PhD in Veterinary Sciences for Animal Health and Food Safety

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Phd thesis

New insights into the diagnosis and treatment of endometritis in the mare

Equine endometritis has been ranked as the third most common medical problem of adult horses and it is the main causes for infertility in mares (Traub-Dargatz et al. 1991). Infectious endometritis is observed in up to 60% of barren mares and results in reduced pregnancy rates and economic losses (LeBlanc & Causey 2009). The traditional approach to diagnosis of endometritis is via bacterial culture from the uterus, but, with the recent developments in metagenomic sequencing, it has been demonstrated that in the uterus there are complex microbial populations of bacterial organisms that can’t be find in the laboratory. The uterine microbiota of women and cows suffering from endometritis differs from that of healthy females, suggesting that endometritis is associated with microbiome dysbiosis (Canisso et al. 2020). Furthermore, considering the increase of the antibiotic resistance phenomenon, a non-traditional therapy, like ozone, thanks to its proved antimicrobial, anti-inflammatory, immunostimulant, and angiogenetic properties is representing an important field of research in the treatment of mare’s subfertility. Finally, considering that many factors are involved in equine infertility, investigation on new potential sexual-transmitted pathogens, like Equine Papillomavirus, could be useful.

Throughbred or Standardbred mares are enrolled in this study.
Every mare is submitted to transrectal ultrasonography, vaginal swab for detection of Equine Papillomavirus, cytological examination , endometrial culture, antibiotic sensitivity test and endometrial biopsy.
Based on the results of the exams, the mares are divided into three groups. 
The first group is made by healthy mares
The second group is constituted by mares with endometritis, diagnosed with a positive cytologic exam but negative to bacteriologic exam.
The third group is constituted by mares with endometritis, diagnosed with a positive bacteriologic exam.

Subsequently, mares of the second group are treated with intrauterine commercial ozone foam spray (Riger, Novagen) for three days. During the subsequent estrous, after at least 14 days from the last medication, sampling for microbiota and uterine biopsy are repeated. On the other hand, mares of the third group are treated with intrauterine antibiotic for three days, according to the result of antibiotic sensitivity test. During the subsequent estrous, after at least 14 days, sampling for microbiota and bacteriological examination are repeated.

Finally, we would like to evaluate efficacy of the different therapies to restore fertility, and, in order to achieve this goal, the mares of group 2 and 3 are inseminated to evaluate the recovery of fertility.

Preliminary results:

Ozone treatment caused a transient inflammation, but it did not cause chronic inflammatory changes in the endometrium and 81,3% (13/16) of the mares recovered fertility.
Regarding the detection of Equus Caballus Papillomavirus Type-2, 40.2% (51 out of 129) of the mares resulted in being positive and we observed a significantly higher EcPV2 genoprevalence in pluriparous vs maiden, in mares subjected to natural mating vs artificial insemination and in barren subfertile mares vs pregnant mares.

Bibliography:

1. Canisso I, Segabinazzi L e Fedorka C (2020). Persistent breeding-induced endometritis in mares – a multifaceted challenge: From clinical aspects to immunopathogenesis and pathobiology. In International Journal of Molecular Sciences 21(4).
2. Leblanc M e Causey R (2009). Clinical and Subclinical Endometritis in the Mare: Both Threats to Fertility. In Reproduction in Domestic Animals 44(3): 10-22.
3. Traub-Dargatz J, Salman L e Voss J (1991). Medical problems of adult horses, as ranked by equine practitioners. In Journal of the American Veterinary Medical Association 198(10): 1745-7.

Research activities

Full articles in ISI journals

• Nervo T, Bertero A, Donato G, Panier Suffat E, Vincenti L. Analysis of factors influencing the transfer of passive immunity in the donkey foal. Italian Journal of Animal Science 2021, 20:1, 1947-1956.
• Cappelli K, De Ciucis CG, Mecocci S, Nervo T, Crescio MI, Pepe M, Gialletti R, Pietrucci D, Migone LF, Turco S, Mechelli S, Passamonti F, Drago C, Donato G, Varello K, Modesto P, Chillemi G, Ghelardi A, Razzuoli E. Detection of Equus Caballus Papillomavirus Type-2 in Asymptomatic Italian Horses. Viruses 2022, 14, 1696.
• De Paolis L, De Ciucis CG, Peletto S, Cappelli K, Mecocci S, Nervo T, Guardone L, Crescio MI, Pietrucci D, Fruscione F, Gabbianelli F, Turco S, Varello K, Donato G, Maurella C, Modesto P, Maniaci MG, Chillemi G, Ghelardi A, Razzuoli E. Equus caballus Papillomavirus Type-9 (EcPV9): First Detection in Asymptomatic Italian Horses. Viruses 2022, 14, 2050.
• Nervo T, Poletto M, Donato G, Bertero A, Bergamini L, Elkhawagah A. Effects of post-freezing addition of relaxin on the fertility parameters of equine semen. Journal of Equine Veterinary Science 2022, 113, 9.

Full articles in non-ISI journals

• Donato G, Nervo T. L’Embryo Transfer nella specie equina. Veterinaria Pratica Equina 2021, 1
• Donato G, Nervo T. L’Endometrite equina. Il Trottatore 2021, Ottobre/Dicembre, 50-51.

Congress abstract 

• Donato G, Nervo T, Bertero A, Varello K, Ponzio P, Bozzetta E, Appino S. Effects of treatment with ozonides and Salix caprea bud-extract on equine endometrium. LXXIIII Convegno SISVET, Virtual edition, 23-26 June 2021.
• Nervo T, Poletto M, Donato G, Bertero A, Bergamini L, Elkhawagah A. Effects of post-freezing addition of relaxin on the fertility parameters of equine semen. 10th ISEET - International
Symposium on Equine Embryo Transfer and Technology - Pisa, 30 June- 2 July 2022.

• Martino NA, Avino A, Donato G, Ricci A, Starvaggi Cucuzza A, Vincenti L. Production of bovine embryos using cysteamine-treated cryopreserved bull semen: an in vitro comparative study. LXXIIII Convegno SISVET, Virtual edition, 23-26 June 2021.
• Malledevarahalli Chandrappa S, Donato G, Alkhawagah A, Meineri G, Vincenti L, Ricci A.
Evaluation of the effect of oxidative stress on uterine pathologies in postpartum dairy cows.
LXXV Convegno SISVET, 15-18 June 2022.
• De Paolis L, De Ciucis M, Cappelli K, Mecocci S, Nervo T, Guardone L, Crescio MI, Fruscione
F, Donato G, Maurella C, Modesto P, Ghelardi A, Elisabetta Razzuoli. First detection of
equus caballus papillomavirus type-9 (ecpv9) in asymptomatic italian horses. LXXV
Convegno SISVET, 15-18 June 2022.
• De Paolis L, De Ciucis CG, Cappelli K, Mecocci S, Nervo T, Crescio MI, Pepe M, Gialletti R,
Migone L, Mechelli L, Donato G, Varello K, Modesto P, Ghelardi A, Razzuoli E. Evaluation of
ecpv2 prevalence in Italy. LXXV Convegno SISVET, 15-18 June 2022.