Agent Of Abortion Research Articles

Development of a set of multiplex standard polymerase chain reaction assays for the identification of infectious agents from aborted bovine clinical samples

The current study describes the development of a set of 5 multiplex polymerase chain reaction (mPCR) assays for the simultaneous detection of abortive infection agents in bovine fetal tissues, including Brucella spp., Leptospira spp., and Campylobacter fetus (mPCR1); Hammondia heydorni, Neospora caninum, and Toxoplasma gondii (mPCR2); Coxiella burnetii and Chlamydophila psittaci (mPCR3); Mycoplasma bovis, Mycoplasma bovigenitalium, and Ureaplasma diversum (mPCR4); and Bovine viral diarrhea virus (BVDV) and Bovine herpesvirus-1 (BoHV-1; mPCR5). The protocol was tested on different tissue samples collected from 50 aborted bovine fetuses, and it showed that out of the 50 fetuses, 7 (14%, mPCR2) were PCR-positive for N. caninum, 4 (8%, mPCR5) were PCR-positive for BVDV, and 2 (4%, mPCR4) were PCR-positive for U. diversum. The results obtained by using each multiplex PCR were 100% concordant with those obtained by using the respective PCR assays targeting single genes on the same specimens. Moreover, all multiplex PCR assays on clinical samples were compared with reference methods, obtaining a perfect accordance in all samples and confirming the validity of the set of multiplex PCR assays. The proposed set of multiplex PCR assays is, therefore, suitable for the simultaneous detection of the main infectious agents responsible for bovine abortion.

Waddlia, Parachlamydia and Chlamydiaceae in bovine abortion

The etiology remains unknown in many cases of bovine abortion in Switzerland. Bacteria of the Chlamydiales order are known abortive agents, therefore cases of bovine abortion from three representative regions of Switzerland were investigated in this study. Particularly Chlamydiaceae as well as the Chlamydia-like organisms Waddlia and Parachlamydia were of interest, especially because of their possible zoonotic potential. Placenta samples (n=343) were tested for these bacteria by different PCR-methods, immunohistochemistry and serology for Chlamydia abortus. Additionally an attempt for the isolation of Waddlia and Parachlamydia was made by co-cultivation in amoebae.In 67.3% of the 343 cases a necrotizing and/or purulent placentitis was found histologically. By real-time PCR, 0.9% (3/343) of the cases were positive for Waddlia, 13.4% (46/343) positive for Parachlamydia and 14.6% (50/343) positive or questionable positive for Chlamydiaceae. Of these samples, confirmation by immunohistochemistry was possible in 2/3 cases for Waddlia, 25/46 for Parachlamydia and 4/50 for Chlamydiaceae. Of the 50 cases positive or questionable positive for Chlamydiaceae, species-identification by ArrayTube Microarray or 16S rRNA PCR resulted in 41 cases positive for C. abortus whereas the presence of Chlamydia suis was confirmed in four and Chlamydia pecorum in one case.This study brought evidence for the importance of different members of Chlamydiales in different regions of Switzerland although Waddlia is not occurring in a high prevalence. On the other hand mixed infections with different Chlamydiales as well as with other abortigenic agents could be found.

Diagnosis by PCR-REA of Chlamydophila species infections in late-term abortions of domestic ruminants

Veterinary RecordVolume 168, Issue 23 p. 619-619 Research Diagnosis by PCR-REA of Chlamydophila species infections in late-term abortions of domestic ruminants M. L. Tavares Clemente DVM, MSc, Corresponding Author M....

Study on ovine abortion associated withToxoplasma gondiiin affected herds of Khorasan Razavi Province, Iran based on PCR detection of fetal brains and maternal serology

Toxoplasma gondii, an obligatory intracellular protozoan parasite, is one of the causative agents of ovine abortion, as reported in many countries. Different techniques are being used to detect this pathogen in infected ovine fetuses. One of the most sensitive and specific diagnostic techniques is Nested-PCR amplification of the B1 target gene of the organism. In total, 200 brain samples of aborted ovine fetuses and maternal sera submitted from different parts of Khorasan Razavi province, Iran were investigated to track the role of Toxoplasma gondii in ovine abortion by a slightly modified Nested-PCR and IFAT assays, respectively. Among all samples, 27 (13·5%) were PCR-positive and 31 (15·5%) were IFAT-positive and the Toxoplasma-induced abortion prevalence calculated was 8·8% to 18·2% with 95% confidence interval. Results show that high levels of congenital transmission may occur in 27/31(87%) of pregnancies with an excellent logical agreement (ĸ=0·9) between 2 different tests. According to the results of this study, the Nested-PCR employed in this investigation could be recommended as an applied routine test for the routine examination and confirmation of Toxoplasma gondii-induced ovine abortion.

Toxicidad del misoprostol sobre la gestación: Revisión de la literatura

Misoprostol, a synthetic analog of prostaglandin E1, is currently used in Chile and other countries as an antiulcer medication, mainly for the prevention of non-steroidal anti-inflammatory-induced gastric ulcers. Due to its uterotonic properties, it is also indicated in obstetrics for induction of labor and termination of pregnancy. In this last case, misoprostol is either used alone or in combination with other oxytocic drugs such as methotrexate or mifepristone. The use of misoprostol as an abortifacient agent is considered to be safe since it rarely causes serious side effects. However up to 15 % of misoprostol-induced-abortions may not be successful, even under medical supervision, leading to in utero exposure to the drug and to the induction of a series of birth defects including limb and joints defects and Moebius syndrome. Reports from the nineties failed to show a strong epidemiological association between in utero drug exposure and induction of defects, a situation that has changed now that the number of cases reported has increased. Since the practice of abortion is illegal in Chile, many women turn to off-medical procedures to interrupt their pregnancy and use misoprostol as an easy and cheap alternative, readily available in the INTERNET. The lack of medical supervision in these cases may lead to situations that favor the induction of congenital defects. Here, we present an updated review of scientific data, to evaluate the risk of birth defects in babies exposed to the drug during pregnancy termination failed attempts.

Developmental toxicity of misoprostol: an update

Misoprostol, a synthetic analog of prostaglandin E1, is currently used in Chile and other countries as an antiulcer medication, mainly for the prevention of non-steroidal anti-inflammatory-induced gastric ulcers. Due to its uterotonic properties, it is also indicated in obstetrics for induction of labor and termination of pregnancy. In this last case, misoprostol is either used alone or in combination with other oxytocic drugs such as methotrexate or mifepristone. The use of misoprostol as an abortifacient agent is considered to be safe since it rarely causes serious side effects. However up to 15 % of misoprostol-induced-abortions may not be successful, even under medical supervision, leading to in utero exposure to the drug and to the induction of a series of birth defects including limb and joints defects and Moebius syndrome. Reports from the nineties failed to show a strong epidemiological association between in utero drug exposure and induction of defects, a situation that has changed now that the number of cases reported has increased. Since the practice of abortion is illegal in Chile, many women turn to off-medical procedures to interrupt their pregnancy and use misoprostol as an easy and cheap alternative, readily available in the INTERNET. The lack of medical supervision in these cases may lead to situations that favor the induction of congenital defects. Here, we present an updated review of scientific data, to evaluate the risk of birth defects in babies exposed to the drug during pregnancy termination failed attempts.

Psychological treatments for pediatric headache

Headache is the most common pain problem in children and adolescents and, in a considerable proportion, a source of suffering and disability. Medical intervention mainly relies on abortive pharmacological agents (analgesics and antimigraine drugs). Psychological therapies aim at the prevention of headache episodes and the modifications of cognitive–emotional and cognitive–behavioral processes influencing pain. Three main forms of therapy have been evaluated in randomized controlled trials and reviewed in meta-analyses: relaxation training, biofeedback and multimodal cognitive–behavioral therapy. So far there is only scarce evidence on hypnosis and acceptance and commitment therapy, although they seem to be promising. Evidence demonstrates that psychological therapies are efficacious, and that clinically relevant improvement is found in approximately 70% of the treated children at follow-up examination. Future research needs to focus on mechanisms of change, and to extend its view of effects induced by therapy beyond headache improvement to indicators of quality of life.

Development of Validated Spectrophotometric Methods for Estimation of Ethacridine Lactate in Pharmaceutical Formulations

Ethacridine lactate (EAL) is an antiseptic in solutions of 0.1%. It is also used as an agent for second trimester abortion. Two simple and sensitive spectrophotometric methods (Method A and Method B) were developed for the estimation of EAL in pharmaceutical formulations. Method A is based on the condensation of the primary aromatic amino group of ethacridine lactate with an aromatic aldehyde (p‐Dimethylamino‐ cinnamaldehyde) to form a chromophore with absorption maximum of 600nm. Method B is based on redox reaction followed by complex formation of EAL with IO3-Metol reagent to form a stable chromogen, which can be estimated at 520 nm. Method A obeys Beer’s law in the concentration range of 2 to 12 μg/mL and method B in the range of 20 to 100 μg/mL. Interference studies were conducted to see the influence of excipients with the proposed methods. The common excipients usually present in dosage forms do not interfere in the proposed method A and method B. The optical characteristics, regression analysis data and precision of the methods were also calculated. The methods were validated for use in routine quality control of EAL in pharmaceutical formulations.

Comparison of sublingual, vaginal, and oral misoprostol in cervical ripening for first trimester abortion.

Objectives:To compare the effectiveness and tolerability of misoprostol as a cervical ripening agent in first trimester abortion through three different routes of administration before surgical evacuation (SE).Materials and Methods:It was a hospital based prospective randomized open labeled parallel study. A total of 150 randomly selected married women were divided in three groups for sublingual (S/L), vaginal and oral 400 μg of misoprostol single dose administration. The drug was administered 3-4 h before SE in the S/L and vaginal groups and 12 h before the procedure in the oral group. Efficacy was assessed on the basis of time taken for ripening, dilatation achieved, duration of the procedure, intra-operative blood loss, and pain score. The tolerability was noted on the basis of side effects.Results:The mean time taken for cervical ripening was less in sublingual administration (3.7±1.2 hr) as compared to the vaginal and oral routes. The S/L group had significant cervical dilatation (P<0.001) and the duration of SE was less as compared to the vaginal and oral routes. However, the mean intraoperative blood loss was more in sublingual as compared to the vaginal and oral groups. The intra-operative pain score of the S/L group was significantly lower (1.9±1.1, P<0.05) as compared to the vaginal (2.6±1.7) or oral route (3.3±1.7). Loose motions and nausea/vomiting were more with the S/L and oral routes while blood loss was more in the vaginal route.Conclusion:Administration of misoprostol by the sublingual route is better than the oral and vaginal routes for cervical ripening.

Detection and characterization of Leptospira spp. isolated from aborted bovine clinical samples

Leptospirais recognized as an important public health problem worldwide, especially in tropical countries, and is a common cause of abortion in dairy and beef herds. The aim of the present study was to detect and characterizeLeptospiraas the causative agent of abortion in cattle using a PCR-RFLP in Chaharmahal va Bakhtiari and Isfahan provinces, Iran. A total of 220 bovine aborted foetuses and 120 vaginal discharges from an aborted calf were collected from 64 commercial dairy herds. After isolation of 60Leptospiraspp. from samples, RFLP analysis was carried out withHindIII andHaeIII restriction enzymes in reference strains and isolated for characterization. In a total of 340 specimens, 46 (20.9%) and 14 (11.66%) were identified positive forLeptospiraspp. from aborted bovine foetuses and vaginal discharges, respectively. The present results also suggest thatL. interrogansserovarhardjohas the highest prevalence in the region under study andL. hardjois a major pathogen causing bovine abortion in Chaharmahal va Bakhtiari and Isfahan provinces of Iran.

Confirmation of Chlamydophila abortus in infected cell culture using Indirect Immunofluorescence technique

Chlamydophila abortus (C. abortus) is an important abortifacient agent in bovines and ovines. Clinical diagnosis of the disease is often difficult. An early diagnosis can be achieved based on direct demonstration of the organism in clinical material and through the cultural recovery of the organism in embryonated chicken egg. For confirmatory diagnosis antigen detection methods or serological techniques can be adopted. The present study is aimed at the confirmatory diagnosis of C. abortus infection by indirect immunofluorescence technique following the isolation of the organism in cell culture. Specific apple green fluorescing inclusions of C. abortus in McCoy cell lines was detected from 72 h to 96 h post infection employing anti-chlamydial group specific monoclonal antibodies. Thus, a confirmatory diagnosis of the infection was possible with this study.

Management of Chronic Cluster Headache

Primary cluster headache (CH) is an excruciatingly severe pain condition. Several pharmacologic agents are available to treat chronic CH, but few double-blind, randomized clinical trials have been conducted on these agents in recent years, and the quality of the evidence supporting their use is often low, particularly for preventive agents. We recommend sumatriptan or oxygen to abort ongoing headaches; the evidence available to support their use is good (Class I). Ergotamine also appears to be an effective abortive agent, on the basis of experience rather than trials. We consider verapamil and lithium to be first-line preventives for chronic CH, although the trial evidence is at best Class II. Steroids are clearly the most effective and quick-acting preventive agents for chronic CH, but long-term steroid use carries a risk of several severe adverse effects. We therefore recommend steroids only if verapamil, lithium, and other preventive agents are ineffective. In rare cases, patients experience multiple daily cluster headaches for years and are also refractory to all medications. These patients almost always develop severe adverse effects from chronic steroid use. Such patients should be considered for neurostimulation. Occipital nerve stimulation is the newest and least invasive neurostimulation technique and should be tried first; the evidence supporting its use is encouraging. Hypothalamic stimulation is more invasive and can be performed only in specialist neurosurgical centers. Published experience suggests that about 60% of patients with chronic CH obtain long-term benefit with hypothalamic stimulation.

Amniotic and allantoic fluids from experimentally infected sheep contain immunoglobulin specific for Chlamydophila abortus

Chlamydophila abortus, the aetiological agent of enzootic abortion of ewes (EAE), replicates in trophoblast cells leading to their destruction and dissemination of the bacterium to foetal organs. To further understand the pathogenesis of EAE, amniotic and allantoic fluids were collected from experimentally infected pregnant ewes at 30 (7 samples from each fluid), 35 (8 samples from each fluid), 40 (10 samples from each fluid) and 43 (6 amniotic fluids and 7 allantoic fluids) days post-infection to determine pathogen numbers and other markers of infection. Whilst experimentally infected ewes had characteristic placental lesions, only two amniotic and seven allantoic fluid samples were positive for C. abortus by real-time PCR. In contrast, all amniotic and allantoic fluids were positive for immunoglobulin. Immunoglobulins were generally detected earlier in allantoic fluid than in amniotic fluid and the numbers of samples containing immunoglobulins increased as infection progressed. IgG in amniotic and allantoic fluids was shown to be specific for C. abortus, and reacted with the major outer membrane proteins, polymorphic outer membrane protein and macrophage infectivity potentiator protein. A comparison of two-dimensional immunoblots using purified IgG from the allantoic fluid, amniotic fluid, ewe serum and foetal serum of a C. abortus infected animal at 40 days post infection indicated a pattern of reactivity intermediate between that of the ewe serum and the foetal serum. Results suggest that a maternal source of immunoglobulin is predominant at 30 days post-infection but that foetal derived antibodies may be contributed at a later stage.

Pathological and parasitological studies on Toxoplasmosis as a causative agent of abortion among goats in Egypt

Pathological and parasitological studies on Toxoplasmosis as a causative agent of abortion among goats in Egypt

Main constituents and cytotoxic activity of the essential oil of Piper artanthe

The identification of novel antitumor agents that demonstrate efficacy is necessary to improve medical treatment based on new drugs. The pharmaceutical industries have been working together in this area for several years to try to find novel compounds with good antitumor activity. Brine shrimp (Artemia salina) larvae have been reported as an organism that can detect bioactive compounds. Since there is a good correlation between the toxicity on Artemia salina and antitumor [1, 2], phototoxicity [3], and anti-Trypanosoma cruzi [4] activity, this test is a good prescreening method for identifying new potential antitumor agents. Piper artanthe C.DC. (Piperaceae) is commonly known as the “cordoncillo.” It is a shrub or small tree growing to 1 to 2 m, and it is widespread in America [5] and used in folk medicine for the treatment of different disease symptoms. Despite phytochemical studies on the Piper species describing the isolation of several bioactive compounds [6], to our knowledge P. artanthe has not been investigated for its biological activity, and no data can be found concerning the chemical composition of this species. The main purpose of this study was not only to evaluate the biological activities using the Artemia salina toxicity model as a prescreening procedure but also to evaluate the composition and structural characterization of the main components of the essential oil from the aerial parts of P. artanthe C.DC. As shown in Table 1, 41 compounds consisting of more than 87% of the total hydrodistilled essential oil were identified. The chromatographic data on both columns, HP-5MS and DB-1, indicate that the major components were apiol (14.52%), -elemene (11.69%), (E)-caryophyllene (10.24%), epi-cubebol (8.85%), myristicin (6.38%), and cubebol (6.30%). Over 44% of these compounds were sesquiterpenes, and 45% of them were oxygenated sesquiterpenes. The evaluation of biological activity using the brine shrimp lethality test showed that the essential oil of the fresh aerial parts of P. artanthe displayed an LC 50 value of 107.1 g/mL ( n-1 : 8.9 g/mL). According to [7], crude extracts resulting in LC 50 values of less than 250 g/mL are considered significantly active. For that reason, this oil can be regarded as a promising candidate for further investigation. Accordingly, our following objective was to isolate and identify the major components of this essential oil in order to evaluate their individual biological activities. Though all constituents could not be isolated, the main component was made pure by means of column chromatography on silica gel as a yellowish oil (980 mg). This compound displayed an LC 50 value of 13.1 g/mL ( n-1 : 0.4 g/mL). Plant polyalkoxybenzenes display a broad spectrum of biological activities. For example, apiol has previously been reported to be a specific inhibitor of the biosynthesis of aflatoxin G1, a calcium antagonist, and a diuretic, abortive, sedative, antioxidant, and insecticidal agent 8–11.

NovelChlamydia-like Organisms as Cause of Bovine Abortions, UK

Novel<i>Chlamydia</i>-like Organisms as Cause of Bovine Abortions, UK

Identification of Immunologically Relevant Proteins ofChlamydophila abortusUsing Sera from Experimentally Infected Pregnant Ewes

Chlamydophila abortus is an intracellular pathogen and the etiological agent of enzootic abortion of ewes (EAE). C. abortus has a biphasic development cycle; extracellular infectious elementary bodies (EB) attach and penetrate host cells, where they give rise to intracellular, metabolically active reticulate bodies (RB). RB divide by binary fission and subsequently mature to EB, which, on rupture of infected cells, are released to infect new host cells. Pregnant ewes were challenged with 2 x 10(6) inclusion forming units (IFU) of C. abortus cultured in yolk sac (comprising both EB and RB). Serum samples were collected at 0, 7, 14, 21, 27, 30, 35, 40, and 43 days postinfection (dpi) and used to identify antigens of C. abortus expressed during disease. Additionally, sera from fetal lambs were collected at 30, 35, 40, and 43 dpi. All serum samples collected from experimentally infected pregnant ewes reacted specifically with several antigens of EB as determined by one-dimensional (1-D) and 2-D gel electrophoresis; reactive antigens identified by mass spectrometry included the major outer membrane protein (MOMP), polymorphic outer membrane protein (POMP), and macrophage infectivity potentiator (MIP) lipoprotein.

Evidence of Chlamydophila abortus vaccine strain 1B as a possible cause of ovine enzootic abortion

Chlamydophila abortus, the agent of ovine enzootic abortion (OEA), is a major cause of lamb mortality worldwide. Disease can be controlled through the use of vaccines based on the 1B temperature-sensitive mutant strain of C. abortus. This study investigated suspected OEA cases across Scotland for the presence of the 1B strain by analysis of recently identified unique point mutations (9). Thirty-five cases were C. abortus-positive and 14 came from vaccinated flocks. Analysis of single nucleotide polymorphisms by PCR-RFLP and sequence analysis revealed the presence of point mutations consistent with the presence of the 1B vaccine strain in 5 of these 14 samples. Quantitative real-time PCR revealed comparable numbers of genome copies of the 1B strain in infected placentas to those present following wild-type infection. This study is the first to demonstrate the presence of the 1B vaccine strain in the placentas of OEA cases and suggests a probable causal role in the disease.

Interruption of Pregnancy by Indigenous Method

The stem from a creeper vine of the species Asclepeadaeae (further identification of the plant has not yet been made) has been used in some areas of Bangladesh as a means of inducing abortion. Although it has not been performed by physicians, many women believe this method of abortion to be safe and effective. In a study conducted to determine the safety and effectiveness of this method, 108 women, between 12 and 24 menstrual weeks' gestation and 15 to 42 years of age, were studied. One of three sizes of stems from the plant (thin, 2–3 mm; medium, 3–4 mm.; thick, 4-4.5 mm.) was inserted through the cervical canal either manually or with a Lippes loop inserter. Subjects were observed a minimum of 72 hours after insertion of the stem. Fifty-four (54.596) of the patients aborted within 72 hours. Insertion-to-abortion time was affected by the size of the stem and by gestational age, but not by parity. Patients aborted with the thick stem and those greater than 16 weeks' gestation aborted more rapidly (mean: 47.2 and 41.4 hours, respectively). Complications consisted of excessive bleeding (13.1%), persistent fever (9.1%), and persistent pain (14.1%). Thirteen (13.1%) of the patients were diagnosed as having pelvic infections. No other potentially serious complications were observed. In this initial series, the method compares favorably with other experimental methods (urea, alcohol, laminaria tents, or Karman coil methods); however, it is not as safe or effective as intra-amniotically administered Prostaglandin F2g Abortion rates and induction-to-abortion times were similar to those for intra-amniotic saline inductions, but the rate of complications for the stem was significantly higher. Further study is needed to determine the mechanism of action for this indigenous method. It would appear that the stem contains an oxytocic agent; however, the oxytocic and/or abortifacient agent has not been identified. Hopefully, when the aborti-facient agent(s) are identified and administered in maximally effective doses under sterile conditions, a practicable method of artificial aobrtion can be defined.

YersiniaSpecies Isolated from Bats, Germany

<i>Yersinia</i>Species Isolated from Bats, Germany