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. 2024 May 8;13(5):391.
doi: 10.3390/pathogens13050391.

Association of Mycoplasma canis with Fertility Disorders in Dogs: A Case Study Supported by Clinical Examination, PCR, 16S Microbiota Profiling, and Serology

Sara Suhadolc Scholten  1 , Brigita Slavec  2 , Primož Klinc  3 , Nataša Tozon  1 , Bojan Papić  4 , Saša Koprivec  5
Affiliations

Association of Mycoplasma canis with Fertility Disorders in Dogs: A Case Study Supported by Clinical Examination, PCR, 16S Microbiota Profiling, and Serology

Sara Suhadolc Scholten et al. Pathogens. .

Abstract

The role of Mycoplasma canis in canine fertility disorders is still poorly understood. As infection is often asymptomatic, there is an increasing need for appropriate diagnostic methods and treatment plans that would allow the reliable detection of M. canis infection and rapid alleviation of infection symptoms in affected dogs. In this study, we included 14 dogs with fertility problems and 16 dogs without fertility disorder signs. We compared clinical examination data and selected laboratory parameters (hematology and biochemistry) between the groups. We performed PCR-based detection of M. canis and 16S rRNA gene-based microbiota profiling of DNA isolated from vaginal and preputial swabs. Dog sera were tested for the presence of M. canis-specific antibodies. Hematological and selected biochemical parameters showed no differences between groups. PCR-based detection of M. canis in the samples was consistent with the results of 16S microbiota profiling. Several other bacterial taxa were also identified that could potentially be involved in different fertility disorders. Serological methods were not accurate enough since high cross-reactivity rates were observed. In the future, more accurate and efficient methods will be needed to determine the role of M. canis and its true role in the pathogenesis of specific fertility disorders in dogs.

Keywords: 16S microbiota profiling; Mycoplasma canis; PCR; canine fertility disorders; preputial swab; specific antibodies; vaginal swab.

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Conflict of interest statement

The authors declare no conflicts of interest.

Figures

Figure 1
Figure 1
Microbiota composition of each sample. Samples are stratified according to the analyzed group: (A) top five bacterial phyla; (B) top eight bacterial families; (C) top eight bacterial genera. The sex of animals is indicated in brackets next to the sample name: F, female; M, male.
Figure 2
Figure 2
Rarefaction curves showing the number of OTUs per sequencing depth. The sex of the animal is indicated in brackets next to the sample name: F, female; M, male.
Figure 3
Figure 3
The presence of M. canis-specific antibodies in selected sera of the analyzed animals. Whole cell lysates of M. canis PG14T (left panel) and M. canis Larissa (right panel) were incubated in sera of animals from the FD and CTRL group. Reactions of serum antibodies with mycoplasma proteins, ranging from 15 to 130 kDa, are clearly visible in all samples, regardless of the results of molecular examination. In particular, in the right panel (M. canis Larissa), antibodies reacted to two proteins, approximately 55 and 40 kDa in size (blue arrows). M: molecular size marker, PageRuler Plus Prestained Protein Ladder), 1: serum of patient 1/FD, 2: serum of patient 6/FD, 3: serum of patient 11/FD, 4: serum of patient 13/FD, 5: serum of patient 16/CTRL, 6: serum of patient 42/CTRL, 7: serum of patient 5/FD after antibiotic treatment.
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