction secondary to the effects of bacterially secreted SCFA in acidotic microenvironments.
Topics: Analysis of Variance; Animals; Bacteroides fragilis; Cattle; Cattle Diseases; Cellulitis; Chromatography, High Pressure Liquid; Fatty Acids, Volatile; Hydrogen Peroxide; Hydrogen-Ion Concentration; In Vitro Techniques; Neutrophils; Phagocytosis; Porphyromonas; Prevotella; Respiratory Burst
PubMed: 17078753
DOI: 10.2460/ajvr.67.11.1901 -
Journal of Clinical Microbiology Sep 2005Porphyromonas levii is an anaerobic, pigmented gram-negative bacillus originally isolated from bovine rumen. We describe 58 human clinical strains of P. levii-like...
Porphyromonas levii is an anaerobic, pigmented gram-negative bacillus originally isolated from bovine rumen. We describe 58 human clinical strains of P. levii-like organisms, isolated from various human clinical specimens that are phenotypically similar to the type strain of P. levii, a rumen isolate (ATCC 29147). Our biochemical, comparative 16S rRNA sequence analyses, and DNAlpha-DNA relatedness studies indicate that the human P. levii-like organisms are similar to each other but genetically different from the P. levii type strain isolated from bovine rumen. We therefore propose the name Porphyromonas somerae to encompass the human P. levii-like organisms. P. somerae was predominantly isolated from patients with chronic skin and soft tissue or bone infections, especially in the lower extremities.
Topics: Animals; Bacterial Typing Techniques; Bacteroidaceae Infections; DNA, Bacterial; DNA, Ribosomal; Humans; Molecular Sequence Data; Nucleic Acid Hybridization; Phenotype; Porphyromonas; RNA, Ribosomal, 16S; Sequence Analysis, DNA
PubMed: 16145091
DOI: 10.1128/JCM.43.9.4455-4459.2005