Equus caballus papillomavirus type 2 (EcPV2)-associated benign penile lesions and squamous cell carcinomas.

BACKGROUND: Squamous cell carcinoma is the most common genital, ocular and gastric tumour in horses. Equus caballus papillomavirus type 2 (EcPV2) DNA has been detected in several studies in equine penile squamous cell carcinomas (SCCs) and precursor lesions providing evidence of a causal role of EcPV2 in equine genital SCCs. Recently, EcPV2 E6/E7 nucleic acids were also detected in equine gastric SCCs, but further studies are required to determine the role of EcPV2 infection in the pathogenesis of gastric SCC. EcPV2 nucleic acids have been rarely described in ocular SCCs and precursor lesions. OBJECTIVES: To investigate the presence of EcPV2 nucleic acids with polymerase chain reaction (PCR) and in situ hybridisation (ISH) in penile hyperplasias, papillomas and SCCs in horses and to determine whether EcPV2 nucleic acids can be detected in SCCs affecting other locations, including the stomach, ocular tissues and larynx. METHODS: Twenty-one archival formalin-fixed paraffin embedded (FFPE) tissue samples, including 12 genital lesions comprising penile hyperplasias, papillomas and SCCs, 6 ocular SCCs, 2 gastric SCCs and 1 laryngeal SCC, were screened by PCR and ISH for EcPV2 E6/E7 DNA and mRNA. Archival FFPE tissue samples (eyelid and penile mucosa and preputium) from six horses without a diagnosis or history of neoplastic or papillomavirus-associated disease were included as controls. RESULTS: EcPV2 nucleic acids were detected by PCR and ISH in all genital lesions (12/12) and gastric SCCs (2/2), in two ocular SCCs (2/6) and in one laryngeal SCC (1/1). In control horses, one eyelid sample was positive in PCR but not in ISH. The remaining control samples were negative for EcPV2 E6/E7 nucleic acids in PCR and ISH. CONCLUSIONS: These results further support the role of EcPV2 infection in the development of equine genital SCCs and suggest that EcPV2 infection may also act as a predisposing factor for other SCCs in horses, including gastric, ocular and laryngeal SCCs.

Accreditation in continuing veterinary education: development of an accreditation system and selection of accreditation criteria.

With the increased supply and demand of veterinary continuing education (CE) and the growing number of CE providers, a clear need has arisen for a multinational accreditation system for veterinary CE. The objective of this document is to describe the current state of veterinary CE accreditation and the development of an accreditation system for veterinary CE, and discuss accreditation criteria and their pedagogical and practical significance. The hypothesis is that a profile of essential, pedagogically grounded, accreditation criteria can be established and utilized effectively in quality assessment. Accreditation criteria for veterinary CE can be created based on four selection principles: educational minimum requirements, coherence, efficacy, and assessability. The selected educational quality criteria are related to needs assessment, correlation of target audience and level of instruction, definition of scope, workload, and number of credits, organizer and instructor qualifications, constructive alignment, assessment of learning, learner engagement, and scientific quality of CE content. The created accreditation criteria and protocols should be regularly re-evaluated and modified in close collaboration with the relevant stakeholders. The desired outcome of CE, including behavior change and improvement of practice and ultimately human and animal health, remains challenging to predict based on course descriptions by the providers, and further research is needed.

RNA in situ hybridization of Escherichia coli in equine endometrial biopsies.

Endometritis is one of the major causes of infertility in mares. Escherichia coli and ß-haemolytic streptococci are among the bacterial species most frequently isolated from the equine uterus. Some bacteria such as ß-hemolytic streptococci, can persist in dormant forms and cause prolonged, latent or recurrent infections. Dormant bacteria may be present despite negative bacterial cultures, and they are resistant to antimicrobial treatment due to their resting metabolic state. The purpose of this study was to study formalin-fixed paraffin-embedded equine endometrial biopsies for the presence and localization of E. coli-bacteria, with a chromogenic RNAscope®-method for detection of E. coli-related 16S ribosomal RNA. Hematoxylin-eosin-stained endometrial biopsies were evaluated to determine the level of inflammation and degeneration. During estrus, samples were taken for endometrial culture and cytology with a double-guarded uterine swab. The samples included eight samples with moderate to severe endometrial inflammation detected in endometrial histopathology, and growth of E. coli in bacterial culture, six samples with moderate to severe endometrial inflammation but negative bacterial culture, and five samples with no endometrial pathology (grade I endometrial biopsy, negative endometrial culture and cytology) serving as controls. Positive and negative control probes were included in the RNA in situ hybridization, and results were confirmed with a fluorescence detection method (fluorescence in situ hybridization). Only unspecific signals of limited size and frequency of occurrence were detected in all samples, with random localization in the endometrium. No samples contained rod-shaped signals corresponding to bacterial findings. In conclusion, there was no evidence of bacterial invasion in the endometrium regardless of the inflammatory status of the biopsy or previous bacterial culture results. According to these findings on a small number of samples, invasion of E. coli is not a common finding in the lamina propria of mares, but these bacteria may also evade detection due to localized foci of infections, or supra-epithelial localization under the cover of biofilm. These bacteria and biofilm covering the epithelium may also be lost during formalin-fixation and processing.

The effects of obesity and insulin dysregulation on mare reproduction, pregnancy, and foal health: a review.

Obesity is a growing welfare concern in modern equine populations and predisposes horses to disturbances in energy metabolism such as insulin dysregulation. However, equine metabolic syndrome has only been recognized in recent decades. Functioning energy metabolism is pivotal to normal body homeostasis and affects essentially all organ systems, including reproduction. Previous literature suggests that obesity has an effect not only on the reproductive processes in mares but also on offspring health, predisposing the offspring to later-onset orthopedic and metabolic problems. This review focuses on the effects of obesity, insulin dysregulation and hyperinsulinemia on the reproductive functions of mares and the implications on foal health before and after birth. The points of interest are the cyclicity and ovarian function, uterine environment, gestation, the postpartum period, and the newborn foal. The aim is to review the current state of knowledge, and identify outstanding questions that could stimulate future research. This topic is important not only from the equine industry and production perspective but is also relevant for the welfare of future populations and individuals.

One Health challenges for pig reproduction.

The current state of the world challenges pig reproduction as an important part of One Health, which involves interrelationships between animal, human and environmental health. The One Health concept underlines a comparative aspect in reproductive physiology and disease occurrence, bridging knowledge from one species to another. Seasonal changes in the environment affect pig reproduction and climate change may further strengthen those effects. Endocrine-disrupting chemicals (EDCs), and specifically phthalates and heavy metals, interfere with endocrine function, and thereby sexual behavior, fertilization capacity and steroidogenesis. Reproductive infections and extended semen storage are important indications for antimicrobial use. Innovative solutions are needed to explore alternatives to antimicrobials. Efforts to ensure reproductive efficiency have prolonged farrowing as litter size has doubled over the past three decades, compromising immune transfer and welfare. Physiological, metabolic and programming related events around parturition are key areas for future One Health research in pig reproduction. In conclusion, climate change challenges reproductive management and breeding. More resilient pigs that can tolerate harsh environment but maintain high reproductive performance are needed. EDCs continue to grow as an environmental challenge for reproductive management and alternatives to antibiotics will be required.

The Effect of Uterine Lavage on Soluble CD14, Chemokine Ligand 2, and Interleukin 10 Levels in Mares With Postpartum Metritis.

Postpartum metritis in mares is a life-threatening condition associated with severe clinical signs due to endotoxemia, and it is often followed by complications such as laminitis. Repeated large-volume uterine lavages are commonly recommended as a part of the treatment protocol to remove endotoxin-laden contents from the uterus. It has, however, also been suggested that lavages may increase the uptake of endotoxin into the circulation, leading to a deterioration of clinical signs. Endotoxemia is associated with the release of a multitude of inflammatory mediators regulating the immune response. The aim of this study was to evaluate if uterine lavage influences serum levels of the inflammation markers soluble CD14 (sCD14), chemokine (C-C motif) ligand 2 (CCL2), and interleukin (IL)-10 in mares with postpartum metritis. Serum samples were collected from eight mares treated for metritis at a university teaching hospital. Mares with fever, tachycardia, and/or leukopenia and hemosanguineous or purulent intrauterine fluid within 1 week of foaling were included in the study. Serum samples were taken before uterine lavage and 15 and 30 minutes after starting the lavage. The concentrations of sCD14, CCL2, and IL-10 were determined with a fluorescent bead-based immunoassay. There were no significant differences between sCD14, CCL2, or IL-10 levels at different sampling times. Heart rate was significantly lower after uterine lavage than before. The differences in body temperature and leukocyte count before and after lavage were not significant. In conclusion, uterine lavage had no immediate effect on the serum concentration of sCD14, CCL2, or IL-10, nor did it affect clinical parameters negatively.

Matrix metalloproteinase (MMP)-2, MMP-9, semen quality and sperm longevity in fractionated stallion semen.

Matrix metalloproteinase (MMP)-2 and MMP-9 are gelatinases that take part in several reproductive processes. The aim of this study was to measure levels of MMP-2 and MMP-9 in fractionated stallion ejaculates, and to evaluate the association between these components and semen quality, and sperm longevity during cooled storage. Semen quality were assessed separately for sperm-rich fractions (HIGH), sperm-poor fractions (LOW), and whole ejaculate samples (WE) from 33 stallions. After cooled storage with SP either present or removed, sperm motility and DFI were determined. The relative activity of the pro-form of MMP-2, active MMP-2 and total MMP-9 were evaluated using gelatin zymography, and all were present in all fractions of the stallion's ejaculate, with higher relative activity of the latent than active forms and the highest relative activity in the HIGH fraction. The relative activities of MMP-2 and MMP-9 were positively correlated to sperm concentration and total sperm count, but only in the HIGH fraction and not in LOW or WE. The relative activities of MMPs were not related to differences in sperm longevity during cooled storage, measured as sperm motility and DFI. There was a harmful effect of SP on DFI during storage, but this effect was not associated with differences in the relative activities of MMPs. In conclusion, the relative activities of MMPs are not useful as markers for semen quality (other than sperm concentration), or sperm survival during storage in horses.

Analysis of factors affecting the pregnancy rate of mares after inseminations with cooled transported stallion semen.

Artificial insemination (AI) with cooled stallion semen has increased markedly during the last decades in all countries, but fertility is often lower than when fresh semen or natural mating is used. The objective of this study was to examine field data (1634 ,cycles 523 Standardbred (SB) mares, 575 Finnhorse (FH) mares, and 90 stallions) using multivariable logistic regression for factors influencing the pregnancy rate (PR) after AI with cooled transported semen from SB and FH stallions. The PR per cycle for the material was 47%: Finnhorses 42% and Standardbreds 53%. When assessed with multivariable logistic regression analyses with a generalized linear mixed model, variables that affected the PR were breed, the number of inseminated estrus cycles, the percentage of progressively motile sperm (PMOT) in the ejaculate/AI dose at the time of shipment, and the number of progressively motile sperm in the AI dose at the time of insemination. In Standardbreds, variables that increased the per cycle PR were the number of AI per estrus cycle (multiple inseminations increasing the probability of pregnancy compared to only one insemination), the number of inseminated cycles, and PMOT in the AI dose at the time of insemination. In Finnhorses, the number of AI per estrus cycle (two and three inseminations increasing the probability of pregnancy compared to only one), the number of spermatozoa in the ejaculate and in the AI dose, and PMOT in the ejaculate/AI dose at the time of shipment increased the per cycle PR. Non-significant factors for the whole material included the type of artificial vagina (open-ended or closed), transport time, place of AI (stud farm or home stable), insemination done by veterinarian or technician, weekday, month, age of the mare (all age classes combined), age of the stallion, ejaculate parameters (sperm concentration, total number of sperm), and insemination dose parameters (volume proportion of seminal plasma, sperm concentration, PMOT, total number of sperm). In conclusion, breed, breeding opportunity in more than one cycle, more than one insemination/estrus, PMOT of the ejaculate/AI dose and the number of progressively motile sperm in the AI dose at the time of insemination are important for the outcome of inseminations with cooled semen.

Components of stallion seminal plasma and the effects of seminal plasma on sperm longevity.

Seminal plasma is a mixture of secretions produced in the testes, epididymides and accessory sex glands, and ejaculated as several consecutive fluid fractions. The composition of seminal plasma and the effects on sperm longevity vary between fractions and individual stallions. This review focuses on the sequence of ejaculation, constituents of seminal plasma and their potential use as fertility markers as well as the influence of seminal plasma on spermatozoa during storage.