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Serotyping of Campylobacter fetus subsp. fetus and C. fetus subsp. venerealis by passive hemagglutination technique based on soluble autoclaved antigens.

Antigens for sensitization of sheep erythrocytes were released from C. fetus subsp. fetus and C. fetus subsp. venerealis suspensions by autoclaving in alkaline and neutral saline and by boiling in alkaline but not in neutral saline. Serotyping with rabbit antisera showed the same two serotypes of O-antigens, A and B, as shown by tube agglutination of heated bacteria. All C. fetus subsp. venerealis strains had the A antigen. The C. fetus subsp. fetus strains tested had either the A or B antigen. All ten human isolates tested were of the A type.

Antigens, Bacterial

Campylobacter fetus subspecies fetus infection.

During a six-year period five patients with Campylobacter fetus subspecies fetus infections were seen at the Mayo Clinic in Rochester, Minnesota. Bacteremia was observed in two patients, one presenting with aortic valve endocarditis and the other with abdominal atherosclerotic aortic aneurysm. C. fetus subsp. fetus was isolated from tibial tissue of a patient with osteomyelitis. Diarrhea was the main complaint of two further patients, and was also mentioned by the patient with the aortic aneurysm. Despite the use of incubation conditions and selective media geared to detect only Campylobacter jejuni, C. fetus subsp. fetus was isolated from stool specimens of the two patients with gastrointestinal symptoms. The fact that three of five C. fetus subsp. fetus infections observed in this study were associated with intestinal symptoms further supports the importance of the gastrointestinal tract in the pathogenesis of C. fetus subsp. fetus infections.

Aged

Thrombophlebitis and cellulitis due to Campylobacter fetus ssp. fetus. Report of four cases and a review of the literature.

Four cases of acute thrombophlebitis and cellulitis due to C. fetus ssp. fetus are reported, with a review of 18 previously reported cases. Vascular infection with thrombophlebitis due to C. fetus ssp. fetus occurred predominantly in adult male patients with underlying debilitating, immunocompromising illnesses resulting in a mortality rate of 32%. Although approximately one-third of the patients had exposure to known reservoirs of C. fetus ssp. fetus, none of the patients presented with diarrhea, and only one of the cases had C. fetus ssp. fetus recovered from stool culture. Diagnosis of C. fetus ssp. fetus thrombophlebitis or cellulitis is based on clinical suspicion and recovery of the agent from blood culture; the latter requires an average incubation period of 8 days. Empiric therapy with erythromycin, and an aminoglycoside or chloramphenicol is recommended in suspect patients pending results of blood cultures.

Adult

Uterine and umbilical blood flows and net nutrient uptake by fetuses and uteroplacental tissues of cows gravid with either single or twin fetuses.

To evaluate the influence of cow breed and number of fetuses on uterine and umbilical blood flows and nutrient fluxes or uterine tissues from gravid cows, surgery was performed on Charolais or Hereford cows with single or twin fetuses at 177 +/- .2 d (mean +/- SEM) after mating. Indwelling catheters were placed in a fetal femoral artery and vein, in an umbilical vein of each fetus, and in a uterine artery and vein of each gravid horn. Deuterium oxide (D2O) was infused into a fetal femoral vein at 183 +/- .3 and 190 +/- .5 d after mating to estimate uterine and umbilical blood flows (liters/minute). Blood oxygen and plasma glucose and lactate concentrations were determined and uterine arterio-venous (A-V) and umbilical venous-fetal arterial (v-a) differences and net uterine and fetal uptakes were calculated. Net utilization by the uteroplacenta was calculated as the difference between uterine and fetal net uptakes. Uterine blood flows were lower (P less than .01) in Hereford (4.80 +/- .28) than in Charolais (7.07 +/- .33) and lower (P less than .01) per fetus in cows with twin fetuses (5.22 +/- .34) than in cows with a single (6.65 +/- .28) fetus. Umbilical blood flows were greater for single than for twin fetuses. Fetal oxygen and glucose net uptakes averaged 57 and 12%, respectively, of net uteroplacental utilization. Lactate was released from uteroplacental tissues to fetal (42%) and maternal circulations (58%). Fetal oxygen uptakes tended to be less for twin fetuses (P = .08) than for a single fetus.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals

Campylobacter infection associated with raw milk. An outbreak of gastroenteritis due to Campylobacter jejuni and thermotolerant Campylobacter fetus subsp fetus.

Raw milk is identified with increasing numbers of outbreaks of gastroenteritis and is an important vehicle for transmission of Campylobacter infection. Unlike Campylobacter jejuni, Campylobacter fetus subsp fetus has not been associated with common-source outbreaks of gastroenteritis. This report describes an outbreak of gastroenteritis involving C jejuni and a thermotolerant strain of C fetus subsp fetus associated with raw milk. Fifteen (39%) of 38 persons who attended a banquet in Wisconsin in June 1982 developed acute gastroenteritis. Stool specimens were obtained from nine ill guests; four yielded C jejuni and three yielded C fetus subsp fetus. The C fetus subsp fetus isolates were identified fortuitously, in part because of unusual thermotolerance (growth at 42 degrees C), permitting isolation at temperature appropriate for C jejuni. Survey results implicated raw milk as the source of the outbreak. Findings provide evidence of a potentially emergent milkborne pathogen contributing to the risk of raw milk consumption and suggest that current diagnostic laboratory techniques may fail to identify significant foodborne agents.

Adolescent

Biochemical and genetic characteristics of atypical Campylobacter fetus subsp. fetus strains isolated from humans in the United States.

During a 2-year period, 14 biochemically atypical Campylobacter fetus subsp. fetus-like strains were received by the Campylobacter Reference Laboratory at the Centers for Disease Control. Sources of the isolates were blood, nine strains; stools, two strains; amniotic fluid, one strain; and abscesses, two strains. Atypical phenotypic characteristics exhibited by one or more strains were growth at 42 degrees C, 10 strains; no H2S by lead acetate paper, 3 strains; resistance to a 30-micrograms cephalothin disk, 2 strains; and nonmotility, 1 strain. By DNA-DNA hybridization, all 14 isolates and the type strain of C. fetus subsp. fetus (ATCC 27374) were 94 to 100% related in reassociation reactions at 50 degrees C, with 0.0 to 0.5% divergence, and were 86 to 100% related in reassociation reactions at 65 degrees C. Thus, all of these atypical strains were C. fetus subsp. fetus. MICs of 11 antimicrobial agents for these 14 strains were variable. All strains were susceptible to chloramphenicol, erythromycin, gentamicin, and tetracycline, and most were susceptible to ampicillin, clindamycin, and penicillin. Eleven strains were resistant to cephalothin (MIC greater than or equal to 16 micrograms/ml), nine were resistant to rifampin (MIC greater than or equal to 8 micrograms/ml), and all were resistant to nalidixic acid (MIC greater than 32 micrograms/ml) and vancomycin (MIC greater than 32 micrograms/ml). One can expect to see biochemical variability in C. fetus subsp. fetus strains and to encounter such strains from a variety of human sources, the most important of which appears to be blood.

Campylobacter Infections

Hormone ontogeny in the ovine fetus. XXVI. A sex difference in the effect of castration on the hypothalamic-pituitary gonadotropin unit in the ovine fetus.

The detection of pulsatile ovine LH (oLH) secretion in the sheep fetus by 81 days gestation (term 147 days), the suppression of fetal gonadotropin secretion by chronic administration of an LH-releasing factor agonist or antagonist, and the capacity of N-methyl; D-aspartate (a neuroexcitatory amino acid analog) to evoke a fetal oLH pulse strongly support a functional LH-releasing factor pulse-generator in the ovine fetus. In light of the sex difference in fetal gonadal function and gonadotropin secretion before day 114, we postulated that fetal castration would have a discordant effect on the pattern of gonadotropin secretion in males and females. Fetal sheep were either castrated (male = 11; female = 9) or sham operated (male = 9; female = 6) at 110-115 days gestation. Chronic indwelling arterial and venous catheters were implanted, and animals were studied for up to 30 days. During each study period fetal arterial blood samples were drawn every 15 min for 5 h and the plasma assayed for oFSH and oLH by specific RIAs. Multiple studies were performed on each fetus. In all fetuses (both intact and castrated) a decrease in oLH pulse frequency occurred after day 130. In female fetuses before day 130, castration had no effect on mean oLH pulse frequency (sham, 0.72 +/- 0.19 pulses/5 h; castrate, 0.50 +/- 0.13 pulses/5 h; P greater than 0.05). After day 130, pulsatile oLH secretion decreased in both intact and castrated female fetuses to undetectable levels during the sampling period. In contrast, castration significantly (P less than 0.001) increased mean oLH pulsatility in males before and after day 130 (less than 130 days, sham, 1.06 +/- 0.24 pulse/5 h; castrate, 2.70 +/- 0.22 pulse/5 h; greater than 130 days, sham, 0.18 +/- 0.12 pulses/5 h; castrate, 1.65 +/- 0.26 pulses/5 h). Mean oLH pulse amplitude was increased by castration only in the male fetuses (sham, 3.89 +/- 0.87 ng/ml; castrate, 6.02 +/- 0.39 ng/ml; P less than 0.05). oFSH pulses were infrequent in both sexes and not influenced by castration. The mean plasma concentration of oFSH was greater in intact female fetuses than in intact males (female, 5.65 +/- 1.15 ng/ml vs. male, 2.07 +/- 0.45 ng/ml; P less than 0.01). Castration increased the mean value for plasma oFSH in males (4.40 +/- 0.43 ng/ml; P less than 0.001) but had no effect in females (3.83 +/- 0.64 ng/ml; P greater than 0.05).(ABSTRACT TRUNCATED AT 400 WORDS)

Animals

Plasma growth hormone concentration in the chronically catheterized ovine fetus during spontaneous term delivery and premature delivery induced by continuous intravascular infusion of low doses of adrenocorticotropin or cortisol to the fetus.

Fetal plasma growth hormone concentrations were measured in 15 pregnant ewes over the last 5 days before delivery. In five chronically catheterized pregnant ewes that underwent spontaneous vaginal delivery 146 +/- 2 days (mean +/- SD) of gestation, fetal plasma growth hormone concentrations fell from 124.6 +/- 44.1 ng X ml-1 5 days before delivery to 35.2 +/- 31.3 ng X ml-1 at delivery. In three fetuses in which premature delivery was induced by the infusion of cortisol to the fetus at 128 days of gestation, fetal plasma growth hormone levels fell from 195 +/- 19.1 ng X ml-1 to 70.7 +/- 25.5 ng X ml-1 over the last 5 days of intrauterine life. In seven fetuses in which premature delivery was induced with infusion of synthetic adrenocorticotropin to the fetus beginning at 120 or 130 days of gestation, the fetal plasma growth hormone level did not fall (175.6 +/- 75.7 to 158.9 +/- 60.1 ng X ml-1). Fetal plasma cortisol concentrations at delivery were significantly higher in the cortisol-infused fetuses (214 +/- 38.4 ng X ml-1) than in both control (94.4 +/- 33.7 ng X ml-1) and adrenocorticotropic hormone-infused fetuses (94.5 +/- 31.9 ng X ml-1). The fall in the fetal plasma growth hormone level in cortisol-induced fetuses may be due to the higher fetal plasma cortisol concentrations achieved in the cortisol-infused compared with the adrenocorticotropic hormone-infused fetuses in which no comparable decrease in growth hormone was observed. These findings suggest that there are significant differences in the fetal response to various experimental regimens used for the induction of premature labor in the sheep.

Adrenocorticotropic Hormone

Comparison of surface antigens of some Campylobacter fetus subsp. fetus strains of ovine origin by polyacrylamide gel electrophoresis and immunoblotting.

Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblotting were used to identify and to compare the surface antigens of eight C. fetus subsp. fetus strains. Seven strains (one of serogroup A and six of serogroup B) were isolated from aborted ovine fetuses, while one strain (serogroup A) originated from an aborted calf fetus. Saline extracts at 56 degrees C and 100 degrees C were used as antigens. Antisera were produced in rabbits. In saline extracts (56 degrees C) of the strains at least 19 fractions were identified by SDS-PAGE, with molecular masses ranging from approx. 4,800 to 205,000. The major bands appeared at 205,000, 66,000, 31,500, 25,000, 21,000 and 17,500. Despite the fact that the strains were cultured from 4 different sheep flocks and belonged to serogroup A or B, the SDS-PAGE profiles of the strains were very similar. When boiled (100 degrees C) extracts were used, a band migrating at 32,500 in sheep strains and a band at 97,500 in the calf isolate were missing. Most of the bands obtained by SDS-PAGE could be identified also by the immunoblot procedure. A or B type specificity of the ovine isolates was due to an LPS fraction, migrating at approx. 21,000, while the other LPS fractions appearing under this region although reacted with antisera did not influence the type specificity. Using alkaline extracts (pH 12) in SDS-PAGE, LPS fractions gave more pronounced profiles. In two of our C. fetus subsp. fetus isolates, plasmids with a molecular mass of 31,500 were identified.

Abortion, Veterinary

Immunisation of cattle against vibriosis with vaccines prepared from Campylobacter fetus subsp fetus.

Subcutaneous administration of vaccines prepared from cells of Campylobacter fetus subsp fetus strain A28 to heifers gave substantial protection against infertility due to C. fetus subsp venerealis strain B6. Strains A28 and B6 had different heat-stable antigens and conformed respectively to serotype B and serotype A of Berg et al (1971). The results suggested that the protective antigens were heat-labile antigens common to both strains. Although vaccines prepared from serotype B strains of C. fetus subsp fetus could be used to immunise cattle against vibriosis, the results did not suggest that their use in preference to those prepared from C. fetus subsp venerealis would offer any advantages.

Abortion, Veterinary

[A case report of Campylobacter fetus subspecies fetus bacteremia].

An unusual case of Campylobacter fetus subspecies fetus bacteremia was presented. A twenty four year old male was admitted to our hospital due to abdominal pain, general malaise, diarrhea, high fever, and hemoptysis. He was alcoholic and fond of eating raw liver. He had a history of partial gastrectomy and disturbance of pancreatic function. He showed pulmonary empyema, pleuritis, thrombophlebitis of lower legs, jaundice, hepatomegaly, diarrhea, pneumothorax, and low T3 low T4 syndrome. C. fetus subsp. fetus was detected from the venus blood and pleural effusion on admission. He was successfully treated by gentamicin, chloramphenicol, and minocycline. This is the fourth case of C. fetus subsp. fetus bacteremia in the Japanese literature. This microanerophilic gram negative curved bacillus has been increasingly associated with human disease and relapsing in nature, so protracted antimicrobial therapy was recommended.

Adult

Colonization and infection of athymic and euthymic germfree mice by Campylobacter jejuni and Campylobacter fetus subsp. fetus.

Human clinical strains of Campylobacter jejuni and Campylobacter fetus subsp. fetus colonized the gastrointestinal tracts of both athymic (nu/nu) and euthymic (+/nu) germfree mice (BALB/c). Viable Campylobacter spp. (10(9) to 10(10) CFU/g [dry weight] of cecum and colon contents) were isolated on day 3 after oral challenge, and similar large numbers of viable cells were evident at several intervals during a 10-month experiment. The stomachs and upper small intestines of nu/nu and +/nu mice that were monoassociated for 224 days with C. jejuni 45100 contained 3 to 4 logs fewer viable bacteria than did their ceca or colons. Athymic mice that were monoassociated for 224 days with C. fetus subsp. fetus had 2 to 3 logs more viable Campylobacter spp. in their upper gastrointestinal tracts than did their +/nu littermates. Large viable populations (approximately 10(9)/g of contents) of C. fetus subsp. fetus were in the ceca and colons of both nu/nu and +/nu mice. All C. jejuni strains used in this study chronically infected the mesenteric lymph nodes of both nu/nu and +/nu mice. C. jejuni strains 24 and INN 73-83, which were cytotoxic for Chinese hamster ovary cells in vitro, were also more frequently isolated from the livers, spleens, and kidneys of nu/nu mice than was the weak cytotoxin-producing strain 45100. Additionally, heat-labile-enterotoxin-producing C. jejuni INN 73-83 was recovered more frequently from the internal organs of monoassociated +/nu mice than were any other Campylobacter spp. tested. Natural gastrointestinal colonization of neonatal nu/nu and +/nu mice (born to Campylobacter-colonized mothers) with Campylobacter spp. appeared to be delayed until approximately 1 to 2 weeks after birth. Conventionalization of C. jejuni 45100-monoassociated BALB/c mice with a complex mouse fecal microflora eliminated viable C. jejuni from the mesenteric lymph nodes by day 14 and from the cecum by day 78. These findings show that the gnotobiotic BALB/c mouse is a new model for studying acute and chronic host-Campylobacter sp. interactions.

Animals

Hormone ontogeny in the ovine fetus: XIX: The effect of a potent luteinizing hormone-releasing factor agonist on gonadotropin and testosterone release in the fetus and neonate.

To investigate further the role of the hypothalamic luteinizing hormone releasing factor (LRF) pulse generator and the pituitary LRF receptor in the regulation of gonadotropin secretion and gonadal steroidogenesis in the ovine (O) fetus and neonatal lamb, we measured the increment (the difference between the concentration of plasma LH at time 0 and peak LH) in oLH (delta oLH) and oFSH (delta oFSH) responses to a potent LRF agonist, D-Trp6Pro9NEt-LRF (LRF-A), after consecutive daily doses in 17 ovine fetuses (six females, 11 males) and in 15 neonatal lambs (six females, nine males). Seven of the lambs had been studied as fetuses. In addition, plasma concentrations of testosterone (T) and androstenedione (delta 4A) were measured in nine male fetuses. After a stimulatory response to the first dose of LRF-A, the mean delta oLH and delta oFSH responses in the 106- to 118-d gestation fetuses of both sexes were significantly suppressed by the fourth dose and in the neonatal lamb by the second dose. Suppression was sustained throughout the duration of LRF-A therapy which included the gestational interval when the fetal pituitary exhibits its greatest responsiveness to an acute dose of synthetic LRF. The duration of oLH and oFSH suppression after cessation of LRF-A therapy was studied by measuring the delta oLH and delta oFSH responses to LRF before and at intervals after LRF-A therapy. In the fetus, the delta oLH and delta oFSH responses remained significantly decreased 7-8 d after the agonist was discontinued.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals

Nosocomial meningitis due to Campylobacter fetus subspecies fetus in a neonatal intensive care unit.

We experienced a nosocomial outbreak of Campylobacter fetus subspecies fetus (C. fetus) meningitis in a neonatal intensive care unit. A cluster of three infants developed meningitis approximately 1-1.5 months after the index patient was admitted. Two asymptomatic intestinal carriers of C. fetus were detected during the outbreak. Our experience indicates that C. fetus can cause nosocomial meningitis in neonates, and asymptomatic carriers may play a role in the transmission of the organism.

Campylobacter Infections

Control of gonadotropin secretion in the ovine fetus. IV. Male-specific entrainment of the hypothalamic control of luteinizing hormone secretion by testosterone in the female ovine fetus.

The role of testosterone (T) in the sex-specific entrainment of hypothalamic LH regulation was studied in five castrate and four sham-castrate chronically catheterized female fetuses androgenized by the prior administration of T cypionate (200 mg, im, every 2 weeks) to pregnant ewes from 30-86 days gestation (term = 147 days). Eight female and three male castrate fetuses served as controls. After a 2-week washout period all fetuses were operated upon in utero between 106-116 days and were studied longitudinally over a 2- to 35-day period. LH pulsatility was determined from blood samples obtained every 15 min over a standard 3-h observation period and assayed for LH by RIA (NIH LH-S16 standard). LH pulse frequency in the castrate androgenized females (41 pulses in 19 observation periods; 1 pulse/1.4 h) was significantly higher than that in non-androgenized female controls (18 pulses in 28 observation periods; 1 pulse/4.7 h observation), but was similar to that in castrate males (23 pulses in 28 observation periods; 1 pulse/1.3 h). Furthermore, LH pulse frequency in the sham castrate androgenized females (26 pulses in 13 observation periods; 1 pulse/1.5 h) was comparable to that in castrate androgenized females as well as that in castrate males. The enhanced LH pulsatility in androgenized female fetuses strongly suggests that T exposure between 30-86 days results in male-specific entrainment of hypothalamic LH regulation. Moreover, the comparable enhancement of LH pulse frequency in both sham castrate and castrate androgenized female groups suggests that in the T-exposed fetus T withdrawal alone is sufficient to result in enhanced LH pulsatility. These findings strongly suggest that T of fetal testicular origin results in male-specific entrainment of hypothalamic function and may be an important feature of male neural organization in this species.

Animals