Application of blood parameters for the early diagnosis of natural ascending placentitis in pregnant mares.

Placental infection is an important cause of late-term pregnancy loss and neonatal diseases in horses. Detection of changes in blood parameters especially during early placentitis could improve the diagnostic accuracy, treatment decision, and potential outcomes. The objectives of this 2-part study were to identify differences in circulating immunological, inflammatory, and hormonal parameters between mares with natural ascending placentitis and control mares; evaluate each and combination of parameters as predictors of placentitis; and determine how these parameters indicate severity of placentitis. Reproductive examination and blood sampling were prospectively performed on pregnant mares in a natural setting. Study 1 enrolled mares diagnosed with early stage of ascending placentitis based on ultrasonographic findings (n = 12), and gestationally age-matched mares with healthy pregnancies as controls (n = 12). Blood samples were classified as pre-onset (before) and early onset (at the time of ultrasonographic changes) of placentitis. There were no detected statistically significant differences between groups and timepoints in immunological and inflammatory parameters, including peripheral lymphocyte subpopulations, cytokine, and serum amyloid A concentrations. The placentitis group showed a reduced (P = 0.01) DHP/20α-DHP ratio when compared to the control group at the early onset timepoint. Plasma estradiol-17ß concentration <359 pg/mL predicted ascending placentitis with acceptable accuracy (area under the curve, AUC = 0.71). Combined albumin <3.7 g/dL, estradiol-17ß < 499 ng/mL, and DHP <33 ng/mL predicted 100 % of cases of ascending placentitis. In study 2, samples were classified according to the presence and severity of the abnormal ultrasonographic findings as mild (n = 11) and moderate-severe (n = 23), and gestationally age-matched with samples from control mares (n = 34). Mares with moderate-severe ascending placentitis had increased (P = 0.03) plasma 20α-DHP concentration and reduced (P = 0.03) DHP/20α-DHP ratio when compared to control mares. Our results suggest that the early events of ascending placentitis detected by ultrasonographic findings include hormonal alterations of feto-placental metabolism measurable in the mare's circulation, yet without obvious systemic immunological and inflammatory changes. Additional studies are warranted to further assess how hormonal markers and cutoff values could guide decisions for timely therapeutic intervention.

Cortisol, progesterone, 17αOHprogesterone, and pregnenolone in foals born from mare's hormone-treated for experimentally induced ascending placentitis.

This study aimed to evaluate steroid hormones in foals born from mares treated for ascending placentitis with different combinations of trimethoprim-sulfamethoxazole (TMS), flunixin meglumine (FM), long-acting altrenogest (ALT) and estradiol cypionate (ECP) for ten consecutive days, starting two days after experimental induction of placentitis with Streptococcus zooepidemicus. Fourty-six pregnant mares and respective foals were assigned as healthy group (Control, n = 8) or treated groups as follows: TMS+FM (n = 8), TMS+FM+ALT (n = 8), TMS+FM+ALT+ECP (n = 6), TMS+FM+ECP (n = 6) and no treatment (NO TREAT n = 10). At delivery, foals were classified as high-risk or low-risk based on clinical and hematologic findings, and survival rates were recorded during the first week of life for comparisons across groups. Cortisol, progesterone, 17αOHprogesterone, and pregnenolone concentrations were determined via immunoassays in 31 of the 46 foals immediately after foaling (0 h), at 12, 24, 48 h, and seven days post-partum (168h). At birth, serum cortisol concentrations were higher in Control and TMS+FM+ECP foals than in remaining groups (p < 0.05). Foals in TMS+FM+ALT and TMS+FM groups had higher 17αOHprogesterone concentrations at 24 h and 48 h, respectively (p < 0.05). Pregnenolone concentrations were higher in TMS+FM than TMS+FM+ALT+ECP foals at 7 days (p < 0.05). High-risk and non-surviving foals had decreased concentrations of cortisol at parturition, but increased concentrations of progesterone from 0 h to 48 h. Pregnenolone and 17αOHprogesterone concentrations were increased and pregnenolone after 12 h in high-risk and non-surviving foals (p < 0.05). In conclusion, adding ECP to the treatment of experimentally-induced placentitis appears to improve foal viability and endocrine response. Cortisol and progestogen profiles were abnormal in high-risk and non-surviving foals, and those treated with ALT or TMS+FM only.

Hematological and biochemical indicators of maturity in foals and their relation to the placental features / Indicadores hematológicos e bioquímicos de maturidade em potros e relação com suas características placentárias

Newborn's health is directly related to gestational conditions and placental efficiency. The aims of this study were: (1) To evaluate hematological and biochemical parameters of foals born from mares with placentitis at birth and at 24h of age, (2) to verify if placental pathology had any influence on neonatal maturity degree through hematological and biochemical response of those foals. According to placental findings (control and placentitis) and neonatal maturity degree (mature and immature), foals were divided into three groups: (1) Control group (n=22), foals born from mares with placentitis and classified as (2) Mature (n=26), and (3) Immature (n=10). The hematocrit and plasma concentration of fibrinogen, total plasma protein, white blood cells count, lactate, glucose, creatinine, urea, albumin, bilirubin, triglyceride, cholesterol, calcium, phosphorus, magnesium, aspartate aminotransferase (AST), creatine kinase (CK), alkaline phosphatase (ALP), and gamma-glutamyltransferase (GGT) were measured. Placental features were significantly different between neonatal maturity degree (P=0.001). Mares that had acute placentitis foaled more immature neonates (n=8/10; 80%). Concentrations of fibrinogen (P=0.003), creatinine (P=0.021), total cholesterol (P=0.014), AST (P=0.001), GGT (P=0.002), total (P=0.001) and unconjugated bilirubin (P=0.010) were higher at birth in the Immature group, whereas albumin levels were lower (P=0.002). Foals born from mares with placentitis presented hyperlactatemia at 24h of age (P=0.002). Acute placentitis had an influence on the neonatal maturity, allowing an accelerated but incomplete fetal maturation. The monitoring of lactate, fibrinogen, creatinine, bilirubin, cholesterol, albumin, AST, and GGT levels, associated with clinical, physical, and behavior evaluation may contribute as indicators of neonatal maturity.(AU)

A saúde do neonato está diretamente relacionada às condições gestacionais e eficiência placentária. Os objetivos deste estudo foram: (1) avaliar parâmetros hematológicos e bioquímicos de potros nascidos de éguas com placentite ao nascimento e com 24h de vida e (2) verificar se a patologia placentária exerceu influência no grau de maturidade através da resposta hemato-bioquímica destes neonatos. De acordo com os resultados histopatológicos placentários (controle e placentite) e grau de maturidade neonatal (maturo e imaturo), os potros foram divididos em três grupos: grupo controle (n=22); e potros nascidos de éguas com placentite classificados como (2) maturos (n=26) e (3) imaturos (n=10). Foi avaliado hematócrito e concentrações sanguíneas de fibrinogênio, proteína plasmática total, leucócitos totais, lactato, glicose, creatinina, uréia, albumina, bilirrubinas, triglicerídeos, colesterol, cálcio, fósforo, magnésio, aspartato aminotransferase (AST), creatina quinase (CK), fosfatase alcalina (FA) e gama glutamiltranferase (GGT). As características placentárias foram significativamente diferentes entre os graus de maturidade neonatal (P=0.001). Éguas com placentite aguda produziram mais potros imaturos (n=8/10; 80%). No nascimento, os potros imaturos apresentaram maiores concentrações de fibrinogênio (P=0,003), creatinina (P=0,021), colesterol total (P=0,0014), AST (P=0,001), GGT (P=0,002), bilirrubina indireta (P=0,010) e total (P=0,001) e menor concentração de albumina (P=0,002). Os potros nascidos de éguas com placentite apresentaram hiperlactatemia com 24h de vida (P=0,002). A placentite aguda exerceu influência na maturidade neonatal, permitindo uma maturação fetal acelerada, porém, incompleta. Mensurações dos níveis sanguíneos de lactato, fibrinogênio, creatinina, colesterol total, AST, GGT, bilirrubinas e albumina, associado à avaliação clínica, física e comportamental, podem contribuir como indicadores de maturidade neonatal.(AU)

Estradiol cypionate aided treatment for experimentally induced ascending placentitis in mares.

The overall goal of this study was to assess the efficacy of various therapeutic combinations of estradiol cypionate (ECP, a long-acting estrogen) and altrenogest (ALT, a long-acting progestin) in addition to basic treatment for placentitis with trimethoprim-sulfamethoxazole (TMS) and flunixin meglumine (FM). Specific outcomes measured in this experiment were (i) time from induction of bacterial placentitis to delivery, and foal parameters (high-risk, survival, and birth weight); and (ii) serum steroid concentrations (progesterone, 17α-hydroxyprogesterone, 17ß-estradiol, and cortisol) in response to treatment. Pregnant mares (∼300 days gestation, n = 46) were randomly assigned into healthy mares (control group, CONT, n = 8) and mares with experimentally induced ascending placentitis (n = 38). Placentitis was induced via intracervical inoculation of Streptococcus equi subspecies zooepidemicus. Thereafter, placentitis induced mares were randomly assigned into: (1) basic treatment, TMS+FM (n = 8); (2) basic treatment with ALT supplementation, TMS+FM+ALT (n = 8); (3) basic treatment with ECP supplementation, TMS+FM+ECP (n = 6); (4) basic treatment with ALT and ECP supplementation TMS+FM+ALT+ECP (n = 6); and (5) no treatment (INOC, n = 10). Treatments were started 48 h after bacterial inoculation and carried out for ten consecutive days. Blood samples were collected daily, and mares were assessed for signs of placentitis until the mare delivered, or for ten consecutive days after onset of treatment. Steroids were analyzed via RIA. Continuous data were analyzed by ANOVA, and categorical data analyzed by Fisher's exact test. Significance was set at p < 0.05. Foal survival at parturition and seven days post-delivery were similar across treated groups (66.7-100%), and to the CONT group. Similar to CONT group, mares in the TMS+FM+ECP group had no high-risk foals while mares in the other groups had higher incidences (50-75%) (p < 0.05). The inclusion of ECP in the treatments resulted in foals with body weight similar to CONT group (p > 0.05). There were no group effects or time by group interactions on concentrations of steroids assessed herein (p > 0.05). In conclusion, in addition to basic treatment TMS+FM, mares with experimentally induced ascending placentitis benefited from ECP supplementation. Conversely, ALT did not appear to make a difference in outcomes. The immunoassays used for measurements of steroid concentrations did not appear useful to assess treatment response.

Histomorphometry of the placental vasculature and microcotyledons in Thoroughbred mares with chronic laminitis.

The objective of this study was to assess the placental vasculature and microcotyledons in pregnant mares with chronic laminitis. Twenty-six pregnant mares were enrolled in the study, 13 had chronic laminitis (Laminitis Group) and 13 were healthy mares (Healthy Group). Arterial systolic pressure and heart rate were measured in the last 30 days of gestation. After foaling, the fetal membranes were grossly evaluated and samples were harvested for histopathologic examination. All mares had digitalized images taken from chorioallantois for histomorphometry analyses (software-NIH ImageJ). Images were assessed for: (i) arterioles from the allantoic region: total and lumen vascular diameter and vascular wall thickness; (ii) microcotiledonary and capillary area/field. Mares in the Laminitis Group showed hypertension, shorter gestational length, lower placental weight and lower birthweight (p < 0.05) foal in comparison with mares in the Healthy Group. Laminitis mares had a reduction of vascular lumen diameters in the uterine body and pregnant horn (p < 0.05), vascular wall thickening in the pregnant horn (p < 0.05) and smaller capillary area/field in the microcotyledons of uterine body and pregnant horn (p < 0.05). In conclusion, pregnant mares with chronic laminitis presented signs of hypertension syndrome, and vascular abnormalitities in placental vessels such as reduction in the vascular lumen and capillary area in the microcotyledones, and thickening of the vascular wall. Foals born from mares with chronic laminitis showed lower birth weight and shorter gestation lengths.

Histological features of the placenta and their relation to the gross and data from Thoroughbred mares / Achados histológicos da placenta e sua relação com seus aspectos macroscópicas e dados de éguas Puro Sangue Inglês

The placenta is a transitory organ that originates from maternal and fetal tissues, the function of which is transporting nutrients from the mother to the fetus. The aim of this study was describe the histological features of placentas in healthy Thoroughbred mares at foaling and evaluate their relation with the gross placental and data of these mares. For this study 188 Thoroughbred mares were used. It was performed clinical observation for signs of placentitis during daily health checks and ultrasonic examination monthly to assess the fetus and placenta. All of the mares that exhibited clinical signs of placentitis were treated during gestation. The parturition was assisted, the placentas were grossly evaluated and samples were collected immediately after expulsion. The following data were considered for each mare: age, gestational age, number of parturition, time for placental expulsion, umbilical-cord length, placental weight and clinical signs of placentitis. Histological evaluation of the placentas revealed extensive cytoplasmic vacuolization of the epithelial areolar cells, presence of inflammatory infiltrates and hypoplasia-atrophy of the microcotyledons. Most of the gross placental findings were consistent with the histological results. In conclusion the mares with a vacuolated placental chorionic epithelium were older and had experienced a larger number of births. Great part of the mares with inflammatory infiltrates did not showed any clinical signs of placentitis during gestation.(AU)

A placenta é um órgão transitório originado do tecido fetal e materno, com função de transportar nutrientes da mãe para o feto. O objetivo deste estudo foi descrever os achados histológicos das placentas de éguas Puro Sangue Inglês (PSI) a termo e avaliar sua relação com a macroscopia da placenta e dados dessas éguas. O estudo utilizou 188 éguas PSI. Foram realizadas observações clinicas diárias para presença de sinais clínicos de placentite e ultrassonografia mensal para avaliar saúde do feto e placenta. As éguas que apresentaram sinais clínicos de placentite durante a gestação foram tratadas. Os partos foram assistidos, as placentas avaliadas macroscopicamente e coletadas imediatamente após sua expulsão. Como dados das éguas foram considerados: idade, tempo de gestação, número de partos, tempo de eliminação da placenta, comprimento de cordão umbilical, peso da placenta e sinais clínicos de placentite. A avaliação histológica das placentas demonstrou extensiva vacuolização citoplasmática das células do epitélio coriônico das regiões areolares, presença de infiltrados inflamatórios e hipoplasia-atrofia de microcotilédones. A maior parte dos achados macroscópicos da placenta foram condizentes com os resultados de histologia. Como conclusão, a vacuolização do epitélio coriônico foi característica de éguas mais velhas e com maior número de partos. Grande parte das éguas com infiltrados inflamatórios não demonstraram sinais clínicos de placentite.(AU)

Verificação da transmissão vertical de Neospora spp. em equinos / Verification of vertical transmission of Neospora spp. in horses.

O gênero protozoário Neospora é reconhecido como causador de desordens reprodutivas e abortos em bovinos. Entre os equinos pouco se sabe sobre os efeitos da infecção por estes protozoários. Atualmente é admitido que os efeitos da infecção por Neospora hughesi em equinos possam ocorrer no sistema nervoso central e, os efeitos provocados pela infecção por Neospora caninum recaiam sobre o sistema reprodutor de éguas. O presente trabalho verificou a presença de imunoglobulinas da classe G no soro sanguíneo de uma população de éguas de cria e, em seus respectivos potros antes da ingestão do colostro. Para execução deste trabalho foi empregada técnica de imunofluorescência indireta (RIFI), utilizando como antígeno taquizoítos de Neospora caninum, a diluição inicial dos soros das éguas foi de 1:50 e a diluição do soro dos potros empregada foi de 1:16. Foram assistidos 78 partos e todos os potros tiveram seu soro sanguíneo coletado imediatamente após o nascimento. A pesquisa de anticorpos contra Neospora spp. apontou que 50 (64%) éguas e 32 (41%) potros foram positivos. Das 50 éguas que apresentaram anticorpos contra Neospora spp. 24 geraram potros positivos. Entre as 28 éguas que não reagiram, oito deram a luz a potros positivos. De posse dos resultados encontrados podemos concluir que ocorreu a transmissão vertical de Neospora spp. nos equinos pesquisados...

The genre protozoan Neospora is recognized as causing reproductive disorders and miscarriages in cattle. Among the horses little is known about the effects of infection by these protozoa. It is currently accepted that the effects of infection by Neospora hughesi in horses may occur in the central nervous system, and effects of Neospora caninum infection occur in the reproductive system of mares. The present study examined the presence of class immunoglobulin G in blood serum of a population of brood mares and their foals before colostrum ingestion. For this assignment was employed indirect immunofluorescence assay (IFA) using as antigen tachyzoites of Neospora caninum, the initial dilution employed in sera of the mares was 1:50 and dilution in the serum of foals was 1:16. Were assisted 78 deliveries and all foals had their blood serum collected immediately after birth. The presence of antibodies against Neospora spp. found in mares was 50 (64%) and 32 (41%) foals were positive. Of the 50 mares that had antibodies to Neospora spp. 24 generated positive foals. Among the 28 mares unreacted eight gave birth to foals positive. Having the results we can conclude that vertical transmission occurred Neospora spp. researched in horses...