Fetal Stomach Contents Research Articles

Comparison of Dot-ELISA with microbial culture for detection of Brucella spp. in clinical specimens

Summary Definitive diagnosis of brucellosis is made by isolation of the causative agents, which is a timeconsuming procedure. To evaluate the efficacy of Dot-ELISA for detecting brucellae in clinical samples, 94 different specimens taken from animal origin were cultured on brucella selective culture media and colonies were identified biochemically. The specimens were also examined after centrifugation by Dot-ELISA using a specific anti-brucella antibody, a suitable peroxidase conjugate and substrate. Of the 94 samples, 5 (5.31%) were positive in Dot-ELISA and 4 (4.25%) had positive cultures. In comparison with culture, the sensitivity and specificity of Dot-ELISA for detection of brucellae in the samples was 80 and 100%, respectively. There was 98.9% agreement between the two tests. The results indicated that Dot-ELISA is a good and rapid test with acceptable sensitivity and specificity for detection of Brucella spp. in aborted fetal stomach contents.

Transplacental Transfer of a Growth Hormone-Releasing Hormone Peptide from Mother to Fetus in the Rat

Previous studies showed that when growth hormone-releasing hormone (GHRH) was administered to either pregnant rats or pigs as a plasmid-mediated therapy, pituitary weight, somatotroph and lactotroph numbers, and postnatal growth rate of the offspring increased. To determine if these responses resulted from direct effects of GHRH on the fetus or were secondary to effects incurred in the mother, we studied in the rat the transplacental transfer of a GHRH analog (HV-GHRH) to the fetus from the maternal circulation. For the in vivo study, HV-GHRH was labeled with 125I and purified by reverse-phase high-performance liquid chromatography (HPLC). At 18 days of gestation, pregnant dams were administered a priming intravenous dose followed by a constant infusion of the labeled peptide. Approximately 2 days later, intact [125I]-HV-GHRH was isolated from the fetal liver, stomach contents, and brain. The amounts of tracer were positively correlated with those present in the corresponding dam's plasma. These data suggest that a GHRH analog of nonplacental origin, even at physiologic concentrations, can cross the placenta and, therefore, has the potential to influence fetal pituitary development directly.

Comparison of polymerase chain reaction and bacteriological culture for the diagnosis of sheep brucellosis using aborted fetus samples

PCR assay has been shown to be a promising option for the diagnosis of brucellosis. However, few studies have been performed with field samples in order to evaluate the assay as a diagnostic tool. In this study, routine use of a species-specific PCR assay previously developed for the identification of Brucella cultures was assessed for the detection of Brucella DNA directly from the stomach contents of aborted sheep fetuses. The assay is based on the insertion sequence IS711 in the Brucella chromosome. In the study, during 3 successive lambing seasons (1998–1999, 1999–2000 and 2000–2001) 126 aborted fetus samples each from different flocks and locations were examined. Brucella strains were isolated from 39 (31%) of the samples and all of the strains were identified as Brucella melitensis by biochemical characteristics, agglutination with monospesific A and M sera and PCR. Thirty-seven of 39 B. melitensis isolates were biotyped as biotype 3, and 2 isolates as biotype 1. From 38 of 39 culture positive fetal stomach contents B. melitensis-specific DNA was detected by PCR. PCR was found negative in all of the culture negative samples. Compared with culture, sensitivity and specificity of PCR were determined as 97.4 and 100%, respectively. The results indicate that this PCR procedure has a potential for use in routine diagnosis of sheep brucellosis.

Rapid diagnosis of ovine Brucella, Campylobacter and Salmonella infections from fetal stomach contents by coagglutination test

A coagglutination test (CT) was developed to detect Brucella, Campylobacter and Salmonella antigens in stomach contents of 58 aborted sheep fetuses and compared with conventional culture techniques. Brucella melitensis, Campylobacter fetus and Salmonella abortus ovis were isolated from 31 (53.4%), 13 (22.4%) and 4 (6.9%) fetuses, respectively. Thirty-five of the fetal stomach contents were positive with the CT test, including 19 (61.3%) with Brucella, 12 (92.3%) with Campylobacter and 4 (100%) with Salmonella CT tests, respectively. Seven samples were negative with all three CT reagents. Cross-reactions were seen with C ampylobacter CT (11 samples) and S almonella CT (six samples). The sensitivity and specificity of the CT tests were 61 and 80% for Brucella, 92 and 80% for Campylobacter, and 100 and 90% for Salmonella, respectively. The CT was found to be technically simple and rapid in diagnosing Salmonella, Campylobacter or Brucella infections by comparison with conventional culture methods.

Bioserogroups of Campylobacter species isolated from sheep in Kaduna State, Nigeria

Sheep Campylobacter isolates from Kaduna State were characterized into their species and bioserogrouped. A total of 1100 samples were collected from Kaduna abattoir and National Animal Production Research Institute (NAPRI), Shika. The samples were from 250 gallbladder, 250 intestinal contents, 100 fetal stomach contents all from Kaduna abattoir while 250 rectal swabs and 250 vaginal swabs were from the NAPRI Small Ruminant Programme. Of a total of 1100 samples, 39 (3.54%) yielded Campylobacter organisms. The highest isolation rate (6.8%) was from samples of intestinal contents followed by those from gall bladders (4.0%). Samples from the vaginal and fetuses had the lowest isolation rates (2.80%) and (0%), respectively. Of the 39 Campylobacter isolates from all the sources, (79%) were characterized as C. fetus subsp jejuni, C. coli (13%) and C. laridis (8.0%). while C. coli and C. laridis were isolated from gall bladder and intestinal contents only. Campylobacter fetus subsp jejuni biotype 1 accounted for 40.3% of the total isolates. C. laridis biotypes I and II were also isolated and accounted for 5% and 3% of the isolates, respectively. 5% of the isolates were not typeable. The serogroups 4 (13%), 36 (10%), 9 (10%), 84 (8%), 29 (5%) and 20 (8%) were the commonest serogroups identified in sheep at two locations surveyed. The isolation of Campylobacter organisms from rectum, vagina, gallbladder, and intestinal contents is a clear indication that sheep serves as a reservoir of this organisms in Nigeria. Similarities between documented human Campylobacter isolates in Nigeria and those in the present study raised the possibility of cross-transmission between sheep and man. It is concluded that biotyping and serotyping can be used for epidemiological study of campylobacteriosis due to Campylobacter jejuni in sheep in Kaduna State of Nigeria.

Tissue-invasive Tritrichomonas foetus in four aborted bovine fetuses.

Bovine trichomoniasis is a venereally transmitted protozoal infection that is usually associated with infertility and occasionally causes pyometra and midto late-term abortions. Some cows can carry the infection throughout gestation and remain infected after parturition. Abortions caused by Tritrichomonas foetus are usually diagnosed by microscopic identification of trichomonads in placental fluids, uterine exudate, or fetal stomach contents or by culture. In addition, characteristic placental and fetal lung lesions have been described, and fluorescent antibody and immunohistochemica1 techniques have been used to identify T. foetus in tissue sections. The purpose of this report is to describe 4 cases of tissue-invasive T. foetus infection in aborted bovine fetuses. A male fetus (fetus 1) was aborted in the seventh month of gestation by an Angus-cross heifer and submitted to the Tulare Branch of the California Veterinary Diagnostic Laboratory System (CVDLS) for postmortem examination. At necropsy, spleen and liver were enlarged, and the lungs were firm with patchy, suffusive hemorrhages, interlobular edema, and pale tan areas. Multifocal mucosal necrosis ranging from 3 mm in diameter to 20 x 15 mm was present in the rumen, and focal mucosal hemorrhages were present in the rumen, omasum, and abomasum. Multifocal hemorrhages and segmental hyperemia were seen in the small intestine, and petechial hemorrhages were present in the meninges. A 7-month female fetus (fetus 2) and a 6-month male fetus (fetus 3) were aborted by Holstein cows from the same dairy and were submitted to CVDLS for necropsy and diagnostic tests. Necropsy of fetus 2 revealed 2 liters of cloudy, yellow, viscous fluid in the rumen and abomasum and rare fibrin flecks adherent to the surface of a mottled liver. Fetus 3 had firm, diffusely yellow lungs with interlobular edema and patchy, swollen, hemorrhagic areas in the cranial lobes. The lungs floated in formalin. The rumen was diffisely hemorrhagic with a granular mucosa and patchy to paintbrush serosal hemorrhages. The omasal wall was diffusely reddened and hemorrhagic. Numerous ecchymotic hemorrhages were visible from the serosal and mucosal surfaces of the small intestine. Mesenteric lymph nodes were mildly enlarged and contained focal hemorrhages. Rare hemorrhages were present in the spiral colon and capsule of the spleen. The liver was mottled and slightly yellow.

Isolation of Pasteurellaceae from Bovine Abortions

A 2-year study was conducted to determine the incidence of Pasteurellaceae in abortion samples submitted for diagnostic evaluation. A total of 687 cases, including 623 with fetal tissues and/or stomach contents and 302 with placenta and/or uterine discharge, were evaluated. Pasteurellaceae were isolated on a nonselective medium from 9 (1.5%), 14 (2.8%), 13 (12.1%), and 42 (17.4%) of the fetal tissues, stomach contents, uterine discharges, and placentas, respectively. A total of 35 (19.9%) of 176 placental samples cultured on both a selective medium for Pasteurellaceae and a nonselective medium were positive for Pasteurellaceae. Fifteen (42.9%) of these isolates were detected only on the selective medium, whereas 5 (14.2%) were detected only on the nonselective medium and 15 (42.9%) grew on both media. Placentitis of different severity was evident in 13 (68.4%) of the 19 placentas from which Pasteurellaceae were isolated in the absence of other known abortifacient agents.

Counter-immunoelectrophoresis for the detection of Brucella antigen and antibodies in the diagnosis of brucellosis in buffaloes.

Counter-immunoelectrophoresis was employed for the detection of Brucella antigen in stomach contents of aborted buffalo fetuses and antibody in aborted as well as apparently healthy in contact buffaloes. Five of 16 aborted cases were serologically positive for brucellosis but isolation of Brucella abortus was successful in only two cases. By counter-immunoelectrophoresis, Brucella antigen was detected in the fetal stomach contents of four serologically positive cases. Of the 68 serum samples from in contact healthy buffaloes, 10 were positive with counter-immunoelectrophoresis: more than were detected by tube agglutination, Rose Bengal plate agglutination, complement fixation and agar gel precipitation test.

Detection of brucella antigen in bovine fetal stomach contents by agar gel precipitation and counter-immunoelectrophoresis

Among 21 fetuses from serologically positive dams soluble brucella antigen was detected in the stomach contents in 14 instances by agar gel precipitation test and in 19 by counterimmunoelectrophoresis. Brucella abortus biotype 3 was isolated from the fetal stomach contents in 15 of these cases. None of 20 fetuses from serologically negative dams revealed brucella antigen in the stomach contents in either test.

SuspectedSarcocystisinfection in an aborted bovine foetus

Abstract Extract Sir:- Samples from a case of sporadic abortion, two weeks before term in a two-year-old heifer, were submitted to the Palmerston North Animal Health Laboratory. They consisted of fresh foetal stomach contents, fixed foetal lung, and maternal urine and serum. No significant pathogens were isolated from the stomach contents or urine and serological tests for brucellosis and leptospirosis were negative.

Formula omitted] sp.—Induced abortion

History and Clinical Signs : Prodromal signs are not consistent. Most abortions occur in the last half of gestation. Gross Lesions : No grossly observable lesions occur consistently in the fetus. Autolysis of the fetus occurs frequently. Placentitis is characteristic but not pathognomonic. Microscopic Lesions : Lesions of inflammation such as placentitis and suppurative bronchopneumonia of the fetus are characteristic but not pathognomonic. Cultural Procedures : Isolation of Brucella sp. from the fetal stomach contents or tissues or placental tissues provides conclusive evidence for a diagnosis. Isolation of Brucella sp. from the uterine fluids, milk or tissues of the aborting cow is absolute evidence of infection and presumptive evidence of the cause of the abortion. Serologic Procedures : A positive card test, or a plate or tube agglutination titer to brucellosis of 1:100 or greater in the cow's serum is absolute evidence of infection and presumptive evidence of the cause of the abortion. Special Procedures : Brucella organisms may be demonstrated in the stomach contents, and fetal or placental tissues by proper application of fluorescent antibody techniques. This procedure should be supplemented by culture. Preferred Diagnostic Methods : 1. 1. Isolation of brucella organisms from the fetus, placenta or uterine fluids of aborting cows. 2. 2. Demonstration of brucella organisms in the fetus or placenta by FA techniques. 3. 3. Demonstration of a plate or tube brucella titer of 1:100 or greater in the cow provides evidence of infection but does not define the cause of the abortion.

Protein composition of the rabbit foetal fluids

The total protein concentration of fluids in different compartments of the rabbit conceptus from 20 to 28 days of age was measured, and concentrates of these fluids analysed electrophoretically in order to determine their protein composition. For comparison, similar analyses of foetal sera, maternal sera and, where possible, uterine fluid were carried out. The allantoic fluid had a protein concentration less than that of the amniotic fluid, whilst the exocoelomic fluid had a protein concentration greater than that of the amniotic fluid. The total protein concentration of these fluids increased from about 23 days onwards. Compared to the other fluids, protein became much more concentrated in the foetal stomach contents. When analysed by fluid agar electrophoresis, the foetal exocoelomic, amniotic and allantoic fluids and stomach contents contained similar electrophoretic components (albumin, α -1- globulin, α -2-globulin, β -globulin and γ -globulin) and resembled each other in the proportion of the components. These proportions (as best exemplified in the amniotic fluid) changed during the period investigated, and at all times were different to those of the foetal and maternal sera. In certain respects these proportions more closely resembled those of the proteins in the uterine fluid. Analysis of the foetal fluids, both by starch gel electrophoresis and immunoelectrophoresis, revealed a paucity of lipoproteins and of α -1- and α -2-macroglobulins. Some samples of uterine fluid showed a similar paucity of these components. An α -2-globulin component, common to the foetal exocoelomic, amniotic and allantoic fluids, stomach contents and serum, but absent from the maternal serum, was detected (with a rat antiserum to the combined foetal amniotic fluid and stomach contents). Additional components were detected in the foetal stomach contents, both by the rat antiserum and by starch gel electrophoresis. Experiments on the permeability of the membranes in vitro were performed. No obvious increase in permeability of the amnion and yolk sac splanchnopleur with increasing gestational age could be found when maternal serum was dialysed across the isolated foetal membranes. The proportions of the protein components in the dialysates were similar, indicating that no alteration in permeability of the membranes had occurred. The paraplacental chorion, as compared to the yolk sac splanchnopleur, showed a greater permeability to protein in vitro . The role which the uterine fluid and foetal serum might play in governing the protein composition of the foetal fluids is discussed. It is suggested that the paraplacental chorion is the major site for the passage of proteins to the foetal fluids other than the blood.

Passage of heterologous serum proteins from mother into fetal compartments in the rabbit

Dutch rabbits were injected with human or bovine serum albumin or human gamma globulin at 19, 24, and 28 days of gestation. Fetal serum, stomach contents, and exocoel, amniotic, and allantoic fluids were collected at intervals from 1 to 48 hr later and titrated with specific antisera. The presence of injected protein in fetal samples was confirmed by electrophoresis and absorption of rabbit protein, followed by the precipitin test. The three nonantibody proteins were detected in fetal serum and exocoel fluid after day 19 and in all fetal compartments after day 22, as soon as 1 hr after injection into the mother and after a maternal dose as low as 10 mg/kg., About 1 mg was found in the entire fetus 24 hr after a maternal dose of 62.5 mg/kg. The proteins passed from maternal circulation to uterine lumen, to exocoel, to allantois and amnion, and from amnion to stomach. They passed from mother to fetal circulation via the vessels of the yolk sac splanchnopleure and not through the allantoidean placenta.

Selection of antitoxins by the foetal membranes of rabbits.

Antibodies pass from the uterine lumen of pregnant rabbits into the foetal body fluids. It has been shown that antibodies formed in rabbits enter the foetal circulation by this route much more readily than those formed in horses or cows. It is the intention of this paper to investigate further this selective admission to the foetal circulation, to determine if there is a differential selection of heterologous antibodies according to the species in which they were produced and if this selectivity changes with foetal age. Antitoxins were employed throughout and were prepared in rabbit, man, guinea-pig, dog, horse and cow, against diphtheria or tetanus toxin or Clostridium welchii α-toxin. Mixtures of sera from two or three of these species, each serum containing a different antitoxin, were administered so far as possible. The antitoxin concentrations observed in the experimental samples were corrected to unit concentration in the serum administered, the term concentration quotient (c.Q.) being used for this quantity. Measured in this way the entry of antitoxin to the foetal serum varied with the species in which the antitoxin was produced; rabbit, man, guinea-pig, dog, horse and cow forming a descending series in that order. The entry of antitoxin into the amniotic fluid, or the foetal stomach contents derived from it, does not vary with the species in which the antitoxin was produced. This is so also for the maternal serum. Thus the species in which an antitoxin is produced affects the concentration at which it will appear in the foetal serum but not that at which it will appear in the other foetal body fluids or in the maternal serum. The entry of antitoxin prepared in rabbits to the foetal circulation increases with foetal age from 20 to 26 days after copulation and decreases thereafter.

Electrophoretic studies of serum proteins of foetal rabbits.

The sera of non-pregnant adult rabbits which had been hyperimmunized to Brucella abortus antigen, the sera of pregnant female rabbits which had not been immunized, and the sera, exocoelomic fluids, amniotic fluids and stomach contents of 25-day-old foetal rabbits were examined electrophoretically and ultracentrifugally. The serum of pregnant rabbits differed both in total protein concentration and in the proportions of the components from that of non-pregnant hyperimmunized rabbits. The foetal sera contained components corresponding to albumin, α-, β- and γ -globulin, but the proportions of these components, as well as the total protein concentrations, differed widely from those of the sera of both pregnant and non-pregnant adults. Foetal exocoelomic fluid, amniotic fluid and stomach contents contain similar electrophoretic components and resemble each other closely in the proportions of the components, though differing in the total concentrations. The components resemble those of sera in mobility, but the proportions of the components differ widely from those of the sera, both foetal and adult. Various experimental procedures to which rabbits were subjected resulted, after 24 h, in a significant increase in the total protein concentrations, without any corresponding change in the proportions of the components, of the foetal sera. No corresponding changes were detected in the maternal sera. The effect on the sera could be explained by withdrawal of water alone from the foetal circulation. Using antibodies as markers, it was shown that both β - and γ -globulin enter the foetal circulation from immune rabbit serum injected into the uterine lumen. The importance of the contribution of maternal serum proteins to the foetus is discussed in the light of the immunological and electrophoretic results.

The accumulation of antibodies in the stomach contents of foetal rabbits.

Antibodies can be detected in the amniotic fluid and in stomach contents of rabbit foetuses exposed to immune serum following injection into the uterine lumen or into the maternal circulation. It is shown that the positive reactions obtained with foetal stomach contents were due to the presence of specific antibodies derived from the immune sera to which the foetuses were exposed. By means of injections of immune rabbit and bovine sera into the embryonic membranes of 24-day rabbit foetuses, it was demonstrated that the antibodies in the stomach contents are derived from the amniotic fluid and not from the foetal circulation. There is a pronounced tendency for antibodies to be concentrated in the stomach, concen­trations of 32 and 64 times the titre of the amniotic fluid being observed. Although anti­bodies of rabbit origin readily pass through the yolk-sac splanchnopleur into the foetal circulation, they do not do so through the stomach wall in detectable amounts.

Monovalent and Polyvalent Antigens for Use in the Diagnosis of Bang's Disease.

The agglutinability of 54 strains of Bact. abortus prepared as monovalent antigens and one polyvalent antigen has been studied. Bovine sera showing a large variation in agglutinin content have been employed. In these studies there was no significant variation in the agglutinability of the monovalent antigens. Any of the cultures would have been suitable for the production of antigen for use in the diagnosis of Bang's disease. There was no advantage or disadvantage in favor of any of the monovalent antigens as compared with the polyvalent antigen. The one polyvalent antigen used was prepared from 8 strains of Bact. abortus, all of which were isolated from cattle. The 54 cultures varied in age of cultivation on artificial media from 13 years to a few days. Thirty-nine were isolated at the Minnesota Experiment Station. Four originated in Pennsylvania. Four were received from Connecticut, 3 from New Zealand, one (No. 80) was isolated in California, and one more was received from California that had been isolated in Oregon but cultivated in California for several years, one from New York, and one from Oregon. Of the 39 isolated at Minnesota, 32 were from cattle, 5 from horses, and 2 from swine. Those from cattle were isolated from whole milk, colostrum, placenta, vaginal discharges, foetal lung, foetal stomach contents and from enlarged joints. In one case 4 cultures were isolated from the same animal but from different tissues or fluids. One of the cultures isolated from a bovine was classified by Huddleson as Para-abortus. In preliminary studies, sera from 97 cattle were used, 14 of which showed high agglutinin content, 15 showed medium agglutinin content, 24 had a very low titre, and 44 negative sera with no agglutinin content.