Technique for guttural pouch bead removal using a novel three-dimensional (3D)-printed instrument.

OBJECTIVE: The aim of the present study was to determine if a three-dimensional (3D)-printed instrument technique would improve lavage removal of plastic beads (guttural pouch [GP] chondroid mimics) through a dorsal pharyngeal recess (DPR) fenestration. We hypothesized that using a 3D-printed instrument placed through the DPR fenestration would remove more beads, reduce lavage time and incur less soft tissue damage than using a lavage tube control or instrument placement through the salpingopharyngeal ostium (SPO). STUDY DESIGN: Experimental cadaveric study. SAMPLE POPULATION: A total of 30 cadaveric equine heads. METHODS: DPR fenestration was performed using transendoscopic laser and 50 plastic 12 mm beads were placed into one GP of horse heads. Four removal procedures using a 3D-printed instrument or lavage tube control placed through the DPR fenestration or the SPO were compared. Number of beads removed and number of 2-min lavage cycles to recover ≥96% of beads or three consecutive no-yield cycles were recorded. Endoscopic soft tissue damage was graded. Data were compared by generalized estimating equations (GEE) model and Fisher's exact test (p < .05). RESULTS: More beads (median 48 beads; range 0-49) were removed faster (median 24 beads/cycle; range 12-50) using the 3D-printed instrument compared to control (median 6 beads; range 0-29, 0.66 beads/cycle, range 0-49). There was no difference between total beads removed or removal speed between placement sites. There was no difference in soft tissue damage between procedures. CONCLUSION: Our 3D-printed instrument enabled efficient plastic bead removal. CLINICAL SIGNIFICANCE: DPR fenestration and use of our 3D-printed instrument represents an alternative to current chondroid removal techniques, warranting investigation in clinical cases.

Multicenter study investigating long-term survival after synovial lavage of contaminated and septic synovial structures in horses presented to 10 UK referral hospitals.

OBJECTIVE: To report the long-term survival of adult horses that were subjected to synovial lavage for treatment of contaminated and septic synovial structures. STUDY DESIGN: Multicenter, prospective observational trial. ANIMALS: Horses (n = 240) presented for synovial sepsis at 10 UK referral centers. METHODS: Data for horses presented for treatment of synovial sepsis were collected over a 15 month recruitment period. Owners were contacted a minimum of 365 days after surgery using a structured client interview to assess long term survival. Descriptive statistics, and univariable and Cox proportional hazards models for postoperative survival time were developed. RESULTS: Survival to discharge was 228/240 (95%) and overall long-term survival was 89.4% (185/207). Unknown cause of injury (p = .017), increasing duration of surgery (p = .003), increasing weight (p = .008), forelimb injuries (p = .027), and type of synovial structure (p = .008) were found to be associated with death using Cox proportional hazards models. CONCLUSION: This study provides information on risk factors associated with survival and death after treatment for synovial sepsis at referral hospitals in the UK. Survival to discharge and long-term survival was excellent. Heavier horses, injuries affecting the forelimbs, tendon sheaths and bursae were associated with poorer long term outcomes. Longer duration of surgery was also found to be associated with a worse prognosis. CLINICAL SIGNIFICANCE: These findings help to provide prognostic information for owners and veterinarians treating horses with synovial sepsis.

Ovarian lavage / artificial insemination method as an alternative approach for controlled reproduction of the northern pike (Esox lucius).

This study aimed to compare the effectiveness of the ovarian lavage / artificial insemination method with the traditional hormonal administration and fertilization methods over the artificial reproduction of the northern pike (Esox lucius). For this purpose, groups of five females were treated as follows: intraperitoneal injection of saline (C1); ovarian lavage with saline (C2); intraperitoneal injection of carp pituitary extract (CPE, T1); ovarian lavage with CPE (T2); intraperitoneal injection of CPE and ovarian lavage with semen after 72 h (T3); ovarian lavage with CPE and ovarian lavage with semen after 72 h (T4). According to the results, no fish ovulated in the control groups (C1 and C2). There were no significant differences (n.s.) among experimental treatments (P > 0.05, n.s.) in the reproductive parameters, such as latency time, ovulation rate, stripped egg amount, and pseudo-gonadosomatic index. The lowest fertilization rate (54.8%) was observed in the T4 treatment and significantly differed from the T1 and T2 treatments (P < 0.05). Moreover, the highest survival at swim-up stage was measured in the T4 treatment with a significant difference compared to the T1 group (P < 0.05). The survival at the eyed-egg stage, hatching rate, and malformations were similar (P > 0.05, n.s.) in all applied strategies. The results demonstrated that the ovarian lavage / artificial insemination method could be applied to control northern pike reproduction, like the traditional fertilization method. Consequently, this novel technique can be suggested as an alternative strategy to facilitate the hatchery operations in the controlled reproduction of this species.

Influence of antiseptic lavage during tibial plateau leveling osteotomies on surgical site infection in 1422 dogs.

OBJECTIVE: To determine the influence of preclosure antiseptic versus saline lavage on surgical site infections (SSI) in dogs following tibial plateau leveling osteotomy (TPLO). STUDY DESIGN: A multicenter retrospective study. SAMPLE POPULATION: Dogs treated with TPLO (n = 1422) between December 2019 and October 2021. METHODS: The medical records of dogs treated with TPLO were reviewed for preclosure antiseptic lavage or saline irrigation. Additional collected data included signalment, unilateral or bilateral TPLO, postoperative administration of antimicrobial medications, antibiotic agent, surgeon, and development of SSI within 90 postoperative days. Eleven factors were analyzed for association with SSI using univariate and multivariate analysis. RESULTS: Data were collected from the records of 519 dogs treated with antiseptic lavage and 903 dogs treated with saline lavage during TPLO. Surgical site infections were diagnosed more frequently in dogs that received preclosure antiseptic lavage (77/519, 14.84%) than those with saline irrigation (80/903, 8.86%) (p = .001). Single session bilateral TPLO increased the odds of SSI by 2.5x (p = .004). The odds of SSI increased by 11% (p = .001) for every 5 kg increase in bodyweight. Postoperative administration of antimicrobials decreased the risk of SSI (p = .008). CONCLUSION: The preclosure antiseptic lavage tested here did not decrease the incidence of SSI after TPLO. CLINICAL SIGNIFICANCE: The results of this study do not provide evidence to support preclosure antiseptic lavage during TPLOs.

Proteomic analysis of uterine lavage fluid of dairy cows at different time after delivery by mass spectrometry.

Endometritis is a common disease in the reproductive system, which is the infection and inflammation of the endometrium. In severe cases, it can affect the myometrium and adversely affect the subsequent fertility of dairy cows. We used a mass spectrometry-based technique to compare proteomics of uterine lavage fluid between healthy cows and cows with cytological endometritis classified according to 100-day postpartum pregnancy results and diagnosis result. The uterine lavage fluid of dairy cows collected at 15 and 30 days after delivery was analyzed. 15 days postpartum, we identified a total of 1129 proteins in the control and cytological endometritis (CEM) groups. Among them, 160 proteins were accurately screened out. 30 days postpartum, we identified a total of 846 proteins in the control and cytological endometritis (CEM) groups. Among them, 186 proteins were accurately cytological endometritis (CEM). Endometritis is a costly reproductive disease in lactating cows, which needs to be diagnosed in time. Using proteomics method based on gel mass spectrometry, we compared the proteome of uterine lavage fluid of dairy cows with and without cytological endometritis to characterize the changes of proteomic characteristics associated with postpartum uterine disease. To provide reference for clinical application and basic research.

Ovarian lavage method as an alternative route for hormonal administration and short-term sperm storage in Clarias gariepinus.

Ovarian lavage is a term used to describe the injection of fish with a catheter through the oviduct into the ovary. In this study, the efficacy of this technique was evaluated as a route for hormone administration and sperm preservation in the African catfish Clarias gariepinus. Firstly, the effects of hormone injection routes (namely, intramuscular, intraperitoneal, and ovarian lavage) were evaluated on breeding and haematological parameters. In the second study, the fish's spermatozoa were stored in the ovaries for 1, 2, 3, and 4 days before stripping, sperm activation with freshwater, and fertilization. The breeding performance was then compared with eggs fertilized using spermatozoa refrigerated for the same duration. The study showed that the administration of synthetic hormone (ovaprim®) through the ovaries was comparable to the intramuscular route, while those injected intraperitoneally had the least values (P < 0.05) for breeding performance. The trend of the haematological parameters also suggests the intraperitoneal administration procedure elicited more stress compared to intramuscular or ovarian lavage routes (P < 0.05). In the second experiment, the use of sperm stored in the ovaries or refrigerated for 24hr didn't affect (P > 0.05) the fertilization (92-93%) and hatching (81-83%) of the eggs when compared to the control (91% and 82%). Beyond this 24hr threshold, breeding performances were significantly reduced in the ovarian lavage treatments compared to those fertilized with refrigerated sperm (P < 0.05). The loss of fertilizing capacity observed herein was justified by the reduction in sperm quality over time (P < 0.05). It was concluded that ovarian lavage is a viable route for hormonal administration and 24hr sperm storage in C. gariepinus.

Preclosure povidone-iodine lavage in total hip replacement surgery: Infection outcomes and cost-benefit analysis.

OBJECTIVE: To report the outcomes and cost-benefit analysis of preclosure povidone-iodine lavage (PrePIL) used to reduce the risk of infection following total hip replacement (THR) surgery. STUDY DESIGN: Retrospective study. ANIMALS: One thousand six hundred ninety-nine dogs, 17 cats. METHODS: The medical records of 2213 consecutive THR cases were reviewed to determine the incidence of infection. The last 102 were treated with PrePIL using a commercially sourced 0.035% povidone-iodine solution. Postoperative infection rates were compared. A cost-benefit analysis was used to calculate if a PrePIL protocol is economically feasible. RESULTS: Twenty-one THRs out of 2111 (0.99%) that did not have PrePIL developed infection. Infection occurred in none of the 102 PrePIL cases. Cost analysis revealed a PrePIL break-even cost at $49.74 and a break-even infection rate of 0.949%. No complications were identified related to the use of PrePIL. CONCLUSION: Preclosure povidone-iodine lavage appeared to be efficacious in lowering THR infection rates, and it appeared to be safe for this use based on our 102 consecutive cases. The cost of the PrePIL was minimal compared to the overall cost to resolve THR infection and the potential effect on hip function prognosis. The math formulas developed can be used by surgeons to calculate cost effectiveness and break-even cost based on their THR infection rate, and to compare to the cost of a THR revision and infection resolution. CLINICAL SIGNIFICANCE: At current costs, PrePIL can be used in 2415 THR cases at a similar cost of a single revision surgery and resolution of a periprosthetic infection.

Subpalpebral lavage placement for remote topical administration of ocular medications in 12 dogs: A retrospective review and assessment of owner perception.

OBJECTIVE: The aim of this study was to describe the placement of subpalpebral lavage (SPL) systems in 12 dogs (15 eyes) intolerant of topical ocular medications to assess the suitability, complications encountered and owner perception of use. ANIMALS STUDIED: Retrospective review of dogs that underwent SPL placement for treatment of ocular disease at the Ophthalmology Department, University of Bristol Small Animal Hospital between 2017 and 2021. PROCEDURE(S): Data recorded included signalment, history, diagnosis, treatment, reason for SPL placement, uni- or bilateral placement, duration of placement, complications, and outcome. Owner perception was assessed using an online questionnaire. Statistical analysis included McNemar and Wilcoxon signed-ranks tests. RESULTS: Twelve dogs (15 eyes) underwent SPL placement. Eleven owners completed the online questionnaire. Corneal ulceration was the most common disease requiring SPL placement (n = 13/15 eyes, 86.7%). Most cases received multimodal topical therapy (n = 9/15 eyes, 60.0%) via SPL. Owners administered medication 6.63 times daily via SPL (range 1-16 applications/day). All dogs requiring ongoing topical medication (n = 8/12, 66.7%) were trained to accept direct administration during SPL treatment. Statistically significant improvements in medication compliance, ease of application, and reduced perceived risk of iatrogenic ocular injury were reported by owners (p-value = .001, .004, and .031 respectively). Minor complications were infrequently reported but an excellent outcome was achieved for all eyes. CONCLUSION: Subpalpebral lavage placement provides a practical and safe solution for the provision of frequent multimodal ocular medication when treating patients with a challenging temperament.

Synovial Fluid Analysis and Microscopic Assessment of Macrophage Quantities and Morphology in Equine Septic Arthritis.

OBJECTIVE: Research and provision of data on macrophages by cytological synovial fluid analysis and light microscopy in horses with septic arthritis MATERIAL AND METHODS: Records of 167 synovial fluid samples were evaluated and subdivided into different groups: (1) non-septic, (2) haematogenous septic arthritis in foals and (3) traumatic/iatrogenic septic arthritis. The effect of joint lavage on synovial fluid cytology and on the occurrence of macrophage phenotypes was investigated. RESULTS: Regardless of aetiology and age of the horse, macrophage concentrations in synovial sepsis are decreased to a median of 5-6 % (unaffected joints: 23.5 %) and further diminished by joint lavage. Microscopic assessment led to the identification of 4 phenotypes. Morphological characteristics of type 1 showed similarities to monocytes and predominated in unaffected and in septic joints after lavage. CONCLUSION AND CLINICAL RELEVANCE: Macrophages are highly versatile by altering their phenotype. A morphological assessment by light microscopy is easily applicable. Type 1 presumably contributes to joint homeostasis.

Strongly acidic gastroesophageal reflux and esophageal lumen pH before and after esophageal lavage with water or two bicarbonate concentrations in anesthetized dogs.

OBJECTIVE: To increase acidic esophageal lumen pH in dogs that developed gastroesophageal reflux (GER) during anesthesia. We compared water and 2 different bicarbonate concentrations. ANIMALS: 112 healthy, nonbrachycephalic dogs presented for ovariectomy. PROCEDURES: Following standard anesthesia and surgery protocols for ovariectomy in all dogs, esophageal lumen impedance and pH were monitored using a dedicated probe. Esophageal impedance indicates the presence of GER whereas pH indicates the acidity level. Dogs with strongly acidic GER and an esophageal lumen pH value < 4.0 were included in the study, and lavage was performed with either tap water, bicarbonate 1%, or bicarbonate 2% until the pH increased to > 4.0. The effect of lavage on esophageal pH was compared using the Kruskal-Wallis and Wilcoxon 2 sample tests. Associations between lavage and pH changes were determined. RESULTS: Of 48/112 dogs with strongly acidic GER, 33% neutralized their esophageal pH during surgery. For the 32 dogs that maintained an esophageal lumen pH value < 4, esophageal lavage with water increased the lumen pH to > 4 in 78.6% of dogs, whereas both bicarbonate concentrations increased it in 100% of the dogs to a more neutral pH (P < .0001). The dogs in the water group were more likely to regurgitate after anesthesia (36% vs 0% in both bicarbonate groups, P = .028). CLINICAL RELEVANCE: Bicarbonate 1% and 2% increased esophageal lumen pH to more than 4 after strongly acidic GER. Lavage with water was mildly effective, but required large volumes and predisposed to further regurgitation after anesthesia.

Changes in the C-reactive protein and 13,14-dihydro-15-keto-prostaglandin F2α concentrations of uterine lavage samples after the administration of povidone-iodine in cows.

Changes in the C-reactive protein (CRP) and 13,14-dihydro-15-keto-prostaglandin F2α (PGFM) concentrations of uterine lavage fluid were examined in cows given an intrauterine povidone-iodine (PI) infusion. The mean polymorphonuclear leukocyte (PMN) ratios (the ratio of PMN to total cells) and CRP concentration of uterine lavage fluid on the day after the treatment were significantly (P<0.05) greater in the PI infusion group (PMN: 53.0 ± 32.7%, CRP: 50.2 ± 32.3 ng/mL) than in the non-treatment control group (PMN: 7.9 ± 21.9%, CRP: 17.2 ± 5.9 ng/mL), whereas there was no significant difference in the mean PGFM concentration between the two groups. The present findings suggest that the uterine CRP level is a useful biomarker of local uterine inflammation in cows.

Outcomes and Complications Associated with Superior and Inferior Subpalpebral Lavage Treatment Systems Placed in 61 Equine Eyes (2004-2021).

Painful eye conditions are a therapeutic challenge in horses. Subpalpebral lavage (SPL) treatment systems allow topical aqueous medications to be conveniently, safely, and frequently administered to the ocular surface. The aim of this retrospective study was to describe the outcomes and the complications associated with the location of the SPL treatment systems, in the superior or inferior eyelid in horses in a university practice. Clinical records of all horses admitted to the National Veterinary School of Alfort between January 2004 and October 2021, in which a SPL treatment system was used to administer the topical ophthalmic medications, were reviewed. Sixty horses were included in the study representing 61 SPL treatment systems. Uneventful outcomes occurred in 53 cases (86.9%) and complications were recorded in 8 cases (13.1%). Seven complications were reported for upper eyelid systems (23.3%) and 1 complication for lower eyelid systems (4.2%). The complication rates were not significantly different between these two locations (P= .06). The complications identified were iatrogenic corneal ulceration, palpebral abscess, overgrowth of conjunctiva over the footplate, and palpebral cellulitis. Median duration of SPL treatment system use was 7.5 days. The easier placement and removal and the lower incidence of complications identified in our study encourage the authors to favor the lower fornix for subpalpebral lavage treatment systems within our equine hospital.

Dorsally placed commercially available subpalpebral lavage systems have low complication rates in horses.

OBJECTIVE: To retrospectively evaluate the complication rate following dorsal placement of a commercially available 1-hole subpalpebral lavage system (SPL) at a veterinary teaching hospital. ANIMALS: 102 client-owned horses with ophthalmic disease. PROCEDURES: Medical records of horses (2010 to 2020) with ophthalmic disease were reviewed to determine whether a commercially available SPL system was dorsally placed. Data collected from the medical record included signalment, presenting complaint(s), diagnosis, ophthalmic procedures performed, SPL laterality, hospital service that placed the SPL, anesthetic technique for placement (general anesthesia or sedation with local nerve blocks), duration of SPL management while hospitalized or at home, type of enclosure for the horse, use of eye protection, duration of time the SPL was in place, location of SPL management (home vs hospital), types and numbers of medications administered, recorded complications, and outcome of the globe. Complications experienced during treatment were categorized as either ocular or nonocular. The χ2 test for independence test and Fisher exact test were performed to examine the relationship between the department that placed the SPL, method of anesthesia, antimicrobial administration, type of facial protection used, and complication type and rate. RESULTS: Overall complication rate for SPL systems was 29.1% (37/127), with 21.2% (27/127) being ocular complications and 7.9% (10/127) being nonocular complications. SPL complication rate was not affected by any variable that was examined. CLINICAL RELEVANCE: Commercially available SPL systems placed dorsally have a low ocular complication rate. These SPL systems may be placed by veterinarians with varied training backgrounds and managed at home without significantly increasing complication rate.

Repeated nasopharyngeal lavage predicts freedom from silent carriage of Streptococcus equi after a strangles outbreak.

BACKGROUND: The value of repeated nasopharyngeal lavage (NPL) to detect silent carriers of Streptococcus equi has not been investigated. HYPOTHESIS/OBJECTIVES: Determine if results of serial testing for S. equi by NPL predicts subsequent true carrier status as determined by both NPL and guttural pouch lavage. ANIMALS: An outbreak of strangles with 100% morbidity in 41 mature Icelandic horses was followed prospectively to investigate development of silent carriers. All were initially positive to S. equi on NPL. The farm was closed to horse movement during the entire study. METHODS: Prospective observational study. Testing for S. equi was performed by NPL at weeks 18, 28, 29, and 30 postindex case and subsequently at week 45 by both NPL and guttural pouch lavage. Carrier status at week 45 was compared to results obtained at weeks 18, 28, 29, and 30. Descriptive statistics were calculated. Comparisons were made using Fisher's exact test or the Freeman-Halton extension with a P < .05 level of significance. RESULTS: Of 24 noncarriers at week 45, only 4 horses were negative on all 3 consecutive weekly NPL samples at weeks 28 to 30. However, 10 of the 11 horses with at least 3 negative NPL obtained from weeks 18, 28, 29, and 30 were S. equi-free at week 45 (P = .03). CONCLUSIONS AND CLINICAL IMPORTANCE: Repeated NPL on at least 3 separate occasions can assist in predicting S. equi carrier-free status in horses after recovery from a strangles outbreak.

Extracellular vesicles in low volume uterine lavage and serum: novel and promising biomarker for endometritis in Arabian mares.

BACKGROUND: Extracellular vesicles (EVs) are a promising biomarker and play a vital role in cell-cell communication. This study aimed (I) to identify and characterize EVs from low volume uterine lavage (LVL) and serum in mares with endometritis, compared to healthy controls and (II) to measure serum levels of interleukin 6 (IL-6), and prostaglandins (PGF2α and PGE2). Mares were divided into 30 sub-fertile (endometritis) and 20 fertile (controls). Serum and LVL was collected for EV isolation, and determination of serum levels of inflammatory mediators. Characterization and visualization of EVs were done by electron microscopy, dynamic light scattering and flow cytometry. RESULTS: Serial ultracentrifugation of LVL and use of a commercial kit for serum were strategies for EVs isolation. Mares with endometritis released higher amounts of larger size EVs. The EVs from mares with endometritis differentially expressed CD9 and CD63, compared to controls. Mares suffering from endometritis evoked higher levels of inflammatory mediators. CONCLUSIONS: Thus, EVs could be used for a better understanding the regulatory mechanisms associated with developing endometritis in mares.

Investigation of synovial fluid lubricants and inflammatory cytokines in the horse: a comparison of recombinant equine interleukin 1 beta-induced synovitis and joint lavage models.

BACKGROUND: Lameness is a debilitating condition in equine athletes that leads to more performance limitation and loss of use than any other medical condition. There are a limited number of non-terminal experimental models that can be used to study early inflammatory and synovial fluid biophysical changes that occur in the equine joint. Here, we compare the well-established carpal IL-1ß-induced synovitis model to a tarsal intra-articular lavage model, focusing on serial changes in synovial fluid inflammatory cytokines/chemokines and the synovial fluid lubricating molecules lubricin/proteoglycan 4 and hyaluronic acid. The objectives of this study were to evaluate clinical signs; synovial membrane and synovial fluid inflammation; and synovial fluid lubricants and biophysical properties in response to carpal IL-1ß synovitis and tarsal intra-articular lavage. RESULTS: Hyaluronic acid (HA) concentrations, especially high molecular weight HA, and synovial fluid viscosity decreased after both synovitis and lavage interventions. Synovial fluid lubricin concentrations increased 17-20-fold for both synovitis and lavage models, with similar changes in both affected and contralateral joints, suggesting that repeated arthrocentesis alone resulted in elevated synovial fluid lubricin concentrations. Synovitis resulted in a more severe inflammatory response based on clinical signs (temperature, heart rate, respiratory rate, lameness and joint effusion) and clinicopathological and biochemical parameters (white blood cell count, total protein, prostaglandin E2, sulfated glycosaminoglycans, tumor necrosis factor-α and CC chemokine ligands - 2, - 3, - 5 and - 11) as compared to lavage. CONCLUSIONS: Synovial fluid lubricin increased in response to IL-1ß synovitis and joint lavage but also as a result of repeated arthrocentesis. Frequent repeated arthrocentesis is associated with inflammatory changes, including increased sulfated glycosaminoglycan concentrations and decreased hyaluronic acid concentrations. Synovitis results in more significant inflammatory changes than joint lavage. Our data suggests that synovial fluid lubricin, TNF-α, CCL2, CCL3, CCL5, CCL11 and sGAG may be useful biomarkers for synovitis and post-lavage joint inflammation. Caution should be exercised when performing repeated arthrocentesis clinically or in experimental studies due to the inflammatory response and loss of HA and synovial fluid viscosity.

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.

Detection of Streptococcus equi subsp. equi in guttural pouch lavage samples using a loop-mediated isothermal nucleic acid amplification microfluidic device.

BACKGROUND: Rapid point-of-care (POC) detection of Streptococcus equi subsp. equi (S. equi) would theoretically reduce the spread of strangles by identifying index and carrier horses. HYPOTHESIS: That the eqbE isothermal amplification (LAMP) assay, and the same eqbE LAMP assay tested in a microfluidic device format, are comparable to a triplex real-time quantitative polymerase chain reaction (qPCR) assay that is commonly used in diagnostic labs. SAMPLES: Sixty-eight guttural pouch lavage (GPL) specimens from horses recovering from strangles. METHODS: Guttural pouch lavage specimens were tested for S. equi retrospectively using the benchtop eqbE LAMP, the eqbE LAMP microfluidic device, and compared to the triplex qPCR, that detects 2 S. equi-specific genes, eqbE and SEQ2190, as the reference standard using the receiver operating characteristic area under the curve (ROC). RESULTS: The 27/68 specimens were positive by benchtop eqbE LAMP, 31/64 by eqbE LAMP microfluidic device, and 12/67 by triplex qPCR. Using the triplex PCR as the reference, the benchtop eqbE LAMP showed excellent discrimination (ROC Area = 0.813, 95% confidence interval [CI] = 0.711-0.915) as did the LAMP microfluidic device (ROC Area = 0.811, 95% CI = 0.529-0.782). There was no significant difference between the benchtop LAMP and LAMP microfluidic device (ROC Area 0.813 ± 0.055 vs 0.811 ± 0.034, P = .97). CONCLUSIONS: The eqbE LAMP microfluidic device detected S. equi in GPL specimens from convalescent horses. This assay shows potential for development as a POC device for rapid, sensitive, accurate, and cost-efficient detection of S. equi.

Effects of intrauterine devices on proteins in the uterine lavage fluid of mares.

Intrauterine devices block luteolysis in cyclic mares, but the underlying mechanism is unknown. To clarify the mechanisms, the protein profile of the endometrial secretome was analyzed using two-dimensional difference gel electrophoresis (2D-DIGE). Twenty-seven mares were classified according to whether they were inseminated (AI) or had an intrauterine device (IUD), a water-filled plastic sphere, inserted into the uterus on Day 3 after ovulation. Uterine lavage fluids were collected on Day 15 from pregnant inseminated mares (AI-P; n = 8), non-pregnant inseminated mares (AI-N; n = 4), and mares with IUD (n = 15). The IUD group was further divided into prolonged (IUD-P; n = 7) and normal luteal phase (IUD-N; n = 8) groups on the basis of ultrasound examinations, serum levels of progesterone and PGFM on Days 14 and 15, and COX-2 results on Day 15. Four mares from each group were selected for the 2D-DIGE analyses. Ten proteins had significantly different abundance among the groups, nine of the proteins were identified. Malate dehydrogenase 1, increased sodium tolerance 1, aldehyde dehydrogenase 1A1, prostaglandin reductase 1, albumin and hemoglobin were highest in pregnant mares; T-complex protein 1 was highest in non-pregnant mares; and annexin A1 and 6-phosphogluconolactonase were highest in IUD mares. The results suggest that the mechanism behind the intrauterine devices is likely related to inflammation.

Isolation and Characterization of Equine Uterine Extracellular Vesicles: A Comparative Methodological Study.

Extracellular vesicles (EVs) have been identified in the uterine fluid in different species and have been pointed as key players in the embryo-maternal dialogue, maternal recognition of pregnancy and establishment of pregnancy. However, little is known about the uterine EVs in the mare. Therefore, the present study aimed at characterizing EVs from uterine lavage of cyclic mares by comparing five EVs isolation methods and the combination of them: (1) ultracentrifugation (UC); (2) concentration of lavage volume by Centricon ultrafiltration (CE); (3) the use of CE with different washing steps (phosphate-buffered saline with or without trehalose); (4) size-exclusion chromatography with iZON-qEV columns, and (5) a combination of the methods with best results based on EVs yield, purity, and protein cargo profiles. Transmission electron microscopy and Western blotting confirmed the isolation of EVs by all methods but with quantitative and qualitative differences. Mass spectrometry provided differences in protein profiles between methods, number of identified proteins, and protein classes. Our results indicate that the combination of CE/trehalose/iZON/UC is an optimal method to isolate equine uterine EVs with good yield and purity that can be applied in future studies to determine the role of equine uterine EVs in embryo-maternal interactions.