Equine abortion associated with placentitis caused by Pasteurella pneumotropica / Aborto equino associado a placentite por Pasteurella pneumotropica

Pasteurella pneumotropica is a bacterium that has so far not been described as a cause of placentitis in animals. Two cases of aborted equine fetuses were sent to the Department of Veterinary Pathology of the "Universidade Federal do Rio Grande do Sul" (SPV-UFRGS) for anatomopathological examination. Both cases presented suppurative placentitis associated with multiple basophilic bacterial cells. After bacterial isolation and biochemical analysis, P. pneumotropica was identified.(AU)

Pasteurella pneumotropica é uma bactéria que até o momento não foi descrita como causa de placentite em animais. Dois casos de fetos equinos abortados foram enviados ao Setor de Patologia Veterinária da Universidade Federal do Rio Grande do Sul (SPV-UFRGS) para exame anatomopatológico. Em ambos os casos se observou placentite supurativa associada a múltiplas colônias bacterianas basofílicas. Após o isolamento bacteriano e análise bioquímica, indentificou-se P. pneumotropica.(AU)

Equine abortion associated with placentitis caused by Pasteurella pneumotropica / Aborto equino associado a placentite por Pasteurella pneumotropica

Pasteurella pneumotropica is a bacterium that has so far not been described as a cause of placentitis in animals. Two cases of aborted equine fetuses were sent to the Department of Veterinary Pathology of the "Universidade Federal do Rio Grande do Sul" (SPV-UFRGS) for anatomopathological examination. Both cases presented suppurative placentitis associated with multiple basophilic bacterial cells. After bacterial isolation and biochemical analysis, P. pneumotropica was identified.

Pasteurella pneumotropica é uma bactéria que até o momento não foi descrita como causa de placentite em animais. Dois casos de fetos equinos abortados foram enviados ao Setor de Patologia Veterinária da Universidade Federal do Rio Grande do Sul (SPV-UFRGS) para exame anatomopatológico. Em ambos os casos se observou placentite supurativa associada a múltiplas colônias bacterianas basofílicas. Após o isolamento bacteriano e análise bioquímica, indentificou-se P. pneumotropica.

Clinical, pathologic, and epidemiologic features of nocardioform placentitis in the mare.

Placentitis is the leading cause of infectious abortion in the horse and contributes to roughly 19% of all abortions in the United States. A type of placental infection, nocardioform placentitis (NP) is associated with gram-positive branching actinomycetes localized within the ventral body of the feto-maternal interface to create a lymphoplasmacytic mucoid lesion. While the etiology of this disease is poorly described, this placental infection continues to cause episodic abortions in addition to weak and/or growth retarded neonates. The goal of the present study was to perform a comprehensive analysis of pregnancies associated with a nocardioform-affected placenta and make inferences into the epidemiology of this elusive disease. To do so, 264 mares were enrolled in the study, with 145 as having suspected disease (n = 145; NP) either based on pregnancy-related complications or postpartum placental evaluation, while an additional 119 were enrolled as healthy pregnancies (n = 119; CON). Following diagnosis as either NP or CON based on gross and histopathology at the University of Kentucky Veterinary Diagnostic Laboratory, information was gathered on the mares and neonates for comparisons between diseased and healthy pregnancies. Clinically, a significant portion of diseased mares had clinical indications of NP, including premature mammary gland development, thickening of the placenta noted on transrectal ultrasonography, and separation between the chorioallantois and endometrium noted on abdominal ultrasonography, while vulvar discharge was not commonly noted. Additionally, NP was correlated with increased mare age, decreased gestational length, and decreased neonatal weight, although neonatal IgG and WBC were comparable to CON. Incidence of NP was not correlated with last breeding date, pre- and post-breeding therapeutics, parity, prophylactic medications, or housing. Additionally, NP did not affect postpartum fertility. While NP was associated with a poor neonatal outcome (abortion and/or growth retarded neonate), this did not appear to be influenced by the bacteria isolated (Amycolatopsis spp. vs. Crossiella equi), and mares diagnosed with NP do not appear to be infectious to other pregnant mares nor have repetitive years of the disease. Interestingly, lesion size was positively correlated with last breeding date, as mares bred later in the breeding season correlating with a larger placental lesion. In conclusion, while the etiology of NP continues to elude researchers, the epidemiology of this disease has gained clarity, providing inferences into the management of suspect mares.

Interleukin-6 pathobiology in equine placental infection.

PROBLEM: Ascending placentitis is the leading cause of abortion in the horse. Interleukin (IL)-6 is considered predictive of placental infection in other species, but little is understood regarding its role in the pathophysiology of ascending placentitis. METHOD OF STUDY: Sub-acute ascending placentitis was induced via trans-cervical inoculation of S zooepidemicus, and various fluids/serum/tissues collected 8 days later. Concentrations of IL-6 were detected within fetal fluids and serum in inoculated (n = 6) and control (n = 6) mares. RNASeq was performed on the placenta (endometrium and chorioallantois) to assess transcripts relating to IL-6 pathways. IHC was performed for immunolocalization of IL-6 receptor (IL-6R) in the placenta. RESULTS: IL-6 concentrations increased in allantoic fluid following inoculation, with a trend toward an increase in amniotic fluid. Maternal serum IL-6 was increased in inoculated animals, while no changes were noted in fetal serum. mRNA expression of IL-6-related transcripts within the chorioallantois indicates that IL-6 is activating the classical JAK/STAT pathway, thereby acting as anti-inflammatory, anti-apoptotic, and pro-survival. The IL-6R was expressed within the chorioallantois, indicating a paracrine signaling pathway of maternal IL-6 to fetal IL-6R. CONCLUSION: IL-6 plays a crucial role in the placental response to induction of sub-acute equine ascending placentitis, and this could be noted in amniotic fluid, allantoic fluid, and maternal serum. Additionally, IL-6 is acting as anti-inflammatory in this disease, potentially altering disease progression, impeding abortion signals, and assisting with the production of a viable neonate.

Equine hydrallantois is associated with impaired angiogenesis in the placenta.

INTRODUCTION: Hydrallantois is the excessive accumulation of fluid in the allantoic cavities during the last trimester of pregnancy, leading to abdominal wall hernias, cardiovascular shock, abortion, and dystocia. It has been postulated that hydrallantois is associated with structural and/or functional changes in the chorioallantoic membrane. In the present study, we hypothesized that angiogenesis is impaired in the hydrallantoic placenta. METHOD: Capillary density in the hydrallantoic placenta was evaluated in the chorioallantois via immunohistochemistry for Von Willebrand Factor. Moreover, the expression of angiogenic genes was compared between equine hydrallantois and age-matched, normal placentas. RESULTS: In the hydrallantoic samples, edema was the main pathological finding. The capillary density was significantly lower in the hydrallantoic samples than in normal placentas. The reduction in the number of vessels was associated with abnormal expression of a subset of angiogenic and hypoxia-associated genes including VEGF, VEGFR1, VEGFR2, ANGPT1, eNOS and HIF1A. We believe that the capillary density and the abnormal expression of angiogenic genes leads to tissue hypoxia (high expression of HIF1A) and edema. Finally, we identified a lower expression of genes associated with steroidogenic enzyme (CYP19A1) and estrogen receptor signaling (ESR2) in the hydrallantoic placenta. DISCUSSION: Based on the presented data, we believe that formation of edema is due to disrupted vascular development (low number of capillaries) and hypoxia in the hydrallantoic placenta. The edema leads to further hypoxia and consequently, causes an increase in vessel permeability which leads to a gradual increase in interstitial fluid accumulation, resulting in an insufficient transplacental exchange rate and accumulation of fluid in the allantoic cavity.

Biomarkers for placental disease in mares.

Placentitis is an important cause of abortion, stillbirth, and neonatal death in horses. The diagnosis of placentitis is based on occurrence of clinical signs (premature mammary gland development and vulvar discharge) and ultrasonography of the caudal placental pole. However, early and subtle cases can be missed. In the last few years, several studies have provided objective means of diagnosing placentitis in mares with single or serial measurements of blood markers. Among the markers evaluated the steroids produced by the fetoplacental unit have been shown to change in association with placentitis. Mares with chronic placentitis have an increase in peripheral progestogens; however, mares acutely infected will display a reduction in peripheral concentrations of progestogens. Estradiol-17ß (free- and conjugated form) concentrations are drastically reduced in plasma of mares with placentitis. Acute-phase proteins, particularly serum amyloid A, are increased in plasma of mares suffering from placentitis, and this increase is due to endometrial and chorioallantoic secretions, and minimally from the fetus. Alpha-fetoprotein, a protein expressed in the fetoplacental unit, was shown to be increased in plasma of mares suffering from placentitis. A plephora of microRNA have been identified in plasma and tissues of mares undergoing experimentally induced placentitis, but have not been tested in spontaneous cases. Unique proteomic signatures were found in the fetal fluids of mares undergoing experimentally induced ascending placentitis, making the fetal fluids potentially useful to diagnose placentitis in mares. However, currently the lack of use of transabdominal fetal fluid sampling prevents wide use of the fetal fluids as diagnostic techniques. This manuscript aimed to discuss recent discoveries regarding biomarkers for placentitis in mares.

Uterine cervix as a fundamental part of the pathogenesis of pregnancy loss associated with ascending placentitis in mares.

Anatomical and molecular changes in the cervical barrier in women are a fundamental part of the pathogenesis of pregnancy loss associated with chorioamnionitis. However, there is little information regarding changes in the cervix associated with ascending infection in pregnant mares. To better characterize morphological and molecular changes in the cervix during placentitis, we examined full thickness histology and mRNA expression for a number of inflammatory and endocrine factors in the mucosa and stroma of the cervix of mares (n = 5) after experimental induction of placentitis via transcervical inoculation with Streptococcus equi ssp zooepidemicus at approximately 290d of gestation. Gestationally age-matched mares (n = 4) served as controls. Target transcripts included steroid receptors (PGR, ESR1 and 2), OXTR, prostaglandins synthases and receptors (PTGS1, PTGS2, PGES, PGFS, PTGER2 and PTGER4), cytokines (IL1b, IL6, CLCX8, IL10 and TNFα) and acute phase proteins (SAA). Histologically, a marked modification in the cervical epithelia and stroma was characterizing cervicitis. Additionally, the mRNA expression of IL1ß, IL6, CXCL8, SAA and PTGS2 was greater (P < 0.05) in both mucosa and stroma of the inoculated mares; whereas TNFα, IL10 and PGES were upregulated (P < 0.05) only in the cervical mucosa. Progesterone receptor, ESR1 and PTGER4 were upregulated in the cervical stroma of placentitis mares. In conclusion, the cervical response to placentitis was characterized by an upregulation of inflammatory cytokines that was accompanied by induction of PTGS2 and PGES. Further, receptors known to be associated with relaxation of the cervix in other species (ESR1 and PTGER4) were upregulated in the cervical stroma of placentitis mares. These findings indicate that the cervix is not only a physical barrier but that it has an active role in the pathogenesis of ascending placentitis.

Messages from the placentae across multiple species: A 50 years exploration.

This review explores eight aspects of placentation in multiple mammalian. 1) Specialities of gestational trophoblastic disease. 2) Clinical significance of single umbilical artery (SUA) syndrome. 3) Pulmonary trophoblast embolism in pregnant chinchillas and DIC in pregnant giant panda. 4) Genetics status and placental behaviors during Japanese serow and related antelopes. 5) Specific living style and placentation of the Sloth and Proboscis monkey. 6) Similarities of placental structures between human and great apes. 7) Similarities of placental forms in elephants, manatees and rock hyrax with different living styles. 8) Specialities of placental pathology in Himalayan mountain people. CONCLUSIONS: It was taught that every mammalian species held on placental forms applied to different environmental life for their infants, even though their gestational lengths were different.

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.

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.

Avaliação hematológica e hemogasométrica de potros nascidos de éguas com placentite ascendente / Hematological and hemogasometric evaluation of foals born from mares with ascending placentitis

A placentite é a principal causa de partos prematuros, aborto e nascimento de potros comprometidos, podendo causar hipóxia e septicemia. A hematologia e a gasometria venosa fornecem informações importantes para o monitoramento da saúde de potros nascidos de éguas com placentite. O objetivo deste trabalho foi descrever os valores hematológicos e hemogasométricos durante as primeiras 24h de vida em potros nascidos de éguas mestiças Crioulas submetidas à indução experimental de placentite, com diferentes graus de maturidade. Foram utilizados 16 potros nascidos de éguas submetidas à indução experimental de placentite, divididos em três grupos de acordo com o grau de maturidade: prematuro (n=8), dismaturo (n=4) e a termo (n=4). Foram realizadas coletas sanguíneas nos momentos 0h, 12h e 24h para realização de hemograma completo e gasometria venosa. No eritrograma, foi observada anemia normocítica normocrômica no grupo prematuro em relação ao grupo dismaturo nas 12h e 24h. O grupo prematuro apresentou menor contagem de leucócitos totais nas 24h em relação ao grupo a termo (P=0,01). Os valores de pH, cHCO3 e SO2 não diferiram entre os grupos, porém os animais prematuros apresentaram acidose respiratória (pH=7,28). A PCO2 nos prematuros foi maior na 0h (P=0,02). Nos três grupos, a PCO2 apresentou uma curva adaptativa com redução dos valores durante as 12h e 24h. Os potros prematuros mostraram menores valores de excesso de base (BE) no nascimento (P=0,02), confirmando o quadro de acidose respiratória. Concluiu-se que as respostas hematológicas e hemogasométricas diferem entre potros com diferentes graus de maturidade. A acidose observada no grupo prematuro ao nascimento, com estabilização e resposta compensatória durante as 12h e 24h, demonstra a necessidade de avaliação hemogasométrica sequencial em potros de risco, o que permite a identificação da resposta clínica ao processo e, assim, auxilia no estabelecimento do tratamento e prognóstico para esses potros.(AU)

The placentitis is a major cause of premature birth, abortion and compromised foal delivery, and may result in hypoxia and sepsis. The blood gas analysis and hematology can provide important information for monitoring the foals born from mares with placentitis, with different degrees of maturity. The aim of this study was to describe the hematological and blood gas values during the first 24 hours of life in foals born from crossbreed mares with experimentally induced placentitis, presenting different degrees of maturity. Sixteen foals, born from mares with experimentally induced ascending placentitis were assigned to three groups according to degree of maturity: premature (n=8), dysmature (n=4), and full-term foals (n=4). Blood samples were collected at birth (0), at 12h and 24h, and hematological evaluation and blood gas variables were measured. In the premature group normocytic normochromic anemia was observed compared to dysmature group at 12h and 24h. The premature group showed lower count of white blood cells at 24h relative to the full-term group (p=0.01). The pH, cHCO3 and SO2 values do not differ among the groups; however the premature group showed respiratory acidosis (pH=7,28). The PCO2 was higher at 0h in the premature foals (p=0.02). In all groups, the PCO2 presented an adaptive curve with reduction between 12h and 24h. The premature foals showed lower base excess (BE) values at birth (p=0.02), confirming the respiratory acidosis in this group. We conclude that the hematological and blood gas response differs between foals with different degrees of maturity. Acidosis in the premature foals at birth, with stabilization and compensation of pH value during the first 12-24h demonstrate the necessity of sequential blood gas analysis in risk foals. This may help identify the clinical response to the process and assist in the establishment of adequate treatment and prognosis for these foals.(AU)

Changes in maternal androgens and oestrogens in mares with experimentally-induced ascending placentitis.

REASONS FOR PERFORMING STUDY: While advanced stages of ascending placentitis can be diagnosed by transrectal ultrasonography and clinical signs, early stages can be missed. Thus, additional tools could enhance assessment of placental health. OBJECTIVES: To characterise peripheral dehydroepiandrosterone sulphate (DHEA-S) and testosterone concentrations in mares carrying normal pregnancies (Study 1) and compare plasma concentrations of DHEA-S, testosterone, oestradiol 17-ß (oestradiol) and oestrone sulphate (OES) in mares with or without placentitis (Study 2). STUDY DESIGN: Longitudinal cohort study of healthy mares (Study 1) and controlled experiment (Study 2). METHODS: In Study 1, mares had serum samples collected from 100 days of gestation to term. In Study 2, pregnant mares (260-280 days gestation) were assigned to a control group or a group with placentitis. Placentitis was induced via intracervical inoculation of Streptococcus equi ssp. zooepidemicus. Blood was collected at inoculation/commencement for control mares (day = 0) and daily for 12 days post inoculation (DPI) or until abortion. Steroid concentrations were determined by immunoassays. Concentrations of steroids in Study 2 were also evaluated relative to days from abortion (DFA -8 days to 0). RESULTS: In Study 1, DHEA-S peaked by 180 days gestation, while testosterone concentrations were progressively increased from Days 100 to 180 with a plateau until ~240 days and a progressive decline until 290 days of gestation. In Study 2, concentrations of DHEA-S and testosterone were not significantly different between groups. There were significant effects of time (oestradiol P = 0.0008, OES P = 0.01) and time-by-group interactions (oestradiol P<0.001, OES P<0.0001) for oestrogen concentrations. For mares with experimental placentitis, concentrations of oestradiol were significantly reduced at -6, -2, -1 and 0 DFA, while OES concentrations were significantly reduced on the day before abortion (0 DFA). CONCLUSIONS: Testosterone and DHEA-S were increased and varied through pregnancy. Oestrogens but not androgens decreased significantly in mares with experimentally-induced ascending placentitis.

Proteinograma sérico de éguas com placentite ascendente e seus respectivos neonatos: dados preliminares / Serum protein concentrationin mares with ascending placentitis and their newborns: preliminary data

Este estudo teve por objetivo estabelecer o proteinograma sérico em éguas com placentite induzida e em seus respectivos neonatos. Foram coletadas amostras de sangue das éguas em oito momentos diferentes e dos potros em quatro momentos. Para obtenção da concentração das frações proteicas, utilizou-se eletroforese em gel de acrilaminada contendo dodecil sulfato de sódio (SDS-PAGE). No método utilizado, foram observadas 23 bandas proteicas, cujos pesos moleculares variaram de 16KDa a 245KDa, sendo possível a identificação das seguintes frações: 175KDa, 102KDa, 83KDa, 63KDa, 50KDa, 41KDa, 39KDa e 28KDa. De todas as bandas proteicas encontradas, somente as de 39KDa e 41KDa apresentaram alteração na cinética nos momentos avaliados. De acordo com a solução marcadora, pode-se sugerir que essas proteínas seriam alfa1-glicoproteína ácida (39KDa) e haptoglobina (41KDa). A concentração de imunoglobulinas nos potros apresentou aumento significativo a partir das 12 horas de nascimento. Não está elucidado se estes níveis refletem a persistência do processo inflamatório placentário ou se são alterações fisiológicas do periparto. Não foram observadas alterações na cinética das proteínas nos potros nas primeiras 48 horas.(AU)

The aim of this paper was to identify the serum acute phase protein concentration in mares with induced placentitis and their neonates. Blood samples were collected from the mares in 8 different moments, and from the foals, in 4 moments. To obtain the concentration of protein fractions acrilaminada gel electrophoresis in sodium dodecyl sulfate (SDS-PAGE) was used. In the used method 23 protein bands whose molecular weights ranged from 16kDa to 245kDa were observed, it is possible to identify the following fractions: 175kDa, 102kDa, 83kDa, 63kDa, 50kDa, 41kDa, 39kDa and 28kDa. Of all the protein bands found only the 39KDa and 41KDa have changes in the kinetics in the evaluated times. According to the marker solution, we would suggest that these proteins are alfa1-acid glycoprotein (39kDa) and haptoglobin (41kDa). The concentration of immunoglobulins in foals increased significantly from 12 hours of birth.(AU)

Placental abnormalities in equine pregnancies generated by SCNT from one donor horse.

Placental changes associated with SCNT have been described in several species, but little information is available in this area in the horse. We evaluated the ultrasonographic, gross, and histopathological characteristics of placentas from three successful and five unsuccessful equine SCNT pregnancies, established using cells from a single donor horse. Starting at approximately 6-month gestation, the pregnancies were monitored periodically using transrectal (TR) and transabdominal (TA) ultrasonography (US) to examine the placentas, fetal fluids, and fetuses. Of the five mares that aborted, one mare did so suddenly without any abnormal signs detected by US and four had enlarged umbilical vessels visible on TA-US before abortion. Placental edema (TR-US) and intravascular thrombi in the umbilical cords were seen (TA-US) in two of these four mares; one mare aborted shortly after acute placental separation was identified on TA-US. In three mares that delivered live foals, TA-US showed engorged allantoic vessels and enlarged umbilical vessels. Two of these mares had placental thickening visible on TR-US, interpreted as a sign of placentitis, that subsided after aggressive medical treatment. Seven of the eight placentas were submitted for gross and histopathological examinations after delivery. All placentas had some degree of edema, abnormally engorged allantoic vessels, and enlarged umbilical vessels. Placentitis, large allantoic vesicles, cystic pouches in the fetal part of the cord, and hemorrhages and thrombi in the umbilical vessels were detected only in placentas from mares that aborted. Equine pregnancies resulting from SCNT may be associated with placental pathologies that can be detected using ultrasonography. However, interpreting their severity is difficult. Although placental abnormalities have been observed in SCNT pregnancies in other species, to the best of our knowledge, placentitis has not been previously reported and may be an important complication of equine SCNT pregnancies, leading to pregnancy loss.

Infecção transplacentária e intrauterina por Brucella abortus em búfalos (Bubalus bubalis) / Placenta and intrauterine infection by Brucella abortus in buffaloes (Bubalus bubalis)

O objetivo deste trabalho foi verificar a presença de Brucella abortus e as lesões causadas por esse agente nos anexos fetais e nos fetos de búfalas. Para isso, 20 búfalas em diversos meses de gestação, sorologicamente positivas para brucelose, foram submetidas ao abate sanitário. A idade fetal foi determinada através de exames ultrassonográficos associados à mensuração dos fetos durante a necropsia. Do útero fechado desses animais foram coletadas amostras para histopatologia e qPCR. A partir do segundo mês de gestação foi possível detectar a presença de DNA de B. abortus em líquido amniótico, líquido alantoide e em útero e, a partir do quinto mês, na placenta, coração, baço, rim, pulmão, intestino, fígado e linfonodos dos fetos. Os principais achados anatomopatológicos foram placentite fibrinopurulenta necrótica e endometrite supurativa crônica...

The objective of this study was to detect Brucella abortus and injuries caused by the bacteria in fetal membranes and fetuses. Twenty buffaloes serologically positive for brucellosis were used and subjected to stamping for collection of material from the closed uterus of several months gestation. Fetal age was determined by ultrasound examination and the size of fetuses was measured at necropsy. The samples were subjected to histopathology and qPCR. From the second month of pregnancy on it was possible to detect the presence of B. abortus DNA in amniotic fluid, allantoic liquid and uterus, and from the fifth month on in placenta, heart, spleen, kidney, lung, intestine, liver and lymph nodes of the fetuses. The main pathological findings were fibrinous suppurative necrotic placentitis, and chronic endometritis...

Maturidade de potros nascidos de éguas com placentite / Maturity of foals from the mares with placentitis

Objetivou-se descrever a maturidade neonatal através da resposta clínica, comportamental e hematológica de potros nascidos de éguas com placentite. Participaram do estudo seis potros nascidos de éguas submetidas à indução experimental de placentite ascendente através da infusão intracervical de Streptococcus equi subespécie zooepidemicus e tratadas com Sulfa-trimetoprim e Flunixin meglumine. A formação dos grupos neonatais foi realizada de acordo com o grau de viabilidade e sobrevivência até 60 horas: Grupo Não Sobreviventes (n=2); Grupo Debilitados (n=2); Grupo Saudáveis (n=2). Foi considerado o tempo de gestação, período de intervalo inoculação-parto, avaliação comportamental, clínica e hematológica. O Grupo dos potros Saudáveis apresentou maior tempo de gestação (320±2 dias) e maior intervalo inoculação-parto (20,5±2,5 dias). Os Grupos Não Sobreviventes e Debilitados apresentaram atraso para decúbito esternal e reflexo de sucção. Foi observada bradicardia e hipotermia com 48h de vida no Grupo Não Sobreviventes. Os potros do Grupo Não Sobreviventes e Saudáveis apresentaram leucopenia no nascimento com discretas variações até as 48h. Os potros nascidos de éguas com placentite ascendente e tratadas demonstraram evolução clínica e respostas neonatais distintas. Conclui-se que, quanto maior o tempo de manutenção da gestação após a injúria placentária, melhor será a maturação fetal, o que refletirá em viabilidade e melhor capacidade de resposta neonatal...

The aim of this study was to describe the neonatal maturity through clinical, behavioral and hematologic response of foals born from mares with placentitis. Were used six foals born from mares subjected to experimentally induced ascending placentitis through intracervical infusion of Streptococcus equi subspecies zooepidemicus and treated with trimethoprim sulfametoxazole and flunixin meglumine. The neonatal groups were performed according to the viability and survival rate up to 60 hours: No Survivors group (n = 2); Debilitated group (n = 2); Healthy group (n = 2). Gestational length, the period between inoculation and delivery, and behavioral, clinical and hematologic evaluations were considered. The Healthy group showed longer gestation length (320±2 days) and longer inoculation-delivery interval (20.5±2.5 days). No Survivors and Debilitated groups showed delay in sternal recumbency and sucking reflex. Bradycardia and hypothermia was observed at 48 hours of life in No Survivors Group. Foals from No Survivors and Healthy groups showed leukopenia at birth with slight variations until 48h. Foals born from mares with ascending placentitis and treated showed distinct clinical and neonatal responses. It is concluded that the longer the maintenance of gestation after placental injury, better is the fetal maturation, which reflects in better viability and ability to neonatal response...

Respuesta humoral y consecuencias reproductivas en ovejas desafiadas con Brucella ovis al final de la gestación / Immune response and reproductive consequences in experimentally infected ewes with Brucella ovis during late pregnancy

La brucelosis ovina por Brucella ovis es una enfermedad de prevalencia alta en Argentina. Para evaluar la patogenicidad de B. ovis y la respuesta serológica durante el último mes de gestación, 6 ovejas se distribuyeron en dos grupos: G1, ovejas preñadas, n = 4 y G2, ovejas no preñadas, n = 2. Tres ovejas del G1 (15 días preparto) y una del G2 fueron inoculadas con B. ovis. Se analizaron muestras de suero mediante diferentes pruebas serológicas. Se realizó aislamiento y PCR a partir de mucus cérvico-vaginal (mcv), placenta y leche. En las muestras de placenta se realizó histopatología. Las hembras del G1 parieron corderos vivos; se detectaron anticuerpos en las ovejas desafiadas del G1 a partir de los 5 días posinoculación. El mcv de las ovejas desafiadas resultó negativo al aislamiento en ambos grupos. Las muestras de leche del G1 fueron positivas por cultivo y PCR a B. ovis. La técnica de PCR resultó positiva en las placentas de las ovejas desafiadas del G1. La histopatología reveló una placentitis necrótica supurativa en una de las ovejas desafiadas. El desafío con B. ovis preparto resultó en la invasión de la placenta y de la glándula mamaria, con la consecuente excreción de la bacteria por leche. La infección con B. ovis indujo una respuesta humoral temprana en las ovejas. La colonización de la placenta por B. ovis y la excreción de la bacteria por la leche sugieren un potencial riesgo de infección activa para los corderos y la posibilidad de que estos se comporten como portadores latentes de la infección.

Ovine brucellosis by Brucella ovis is a highly prevalent disease in Argentina. This study aimed to evaluate the pathogenicity of B. ovis and the serological response in ewes during late pregnancy and in their offspring. Six adult ewes were distributed in two groupsGI (pregnant females, n = 4) and G2 (nonpregnant females, n = 2). Three pregnant ewes at 15 days prepartum and one nonpregnant eve were inoculated with B. ovis. Sera of sheep and their offspring were analyzed by different serological tests. Samples of cervicovaginal mucus, placenta and milk were studied by bacteriology. A Brucella genus-specific PCR assay was carried out in placenta and milk samples. Placenta samples were hystopathologically processed. G1 females gave birth to live lambs, but one died hours postpartum. Serological techniques employed detected antibodies in serum of inoculated pregnant animal 5 days postchallenge. Sera of female controls G1 and G2 remained negative throughout the study. Cervicovaginal mucus of infected ewes in G1 and G2 yielded negative results to bacteriology, but B. ovis was isolated from milk. The PCR assay was positive for the placenta and milk from inoculated pregnant ewes. Histopathology revealed necrotic suppurative placentitis in one placenta. However, although results demonstrated that B. ovis can invade the placenta and mammary gland, this bacterium did not cause abortion when it was inoculated intravenously at 15 days prepartum. B. ovis infection induced an early humoral response in pregnant ewes, but their lambs remained seronegative, indicating that there was no transfer of antibodies in infancy. Placenta colonization and milk excretion of B. ovis involves a potential source of infection for lambs, which could play a role as latent carriers of infection.

Relationship between infection, inflammation and premature parturition in mares with experimentally induced placentitis.

REASONS FOR PERFORMING THE STUDY: Ascending placentitis results in premature birth and high foal mortality. By understanding how placentitis induces premature delivery, it may be possible to develop diagnostic markers and to delay premature delivery pharmacologically, thereby decreasing perinatal foal mortality. OBJECTIVE: To identify relationships between bacterial infection, inflammation and premature parturition in mares with experimentally induced placentitis. MATERIALS AND METHODS: Experiment 1: Concentrations of allantoic fluid prostaglandins (PGs) F2alpha and E2 were measured in 8 mares after intracervical inoculation with Streptococcus equi ssp. zooepidemicus (at Days 285-291 of gestation) until parturition and compared with controls (n = 4). Experiment 2: mRNA expression of interleukin (IL)-1beta, IL-6, tumour necrosis factor (TNF)-alpha and IL-8 in the chorioallantois from inoculated mares in Experiment 1 were compared with 7 mares that foaled normally. RESULTS: Bacterial inoculation resulted in 7 aborted fetuses and birth of one premature, viable foal. Infection was associated with inflammation of the chorioallantois in the region of the cervical star, isolation of bacteria and high concentrations of PGE2 and PGF2alpha in allantoic fluid obtained within 48 h of delivery (P = 0.04). Chorioallantois from all mares expressed mRNA for IL-8, TNF-alpha, IL-6 and IL-1beta. Experimentally infected mares expressed more mRNA for IL-6 (P = 0.003) and IL-8 (P = 0.009) in the cervical star region and more mRNA for IL-6 (P = 0.004) in tissues from placental horns than control mares. CONCLUSIONS AND CLINICAL RELEVANCE: Bacterial placentitis may result in liberation of cytokines from the chorioallantois and prostaglandin formation leading to abortion or birth of a precociously mature foal.

Infection of an equine placenta with a novel mycobacterial species leading to abortion.

A 25-year-old pregnant American Quarter Horse mare presented with a 1-week history of progressively worsening vaginal discharge. Transrectal ultrasound revealed increased thickness of the combined uterus and placenta with evidence of chorioallantoic edema but no placental separation. A thickened amnion was visible on transabdominal ultrasound. Abortion occurred 2 days after presentation despite medical treatment. At necropsy, the chorioallantois had variable but diffuse thickening with focally extensive browning of the chorionic surface in the right horn and adjacent body. There were fluid-filled sacculations on the allantoic surface of the umbilical cord, allantoamnion, and chorioallantois associated with diffuse perivascular fluid microscopically. A nonbranching acid-fast bacterium identified as belonging to the genus Mycobacterium Runyon group IV was isolated from the chorioallantois and uterine fluid. Ziehl-Neelsen stain confirmed the presence of intracellular acid-fast bacilli in trophoblasts of the gravid horn and the cervical star area. The current case is unique in that the mycobacteria did not initiate a significant granulomatous inflammatory response in the chorion unless villar necrosis occurred. Sequence analysis of the 16S ribosomal RNA gene and the rpoß gene, encoding the ß subunit of RNA polymerase, indicated that the strain of mycobacteria isolated in this case belonged to a novel species of rapidly growing mycobacteria and not to an established species. Mycobacteria are an uncommon and sporadic cause of placentitis and abortion, but should be suspected in cases of chronic placentitis that are not restricted to the cervical star area.

Benign placental mass with fetal growth retardation in a bull mastiff.

A 4-year-old bull mastiff presented due to premature labour. The referring veterinarian elected to perform a caesarian delivery and at the time of surgery a 4×4×2 cm round, smooth, red to tan, lobulated soft mass was identified attached to the allantoic surface of the zonary placenta of one pup. Microscopically, this mass was composed of loosely arranged confluent undulating cords of polygonal to columnar epithelioid cells separated by a fine fibrovascular stroma resembling the placental labyrinth. The labyrinthine structure and epithelioid nature of the cells suggested that the mass was of trophoblastic origin. Due to the non-invasive nature of the mass and relatively low mitotic activity, this proliferative trophoblastic mass was considered to be benign. The absence of morphological features supporting malignant behaviour and the recapitulation of the normal labyrinthine architecture led to the diagnosis of a trophoblastic hamartoma. To the authors' knowledge this is the first report of a placental hamartoma in the dog.