Comparative histopathology and antibody responses of non-Tritrichomonas foetus trichomonad and Tritrichomonas foetus genital infections in virgin heifers.

The potential pathogenicity of non-Tritrichomonas foetus trichomonads (NTfTs) recently isolated from the prepuce of virgin bulls is not known. The purpose of this study was to determine the ability of these NTfTs to cause disease in the female reproductive tract relative to T. foetus. Forty-four virgin heifers were experimentally infected intravaginally with either one of two NTfTs (Pentatrichomonas hominis or Tetratrichomonas spp.), T. foetus, or sterile media and cultured weekly from 0 time until slaughter at 8 weeks. Serum and vaginal antibody responses during infection were assessed, and the reproductive tracts were histologically examined, scored, and compared based on numbers of neutrophils, eosinophils, lymphocytes, and plasma cells as well as the qualitative appearance of the reproductive tract. The NTfTs did not persist in the reproductive tract, while T. foetus persisted for at least 6-8 weeks. Further, no vaginal IgA response to infection was found in NTfT-infected and control heifers, but a vaginal IgA response was present in the T. foetus-infected group. Heifers infected with NTfT or controls showed little mucosal inflammatory response compared to T. foetus-infected heifers. Among the trichomonads studied, persistent infection by T. foetus alone seems responsible for uterine inflammatory lesions usually associated with pregnancy loss. The NTfTs studied in this work only transiently infected the vagina and were associated with strictly mild inflammatory changes, which probably do not cause significant disease, i.e., pregnancy loss.

Selected diseases and conditions associated with bovine conceptus loss in the first trimester.

The outcomes of insults to the bovine conceptus depend on the predilection of the insulting agent for the gravid reproductive tract, the virulence of the insult, and the developmental maturity/immune competence of the conceptus at the time of the insult. Agents that are lethal at one time during gestation may be harmless at another, or may have completely different effects (some not so harmless) at different gestational ages. This review discusses some of the known physical-mechanical, physiological, and infectious causes of first trimester bovine conceptus losses, including three infectious agents that have been the subject of recent studies for their potential to transmit disease via embryo transfer.

Sensitivity and specificity of culture and PCR of smegma samples of bulls experimentally infected with Tritrichomonas foetus.

The sensitivity (Se) and specificity (Sp) of different testing schemes were estimated for detecting Tritrichomonas foetus (T. foetus) in smegma samples from experimentally infected bulls. Culture and polymerase chain reaction (PCR) on smegma samples were evaluated alone and in parallel testing. Mature dairy bulls (n=79) were intrapreputially inoculated with T. foetus (n=19); Campylobacter (C.) fetus venerealis (n=13); both T. foetus and C. fetus venerealis (n=11); Tetratrichomonas spp. (n=9); C. fetus fetus (n=8); or were not inoculated (n=19). For each bull, smegma samples were collected for 6 week post-inoculation and tested for T. foetus by In Pouch TF culture and PCR. Most T. foetus-inoculated bulls became infected, according to culture (86.7%), PCR (90.0%), and both tests together (93.3%). In T. foetus-inoculated bulls, both tests combined in parallel on a single sample had a Se (78.3%) and Sp (98.5%) similar to two cultures (Se 76.0%, Sp 98.5%) or two PCR (Se 78.0%, Sp 96.7%) sampled on consecutive weeks. The PCR on three consecutive weekly samples (Se 85.0%, Sp 95.4%) and both tests applied in parallel on three consecutive weekly samples (Se 87.5%, Sp 95.6%) were similar to the current gold-standard of six weekly cultures (Se 86.7% and Sp 97.5%). Both tests used in parallel six times had the highest Se (93.3%), with similar Sp (92.5%). Tetratrichomonas spp. were only sporadically detected by culture or PCR. In conclusion, we have proposed alternative strategies for T. foetus diagnostics (for the AI industry), including a combination of tests and repeat testing strategies that may reduce time and cost for bull surveillance.

Importation of in vitro-produced Bubalus bubalis embryos from Italy into the United States: a case report.

On December 19, 2005, 14 in vitro-fertilized water buffalo (Bubalus bubalis) embryos, which had been cryopreserved by vitrification, were thawed and transferred into B. bubalis recipients in California. The embryos had been produced in Italy, following transvaginal oocyte pickup (TVOPU), with subsequent in vitro maturation, insemination, and culture. This case study relates our experience in meeting the regulatory criteria, established by the Animal Import/Export Office of the USDA-Animal and Plant Health Inspection Service (USDA-APHIS), in order to successfully import these embryos into the USA.

Effect of corn residue harvest method with ruminally undegradable protein supplementation on performance of growing calves and fiber digestibility.

Two experiments evaluated the effects of corn residue harvest method on animal performance and diet digestibility. Experiment 1 was designed as a 2 × 2 + 1 factorial arrangement of treatments using 60 individually fed crossbred steers (280 kg [SD 32] initial BW; = 12). Factors were the corn residue harvest method (high-stem and conventional) and supplemental RUP at 2 concentrations (0 and 3.3% diet DM). A third harvest method (low-stem) was also evaluated, but only in diets containing supplemental RUP at 3.3% diet DM because of limitations in the amount of available low-stem residue. Therefore, the 3 harvest methods were compared only in diets containing supplemental RUP. In Exp. 2, 9 crossbred wethers were blocked by BW (42.4 kg [SD 7] initial BW) and randomly assigned to diets containing corn residue harvested 1 of 3 ways (low-stem, high-stem, and conventional). In Exp. 1, steers fed the low-stem residue diet had greater ADG compared with the steers fed conventionally harvested corn residue ( = 0.03; 0.78 vs. 0.63 kg), whereas steers fed high-stem residue were intermediate ( > 0.17; 0.69 kg), not differing from either conventional or low-stem residues. Results from in vitro OM digestibility suggest that low-stem residue had the greatest ( < 0.01) amount of digestible OM compared with the other 2 residue harvest methods, which did not differ ( = 0.32; 55.0, 47.8, and 47.1% for low-stem, high-stem, and conventional residues, respectively). There were no differences in RUP content (40% of CP) and RUP digestibility (60%) among the 3 residues ( ≥ 0.35). No interactions were observed between harvest method and the addition of RUP ( ≥ 0.12). The addition of RUP tended to result in improved ADG (0.66 ± 0.07 vs. 0.58 ± 0.07 for supplemental RUP and no RUP, respectively; = 0.08) and G:F (0.116 ± 0.006 vs. 0.095 ± 0.020 for supplemental RUP and no RUP, respectively; = 0.02) compared with similar diets without the additional RUP. In Exp. 2, low-stem residue had greater DM and OM digestibility and DE ( < 0.01) than high-stem and conventional residues, which did not differ ( ≥ 0.63). Low-stem residue also had the greatest NDF digestibility (NDFD; < 0.01), whereas high-stem residue had greater NDFD than conventional residue ( < 0.01). Digestible energy was greatest for low-stem residue ( < 0.05) and did not differ between high-stem and conventional residues ( = 0.50). Reducing the proportion of stem in the bale through changes in the harvest method increased the nutritive quality of corn residue.

Production and processing of milk from transgenic goats expressing human lysozyme in the mammary gland.

The potential for applying biotechnology to benefit animal agriculture and food production has long been speculated. The addition of human milk components with intrinsic antimicrobial activity and positive charge to livestock milk by genetic engineering has the potential to benefit animal health, as well as food safety and production. We generated one line of transgenic goats as a model for the dairy cow designed to express human lysozyme in the mammary gland. Here we report the characterization of the milk from 5 transgenic females of this line expressing human lysozyme in their milk at 270 microg/mL or 68% of the level found in human milk. Milk from transgenic animals had a lower somatic cell count, but the overall component composition of the milk and milk production were not different from controls. Milk from transgenic animals had a shorter rennet clotting time and increased curd strength. Milk of such nature may be of benefit to the producer by influencing udder health and milk processing.

An improved molecular assay for Tritrichomonas foetus.

Tritrichomonas foetus (T. foetus) is the causative agent of bovine trichomonosis, a sexually transmitted disease leading to abortion (from 1 to 8 months gestation), infertility, and occasional pyometra. The annual losses to the U.S. beef industry are estimated to be in the hundreds of millions of dollars. Currently, the "gold standard" diagnostic test for trichomonosis in most countries is the cultivation of live organisms from reproductive secretions. The cultured organisms can then be followed by PCR assays with primers that amplify T. foetus to the exclusion of all other trichomonad species. Thus, negative results present as null data, indistinguishable from failed PCR amplification during T. foetus specific amplification. Our newly developed assay improves previously developed PCR based techniques by using diagnostic size variants from within the internal transcribed spacer 1 (ITS1) region that is between the 18S rRNA and 5.8S rRNA subunits. This new PCR assay amplifies trichomonad DNA from a variety of genera and positively identifies the causative agent in the bovine trichomonad infection. This approach eliminates false negatives found in some current assays as well as identifying the causative agent of trichomonad infection. Additionally, our assay incorporates a fluorescently labeled primer enabling high sensitivity and rapid assessment of the specific trichomonad species. Moreover, electrophoretic separation of amplified samples can be outsourced, thus eliminating the need for diagnostic laboratories to purchase expensive analysis equipment.

Temporal patterns of delta 4 C-21 steroids in coexisting, genetically dissimilar twin lamb fetuses throughout late gestation.

We have examined the relationships among fetal maturity, adrenal steroidogenesis, and initiation of parturition using embryo transfer procedures. Ovine embryos, genetically coded for relatively short [145.5 +/- 1.4 days; Finnish Landrace (Finn)] and long [150.7 +/- 1.3 days; Rambouillet (Ra)] gestational periods, were transferred to a common recipient, thereby facilitating comparative endocrine studies in genetically dissimilar fetuses with a common intrauterine environment. Maternal and fetal plasma samples were obtained daily from chronic fetal lamb preparations in six mixed breed (MB), three Finn, and three Ra single breed (SB) pregnancies during the last trimester and assayed for cortisol, desoxycortisol, and progesterone by RIA. Mean +/- SEM gestational lengths of the forenamed pregnancies were 141.5 +/- 0.7, 141.7 +/- 0.7, and 149.3 +/- 0.9 days, respectively. Time-trend analysis of C-21 steroid data showed an increase in Finn and Ra SB fetal plasma cortisol of 1.0 ng/ml X day at 9.03 +/- 1.0 and 9.58 +/- 1.10 days prepartum, respectively. Significant (P less than or equal to 0.05) differences in the number of days of incremental change (T*) were noted between Finn (10.56 +/- 0.67) and Ra (4.82 +/- 1.08) siblings in MB pregnancies at increment rates of 1.0 ng/ml X day. Similarly, significant (P less than or equal to 0.05) differences in T* values were observed between Finn (8.31 +/- 0.38 days) and RA (4.63 +/- 1.0 days) siblings at 2.0 ng/ml X day. While Finn and Ra SB fetal desoxycortisol T* values were similar at 0.5 and 0.1 ng/ml X day, significant differences (P less than or equal to 0.05) were noted in Finn and Ra cofetuses in the MB pregnancy. Prepartal changes in maternal and fetal progesterone concentrations were correlated, and a direct parallelism was noted between fetal sibling progesterone concentrations. These results are consistent with the view that the fetal genome has a principal role in the timing of parturition.

Tolerance of sheep-goat chimeras to their component cells.

Four sheep-goat chimeras with a goat or sheep sibling having an identical genotype to one of the two component species of cells were tested for tolerance through mixed lymphocyte response (MLR) and skin grafts. None of the four chimeras showed a response to its sib in MLR and three of the four accepted sib skin grafts. This demonstrates that the chimerism exhibited by these animals was sufficient to render the chimera tolerant to antigens expressed by the sib. Two of the four sibs showed positive responses to their chimeric sibs in MLR and two did not; one negative response was expected because the chimera's lymphocytes were essentially all the same species as the sib's. Chimeric skin grafts were partially accepted by two of the four sibs, suggesting the presence of both sheep and goat cells in the skin grafts derived from the chimeras. Two of the four sibs did not accept chimeric skin grafts, possibly due to lack of compatible cells in the graft. Neither differences between an allo- and xenoresponse in MLR nor time differences in the rejection of allo- and xenografts were observed.

Detection of histocompatibility-Y antigen: identification of sex of pre-implantation ovine embryos.

Indirect immunofluorescence was used to detect the presence of a male-specific protein (H-Y antigen) on pre-implantation ovine embryos. Eight-cell to blastocyst stage embryos were surgically collected from superovulated ewes, classified as fluorescent (H-Y positive) or non-fluorescent (H-Y negative) and either transferred to recipient ewes or karyotyped to confirm embryonic sex. H-Y antigen was detected on eight-cell through blastocyst stage embryos. Overall, 88% (50/57) of the embryos (eight-cell to early blastocyst stage) classified as H-Y positive or H-Y negative were male and female, respectively. Survival after transfer of embryos subjected to the H-Y antigen assay was high (63%), which supports the use of this procedure in conjunction with embryo transfer in sheep to produce pregnancies in which sex of the fetus is known.

Rapid enzyme immunoassay for measurement of bovine progesterone.

Reproductive management is a primary financial concern of the dairy industry with missed estrus detection one of the major causes of lost income. A rapid enzyme immunoassay (EIA) was developed for on-line measurement of progesterone in bovine milk with a biosensor for detection of estrus. The EIA was developed using covalent binding microtiter wells, monoclonal antibody, horseradish peroxidase, and 3,3',5,5'-tetramethylbenzidine (TMB). The EIA took 8 min and had a dynamic response for progesterone in buffer and milk between 0.2 and 20 ng/ml.

Sexual behavior of male dairy goats: effects of deafferentation of the genitalia.

The following study examines the effects of deafferentation of the penis, scrotum, prepuce and adjacent cutaneous surfaces on the development of mounting behavior in male dairy goats. Ten males underwent surgical deafferentations and 10 males received sham operations at 8 to 12 weeks of age. Mounting and other sexual behaviors were monitored while subjects were cohabiting with male and female penmates and when individually exposed to estrous females. Denervated males mounted penmates less frequently than sham controls and lacked vigorous thrusting behavior. When exposed to estrous females denervated goats did not exhibit the normal progression from shallow to deep thrusting with successive mounts and failed to attain intromissions and ejaculations. However, frequency of mounting was similar to that of controls when the latter were prevented from attaining intromissions. It was concluded that sensory feedback from the genitalia and surrounding cutaneous areas is not required for the initiation of leg-kicking behavior in male dairy goats but plays an important role in the development and maintenance of normal patterns of thrusting behavior. Because it was not possible to determine the rate at which reinnervation occurred, the extent to which mounting behavior depends on sensory feedback could not be determined.

Effects of endotoxin infusion on circulating levels of eicosanoids, progesterone, cortisol, glucose and lactic acid, and abortion in pregnant cows.

The effects of Escherichia coli endotoxin infusions (1.0 or 2.5 micrograms kg-1 over 6 h) on pregnancy were investigated in cows in the first, second and third trimester of gestation. Endotoxin increased the plasma levels of prostaglandins (PGs), thromboxane B2 and cortisol, and decreased progesterone. The severity of the clinical signs and the magnitude of the increases in plasma PGs, thromboxane B2 and cortisol tended to depend on the dose of endotoxin, but were independent of the gestation period. There was hyperglycemia followed by hypoglycemia and lactic acidemia. Hyperglycemia and lactic acidemia were significant only at the high dose of endotoxin. Endotoxin infusion at both doses caused a preferential mobilization of oleic acid from adipose tissue, and also had some effects on the mobilization of palmitic and stearic acids during the post-infusion period. The cows in the first trimester of gestation were more sensitive to the abortifacient effect of endotoxin than cows in the second and third trimester of gestation. The results of this study indicate that the mechanism of endotoxin-induced abortion in cows initially involves a prolonged release of PGF2 alpha and its subsequent stimulant effect on uterine smooth muscle contraction and luteolytic effect leading to a gradual decline in the plasma levels of progesterone. It was concluded that pregnancy terminates in the absence of an adequate level of progesterone, especially during the first trimester of gestation, when progesterone of extraluteal origin is not yet available, coupled with the PGF2 alpha-induced propulsive contraction of the uterus. In addition, the metabolic and circulatory failures in severe cases of endotoxemia, especially at the high dose of endotoxin, resulting either directly or indirectly via the release of various autacoids, catecholamines and cortisol, may also contribute to the termination of pregnancy at any stage of gestation.

Degradation of bovine complement C3 by trichomonad extracellular proteinase.

Bovine trichomoniasis is a local infection of the reproductive tract making interaction with mucosal host defenses crucial. Since the parasite is susceptible to killing by bovine complement, we investigated the role of the third component of complement (C3) in host parasite interactions. Bovine C3 was purified by anionic and cationic exchange chromatography. The purified protein was characterized by immunoreactivity, sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and peptide sequencing of the amino terminus of the beta chain. When purified bovine C3 was incubated for varying time periods with trichomonad extracellular proteinases, SDS-PAGE gels revealed digestion of the alpha chain to small fragments. Such degradation in vivo would prevent formation of C3b and completion of the complement cascade, resulting in evasion of killing. To evaluate the relevance of this data, we determined whether C3 was present in bovine genital secretions. With a quantitative ELISA assay, C3 could be demonstrated in both uterine and vaginal washes. To our knowledge, this is the first demonstration of bovine C3 in genital secretions. The C3 concentration increased significantly in vaginal secretions by 8 and 10 weeks in heifers infected with Tritrichomonas foetus. An increase was also seen in uterine secretions of infected heifers, but sample numbers were insufficient for statistical analysis. Transcription of the major extracellular cysteine proteinase (TFCP8) was demonstrated in T. foetus cells from uterine secretions of infected heifers by RT-PCR and Southern blotting. The results indicate that C3 may be important in genital defense and that trichomonad extracellular proteinases may play a role in evasion of complement-mediated killing.

Reproductive cycles and pregnancy in interspecific sheep<==>goat chimaeras.

The objectives of the current study were to determine whether interspecific sheep<==>goat chimaeras exhibited reproductive cycles of their component species and were capable of maintaining ovine and caprine pregnancies to term. All chimaeras had oestrous cycles and several exhibited characteristics of both ewes and does, including short, 6-7-day cycles. Sixteen caprine pregnancies were confirmed in eight sheep<==>goat and one hybrid<==>sheep chimaera from 21 embryo transfers; of these, six appeared normal by ultrasonographic examination during Weeks 5 or 6, but none progressed beyond Week 8. Three apparent pseudopregnancies developed in two animals. In contrast, eight of 11 pregnancies in chimaeras resulted in term ovine offspring after transfer of ovine embryos or natural matings with rams. By comparison, interspecific (caprine or hybrid) pregnancies in ewes were lost in Week 4 (n = 8) or Weeks 5-6 (n = 2). First interspecific (ovine or hybrid) pregnancies in does were maintained longer (Weeks 6-12, n = 7) than second interspecific pregnancies (Weeks 4-5, n = 5) (P < 0.05) or interspecific pregnancies in ewes (P < 0.05). The results suggest that abnormal fetomaternal interactions during the early stages of implantation are responsible for termination of caprine pregnancies in the ovine or chimaeric uterus, whereas ovine conceptuses are able to implant successfully in the chimaeric uterus. All chimaeras were fertile, since each carried at least one ovine pregnancy to term following natural matings with rams.

Experimental infection of pregnant cattle with bluetongue virus serotype 11 between postbreeding days 21 and 48.

Four bluetongue virus (BTV)-seronegative heifers and 2 BTV-seropositive heifers were inoculated with the virulent strain UC-8 of BTV-11 between postbreeding days (PBD) 21 and 30. The heifers were observed for 10-18 days after inoculation for clinical signs, and pregnancy was monitored by ultrasound examination of the uterus and by plasma progesterone levels. Blood samples were collected daily after inoculation and processed for virus isolation and titration. Heifers were euthanized between PBD 31 and PBD 48, and tissues were collected for virologic and pathologic examination. All but 1 heifer inoculated on PBD 21 remained pregnant after BTV inoculation. A cystic corpus luteum was found in the ovary of the nonpregnant heifer, but BTV was not isolated from the reproductive tract of this heifer. Three of the inoculated heifers that remained pregnant showed mild multifocal areas of perivascular lymphocytic infiltration in the ovary. BTV was reisolated from spleen and prescapular and peribronchial lymph nodes 10 days after inoculation from 3 of the 4 BTV-seronegative heifers. BTV was also reisolated from the uterus of 1 of the heifers that remained pregnant, but microscopic lesions were not found in this organ.

Comparison of the 5.8S rRNA gene and internal transcribed spacer regions of trichomonadid protozoa recovered from the bovine preputial cavity.

Sequence analysis of the 5.8S rRNA gene and the internal transcribed spacer regions (ITSRs) was used to compare trichomonadid protozoa (n = 39) of varying morphologies isolated from the bovine preputial cavity. A multiple sequence alignment was performed with bovine isolate sequences and other trichomonadid protozoa sequences available in GenBank. As a group, Tritrichomonasfoetus isolates (n = 7) had nearly complete homology. A similarity matrix showed low homology between the T. foetus isolates and other trichomonads recovered from cattle (<70%). Two clusters of trichomonads other than T. foetus were identified. Eighteen isolates comprised 1 group. These isolates shared >99% homology among themselves and with Pentatrichomonas hominis. The other non-T. foetus cluster (n = 14) did not exhibit a high degree of homology (<87%) with other bovine isolates or any of the trichomonad sequences available in GenBank. The sequence homology among isolates in that cluster was >99%, except for 1 isolate that varied from the others in both ITSRs (approximately 2% dissimilarity). Sequence analysis of the 5.8S rRNA gene and ITSRs was useful for comparing trichomonadid protozoa isolated from the bovine preputial cavity and demonstrated that 2 distinct types of trichomonads constituted the non-T. foetus isolates recovered from the bovine preputial cavity.

Experimental reproduction of bovine fetal Neospora infection and death with a bovine Neospora isolate.

Studies were conducted to determine the pathogenic potential of the recently isolated bovine Neospora protozoa (BPA-1) for the bovine fetus. Cows chosen for study had Neospora titers < 160 using an indirect immunofluorescent antibody (IFA) test. Four experimental groups were studied. In group 1, 2 fetuses were inoculated in utero at 118 days gestation with culture-derived Neospora tachyzoites. A pregnant control cow was housed in the same pen, observed daily and screened serologically for evidence of exposure to Neospora. In group 2, 2 cows were infected with Neospora tachyzoites at 138 or 161 days gestation, and 1 control cow was given uninfected cell culture suspension simultaneously at 154 days gestation. Groups 3 (85 days gestation) and 4 (120 days gestation) each consisted of 2 cows infected with Neospora tachyzoites and 1 control cow given uninfected material at the same stage of gestation. Dead fetuses were surgically removed from the infected cows in group 1 on postinfection day (PID) 17. The histopathology was compatible with protozoal fetal infection, and protozoa were identified by immunohistochemistry. Viable fetuses were removed surgically from cows in group 2 on PID 28-30. The histopathology was compatible with protozoal fetal infection, protozoa were identified by immunoperoxidase techniques, and Neospora tachyzoites were reisolated in vitro from tissues of the 2 infected fetuses. In groups 3 and 4, the control fetus and 1 infected fetus were removed surgically between PID 26 and PID 33. The remaining infected cows were observed until fetal death or abortion occurred.(ABSTRACT TRUNCATED AT 250 WORDS)

Detection of serum antibody responses in cattle with natural or experimental Neospora infections.

Parasite-specific antibody responses were detected using an indirect fluorescent antibody (IFA) test in cattle that were naturally or experimentally infected with Neospora parasites. The test was developed using Neospora tachyzoites isolated from an aborted bovine fetus and grown in bovine cell cultures (isolate BPA1). In all cases, infections were confirmed by the identification of Neospora tachyzoites and/or bradyzoite cysts in fetal or calf tissues using an immunoperoxidase test procedure. Fifty-five naturally infected cows that aborted Neospora-infected fetuses had titers of 320-5,120 at the time of abortion. The titer of 6 cows that were serologically monitored over a prolonged period decreased to 160-640 within 150 days after they aborted infected fetuses. Two of the cows showed an increase in their Neospora titers during their subsequent pregnancy, and they gave birth to congenitally infected calves that had precolostral titers of 10,240-20,480. Postcolostral titers of these calves and of 4 other calves with congenital Neospora infections were all > or = 5,120, whereas calves with no detectable parasites had titers < or = 160. Two pregnant heifers that were experimentally infected with the BPA1 isolate at approximately 120 days gestation seroconverted to Neospora antigens within 9 days and developed peak titers of 5,120 and 20,480 within 32 days of infection. The fetus taken by caesarean section 32 days postinfection from 1 heifer and the full-term calf born to the other had Neospora titers of 640 and 10,240, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)

Development and preliminary assessment of a polyclonal antibody-based enzyme immunoassay for the detection of Tritrichomonas foetus antigen in breeding cattle.

More sensitive tests are required for the diagnosis of Tritrichomonas foetus infection in cattle and an antigen-detecting enzyme immunoassay (EIA) has been applied to this purpose. An affinity purified immunoglobulin fraction obtained from rabbits immunised with cultured T. foetus served as both capture antibody and as biotinylated indicator antibody. While highly sensitive in the detection of antigen derived from cultured organisms, the assay showed poor sensitivity in the detection of antigen in the cervico-vaginal mucus of artificially infected heifers, with only 75% of culture-positive samples being considered positive for antigen. In a direct comparison, 23/122 samples from a naturally infected dairy herd gave positive cultures, while only 10/122 samples were considered antigen positive by EIA.