alterado para: “Cytological endometritis in dairy cattle: news insights into pathogenesis, diagnosis and therapy”. Effects of Bovine Venereal Campylobacteriosis and Bovine Viral Diarrohea interaction on beef herd reproductive efficiency

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Cytological endometritis in dairy cattle : new insights into pathogenesis, diagnosis and treatment

Publication . Pereira, Gonçalo da Silva; Silva, Maria Elisabete Tomé Sousa; Costa, Luís Filipe Lopes da

ABSTRACT - In postpartum dairy cows, subclinical endometritis (SCE) is characterized by persistent endometrial inflammation, with profound detrimental effects on subsequent fertility. Despite the known role of adipokines regulating metabolism and inflammation, the association of adipokine signaling with SCE is still poorly understood. Moreover, the effect of SCE on endometrial transcription was mainly determined from biopsy-derived whole tissue, thus masking effects on cell type-specific gene transcription. In addition, despite the recognized improvement in immune function following n-3PUFA supplementation, the therapeutic potential of these fatty acids for SCE control is still to be determined. Therefore, the main objectives of this work were to assess the relationship between adipokines and SCE (Chapter III), to elucidate the effects of progesterone and SCE in the transcription profiles of endometrial glandular, luminal and stromal cells (Chapters IV, V), and the effects of feeding rumen-protected n-3PUFA on endometrial homeostasis and fertility in postpartum dairy cows (Chapter VI). The results showed that adiponectin in plasma and uterine fluid in addition to chemerin uterine fluid have high discriminatory power for the diagnosis of SCE. This work additionally evidenced that progesterone and SCE affect endometrial transcriptomic profiles in a cell type-specific manner. This work also evidenced that the previous presence of immune cells is still impacting the transcriptome of endometrial cells at the end of the voluntary waiting period and that recovery or persistence of inflammation is associated with transcription patterns involved not only in immune function but also in tissue remodeling, cell adhesion, and uterine receptivity. Furthermore, despite having no apparent effect on the endometrial inflammatory status at the end of the voluntary waiting period, dietary supplementation with n-3PUFA decreased calving to conception interval. Overall, this thesis establishes adiponectin as a suitable biomarker of SCE, able to predict the risk of persistence of inflammation in postpartum dairy cows, provides new insights into the persistence and recovery of SCE, thus presenting putative alternative therapeutic strategies. Moreover, this work substantiates dietary n-3PUFA as valid nutraceuticals to improve reproductive parameters in postpartum dairy cows.

2022-03-28Doctoral thesis

Open access

Genomic and phenotypic characterization of Campylobacter fetus subsp. venerealis strains

Publication . Silva, Marta Filipa Serra da; Pereira, Ana L.; Fraqueza, M. J.; Pereira, Gonçalo; Mateus, Luisa; Lopes-da-Costa, Luís; Silva, E

The pathogenesis mechanisms of Campylobacter fetus subsp. venerealis (Cfv), the etiologic agent of Bovine Genital Campylobacteriosis remain elusive. This study evaluated the virulence potential and biovar characteristics of Cfv isolates (n = 13) by PCR screening of putative virulencefactor (VF) genes, Multilocus Sequence Typing (MLST) analysis, antimicrobial susceptibility to tetracycline, penicillin, enrofloxacin and streptomycin testing and whole-genome sequencing (WGS; n = 5), also comparing the latter with 26 other whole-genome sequences of Cfv strains. The putative VF genes encoding type IV secretion system of Cfv (virB2-virB11/virD4) were absent in 92% of isolates, including isolates from aborted foetuses, evidencing that these VF genes are not essential for Cfv pathogenicity. The parA gene, used as a Cfv diagnostic molecular target, was detected in only 3 of 13 isolates, invalidating its use for diagnosis purposes. Three novel sequence types were identified by MLST. Although no in vitro antimicrobial resistance was detected, WGS identified antimicrobial resistance-related genes, including those encoding the multidrug efflux pumps CmeABC and YkkCD, indicating that their presence is not enough to provide antimicrobial resistance. The SNP and accessory protein families analysis segregated the Cfv and Cfv biovar intermedius (Cfvi) strains into different clusters. In conclusion, this study evidenced virulence potential and biovar characteristics of Cfv and Cfvi, which are of relevance for the control of Bovine Genital Campylobacteriosis.

2021-02-09Journal article

Open access

Progesterone differentially affects the transcriptomic profiles of cow endometrial cell types

Publication . Pereira, Gonçalo; Guo, YZ; Silva, E; Bevilacqua, C; Charpigny, G; Lopes-da-Costa, Luís; Humblot, P

Background: The endometrium is a heterogeneous tissue composed of luminal epithelial (LE), glandular epithelial (GE), and stromal cells (ST), experiencing progesterone regulated dynamic changes during the estrous cycle. In the cow, this regulation at the transcriptomic level was only evaluated in the whole tissue. This study describes specifc gene expression in the three types of cells isolated from endometrial biopsies following laser capture microdissection and the transcriptome changes induced by progesterone in GE and ST cells. Results: Endometrial LE, GE, and ST cells show specifc transcriptomic profles. Most of the diferentially expressed genes (DEGs) in response to progesterone are cell type-specifc (96%). Genes involved in cell cycle and nuclear divi sion are under-expressed in the presence of progesterone in GE, highlighting the anti-proliferative action of pro gesterone in epithelial cells. Elevated progesterone concentrations are also associated with the under-expression of estrogen receptor 1 (ESR1) in GE and oxytocin receptor (OXTR) in GE and ST cells. In ST cells, transcription factors such as SOX17 and FOXA2, known to regulate uterine epithelial-stromal cross-talk conveying to endometrial receptivity, are over-expressed under progesterone infuence. Conclusions: The results from this study show that progesterone regulates endometrial function in a cell type-spe cifc way, which is independent of the expression of its main receptor PGR. These novel insights into uterine physiol ogy present the cell compartment as the physiological unit rather than the whole tissue.

2022-01-27Journal article

Open access