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.

Inoculation of female American black bears (Ursus americanus) with partially purified porcine zona pellucidae limits cub production.

The present 2-year study investigated the feasibility of using porcine zona pellucidae (pZP) as antigen for immunocontraception in American black bears. Sows, 3-6 years of age, were administered either two doses of 250 microg pZP with Freund's adjuvant (n = 10) or adjuvant alone (n = 5), one in April and one in May, and were kept away from the boars until June. Serum samples were collected before injections and before denning (November). The presence of sows with cubs at side was observed during premature emergence from denning. First-year results indicated that anti-pZP antibody titres in vaccinated sows were 2.5-9.0-fold (range) higher compared with non-vaccinated sows and that the vaccinated sows were threefold less likely to become pregnant (P = 0.167). Control and vaccinated bears produced 1.6 and 0.2 cubs per sow, respectively (P = 0.06). The second-year study investigated the feasibility of using pZP sequestered in a controlled-release pellet and a water-soluble adjuvant (QS-21) to avoid regulatory problems associated with Freund's adjuvant. Sows in the treatment group (n = 22) were administered a single dose of an emulsion of 250 microg pZP and 150 microg QS-21 plus a pellet containing 70-90 microg pZP for delayed release as booster dose. Control sows (n = 5) received the QS-21 adjuvant in pellet alone. Serum samples were collected before inoculations (April) and before denning (November). Seven cubs were born to the five control sows, but none was born to the 22 vaccinated sows (P < 0.001). Anti-pZP antibody mean absorbance ratios in control sows remained at background levels, whereas vaccinated sows had ratios fourfold higher than controls. Two-dimensional polyacrylamide gel electrophoresis and immunohistochemical localisation confirmed immunoreactivity of sera from inoculated bears. We conclude that cub production in the American black bear can be effectively limited with either two injections of 250 microg pZP or a single inoculation of partially purified pZP sequestered in controlled-release pellets.

Persistence of anti-zonae pellucidae antibodies following a single inoculation of porcine zonae pellucidae in the domestic equine.

In this study of equids, we investigated the antibody response and the effect on the estrous cycle following a single inoculation of porcine zonae pellucidae (pZP) employing controlled-release methodology. We also investigated the use of two different water-soluble adjuvants as an alternative to oil-based adjuvants. Twenty-seven domestic mares were inoculated with various formulations of pZP and adjuvant. We showed that the anti-pZP antibodies generated as a result of the inoculations persisted for at least 43 weeks (length of the study). Of the various formulations used in the study, pZP and QS-21 water-soluble adjuvant, administered in combination with an emulsified preparation of pZP and Freund's Complete Adjuvant generated a significantly (P < 0.05) higher titer of anti-pZP antibodies when compared with other formulations employing the water-soluble adjuvant, Carbopol. Hormone analyses for cyclicity indicated a high incidence and extended duration of persistent corpora lutea among the treated mares. The positive control group of mares receiving two standard inoculations of pZP and Freund's Complete and Incomplete Adjuvants, as well as the placebo group of mares injected with QS-21 only, also exhibited high incidences of persistent corpora lutea. However, all mares eventually returned to normal cyclicity. The basis for the high incidence and extended duration of persistent corpora lutea was unexplained. The results demonstrate for the first time the persistent generation of anti-pZP antibodies following a single inoculation of pZP incorporated into a controlled-released preparation in the horse. This study further suggests that a single inoculation of pZP sequestered in a controlled-release lactide-glycolide polymer may serve as an alternative to traditional two-inoculation protocols for contraception investigations in the equine.

Intravenous and subcutaneous pharmacokinetics of florfenicol in sheep.

Pharmacokinetic parameters of florfenicol were determined in 10 adult sheep (five wethers and five ewes) after a single 40 mg/kg intravenous (i.v.) dose, and three daily subcutaneous (s.c.) doses of 40 mg/kg of a commercial preparation (Nuflor((R))). The concentration of florfenicol in serum samples was assayed using a proprietary HPLC assay method, and pharmacokinetic parameters derived for individual animal data by each route using compartmental and noncompartmental approaches. Two animals (one male and one female) were excluded due to observed i.v. dosing problems, and a biexponential model was found to fit the i.v. data well for six of the other eight animals. Data from two males showed prolonged low concentrations of florfenicol in serum and were better fit by a three-compartment model. The mean +/- SD for the half-lives of the distribution and elimination phases for the six sheep best fit with a two-compartment model were 0.069 +/- 0.018 and 1.01 +/- 0.09 h respectively, and for the V(d(ss)) and clearances were 0.503 +/- 0.035 L/kg and 366 +/- 53 mL/h/kg respectively. The data collected during the s.c. multiple dose study were analyzed using noncompartmental methods only. The bioavailability (F%) after s.c. dosing was calculated in three ways to compare estimation methods as steady-state had not been reached and single dose s.c. data were not obtained past 24 h. Using the AUC(0--24) and AUC(0--> infinity ) from the first dose, the F% values averaged 27 and 40% respectively. Using the AUC(0--> infinity ) for all doses, the F% was 65%. Calculations of the mean time during which the serum concentration exceeded 0.5 and 1.0 microg/mL were 105 +/- 3.9 and 74.7 +/- 12.2 h respectively.

Serologic survey for infectious pathogens in free-ranging American bison.

From November 1991 through March 1992, we evaluated 101 free-ranging American bison (Bison bison) from Yellowstone National Park, Wyoming (USA) for exposure to infectious organisms that commonly infect cattle. No titers were detected for bluetongue virus, bovine leukemia virus, or Campylobacter fetus in these 101 bison. Detectable antibodies occurred against Anaplasma marginale (eight of 76, 11%), bovine respiratory syncytial virus (31 of 101, 31%), bovine viral diarrhea (31 of 101, 31%), bovine herpesvirus 1 (29 of 76, 38%), Leptospira interrogans icterohaemorrhagiae (four of 101, 4%), L interrogans hardjo (seven of 101, 7%), L interrogans autumnalis (one of 101, 1%), L interrogans bratislava (seven of 101, 7%), L interrogans australis (one of 101, 1%), and parainfluenza 3 virus (27 of 75, 36%). The low antibody titers and the lack of gross lesions are evidence that while previous exposure to infectious organisms may have occurred, none appeared to have active infections.

Hemochromatosis in Salers cattle.

Two 2-year-old Salers cattle from different herds raised on pasture were evaluated for retarded growth and diarrhea. Increase of liver enzyme activities and prolonged sulfobromophothalein (BSP) half life (T1/2) indicated liver disease with impaired liver function. Histopathologic examination of liver biopsies revealed a micronodular cirrhosis with marked deposition of hemosiderin in hepatocytes, Kupffer cells, and arterioles. Transferrin saturation (TS) and liver iron content were markedly increased, consistent with a diagnosis of hemochromatosis. Both animals were euthanatized due to deterioration in their condition. Necropsy findings included hepatomegaly and hemosiderin accumulation in the liver, lymph nodes, pancreas, spleen, thyroid, kidney, brain and other glandular tissue. Continued surveillance of the second herd (serum iron, total iron binding capacity [TIBC], unsaturated iron binding capacity [UIBC], and TS), identified a heifer as a hemochromatosis suspect in a subsequent generation. Liver biopsies from that animal revealed the same histopathologic changes as the previous 2 animals, and similar increases in liver iron content (8,700 ppm, normal range 45 to 300 ppm). The 3 affected cattle were all products of line breeding programs and shared a common ancestor. The absence of dietary iron loading in conjunction with the histopathologic and metabolic findings were consistent with a diagnosis of primary hemochromatosis. The reported disease is similar to idiopathic hemochromatosis in human beings in which there is a hereditary defect in iron metabolism.

Efficacy of buparvaquone as a therapeutic and clearing agent of Babesia equi of European origin in horses.

We evaluated the efficacy of buparvaquone in eliminating infection with Babesia equi of European origin in carrier horses and in splenectomized horses with experimentally induced acute infection. When administered at the rate of 5 mg/kg of body weight, IV, 4 times at 48-hour intervals, buparvaquone prompted rapid abatement of parasitemia. However, secondary and tertiary recrudescent parasitemias invariably returned with establishment of the carrier state. Buparvaquone, at the dosage evaluated, had transitory therapeutic efficacy against acute B equi infection in splenectomized horses, but was unable alone to clear carrier infection.

Pharmacokinetics, safety and tissue residues of sustained-release sulfamethazine in sheep.

Concentration-time profiles and the rates of absorption, extent of distribution and half-lives of sulfamethazine (SMZ), administered intravenously, orally as a water solution and as a sustained-release formulation (CalfSpan) were determined in 10 healthy sheep. The geometric mean half-life of elimination of i.v. SMZ was 10.8 h, compared to 14.3 h for the sustained-release preparation (CalfSpan) and 4.3 h for the oral water solution. Blood levels of SMZ were at or above 50 micrograms/ml for more than 48 h for CalfSpan, for 24 h after i.v. SMZ (100 mg/kg body wt), and for less than 24 h after p.o. SMZ (100 mg/kg body wt). The mean bioavailability of the oral SMZ solution was 58.3% (AUCp.o./AUCi.v.). The estimated bioavailability of the CalfSpan preparation was 52.5%. The safety of the sustained-release preparation was tested by dosing sheep with multiples (one, three and five times) of the recommended dose (one tablet, 8 g SMZ, per 20 kg body wt), once a day for 3 days. Clinical blood chemistries showed a significant increase in serum iron, and a decrease in serum phosphorus in animals treated at the 3x and 5x dose levels. Necropsies of the 5x dose animals did not show any gross signs that could be attributed to SMZ, and histological examination of tissues from the 5x animals revealed no organ pathology. Residues of SMZ in liver, fat, kidney and skeletal muscle were measured in 20 animals that received one bolus per 20 kg body wt. The results indicate that SMZ residues are cleared rapidly, and are at or below the tolerance level of 0.1 mg/kg within 8 days after dosing so that the 18-day withdrawal time used in cattle would provide an appropriate margin of safety if used in sheep.

Serum ionized calcium concentration in clinically normal dairy cattle, and changes associated with calcium abnormalities.

Serum ionized calcium (ICa) concentration was determined in 141 clinically normal dairy cattle by use of a direct-measuring calcium ion-selective electrode instrument. Mean serum ICa concentration 2 hours after blood withdrawal was 4.59 mg/dl; range varied from 3.79 to 5.25 mg/dl. Regression analysis indicated a high degree of correlation between ICa and serum total calcium concentrations if serum stored at 23 C was analyzed within 12 hours after blood withdrawal. Abnormal ICa concentration was detected in 19 of 85 dairy cows that were affected with various pathologic conditions. All 19 cows had hypocalcemia (n = 13 with parturient hypocalcemia, 4 with hypomagnesemic tetany, and 2 with renal disease). In all cases, the ICa concentration clearly related to the clinical manifestation of disease and the functional status of the cow's calcium metabolism.

Evaluations of buparvaquone as a treatment for equine babesiosis (Babesia equi).

We evaluated the efficacy of buparvaquone in eliminating Babesia equi of European origin in carrier horses and in experimentally infected splenectomized ponies. When administered at the rate of 2.5 mg/kg of body weight, IM, 4 times at 96-hour intervals, buparvaquone was effective in eliminating B equi carrier infection in 1 horse. Such results could not be repeated at the same dosage or at 3.5 or 5 mg/kg, IM. Buparvaquone given at the rate of 4 to 6 mg/kg IV and/or IM was therapeutically effective in 4 of 5 acute B equi infections in splenectomized ponies. The treated ponies became carriers.

Detection of rotavirus and coronavirus shedding in two beef cow herds in Idaho.

Fecal samples were taken at the time of pregnancy examinations and at parturition from two beef herds. They were also taken from sick calves at the onset of disease, and from 25% of the healthy calves at 15 days of age. All fecal samples were examined by electron microscopy for viruses.Four cows in herd A were detected excreting coronavirus, one at the time of the pregnancy examinations and three at parturition. The first cow was removed from the herd and the others calved at the end of the season. There were no sick calves.No cows in herd B were detected excreting virus at the time of pregnancy checks, but fourteen coronavirus and two rotavirus carrier cows were found at parturition. All but two calves sampled had large numbers of virus particles in their feces. Clinical illness was associated with dams shedding virus and with nightly low temperatures.

Ovariectomy as treatment for granulosa cell tumor in a heifer.

A granulosa cell tumor in a 15-month-old heifer was associated with abnormal udder development, relaxation of the pelvic ligaments, and nymphomanic behavior. After surgical removal of the tumor, the anatomic changes were reversed, and the nymphomanic behavior subsided. The heifer conceived 3 times thereafter and became a productive dairy cow. In previously reported cases of granulosa cell tumor, ovariectomy was unsuccessful in returning the cow to normal breeding.

Genetic-associated resistance to foot rot in selected Targhee sheep.

Three Targhee rams obtained from the Ohio Agricultural and Experimental Station had been identified as foot rot resistant on the basis of results of challenge exposure. In the first breeding trial, when rams were bred to 20 foot rot-susceptible ewes, the percentages of foot rot-resistant offspring from the 3 foot rot-resistant rams were 68, 82, and 100, compared with 55 and 60 for foot rot-resistant offspring from 2 known foot rot-susceptible rams. In the second year, the foot rot-resistant rams were mated with ewes of unknown foot rot status. The foot rot-resistant status of their lambs was compared with that of range-raised lambs whose parents' foot rot status was unknown. During the first year, challenge exposure to the disease consisted of confinement of the lambs in moist or wet pens with sheep affected with the naturally acquired disease. This protocol was repeated for lambs born during the second-year breeding trial. In addition, the right front foot of each lamb was inoculated with a broth culture of Bacteroides nodosus. During the second year, when data that included infected feet from all lambs were analyzed, 41% of the progeny of the foot rot-resistant rams and 17% of the offspring of parents of unknown foot rot status were unaffected by the disease. When the B nodosus-inoculated foot was not included in the analysis, however, 61% of the progeny of the foot rot-resistant rams and 29% of the others were unaffected. The resistance to foot rot undoubtedly is hereditary. The mechanism of resistance may be in the interdigital skin.(ABSTRACT TRUNCATED AT 250 WORDS)

Comparison of treatment methods for the control of contagious ovine foot rot.

Four methods of treatment for ovine foot rot were evaluated: foot trimming and topical treatment (brief, daily foot bathing in 10% zinc sulfate); vaccination with an oil-alum adjuvanted, multivalent, Bacteroides nodosus bacterin, vaccination with the bacterin plus foot trimming and brief foot bathing, and 1-hour 10% zinc sulfate foot soaks. Compared with results of sheep in a nontreated control group, vaccination reduced the number of infected feet by 8%. Daily walk-through foot bathing reduced the number of infected feet by 16%, and vaccination and foot bathing reduced the number by 27%. In another experiment, treated but nonresponsive sheep were subjected to 2 one-hour foot soaks in 10% zinc sulfate. Compared with results in nontreated controls, the soaks reduced the number of affected sheep by 58%. Previously nontreated sheep responded with a 69% reduction of affected animals over controls.

Death associated with parenteral administration of copper disodium edetate in calves.

Copper disodium edetate in recommended doses was apparently responsible for the deaths of one calf and clinical signs of toxicosis in 5 others on one farm, and 7 deaths and clinical signs of toxicosis in a number of others on another ranch. Signs of hyperexcitability, hypermetria, hindlimb weakness, head pressing, depression, and opisthotonos occurred 6 to 24 hours after injections and preceded death by 1 to 2 days. Necropsy and histologic examination revealed massive liver necrosis. High blood concentrations of liver enzymes in affected cattle that did not die indicated that they had liver damage. High blood concentration of iron in cattle that died indicated possible interaction of copper and iron.

Changes in urea nitrogen and creatinine concentrations in the vitreous humor of cattle after death.

The mean urea nitrogen concentration in vitreous humor (VUN) in 97 healthy steers after death was 15.7 mg/dl. The mean serum urea nitrogen concentration in the same cattle was 20.0 mg/dl. The mean vitreous creatinine (VC) concentration was 0.7 mg/dl, and the serum creatinine value was 1.5 mg/dl. The VUN and VC were both significantly lower than serum urea nitrogen and serum creatinine, respectively, but varied in a consistent, predictable manner. In tests with 10 cows, postmortem intervals as long as 36 hours and ambient temperatures up to 30 C had no significant effects on VUN and VC. Evaluation of 8 animals with uremia confirmed that the vitreous humor is a stable fluid compartment after death, usable as an indicator of serum urea nitrogen and creatinine.

Postmortem magnesium concentration in bovine vitreous humor: comparison with antemortem serum magnesium concentration.

The range for postmortem vitreous humor Mg2+ concentration in 97 healthy cattle was 1.8 mg/dl to 2.72 mg/dl at 23 C for a 48-hour postmortem interval. The postmortem vitreous Mg2+ concentration closely paralleled the antemortem serum Mg2+ concentration at 23 C. Low environmental temperature (4 C) had no effect on postmortem vitreous concentration. However, high environmental temperatures (30 C) significantly (P less than 0.05) reduced postmortem vitreous Mg2+ concentration at the 36-hour postmortem interval. It was concluded that postmortem vitreous humor Mg2+ determination could be a useful diagnostic aid in cattle for detecting Mg2+ imbalances for at least 48 hours after death, provided the postmortem environmental temperature did not exceed 23 C after 24 hours.