'Flexispira rappini' strains represent at least 10 Helicobacter taxa.

'Flexispira rappini' is a provisional name given to gram-negative, microaerophilic, motile, spindle-shaped micro-organisms with spiral periplasmic fibres and bipolar tufts of sheathed flagella. Several investigators, including Kirkbride, Romero, and Archer isolated strains possessing this morphology. Previously, the phylogenetic position of three 'Flexispira rappini' strains was determined by 165 rRNA sequencing, which indicated that flexispira were members of the genus Helicobacter. As more organisms with 'F. rappini' morphology were isolated, it became apparent that there were multiple Helicobacter taxa with this distinctive morphology. The purpose of this study was to examine a collection of 36 'F. rappini' strains from diverse habitats by using 165 rRNA sequence analysis. The strains fell into 10 taxa, each possibly representing a novel Helicobacter species. Two of these flexispira taxa were previously named, by us, Helicobacter bilis and Helicobacter trogontum. Currently, none of the flexispira taxa contains enough phenotypically and genotypically characterized strains to be formally named 'Helicobacter rappinii'.

Diagnoses in 1,784 ovine abortions and stillbirths.

Of 1,799 accessions of ovine abortions and stillbirths by the South Dakota Animal Disease Research and Diagnostic Laboratory during a 10-year period, 1,784 were suitable for examination. Etiologic diagnoses were made in 786 (44%). Infectious agents were found responsible in 702 accessions (39%), and noninfectious causes were involved in 84 (5%). No diagnosis was made in 998 accessions (56%), 61 of which were mummified fetuses. Lesions, most of which indicated the presence of an infectious agent, were found in 274 cases (15%) in which no agent could be identified. Together, Toxoplasma gondii, Campylobacter sp., and Chlamydia psittaci caused approximately 25% of all abortions and stillbirths examined.

Bacterial agents detected in a 10-year study of bovine abortions and stillbirths.

In a 10-year survey started in 1980, specimens from 8,995 bovine abortions and stillbirths were submitted to the South Dakota Animal Disease Research and Diagnostic Laboratory. Of these, 8,962 were suitable for some type of examination. Bacteria were determined to be the cause of 1,299 (14.49%). The 5 bacteria most commonly associated with bovine abortion or stillbirth were Actinomyces pyogenes, 378 (4.22%); Bacillus spp., 321 (3.58%); Listeria spp., 121 (1.35%); Escherichia coli, 98 (1.09%); and Leptospira interrogans, 79 (0.88%). Twelve other genera of bacteria were associated with > or = 10 abortions or stillbirths, and 12 more species were associated with < or = 10 abortions or stillbirths.

Effect of feeding lasalocid to pregnant ewes experimentally infected with Toxoplasma gondii.

Sixteen 50 day gestational ewes were fed lasalocid at the rate of 30 g t-1 and were orally inoculated with 100 infective Toxoplasma gondii oocysts 5 days after beginning feeding of lasalocid. Seventeen control ewes were similarly inoculated with T. gondii and were not fed lasalocid. The rate of abortion and neonatal mortality in both treated and untreated ewes was similar, indicating that feeding lasalocid was not effective in preventing T. gondii abortion in sheep.

Viral agents and associated lesions detected in a 10-year study of bovine abortions and stillbirths.

In a 10-year survey started in 1980, specimens from 8,995 bovine abortions and stillbirths were submitted to the South Dakota Animal Disease Research and Diagnostic Laboratory. Of these, 8,962 were suitable for some type of examination. Viruses were associated with 948 (10.58%). Bovine herpesvirus-1 (IBR) was detected in 485 (5.41%), and bovine viral diarrhea virus (BVDV) was detected in 407 (4.54%). In 1 year of the survey, BVDV was detected in 8/32 fetuses that had lesions of passive congestion. Bovine herpesvirus-4 was isolated from 47 specimens (0.52%), parvovirus and enterovirus were each isolated from 2, and adenovirus, parainfluenza virus, and pseudorabies virus were each isolated from 1. Malignant lymphoid neoplasia was present in 2 fetuses, and their abortion was assumed to have been caused by the bovine leukosis virus.

Etiologic agents detected in a 10-year study of bovine abortions and stillbirths.

During a 10-year period, specimens from 8,995 bovine abortions were submitted to the South Dakota Animal Disease Research and Diagnostic Laboratory; 8,962 of these specimens were suitable for examination. A supportable diagnosis was made in 2,942 (32.82%) of the abortions examined. An infectious cause was determined for 2,723 (30.38%) and a noninfectious cause for 219 (2.44%). Bacteria caused 1,299 (14.49%), viruses caused 948 (10.57%), and fungi caused 476 (5.31%). The infectious bovine rhinotracheitis virus was associated with 485 (5.41%) and the bovine viral diarrhea virus with 407 (4.54%). The most common noninfectious causes of abortion or stillbirth were anomalous development, 112 (1.25%); dystocia, 56 (0.62%); and twinning, 37 (0.41%). The cause of 6,020 (67.17%) abortions was not determined. Lesions, many suggesting the presence of infection, were present in 1,554 (17.34%) fetuses or placentas in which no infectious agent could be demonstrated.

Fungi associated with bovine abortion in the northern plains states (USA).

Mycotic infection was diagnosed in 6.8% of 6,858 cases of bovine abortion and stillbirth examined during a 9-year period. Aspergilli were associated with approximately 5% of all abortion cases and 71% of 446 cases that were cultured for fungi and diagnosed as mycotic abortion. Aspergillus fumigatus was the most frequent isolate (62%), followed by A. terreus (6.7%), Emericella (Aspergillus) nidulans (3.0%), A. flavus (2.9%), and E. rugulosus (less than 1.0%). Zygomycetes (Absidia, Mortierella, Rhizomucor, Rhizopus) accounted for 21% of the cases. Pseudallescheria boydii and yeasts (Candida, Torulopsis) were each identified in 2% of the cases. Fungi that uncommonly cause infection accounted for 2% of the cases and included Curvularia geniculata, Exophilia jeanselmei, Hendersonula toruloidea, Lecythosphora hoffmannii, Talaromyces flavus var. flavus (Penicillium vermiculatus), T. (Penicillium) thermophilus, and Wangiella dermatitidis. About 10% of the mycotic cases were mixed fungal infections involving A. fumigatus (87%), A. flavus (12.5%), or E. nidulans (12.5%) coexisting with Absidia corymbifera (72%), Rhizomucor pusillus (4.3%), or Rhizopus arrhizus (4.3%). In each mixed infection, both septate and nonseptate hyphae were observed in placental tissues. Twelve percent of the mycotic abortion cases were diagnosed by histologic examination alone because isolation attempts were negative or only formalin-preserved tissues were available.

Toxoplasmosis and other causes of abortions in sheep from north central United States.

Between 1983 and 1989, we examined 1,201 aborted fetuses and dead lambs from the north central United States. Toxoplasmosis was diagnosed in 17.5%, campylobacteriosis in 9.9%, chlamydiosis in 4.7%, and miscellaneous infections in 15.1%. Inflammatory lesions suggestive of infectious causes were seen in 13%. Noninfectious causes were identified in 6.1%, and a diagnosis was not reached in 33.3%. An agglutination test was used to detect Toxoplasma gondii-specific antibodies in ovine fluids. Toxoplasma gondii antibodies were detected in 223 of 1,064 (20.9%) fluids from fetuses and dead lambs. Of 201 seropostive (greater than or equal to 16) fetuses and lambs, T gondii antibody titers (reciprocal) were 16 (21 fetuses and lambs), 32 (10 fetuses and lambs), 64 (2 fetuses and lambs), 128 (7 fetuses and lambs), 256 (9 fetuses and lambs), 512 (5 fetuses and lambs), 1,024 (15 fetuses and lambs), 2,048 (13 fetuses and lambs), 4,096 (13 fetuses and lambs), 8,196 (13 fetuses and lambs), 16,392 (19 fetuses and lambs), and greater than or equal to 32,784 (74 fetuses and lambs).

Economic and public health considerations of congenital toxoplasmosis in lambs.

Congenital toxoplasmosis was diagnosed in a flock of Hampshire sheep in South Dakota. The 80 ewes produced 144 lambs, 30 of which were born dead; toxoplasmosis was diagnosed in 11 of the dead lambs. The remaining 114 lambs grew normally, but 68 (40.3%) had agglutinating Toxoplasma gondii antibodies. Modified agglutination test T gondii antibody titers for 114 lambs were: less than 100 (n = 46), 64 (n = 2), 256 (n = 1), 1,024 (n = 12), and greater than or equal to 4,096 (n = 53). Tissues of 8 seropositive lambs were bioassayed for T gondii tissue cysts. Toxoplasma gondii was isolated from the tongue and lamb chops of 7, heart of 3, and legs of lamb of all 8 lambs.

Enzootic toxoplasmosis in sheep in north-central United States.

Of 1,564 serum samples from adult ewes from 33 farms in Iowa, Minnesota, South Dakota, Kansas, and Nebraska where toxoplasmosis-induced ovine abortions had been diagnosed, 65.5% were found positive for Toxoplasma gondii antibodies using the modified agglutination test. Toxoplasma gondii antibody titers of ewes were: less than 64 (34.5%), 64 (14.9%), 256 (22.0%), 1,024 (14.5%), and greater than 4,096 (13.8%). Thus, 28.3% of sheep had high titers (greater than 1,024) indicating recently acquired T. gondii infection. On certain farms, up to 95% of ewes were seropositive. Prevalence of T. gondii antibodies increased with age of the ewe. Of 665 ewes, 53.6% of 1-yr-old ewes were seropositive (titers greater than 64) versus 75% of 5-yr-old ewes. Results indicate that T. gondii infection in sheep in the United States is widespread.

Serologic examination of aborted ovine and bovine fetal fluids for the diagnosis of border disease, bluetongue, bovine viral diarrhea, and leptospiral infections.

Fetal serum from most of 994 bovine and 553 ovine aborted fetuses was tested serologically for antibodies to border disease (BD), bovine viral diarrhea (BVD), and bluetongue (BT) viruses, and to Leptospira sp., and the results were compared with the results of isolation procedures, fluorescent antibody tests (FAT), and histologic examinations of the same fetuses. Antibodies to BT virus were not found in any of the 994 bovine and 553 ovine fetuses. Antibody titers to BVD virus were present in 39 of 966 bovine fetuses tested, and BVD virus was detected in 4 of the 39. Four of 74 fetuses in which the BVD virus was detected by FAT or isolation had titers to BVD virus. Microagglutination (MAT) titers to 1 or more of 5 serovars of leptospires were present in 52 of 773 bovine fetal sera tested. Leptospires were not detected by FAT in any bovine fetuses that had leptospiral antibody titers. Leptospires were detected by FAT in 15 aborted calves, and none of these had MAT titers. Antibody titers to BD virus were present in 80 of 486 fetal lamb sera tested, and the virus was detected by FAT or isolation in 3 of the 80 fetuses. Border disease virus was detected in 14 of 486 fetal lambs tested. Twelve of the 14 were tested serologically and 3 had titers to BD virus. Leptospiral antibody titers were present in 27 of 326 ovine fetal sera tested. Leptospires were not detected in any of the 326 ovine fetuses tested by FAT.

Comparison of enzyme-linked immunosorbent assay, indirect fluorescent antibody test, and direct agglutination test for detecting Toxoplasma gondii antibodies in naturally aborted ovine fetuses.

Results obtained in an enzyme-linked immunosorbent assay (ELISA), an indirect fluorescent antibody test (IFA), and a modified direct agglutination test (MAT) for Toxoplasma gondii antibodies from examination of fetal fluids from 377 aborted ovine fetuses were compared. Sixty-seven samples were positive by MAT (titers 1:16 to greater than 1:65,536), 58 were positive by ELISA, and 62 were positive by immunoglobulin G-IFA. The MAT was preferred because it required less time, labor, and special equipment. It was simple to run, could be done on serum from any species without modification, and it was more effective than the IFA for detecting toxoplasma antibodies in severely autolyzed fetuses. No advantage was found in determining immunoglobulin M antibodies in ovine fetal sera.

Evaluation of multivalent Leptospira fluorescent antibody conjugates for general diagnostic use.

Four lots of conjugate were evaluated for optimal dilution and degree of fluorescence produced with reference cultures and bovine and porcine leptospira isolates. One lot that uniformly produced better fluorescence was evaluated for sensitivity and specificity with reference cultures, isolates, culture-positive tissues, and 13 other bacterial species. Further evaluation of the conjugates was done with bovine, porcine, and ovine specimens submitted to a diagnostic laboratory. Leptospires were detected with the fluorescent antibody test (FAT) in 9 of 21 culture-positive bovine kidneys and were detected in diluted cultures when present at concentrations of 10(2)-10(3) organisms/ml. With the exception of Treponema hyodysenteriae, FAT's of other bacterial cultures produced minimal fluorescence or were negative. Positives were characterized by moderate to brilliant fluorescence of typical cell forms, and most nonspecific fluorescence was eliminated with a flazo-orange counterstain. The results indicated that the FAT utilizing multivalent conjugates could be used successfully as an additional method for diagnosis of leptospira infections.

Experimental infection and abortion of pregnant guinea pigs with a unique spirillum-like bacterium isolated from aborted ovine fetuses.

Study was made of the pathogenicity of a spirillum-like, anaerobic, gram-negative bacterium, originally isolated from aborted lambs, for pregnant guinea pigs. Reproducible conditions for propagation and preservation of the bacterium were determined as requisite for the preparation of cultures for animal inoculation. A preliminary experiment was done with 10 pregnant guinea pigs to test for an infective dose of organisms that would produce abortion. High-passage cultures (n = 50) were used to inoculate these guinea pigs intraperitoneally. Six of 10 guinea pigs aborted, and the organism was cultured from fetal tissues of 5 guinea pigs. Isolates from 3 of the 6 guinea pigs were propagated through 4 passages on blood agar and used to infect 3 groups, each of 5 guinea pigs. A 4th group of 5 guinea pigs was inoculated with the original culture. Three of 5 animals in the first 3 groups, which had been given the low-passage cultures from the preliminary trial, and 2 of 5 guinea pigs in the 4th group, which had been given the original culture, aborted. Antibody against the spirillum was detected in 19 of 30 inoculated guinea pigs. The major microscopic lesions were acute suppurative placentitis and splenitis. This bacterium retained pathogenic properties sufficient to cause infection, abortion, and microscopic lesions in two-thirds of the guinea pigs, in spite of high in vitro passage. The organism has unique ultrastructures, and its genus and species are yet to be determined.

Porcine abortion caused by Bacillus sp.

Bacillus sp is known to cause abortion in cattle and sheep, but it is rarely reported to cause abortion in swine. Bacillus sp was isolated from aborted fetuses, and lesions compatible with a bacterial infection were detected in 4 of 962 aborted porcine fetuses examined. The infection apparently results in sporadic abortions and is not likely to cause economically significant reproductive failures within a herd.

Examination of bovine and ovine fetuses.

Diagnosis of the causes of bovine and ovine abortion is difficult and frustrating and requires a systematic and thorough approach. Laboratory assistance is required in all cases. Herd and individual histories occasionally help, as does knowledge of the gestational age and autolytic condition of the fetus when aborted. Tissues from mummified fetuses should be cultured and examined by fluorescent antibody techniques for viruses. The placental tissue from mummified fetuses should be examined for fungi and lesions. Lung, liver, kidney, spleen, abomasal content, body cavity fluid or serum, and fetal placenta from all but completely mummified fetuses should be submitted to a diagnostic laboratory.

Abortion in sheep caused by a nonclassified, anaerobic, flagellated bacterium.

Twenty-eight pregnant ewes were inoculated IV with approximately 6 X 10(8) nonclassified, anaerobic, flagellated bacteria (NAFB) that had been isolated from an aborted lamb. Abortion occurred in 3 of the ewes and 1 ewe gave birth to a weak lamb. The remaining 24 ewes and 3 other ewes inoculated orally with NAFB did not develop clinical signs of illness. Suppuration and vasculitis were seen in the placentas of the 3 aborted lambs, 1 of which had necropurulent hepatitis indistinguishable from that usually attributed to Campylobacter fetus infection. The NAFB was isolated from fetal placenta, abomasal content, or internal organs of 2 aborted lambs and the weak lamb. A morphologically similar organism was seen in the abomasal content of the other aborted lamb, but the organism did not grow on bacteriologic culture medium. Therefore, in susceptible pregnant ewes, NAFB can cause fetal placentitis and hepatitis and subsequent birth of weak lambs or abortion.