Changes in the rumen bacterial microbiome of cattle exposed to ponderosa pine needles.

Consumption of ponderosa pine needles, as well as needles and bark from a number of other trees, can cause abortions in cattle. The abortifacient compounds in these trees are labdane resin acids, including isocupressic acid and agathic acid. Previous research has demonstrated that cattle conditioned to pine needles metabolize the labdane resin acids more quickly than naïve cattle. The results from that study indicated that changes had occurred in the rumen of conditioned cattle. Therefore, in this study, the changes that occurred in the rumen bacterial microflora of cattle during exposure to ponderosa pine needles were evaluated. Cattle were dosed with ground pine needles twice daily for 7 d. Rumen samples were collected on d 0, 3, 7, and 14 (7 d after treatment stopped) and ruminal bacterial microbiome analyses were performed. There were 372 different genera of bacteria identified in the rumen samples. Principal coordinate analysis indicated that there was a significant difference in the rumen bacterial composition between the time points. There were 18 genera that increased in abundance from d 0 to d 7. Twenty three genera decreased in abundance from d 0 to d 7. The results from this study demonstrated that exposure of cattle to pine needles caused a clear shift in the rumen microbiome composition. In general, this shift lasted less than 1 wk post exposure, which indicates that any prophylactic treatment to manipulate the ruminal metabolism of the abortifacient compounds in pine needles would need to be continuously administered to maintain the necessary microbial composition in the rumen.

Píldora poscoital: un debate científico necesario / The morning after pill, a necessary scientific discussion

No disponible

Intoxicação aguda e abortos em cobaias pelas favas de Enterolobium contortisiliquum (Leg. Mimosoideae) / Acute poisoning and abortions in guinea pigs by the pods of Enterolobium contortisiliquum (Leg. Mimosoideae)

Os objetivos deste trabalho foram determinar a toxicidade aguda e o efeito abortivo de favas de Enterolobium contortisiliquum. Em um experimento frutos da árvore foram administrados a 4 grupos de 3 cobaias cada. Outro grupo de 3 cobaias foi utilizado como controle. Para os Grupos 1 e 2 foram administradas 5 e 10g de favas por kg de peso vivo, respectivamente, em doses únicas. Para os Grupos 3 e 4 foram administradas 10 e 15g/kg, respectivamente, divididas em doses diárias de 5g/kg. Uma cobaia do Grupo 2 e uma do Grupo 4 morreram 12 e 18 horas após o final da administração da planta. As lesões macroscópicas consistiam hemorragias no estômago e intestinos delgado e grosso, fígado aumentado e vesícula biliar distendida. Histologicamente, o fígado apresentava severa vacuolização e necrose dos hepatócitos periportais. Em outro experimento, ração contendo 4% de favas de E. contortisiliquum foi administrada a dois grupos de 4 cobaias 35 dias após o acasalamento. Das 8 cobaias, 4 abortaram 6-15 dias após o início da ingestão. As outras 4 cobaias não estavam prenhes. Duas cobaias controle pariram normalmente. Todas as cobaias foram eutanasiadas no final do experimento. Histologicamente, as cobaias que ingeriram as favas apresentaram, no fígado, necrose hemorrágica periportal de moderada a severa. Todos os fetos apresentavam graus variáveis de autólise. Em 4 fetos que foram estudados histologicamente não foram observadas lesões significantes. As lesões agudas observadas nas cobaias são semelhantes às observadas na intoxicação por favas de E. contortisiliquum em bovinos e às da intoxicação experimental com saponinas da árvore.Os resultados do experimento em cobaias prenhes demostram o efeito abortivo de E. contortisiliquum e sugerem que esta espécie pode ser utilizada para estudar o efeito abortivo da planta ou de princípios ativos identificados na mesma.(AU)

The objective was to study the acute toxicity and the abortive properties of Enterolobium contortisiliquum pods in guinea pigs. Pods of E. contortisiliquum were administered orally to 4 groups of 3 guinea pigs each. Another group of 3 guinea-pigs was used as control. Group 1 and 2 were fed with one dose of 5 and 10g of pods for kg body weight, respectively. The guinea pigs of Group 3 and 4 received 10 and 15g/kg, respectively, divided into daily doses of 5g/kg. One guinea pig from Group 2 and one from Group 4 died 12 and 18 hours after the end of the administration. Gross lesions were hemorrhages of the stomach and of the large and small gut, enlarged liver, and dilated gall bladder. Histologically, the liver had severe vacuolation and necrosis of periportal hepatocytes. In another experiment a ration containing 4% of pods of E. contortisiliquum was fed to 2 groups of 4 guinea-pigs, 35 days after mating. Four of the 8 guinea pigs aborted 6-15 days after the beginning of ingestion. The other 4 guinea pigs were not pregnant. All guinea pigs were euthanized after abortion or at the end of the experiment. Histologically all animals had mild to severe periportal hemorrhagic necrosis. All fetuses had variable degree of autolysis. In 4 fetuses studied no significant histologic lesions were observed. The acute lesions observed in guinea-pigs are similar than those observed in the spontaneous poisoning by Enterolobium spp. in cattle. Similar lesions are observed in guinea-pigs poisoned experimentally with saponins from E. gummiferum. The results of the experiments in pregnant guinea pigs suggest that E. contortisiliquum can be used to study the abortive effect of its pods or its toxic compounds.(AU)

Abortifacient effect of Prangos ferulacia on pregnant rats.

PURPOSE: Prangos ferulacea grows in southern Iran and used in Iranian herbal medicine for gastrointestinal disorders, but it seems it has an abortifacient effect on pregnant women. To verify its potential as an abortifacient agent, we administered the leaves of this plant to pregnant rats. MATERIAL AND METHODS: Hydroalcoholic and aqueous extract of the leaves was administered orally at different doses to 60 rats on the first 18 days of pregnancy. Group 1 (G1) was considered as control group and was given only water. Groups 2-5 (G2-G5) received 25, 50, 100 mg/g per day and Groups 6-8 (G6-G8) received 300, 500 and 1000 mg/g per day, respectively. On Day 18 of pregnancy, they were killed and laparotomized. The uterine horns of each group were opened to see whether they contained any live and degenerated/dead fetuses. We used Student's t-test to analyze the data (p < or = .05 was considered significant). RESULTS: Of the total 504 fetuses in the studied groups, 13 fetuses (2.57%) were aborted. The abortion rate in the control group was 2 (1.94%) of 103 fetuses; the abortion rate was higher in the treated groups but not statistically significant. There was no relationship between the dose and type of extract and abortion rate in all studied groups. CONCLUSION: This study shows that the aqueous or hydroalcoholic extract of P. ferulacea is ineffective on the rate of abortion of pregnant rats. Future studies should be performed with higher doses to test the efficacy of this agent on other animals.

Development of enzyme-linked immunosorbent assays for isocupressic acid and serum metabolites of isocupressic acid.

The consumption of ponderosa pine (Pinus ponderosa), lodgepole pine (Pinus contorta), common juniper (Juniperus communis), and Monterey cypress (Cupressus macrocarpa) causes abortions in pregnant cattle. Recent studies have identified isocupressic acid (1) as the primary abortificient compound in these plants. In vitro and in vivo studies using rumen and blood have shown isocupressic acid (1) is rapidly metabolized to agathic acid (3), dihydroagathic acid (4), and tetrahydroagathic acid (5). Rapid and sensitive diagnostic techniques are needed to identify poisoned animals, to study toxicokinetics, and to elucidate the mechanism of isocupressic acid-induced abortion in cattle. In this study, four competitive inhibition enzyme-linked immunosorbent assays for isocupressic acid and its sera metabolites were developed using polyclonal antibodies. One assay is specific to 1, whereas the other three assays show cross-reactivity to 3-5 in addition to 1. The assay specific to 1 had a limit of detection of 44.1 pg. The other assays which demonstrated cross-reactivity to the isocupressic acid blood metabolites also had comparably low limits of detection. One assay was used to follow the absorption and elimination profile of isocupressic acid metabolites in both cow serum and urine after oral dosage of a cow with common juniper.

The toxic and abortifacient effects of ponderosa pine.

Ponderosa pine needles cause abortion and a poorly described toxicosis when eaten by cattle. In previous trials, the abortifacient compound of pine needles was identified as isocupressic acid. At abortifacient doses, isocupressic acid caused no other toxicosis. However, other pine needle fractions, similar in composition to several commercially available rosin products, caused no abortion but were very toxic. The purpose of this study was to describe the toxicoses of ponderosa pine, compare its toxicity with other rosin and related pine products, and identify the toxin. Four groups of three pregnant beef cows each were treated with either ponderosa pine tips, rosin gum, dehydroabietic acid, or ground alfalfa. The cows treated with pine tips aborted, had retained placentas with endometritis, and developed both renal and neurologic lesions. The cattle treated with rosin gum or dehydroabietic acid did not abort but developed similar signs and lesions of intoxication. Clinical signs of intoxication included anorexia, mild rumen acidosis, dyspnea, paresis progressing to paralysis, and death. Clinical biochemical results, suggestive of renal, hepatic, and muscular disease, included azotemia, hypercreatinemia, hyperphosphatemia, proteinuria, and marked elevations of various serum enzymes. Histologically, all poisoned animals had nephrosis, vacuolation of basal ganglia neuropil with patchy perivascular and myelinic edema, and skeletal myonecrosis. The alfalfa-treated controls were normal. These findings suggest that ponderosa pine needles and tips are both abortifacient and toxic. Because the lesions caused by pine tips, rosin gum, and dehydroabietic acid are similar, toxicosis is most likely due to the diterpene abietane acids, common in all three.

[The determination in cadaveric material of drugs used for the interruption of pregnancy]. / Opredelenie v trupnom materiale lekarstvennykh sredstv, primeniaemykh dlia preryvaniia beremennosti.

A method for detection and measurements in cadaveric material of some drugs used to induce abortions has been developed. The drugs were measured by spectrophotometry, photo-electro-colorimetry, and argentometry. Detection threshold by the said methods was 0.5 to 3 mg%.

[The detection of abortion agents in biological material]. / Obnaruzhenie nekotorykh abortivnykh sredstv v biologicheskom materiale.

Methods of extraction and detection of ascorbic acid, ergot alkaloids, cotarnine, microfollin, tinctures of marigold, peony, sage-brush, iodine, benzestrol, pachycarpine, hexestrol, folliculin, quinine, ethoxydiaminoacridine lactate were developed. Detection limits are 0.1-1 mg/100 g of object. Methods of manganese, iodine and soap detection are developed. Detection limits are 0.05-2 mg/100 g of object.

Levels of the antiprogestin RU 486 and its metabolites in human blood and follicular fluid following oral administration of a single dose.

Using high-pressure liquid chromatography (HPLC) the antiprogestin RU 486 and two of its metabolites (N-monodemethyl RU 486 and propargyl RU 486) were measured in plasma and follicular fluid of 21 women requesting laparoscopic sterilization. Pretreatment of the women involved ovulation induction with clomiphene and HCG. RU 486 (100 mg) was administered orally and 1 h later blood samples were withdrawn. Thirty-four hours later, at laparoscopy, samples of both blood and follicular fluid were collected. During the 34-h period the average plasma level of RU 486 decreased from 1.93 mumol/l to 0.91 mumol/l, i.e. by -50%. The latter concentration of RU 486 was not significantly different from that found in follicular fluid (0.79 mumol/l). The monodemethyl metabolite exhibited significantly higher plasma levels (3.09 mumol/l) than RU 486 1 h after administration. Thirty-four hours later these levels had decreased to 0.92 mumol/l, i.e. by 70%. In follicular fluid, the levels of the monodemethyl metabolite (1.76 mumol/l) were significantly higher than those of RU 486 (0.79 mumol/l). Because of background noise, only approximate values were established for the propargyl metabolite. These were 0.67 and 0.40 mumol/l, respectively, in plasma and 0.42 mumol/l in follicular fluid. The results indicate that RU 486 and two of its major metabolites can readily cross the blood-follicle barrier of human pre-ovulatory follicles.

Purification and characterization of trichosanthin. Homology to the ricin A chain and implications as to mechanism of abortifacient activity.

Trichosanthin, a protein from the Chinese medicinal herb Trichosanthes kirilowii, was purified in two essentially quantitative steps involving CM-Sephadex chromatography and reverse-phase high performance liquid chromatography. The protein was found to have a molecular mass of 25-26 kDa, to contain no cysteine, and to contain no glycosidic linkages. Pure trichosanthin was found to have potent abortifacient activity in pregnant mice. In order to understand the molecular basis of this unique biological activity, we have examined the amino acid sequence of the protein. As purified, trichosanthin was found to contain two amino-terminal sequences which differed only in the absence or presence of a tyrosine at residue 1. Sequence analysis of trichosanthin has allowed for determination of the NH2-terminal 38-amino acid residues. Comparison of this sequence to those present in a data base revealed homology with the ricin A-chain. Consistent with this structural homology, we have found that trichosanthin is a potent inhibitor of protein synthesis in a reticulocyte lysate system.

An X-ray analysis of the orthorhombic crystal form of trichosanthin at 5 angstrom.

Trichosanthin crystals grown at pH 5.4 are orthorhombic P212121 with cell dimension a = 38.23 angstrom, b = 76.18 angstrom and c = 79.12 angstrom. Each asymmetric unit contains one protein molecule of 234 amino acid residues (M. W. 26,000). The structure has been solved at 5 angstrom resolution by conventional MIR method with two heavy-atom derivatives. The preliminary result suggests that the protein molecule in acidic solution does not have dramatic conformational change, compared with that in alkali solution.

The three-dimensional structure of trichosanthin molecule.

A model completed recently of two trichosanthin molecules in an asymmetric unit belonging to the monoclinic system is reported. It can be seen that the molecular structure consists of two domains, one large and one small, and that there are some features in the secondary structure. The dissimilarity between two molecules as well as their interactions in an independent unit correlated in a non-symmetric way has been described, moreover, the binding sites of the heavyatom position in the derivatives are also discussed.