In vitro antimicrobial susceptibilities of three Porphyromonas spp and in vivo responses in the oral cavity of cats to selected antimicrobial agents

OBJECTIVES: To determine in vitro susceptibility of Porphyromonas gingivalis, P salivosa and P circumdentaria to seven antimicrobial agents by agar dilution and Epsilometer test methods and to assess the effectiveness of these antimicrobial agents in reducing the numbers of each Porphyromonas spp in the oral cavity of 16 domestic cats. DESIGN: A two-part prospective study involving in vitro antimicro-bial studies using Porphyromonas spp obtained from naturally occurring feline infections and in vivo antimicrobial response studies using client-owned cats with naturally occurring periodontal disease. PROCEDURE: Isolates (n = 25) of three feline Porphyromonas spp from the oral cavity and oral-associated disease were tested for their in vitro susceptibility to amoxycillin, amoxycillin-clavulanate, benzylpenicillin, clindamycin, doxycycline, erythromycin and metronidazole, using agar dilution and Epsilometer test methods. Digoxigenin-labelled whole chromosomal DNA probes directed against P gingivalis VPB 3492, P circumdentaria NCTC 12469T and P salivosa VPB 3313 were used to quantify organisms taken from two sample sites at the gingival margins of these cats prior to, and 5 days after, treatment with one of four commonly used antimicrobial products (amoxycillin-clavulanate, clindamycin, doxycycline or spiramycin-metronidazole). The response to treatment was assessed clinically for each cat. RESULTS: All isolates were susceptible in vitro to all seven antimicrobial agents using both methods. The numbers of P gingivalis were not reduced at the gingival sample sites by administration of amoxycillin-clavulanate for 5 days, although this treatment reduced the numbers of P salivosa and P circumdentaria to below detection levels in six of eight and two of three of sample sites, respectively; clinical improvement was not observed in cats treated with amoxycillin-clavulanate. Treatment with clindamycin, doxycycline or spiramycin-metronidazole resulted in clinical improvement and a marked reduction of all Porphyromonas isolates at the sample sites. CONCLUSION: The Epsilometer test is a simple and accurate method for determining the minimum inhibitory concentration for P gingivalis, P salivosa and P circumdentaria. All strains were susceptible in vitro to all the antimicrobial agents tested although clinical improvement of gingival disease was not noted with amoxycillin-clavulanate when given for 5 days at usual doses. This appears to be the first report of the disparity between the in vivo and in vitro susceptibility of oral bacterial strains to amoxycillin-clavulanate in the veterinary dental literature. This also appears to be the first report in which clinical and microbiological responses to commonly used antimicrobial agents for periodontal disease in cats has been documented and quantified. It was shown that treatment with clindamycin, spiramycin-metronidazole or doxycycline not only produced a substantial reduction in the number of Porphyromonas spp (in the majority of cases to below detection levels), but also resulted in substantial clinical improvement. This would indicate that these antimicrobial agents are useful adjunctive therapy to mechanical debridement in domestic cats.     

Endogenous lipid pneumonia in cats: 24 cases (1985-1998)

OBJECTIVE: To determine clinical signs, radiographic and histologic abnormalities, and concurrent diseases in cats with endogenous lipid pneumonia (EnLP) and to determine the pathologic importance of EnLP in cats. DESIGN: Retrospective study. ANIMALS: 24 cats. PROCEDURE: Medical records of cats in which EnLP was confirmed by histologic examination of necropsy specimens were reviewed. Information collected from the medical records included signalment, body weight, clinical signs, and results of clinicopathologic tests. Thoracic radiographs were reviewed by a radiologist; histologic specimens were reviewed by a pathologist. RESULTS: All cats had nonspecific clinical abnormalities, such as lethargy, anorexia, or weight loss; 16 had signs of respiratory tract disease. All cats had concurrent systemic diseases, and clinicopathologic abnormalities were reflective of these conditions. Nonspecific abnormalities were detected on thoracic radiographs from 9 of 11 cats. Most cats had macroscopic, multifocal, subpleural lesions; inflammatory infiltrates, cholesterol clefts, and multinucleated giant cells were common. Ten cats had an underlying obstructive pulmonary disease that was the likely cause of EnLP. Lesions of EnLP were not considered to be severe enough or extensive enough to be the cause of death in any of these cats. CONCLUSIONS AND CLINICAL RELEVANCE: EnLP is an uncommon respiratory tract disorder of cats with no pathognomonic clinical, laboratory, or radiographic findings. Although EnLP was not the cause of death in any of these cats, results of the present study do suggest that EnLP may be a marker for potentially severe underlying obstructive pulmonary disease.     

Serum antibody responses of cats to soluble whole cell antigens of feline Porphyromonas gingivalis

The whole cell soluble antigens of two strains (VPB 3457 and VPB 3492) of feline Porphyromonas gingivalis were analysed by Western blotting using serum taken from 40 domestic cats with various grades of periodontal disease. Five strongly immunogenic protein bands (70, 34, 27, 24 and 19kDa) from VPB 3457 and seven from VPB 3492 (58, 44, 34, 27, 25, 24 and 21kDa) were selected for further study. A significant positive correlation was found between the serum antibody response to the 70, 34, 27, 24 and 19kDa bands of VPB 3457 and the 58, 44, 25, 24 and 21kDa bands of VPB 3492 and the overall periodontal grade. A significant positive correlation was also found between the serum antibody response to the 24kDa band of VPB 3457 and the total colony forming units of P. gingivalis. N-terminal sequencing of the 44kDa band of VPB 3492 showed 75% identity with the translated amino acids from the hag A (haemagglutinin) gene of a human strain of P. gingivalis and N-terminal amino acid sequence of the 27kDa band of VPB 3457 showed 88% identity with the amino acid sequences translated from DNA of purported genes coding for variously named proteinases of human strains of P. gingivalis.     

Compositional stratification in the deep mantle

A boundary between compositionally distinct regions at a depth of about 1600 kilometers may explain the seismological observations pertaining to Earth's lower mantle, produce the isotopic signatures of mid-ocean ridge basalts and oceanic island basalts, and reconcile the discrepancy between the observed heat flux and the heat production of the mid-ocean ridge basalt source region. Numerical models of thermochemical convection imply that a layer of material that is intrinsically about 4 percent more dense than the overlying mantle is dynamically stable. Because the deep layer is hot, its net density is only slightly greater than adiabatic and its surface develops substantial topography.     

In vivo imaging of quantum dots encapsulated in phospholipid micelles

Fluorescent semiconductor nanocrystals (quantum dots) have the potential to revolutionize biological imaging, but their use has been limited by difficulties in obtaining nanocrystals that are biocompatible. To address this problem, we encapsulated individual nanocrystals in phospholipid block-copolymer micelles and demonstrated both in vitro and in vivo imaging. When conjugated to DNA, the nanocrystal-micelles acted as in vitro fluorescent probes to hybridize to specific complementary sequences. Moreover, when injected into Xenopus embryos, the nanocrystal-micelles were stable, nontoxic (<5 x 10(9) nanocrystals per cell), cell autonomous, and slow to photobleach. Nanocrystal fluorescence could be followed to the tadpole stage, allowing lineage-tracing experiments in embryogenesis.