Does the Uterine Ozone Therapy Alter the Transcript Profile of Anti- and Proinflammatory Genes in Mares With Endometritis?

This study aimed to evaluate the localised effects of intrauterine ozone therapy on endometrial recovery in mares with endometritis. Our investigation assessed changes in gene expression profiles of anti-inflammatory (IL-1RA and IL-10), proinflammatory (IL-R1B3i and TNFα) and pleiotropic (IL-6) cytokines, along with detailed histological measurements of epithelial and endometrial thickness and the glandular area ratio. Twenty mares were assigned to a 2 × 2 factorial design based on endometritis diagnosis and treatment (control or 42 µg/mL ozone insufflation), resulting in four groups: NC (negative for endometritis/control), NO (negative/ozone), PC (positive/control) and PO (positive/ozone). Oestrus was induced with 2 mg of oestradiol benzoate on Days -1, 1 and 3, plus 1 mg on Day 5. Day 0 marked the initial uterine treatment, followed by insufflations on Days 1 and 2 with O3 (ozone) or O2 (control). Uterine biopsies were taken before treatment on Day 0 and Day 6 for histological analysis and gene expression assessment. Data were analysed using a statistical model that included endometritis status, treatment type, biopsy times (D0 and D6) and their interactions, analysed with Proc Glimmix. Regardless of treatment or endometritis status, significant biopsy effects (p < 0.01) indicated increased epithelial height and endometrial thickness in Day 6 samples. Analysis of IL-1 and TNFα revealed a significant interaction (p < 0.05) among endometritis, treatment and biopsy, with higher IL-1B3i expression on Day 6 in the PC group. The treatment effect (p < 0.04) showed a higher frequency (p < 0.01) of animals with positive modulation in the PC group (66.7%) versus the PO group (0.0%). An interaction effect (p = 0.08) between endometritis and treatment resulted from higher IL-1RA expression on Day 6 in the PC group compared to the PO group. Biopsy effect was significant for IL-10 (p < 0.01), indicating higher values in the second sample associated with tissue repair. In the short-term evaluation, ozone therapy did not influence endometrial morphology and may modulate cytokine expression, specifically the reduction in IL-1 and TNFα levels. Therefore, this therapy appears to be a safe and potentially effective treatment for modulating the inflammatory response in mares with endometritis.

Equine uterine sanitizer: in vitro inhibition of endometritis-causing microorganisms and its stability to dilute antibiotics.

Endometritis is the leading cause of mare subfertility. Most mares respond to standard therapy, but alternative therapies have been developed for mares failing to respond. This study aimed to investigate a commercially available, yet unassessed, product labeled as a uterine sanitizer to determine the in vitro antimicrobial activity against microorganisms associated with endometritis and its in vitro stability to dilute antibiotics. In experiment 1, the microdilution broth technique and antimicrobial effects were assessed against Escherichia sp, Staphylococcus sp., Klebsiella sp., Pseudomonas sp., and Candida sp. Percentage inhibition was calculated by comparing the optical density. The minimum inhibitory concentration (MIC) 100% was determined using the resazurin dye technique. MIC 50% and 90% were determined using a dose-response non-linear regression. In experiment 2, the uterine sanitizer was used to dilute commonly used antibiotics achieving a final volume of 90 mL at 5°C, 21°C, and 37°C. The pH was measured at 0, 1, 3, 6, and 24 h after dilution. The uterine sanitizer had inhibitory properties against all microorganisms; Escherichia sp. being the most susceptible, and Pseudomonas sp. the most resistant. The uterine sanitizer had an acidic pH=4; however, when combined with the antibiotics, the pH of the antibiotic remained unchanged with the different temperatures and did not precipitate. In conclusion, the uterine sanitizer showed antimicrobial effects against endometritis-causing microorganisms. The dilution of antibiotics in the uterine sanitizer was stable and this association could potentiate the antimicrobial effects. Uterine sanitizer's safety and clinical efficacy in vivo remain to be tested.

Association between non-esterified fatty acids and calcium concentrations at calving with early lactation clinical diseases, fertility and culling in grazing dairy cows in Uruguay.

This study describes the association of non-esterified fatty acids (NEFA) and calcium concentrations at calving with early lactation disease, reproductive performance and culling in 646 dairy cows from 13 commercial grazing dairy herds in Uruguay. During one year, health events were recorded from calving to 30 days in milk (DIM). The first author visited each farm every 20 days. During each visit, body condition score (BCS) was recorded (scale 1-5), defining BCS < 3 as suboptimal and BCS > 3 as optimal, and a blood sample was taken from cows between 0 and 4 DIM for metabolite determination. To evaluate the association between health events (i.e., retained placenta-metritis and clinical mastitis) and risk factors (parity, BCS, high NEFA (> 0.6 mmol/L) and subclinical hypocalcemia (SCH) (< 2.10 mM)) data were analysed using multivariable logistic regression models. To evaluate the association of health events and risk factors with reproductive performance and culling, data were analysed using Cox proportional hazard regression models. A risk factor and an outcome of interest were assumed to be associated at P < 0.05 and a tendency to be associated was defined at P < 0.10. Overall, 47 % (n = 303) of the cows showed elevated NEFA concentration and 77 % (n = 499) had SCH. In addition, 21.5 % (n = 139) of the cows recorded at least one clinical disease. Cumulative incidence was 17 % (n = 109) for clinical mastitis, 4.2 % (n = 27) for retained placenta (RP)-metritis and 1.4 % (n = 7) for lameness. Clinical mastitis was associated with parity, with lower odds in primiparous (PP) cows (OR = 0.42, P < 0.01). Cows in an optimal BCS also tended to have lower odds (OR = 0.66, P = 0.07). Moreover, high NEFA and SCH cows had higher odds of CM (OR = 4.5, P = 0.01 and OR = 1.75, P = 0.04, respectively). Retained placenta-metritis tended to be associated with high NEFA concentration (OR = 2.2, P = 0.06). Primiparous cows with suboptimal BCS showed an increased first insemination rate (HR = 2.34; P < 0.01). The risk of culling was lower in PP cows (HR = 0.19; P < 0.01) and in cows with optimal BCS and low NEFA concentration (HR = 0.38; P = 0.03). Our data show that metabolic challenge (defined as peripartum suboptimal BCS, high NEFA or SCH) is associated with increased odds of clinical mastitis and RP-metritis, decreased probability of insemination and increased hazard of culling. Under grazing conditions, we suggest that farm management to improve the metabolic adaptation to lactation represents an opportunity to enhance cow performance in terms of health, fertility and longevity.

Short communication: Association between prepartum subclinical magnesium imbalance and postpartum diseases in grazing dairy cows in Southern Chile.

This study aimed to evaluate the relationship between prepartum subclinical hypomagnesemia (pre-SHMg) and the occurrence of dystocia, metritis, clinical mastitis, lameness, and subclinical hypomagnesemia postpartum (post-SHMg) in pasture-based dairy cows. Also, the difference in means of prepartum magnesium (Mg) concentration by postpartum health events was evaluated. A total of 890 dairy cows from 32 commercial farms located in southern Chile were enrolled. Cows were examined twice, once between 30 and 3 days before and once between 3 and 30 days after calving. Blood samples were collected on both assessments, and cows were considered as having SHMg if serum total Mg < 0.65 mmol/L. On the postpartum visit, cows were evaluated for metritis and lameness. Information about clinical mastitis and dystocia was collected from on-farm records. Data were analyzed using multivariable mixed linear models and multivariable mixed logistic regression models. The overall prevalence of pre-SHMg was 9.9%, and its presence was associated with the occurrence of post-SHMg (odd ratio [OR] = 5.7; P < 0.0001) and metritis (OR = 3.1; P = 0.04). However, we did not detect an association between pre-SHMg and dystocia, clinical mastitis, or lameness after calving. Prepartum serum Mg concentrations were lower in cows that developed post-SHMg than those that did not (LSM ± SE = 0.75 ± 0.02 mmol/L vs. 0.83 ± 0.02 mmol/L; P < 0.0001). In conclusion, pre-SHMg was associated with a higher risk of post-SHMg and metritis in grazing dairy cows but not other postpartum health events.

Antimicrobial efficiency of bromhexine hydrochloride against endometritis-causing Escherichia coli and Trueperella pyogenes in bovines.

In this study, the main agents associated with endometritis in cows in the state of Santa Catarina, Brazil, were identified and the resistance profile and virulence mechanisms of the bacterial isolates were evaluated. Isolates of Escherichia coli and Trueperella pyogenes were tested for their biofilm forming ability and the antimicrobial action of bromhexine hydrochloride in combination with other antimicrobials. A total of 37 uterine lavage samples were collected from cows with endometritis. Of the 55 bacteria isolated, 25.4% were identified as T. pyogenes and 16.3% as E. coli. The bacterial isolates showed greater resistance to sulfamethoxazole + trimethoprim (58.2%) and tetracycline (56.3%). Among the species, E. coli showed the highest resistance rates, with 100% of isolates showing resistance to amoxicillin, streptomycin, and gentamicin. The results of the minimum inhibitory concentration for the T. pyogenes isolates showed that 91.6% of the isolates were resistant to enrofloxacin and tetracycline, and 75% were resistant to ceftiofur and sulfamethoxazole + trimethoprim. All E. coli and T. pyogenes isolates showed biofilm forming ability. The plo, fimA, and nanH genes were identified in 100% of T. pyogenes isolates. In parallel, 100% of E. coli isolates had the fimH gene, and 11.1% had the csgD gene. Bromhexine hydrochloride showed antimicrobial activity against 100% of E. coli isolates and 66.6% of T. pyogenes isolates. Furthermore, when associated with antimicrobials, bromhexine hydrochloride has a synergistic and additive effect, proving to be an option in the treatment of endometritis in cows and an alternative for reducing the use of antimicrobials.

Streptococcus equi subsp. zooepidemicus: High molecular diversity of Argentinian strains isolated from mares with endometritis.

Streptococcus equi subsp. zooepidemicus (S. zooepidemicus) is a mucosal commensal of the lower genital tract in horses and is the most isolated bacterium causing endometritis in mares. The aim of this study was to determine the molecular diversity of S. zooepidemicus obtained from endometritis in mares in Buenos Aires province, Argentina. Thirty isolates obtained from the uterus of mares in 2005 and 2017 were studied. The MLST scheme was applied to identify the Argentinian genotypes and the clonal relationships and patterns of evolutionary descent were identified using the eBURST algorithm - goeBURST. Twenty six different Sequence types (STs) were identified, being only 11 of them previously reported in horses and also, from several host species and tissues. The other 15 STs were reported in Argentinian reproductive strains of mares in our study for the first time. The genotypes obtained from uterus in Argentina were not evenly distributed when all the published S. zooepidemicus STs were analysed, thus, it was not possible to establish that the same lineage circulates in our equine population. The fact that the identified genotypes were also reported in other countries, diverse samples and host species suggest that there is not a host, and an anatomical niche adaptation. Finally, the isolation of the same genotype in the vagina/clitoris and the uterus of the same mare highlights the versatility of S. zooepidemicus and its role as an opportunistic pathogen.

Isolation of lactic acid bacteria from the reproductive tract of mares as potentially beneficial strains to prevent equine endometritis.

Endometritis, the inflammation of the endometrium, is the leading cause of subfertility in mares, and therefore responsible for major economic losses in the horse industry worldwide. It is generally treated with uterine lavages combined with ecbolic agents and local or systemic antibiotics. However, since antibiotic overuse has been associated with antimicrobial resistance in mares with persistent endometritis, new prevention and treatment alternatives are needed. One such alternative could be the use of probiotic lactic acid bacteria (LAB) isolated from the host. Thanks to their species specificity, resident microbiota may restore ecological equilibrium within the host, and therefore, help prevent infections and improve physiological functions. In the present study, 257 bacterial strains were isolated from 77 healthy mares, and 88.76% (n = 228) of them were phenotypically classified as LAB. Within this group, 65.79% were able to inhibit at least one strain from each of the genera that most commonly cause equine endometritis (Streptococcus equi subsp. zooepidemicus, Escherichia coli, and Staphylococcus spp.). Five strains (RCE11, RCE20, RCE91, RCE99, and RCE167) were selected on the basis of their beneficial properties: ability to autoaggregate and adhere to equine epithelial cells, high inhibition of and co-aggregation with all the bacteria isolated from clinical cases of endometritis evaluated, and negative co-inhibition between one another. All five were finally identified as Enterococcus spp., namely E. faecium (two strains), E. hirae (two strains), and E. gallinarum (one strain). Further studies will assess their safety and biotechnological potential for the design of a multi-strain probiotic formula to prevent equine endometritis.

Standardization of chronic uterine infection of hamsters by Leptospira santarosai serovar Guaricura, from serogroup Sejroe.

Leptospirosis is an important zoonosis that in cattle is characterized as a reproductive disease. It is well reported that the main agent of bovine leptospirosis worldwide is Sejroe serogroup serovar Hardjo. Reproductive disease in cattle has several gaps in its knowledge and studies with Golden Syrian hamsters, experimentally infected, are limited. Therefore, a protocol that could reproduce the chronic genital disease in hamsters would be extremely valuable for the advance of the knowledge of that syndrome. The aim of this study was to establish an experimental protocol for chronic non-lethal genital infection of female hamsters by L. santarosai serovar Guaricura (Sejroe serogroup), strain 2013_VF52. For this, two concentrations (1.0 × 108 leptospires/mL and 1.0 × 104 leptospires/mL) were used intraperitoneally in female hamsters of 06-08 weeks of age. Hamsters that survived for up to forty days after inoculation were euthanized. Uterine and renal tissues were collected to evaluate leptospires' presence by PCR and culture. The protocol demonstrated that 1.0 × 104 leptospires/mL of the strain determined chronic genital leptospirosis in the hamster model. The standardization of a protocol for chronic genital leptospirosis in hamsters can be extremely useful for the understanding of the physiopathology of the infection, as the distribution of leptospires in the uterus and the agent-host interactions.

Evaluation of clinical and subclinical endometritis impacts on the reproductive performance and milk production of dairy cows in Brazilian herds.

Clinical endometritis (CE) and subclinical endometritis (SE) are two of the main uterine diseases in dairy cows during the puerperium period that can directly affect milk production and fertility in dairy herds. The first objective of this study was to investigate the effects of the CE and SE on the subsequent reproductive performance and milk production of dairy cows in Brazilian herds, and the second objective was to evaluate the reproductive performance and milk production of dairy cows according to the positive or negative isolation of bacteria in the uterine environment. A total of 279 lactating Holstein dairy cows (28-33 days in milk) from six commercial farms were studied. These were classified into three groups: healthy cows (without CE and SE, n = 161), cows with CE (vaginal discharge score = 3 and ≥18% PMNL, n = 83) and cows with SE (absence of signs of CE and >18% PMNL, n = 35). According to the presence of bacteria isolated from the uterine environment, the animals were classified as healthy cows (n = 161), CE cows with positive (n = 68) and negative (n = 15) bacterial isolation; and SE cows with positive (n = 21) and negative (n = 14) bacterial isolation. Cows with CE (20.5%) and SE (31.4%) had a lower (p < .05) conception rate at first AI than healthy cows (47.8%). Furthermore, both cows with CE (3.1) and SE (2.8) needed more service to become pregnant than healthy cows (1.7) and more days to be pregnant (50.3 days in CE and 43.1 days in SE than healthy group; p < .001). Cows with CE (23.8 kg) and SE (23.7 kg) produced less milk than healthy cows on the day of sampling (28.2 kg); however, milk production during 305 DIM of cows with CE and SE was not impaired when compared to healthy cows. The logistic regression showed that parity, body condition score, 305 days milk production, last calving (eutocia or dystocia), twin birth and stillbirth were not associated with the incidence of CE and SE. In addition, we highlight that cows with CE and SE, but with negative bacteria isolation, did not have reproductive performance and milk production impaired during the postpartum period. These results support that the use of antibiotics would not be beneficial in these cows, contributing to the strategic and rational use of antibiotic therapy on dairy farms.

Endometrial expression of members of the IL-1 family: their involvement in delayed conception of dairy cows.

The cytokines of the interleukin-1 (IL-1) family are closely involved in the resolution of inflammation in cows with metritis and endometritis. However, little is known about the role of these cytokines beyond uterine regression in the absence of disease, especially around conception. Thus, the aim of this study was to examine the gene and protein expression of IL-1α, IL-1ß, IL-1RI, IL-1RII and IL-1Ra in endometrial biopsies previous to conception, to evaluate the possible association of these cytokines with delayed conception in dairy cows. Gene and protein expression levels were evaluated by real-time PCR and immunohistochemistry, respectively. The gene expression levels of cytokines were not associated with the duration of the period to conception following parturition. However, high protein expression of IL-1ß and low protein expression of IL-1Ra were significantly associated with early conception. These results suggest that an imbalanced protein expression of IL-1ß and IL-1Ra in the endometrium of dairy cows could be part of the maternal immune response mechanism necessary to propitiate early conception and probably to maintain pregnancy.

Addition of clinoptilolite in the diet reduces uterine PMN leukocytes and open days in multiparous lactating dairy cows managed in a mountain tropical pasture-based system.

This study aimed to assess the effect of adding clinoptilolite in the diet on uterine health and reproductive performance in multiparous lactating dairy cows managed in a tropical pasture-based system above 2500 m of altitude. Seventy-seven multiparous Holstein crossbred cows from two farms were allocated randomly into two groups: clinoptilolite supplemented cows (CLG, n = 42) and non-supplemented cows as control (CG, n = 35). Cows from CLG were supplemented with clinoptilolite from 30 days (50 g/cow/day) before to 60 days after calving (200 g/cow/day). In CLG cows, percentages of uterine PMN leukocytes (P < 0.0001) and proportion of subclinical endometritis (P = 0.0187) were lower than in CG. The interval calving to first corpus luteum was shorter (P = 0.0759) in CLG than CG, and calving to first service interval was similar between treatments. Cows from CLG became pregnant 35 days earlier than CG cows (P = 0.0224). On farm A, calving to conception interval was 18.1 days longer in CLG than in CG (P = 0.3750); in farm B, this interval was 86.2 days shorter in CLG than in CG (P = 0.0002). In conclusion, daily addition of clinoptilolite in the diet decreased the percentage of uterine PMN leukocytes, the proportion of cows with subclinical endometritis, and shortened the calving-conception interval in multiparous lactating dairy cows.

Utilização da ozonioterapia como tratamento da endometrite em égua / Use of ozone therapy as endometritis treatment in mare

Conformações perineais inadequadas estão frequentemente associadas à pneumovagina, predispondo animais a processos inflamatórios vaginais e uterinos. Dentre as patologias reprodutivas que acometem éguas, a endometrite é a que ocorre com maior frequência, com forte impacto negativo sobre os índices reprodutivos de um criatório. Para essa afecção, a ozonioterapia se apresenta como tratamento em potencial. Assim, este estudo objetiva relatar um caso de endometrite em égua Quarto de Milha, multípara, de 17 anos de idade, atendida no Hospital Veterinário Sylvio Barbosa Cardoso, da Universidade Estadual de Ceará. Durante a avaliação ginecológica, observou-se alterações na conformação vulvar e teste de Windsucker positivo, indicando pneumovagina. Foram coletadas amostras endometriais para exames laboratoriais, os quais apresentaram citologia com contagem de neutrófilos polimorfonucleares ≥30% e cultura negativa para fungos e bactérias. O exame histopatológico revelou fibrose intersticial endometrial crônica acentuada. Diante do diagnóstico de endometrite, iniciou-se o tratamento com lavagem uterina usando solução ringer lactato ozonizada, seguido por insuflação uterina com ozônio. O animal foi submetido à técnica cirúrgica de Caslick para correção da má conformação vulvar. Após o tratamento, a égua foi inseminada, resultando na coleta de dois embriões viáveis. Diante do exposto, o caso reforça a efetividade da ozonioterapia associada à Caslick para o tratamento de endometrite em éguas.

Dysfunctional perineal conformations are often associated with pneumovagina, predisposing animals to vaginal and uterine inflammatory processes. Among the reproductive pathologies that affect mares, endometritis is the most frequent, with a strong negative impact on the reproductive indices of a farm. For this condition, ozone therapy presents itself as a potential treatment. The study aimed to report a case of endometritis in a 17-year-old multiparous quarter-mile mare, which was attended at the Veterinary Hospital Sylvio Barbosa Cardoso located in the State University of Ceará. During the gynecological evaluation, changes in vulvar conformation and a positive Windsucker test were observed, indicating pneumovagina. Endometrial samples were collected for laboratory exams, resulting in cytology with polymorphonuclear neutrophil count ≥30% and negative culture for fungi and bacteria. The histopathological examination revealed severe chronic endometrial interstitial fibrosis. Regarding the diagnosis of endometritis, the treatment with uterine lavage using ozonated ringer lactate solution was initiated followed by uterine insufflation with ozone. The animal was submitted to Caslick's surgical technique to correct the vulvar malformation. After treatment, the mare was inseminated, resulting in the collection of two viable embryos. In view of the above, the case reinforces the effectiveness of ozone therapy associated with Caslick`s technique for endometritis treatment in mares.

Effect of retained placenta and clinical mastitis on reproduction parameters, immune response, and steroidogenic receptors gene expression in postpartum crossbred dairy cows.

The purpose of this study was to evaluate the effect of retained placenta (RP) and clinical mastitis (CM) on the reproductive efficiency of crossbred dairy cows during the postpartum period and the effect in some innate immune system indicators. For this, two experiments were carried out. In the first, a total of 232 cows were evaluated and divided as: healthy control (n = 184), RP (n = 22), and CM (n = 26) groups. The RP and CM was evaluated until 30 days postpartum (DPP) and reproductive rates were measured. In experiment 2, cows were divided in control (n = 10), RP (n = 10), and CM (n = 30) groups. Between 40 and 50 DPP, clinical, gynecological examination and endometrial cytobrush were performed to evaluate subclinical endometritis (SE) and gene expression of interleukins 1ß (IL-1ß) and 6 (IL-6), chemokine ligand 5 (CCL5), estrogen α (ESR1), and progesterone (PGR) receptors by qRT-PCR analysis. In experiment 1, the conception rate at 1st artificial insemination (AI) was lower in RP and CM groups and pregnancy rate at 150 days decreased in CM group. Calving-to-1st AI interval and days open were shorter in healthy cows. In experiment 2, the occurrence of SE was 26.7% and higher in RP and CM groups. The expression of IL-1ß increased in RP and CM groups, while IL-6 was less expressed in RP group. The CCL5, ESR1, and PGR were similar between groups. In conclusion, cows with RP and CM had their reproductive efficiency negatively affected and had they initial pro-inflammatory response improved by the increase of IL-ß.

Vascular endothelial growth factor (VEGF) expression in histological endometritis in the bitch: An immunohistochemical study.

This work aimed to study the immunolocalization of vascular endothelial growth factor (VEGF) and its FLT-1 receptor (VEGFR1) in systemically healthy bitches with and without endometritis (END). Thirty-one uterine samples from mixed-breed, systemically healthy diestrous bitches, were included in the study. The bitches were divided into three groups after histological evaluation (normal endometrium [n = 8]; acute-END [n = 8]; subacute-END [n = 5]; chronic-END [n = 10]). The immunohistochemical localization of VEGF and VEGFR1 were detected following standard procedures. Data were analysed using SPSS 26.0. Immunoreactions were detected in cells from the luminal epithelium (LE), epithelium of the superficial glands (ESG) and epithelium of the deep glands (EDG), and some cells localized in the stroma. Immunostaining of VEGF in acute-END was higher than chronic-END (p < .05) and was higher in LE than EDG (p .017). Marked-area was higher in END than normal endometrium in VEGFR-1 (p < .004). The current results provide new information on the END in systemically healthy females. Data reported here indicate that VEGF could be involucrated in the pathogenesis of END. Future studies could provide more information, support our results and elucidate the role of VEGF in the pathogenesis of END.

Association between serum calcium levels and the presentation of postpartum endometritis in housed dairy cows.

The present work aims to identify the minimum threshold of serum calcium (SC) values in cows during the first week postpartum and evaluate their relationship with the presentation of endometritis in housed dairy cows. In this prospective longitudinal observational study, 467 cows from 3 farms in Lima-Peru were studied. Blood samples were collected from cows during the first week postpartum. Endometritis was diagnosed on day 35 ± 3 days postpartum by evaluation of vaginal discharge samples. The samples were obtained using the MetricheckTM device (Simcro, New Zealand). Two cut-off points were used to diagnose endometritis: a) endometritis metricheck score ≥ 3, and b) endometritis metricheck score ≥ 2. In the univariate model that considers a SC threshold of 5.25 to 8.75 mg/dL (1.31 to 2.18 mmol/L), a significant relationship (p < 0.05) was found for endometritis metricheck score ≥ 3, while no significant relationship was found (p = 0.12) with endometritis metricheck score ≥ 2. In both cases, the optimal SC threshold for the appearance of endometritis was determined to be a value ≥7.0 mg/dL (≥1.75 mmol/L). In the logistic regression models (parity, calving season, calcium level, and their interaction), only two variables were retained, parity and calcium level (p < 0.10). It was found that the probability of having endometritis metricheck score ≥ 3 was 1.9 (95% CI: 1.1 to 3.1), whereas the probability of having endometritis metricheck score ≥ 2 was 1.5 (95% CI: 1.0 to 2.5) in cows with calcium levels <7.0 mg/dL (<1.75 mmol/L). In conclusion, cows with calcium levels <7.0 mg/dL (<1.75 mmol/L) have a 1.9-fold greater risk of developing endometritis metricheck score ≥ 3 in the first week postpartum.

In Vitro Antimicrobial Activity of Selected Essential Oils Against Endometritis-Causing Microorganisms in Mares.

This study aimed to evaluate the in vitro antimicrobial activity of essential oils (EO) from Ocimum basilicum (basil), Rosmarinus officinalis (rosemary), and Cymbopogon citratus (lemongrass) on endometritis-causing microorganisms in mares. Serial concentrations of the EO from 30.00 mg/mL to 0.47 mg/mL were tested. The major compounds of O. basilicum EO were linalyl acetate (33.32 wt.%) and citronellal (25.06 wt.%); of R. officinalis EO were borneol (26.48 wt.%), trans-ß-ocimene (16.76 wt.%), camphene (12.45 wt.%), and α-phellandrene (11.08 wt.%); and of C. citratus EO were geranial (45.96 wt.%) and neral (32.62 wt.%). Regarding antimicrobial activity, C. citratus EO has had the highest inhibition percentage (73.9%), followed by O. basilicum (67.2%) and R. officinalis (58.7%). P. aeruginosa was the only pathogen unable to establish the minimum inhibitory concentrations (MIC) and minimum bactericidal concentration (MBC) values for the studied EO. The EOs were effective against all other microorganisms (S. equi, S. aureus, K. pneumoniae, E. coli, and C. Albicans). In conclusion, the EOs of O. basilicum, R. officinalis, and C. citratus have presented in vitro antimicrobial activity against microorganisms causing endometritis in mares.

Oxidative stress, ceruloplasmin and neopterin biomarkers in dromedary camels with clinical endometritis

The purpose of this study is to investigate the role of some oxidative stress (OS), ceruloplasmin (Cp), and neopterin (NPT) as diagnostic biomarkers for dromedary camels endometritis as well as to explore the impact of ceftiofur treatment on endometritis. Camels were categorized into two groups; healthy control group (n = 20) and endometritis group (n = 60). She-camels with clinical signs of endometritis (CE) received 6.6 mg/kg BW of ceftiofur (i/m). On days 7, and 14, she-camels were evaluated and clinical cure or failure to cure was determined. The comparison of the groups for OS demonstrated that endometritis caused an increase in serum malondialdehyde (sMDA), Cp, and NPT levels (P<0.05), but decreased serum levels of superoxide dismutase (SOD), catalase (CAT), and glutathione (GSH) (P<0.05). The most prevalent pathogens involved in the etiology of CE are Arcanobacterium pyogenes, Streptococcus pyogenes, and Staphylococcus aureus. All examined biomarkers demonstrated a high degree of recognition between CE camel and healthy controls (the area under the curve (AUC) was 95.9 for NPT). A higher proportion of camels with CE that were treated with ceftiofur (90%, P<0.0001) showed clinical cure by the first dose, while 10% required a second dose. In conclusion, CE causes increased oxidative reactions and decreased antioxidant defense competence. Subsequently, the alteration in that balance that was represented by the biomarkers of OS could be beneficial for clinical practice and basic clinical research. Additionally, all trials demonstrated the efficacy of ceftiofur for the treatment of CE in she-camel.(AU)

Clinical endometritis with Trueperella pyogenes reduces reproductive performance and milk production in dairy cows.

The aim of this study was to identify the impact of Trueperella pyogenes in cows with clinical endometritis (CE) on reproductive performance and milk production in affected cows. In total, 230 lactating Holstein dairy cows from six commercial dairy herds were sampled once between 28 and 33 days post-partum. Cows included in the present study did not receive antibiotic or anti-inflammatory treatments prior to the experimental period. Clinical endometritis (CE) was characterized as cow with vaginal mucus score = 3 (>50% of purulent vaginal discharge) and >18% polymorphonuclear neutrophilic leukocyte (PMNL). The body condition scores (BCS) and milk production were evaluated at the time of enrolment. The identification of isolated bacteria was carried out through the analysis of MALDI-TOF MS (matrix-assisted laser desorption ionization time-of-flight mass spectrometry). According to uterine health, three groups of dairy cows were formed: healthy control cows without T. pyogenes (n = 147), CE cows with T. pyogenes (n = 22) and CE cows without T. pyogenes (n = 61). CE cows with T. pyogenes had lower BCS, milk production and conception at first AI (p < .01) than CE cows without T. pyogenes and control cows. Furthermore, CE cows with T. pyogenes had higher (p < .01) service per pregnancy and had greater (p < .01) days to get pregnant than CE cows without T. pyogenes and control cows. This study demonstrates that CE cows with T. pyogenes had impaired reproductive performance and milk production when compared to cows without CE and CE cows without T. pyogenes. This information can contribute to a strategic treatment in cows affected by clinical endometritis, favouring the rational use of antibiotics on dairy farms.

Incidence, clinical patterns, and risk factors of postpartum uterine diseases in dairy cows from high-altitude tropical herds.

The cow's uterus sustains bacterial contamination and active inflammation after calving as part of the normal physiological process of uterine involution. Although the definition, incidence, and risk factors for postpartum uterine diseases (PUD) have been documented in annual calving systems with high-producing cows on total mixed ration, the literature contains little information for other production systems. The objective of this study was to quantify the incidence risk and factors associated with metritis, clinical endometritis, and cytological endometritis in high-altitude tropical dairy herds in a pasture-based system. A total of 248 cows from 5 commercial dairy herds in northern Antioquia, Colombia, were enrolled in this prospective observational longitudinal cohort study. Data collection and PUD diagnosis were performed at d 0, 3, 10, 17, 24, 31, 38, 45, and 52 (±1) after parturition. Between 60 and 70 d, cows were systematically enrolled in a synchronization protocol, and herds were visited monthly for pregnancy diagnosis until 180 ±15 d. The multivariable logistic regression model of variables associated with PUD occurrence included (a) parity, season of calving, transition diet, and body condition score at calving; (b) blood calcium, phosphorus, and magnesium concentrations, and milk urea nitrogen, ß-hydroxybutyrate and fat-to-protein ratio; and (c) dystocia, retained placenta, and delayed uterine involution. The incidences of puerperal metritis, clinical metritis (MET), clinical endometritis (CE), and cytological endometritis (CYTO) were 2.8, 25, 29, and 26%, respectively. Retained placenta and dystocia were associated with MET, and MET was associated with CE. Low blood calcium concentration at calving was associated with CYTO. Overall, 51.2% of the cows suffered at least 1 category of PUD during the first 60 d postpartum (DPP). However, cows also suffered from different consecutive clinical events of PUD (MET, CE, and CYTO) during the follow-up period (0 to 180 ±15 DPP). These events could be grouped into 4 types (which were referred to as "clinical patterns") based on interactions between MET, CE, and CYTO.

Allogenic mesenchymal stem cell-conditioned medium does not affect sperm parameters and mitigates early endometrial inflammatory responses in mares.

This study aimed to evaluate the effects of mesenchymal stem cell-conditioned medium (MSC-CM) on sperm parameters, intrauterine polymorphonuclear neutrophils (PMN), intrauterine fluid accumulation (IUF), and fertility in mares. In experiment 1, two ejaculates from ten stallions were extended to 50 million sperm/mL using a milk-based extender. Thereafter, 20 mL of extended semen was added of MSC-CM as follows: 0, 5, 10, 15, and 20 mL. Sperm kinetics and plasma membrane integrity were evaluated immediately after dilution (T0) and 2 h post-incubation at 37 °C (T2). In experiment 2, mares characterized as resistant (n = 13) or susceptible (n = 7) to endometritis were inseminated with fresh semen 24 h post-induction of ovulation in two (Control and CM-1) and three (Control, CM-1, and CM-2) cycles in a crossover, as follows: control, no pharmacological interference; CM-1, supplementation of semen insemination dose at 3:4 (v:v, MSC-CM:semen); CM-2, 30 mL of MSC-CM was infused into the uterus 24 h before insemination. Endometrial cytology and uterine fluid were collected 6 and 24 h after insemination to evaluate the number of PMNs and concentrations of interleukins IL6, IL10, and TNFα. IUF was determined by ultrasonography 24 and 48 h after insemination. Pregnancy status was diagnosed 14 days after ovulation. The addition of MSC-CM to semen did not influence sperm parameters at T0 and T2 (P > 0.05) and reduced (CM-1; P < 0.05) the number of PMNs at 6 h post-insemination in resistant mares. In susceptible mares, PMNs at 6 and 24 h post-insemination, as well as IUF were reduced (P < 0.05) in both treated cycles (CM-1 and CM-2). In addition, MSC-CM downregulated IL6 and upregulated IL10 concentrations in the uterus of susceptible mares after insemination. There were no differences in fertility rates among groups both in resistant (Control, 77%, 10/13; CM-1, 62%, 8/13) and susceptible mares (Control, 42.8%, 3/7; CM-1, 57.1%, 4/7; CM-2, 85.7%. 6/7). In conclusion, MSC-CM did not affect sperm parameters when mixed with diluted semen, and reduced post-insemination inflammatory responses in mares.