Elsevier

Veterinary Microbiology

Volume 85, Issue 3, 22 March 2002, Pages 285-292
Veterinary Microbiology

Short communication
Neospora caninum and Waddlia chondrophila strain 2032/99 in a septic stillborn calf

https://doi.org/10.1016/S0378-1135(01)00510-7Get rights and content

Abstract

Characteristics of an intracellularly growing micro-organism isolated from an aborted bovine foetus are described. The organism replicated within cytoplasmic vacuoles, was resistant to penicillin and exhibited structural characteristics compatible with Waddlia chondrophila. An ELISA specific for Chlamydia spp., immunofluorescence tests using antibodies directed against Chlamydia spp. or Simkania negevensis, and PCR using Chlamydia-specific primers showed that the agent was distinct from Chlamydiae or S. negevensis. Determination of 16S and partial 23S ribosomal RNA gene sequences in combination with the PCR results and the morphological, antigenic and developmental characteristics provided evidence that the isolate 2032/99 can be classified as W. chondrophila or a closely related organism.

Introduction

Abortions and stillbirths in cattle are often caused by chlamydial or rickettsial pathogens (Ahlers and Grunert, 1997). In 1990, a new obligate intracellular bacterium isolated from pooled lung and liver samples of an aborted bovine foetus was described. The agent (WSU 86-1044) appeared to be a new member of the Rickettsiae because it showed cross-reactions with antisera directed against Cowdria ruminantium (Dilbeck et al., 1990). The organism replicated within intracytoplasmatic vacuoles and had a development cycle comparable to that of Chlamydiae and Ehrlichiae (Kocan et al., 1990). In contrast to Chlamydiae, replication was not inhibited by penicillin (Dilbeck et al., 1990). Phylogenetic relationship to Chlamydiae was shown by analysis of the 16S ribosomal (r)DNA. In 1999, agent WSU 86-1044 was proposed as the new species Waddlia chondrophila, the only member of the novel family Waddliaceae (order Chlamydiales). Currently the species W. chondrophila includes only the type strain (Rurangirwa et al., 1999). Here we report the isolation of a Chlamydia-like agent from an aborted bovine foetus with genetic and morphological characteristics of W. chondrophila.

Section snippets

Tissue sampling

A bovine foetus (Holstein) was obtained from a herd located in the German federal state Rhineland Palatinate. The abortion was part of an endemic event which was related to Neospora caninum infections. The foetus was aborted on day 228 of gestation. A complete post-mortem examination was carried out. Tissue samples of brain, lungs heart, liver, spleen, kidney, skeletal muscle, and placenta were fixed in 4% buffered formol saline and processed using standard techniques. Sections were cut at 5 μm

Pathological findings

The foetus showed moderate to advanced signs of autolysis. Gross lesions were restricted to the brain. The meninges were opaque and velvet like caused by a severe, mainly lymphohistocytic meningitis with fibrinous exudate and few neutrophils only. There was a focal leukoencephalomalacia in the vicinity of the lateral ventricles with mild lymphohistocytic perivascular cuffing. The lungs were voluminous with moderate interstitial oedema and multifocal acute suppurative pneumonia and

Discussion

Here we report the isolation and first characterisation of an obligate intracellular micro-organism from an aborted bovine foetus which at first seemed to be related to Chlamydia because of its phenotypic similarity. Evidence for this was the formation of vacuoles filled with particles as observed for C. trachomatis (Gordon and Quan, 1965, Kahane et al., 1993). Inclusions could be stained using Giemsa or Giménez stain. The heterogeneity in dye uptake of the Giménez stained particles suggested

Acknowledgements

We thank Roswitha Wehr and Petra Meyer for their excellent technical assistance.

References (17)

  • S Kahane et al.

    Description and partial characterization of a new Chlamydia-like micro-organism

    FEMS Microbiol. Lett.

    (1993)
  • G Schares et al.

    The efficiency of vertical transmission of Neospora caninum in dairy cattle analysed by serological techniques

    Vet. Parasitol.

    (1998)
  • D Ahlers et al.

    Aborte beim Rind—diagnostische Maßnahmen und Forensik

    Prakt. Tierarzt

    (1997)
  • P.M Dilbeck et al.

    Isolation of a previously undescribed rickettsia from an aborted fetus

    J. Clin. Microbiol.

    (1990)
  • K.D.E Everett et al.

    Identification of nine species of the Chlamydiaceae using RFLP–PCR

    Int. J. Syst. Bacteriol.

    (1999)
  • K.D.E Everett et al.

    Emended description of the order Chlamydiales, proposal of Parachlamydiaceae fam. nov. and Simkaniaceae fam. nov., each containing one monotypic genus, revised taxonomy of the family Chlamydiaceae, including a new genus and five new species, including a new genus and five new species, and standards for the identification of organisms

    Int. J. Syst. Bacteriol.

    (1999)
  • D.F Giménez

    Staining Rickettsiae in yolk sac cultures

    Stain Technol.

    (1964)
  • F.B Gordon et al.

    Isolation of the trachoma agent in cell culture

    Proc. Soc. Exp. Med.

    (1965)
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