Babesia equi erythrocytic stage continuously cultured in an enriched medium.

Babesia equi was continuously cultured through 90 passages in an enriched chemically defined basal medium (HL-1) supplemented with 20% fetal bovine serum and serum replacement factors, including lipid-rich bovine serum albumin, bovine insulin, and human transferrin. Cryopreservation and subsequent recovery of B. equi were easily achieved. Inoculation of a splenectomized and an intact horse with cultured infected erythrocytes resulted in parasitemias and B. equi in vitro reisolation from both animals. In vitro forms of the parasite resembled in vivo forms. After establishment, parasitemias of 10-15% were commonly observed.

Evaluation of agar gel immunodiffusion and indirect fluorescent antibody assays as supplemental tests for dourine in equids.

The agar gel immunodiffusion (AGID) and indirect fluorescent antibody (IFA) assays were evaluated as supplemental tests to the complement-fixation (CF) test, the official US importation certification test for dourine in equids. The American stabilate (n = 10 animals) or the Canadian stabilate (n = 6 animals) of Trypanosoma equiperdum cultured in rat blood was administered by catheterization and infusion in the urogenital tract of 16 equids. To assess parasitemia and serologic responses by use of the CF, AGID, and IFA tests, a total of 787 serum and blood samples were obtained from equids before exposure and 3 times a week after exposure to T equiperdum. Results of the IFA and AGID tests were compared with the CF test results. The disease was diagnosed earlier by the IFA test than by the AGID test, regardless of antigen preparation or exposure group. The mean number of days between exposure and positive result by the CF and IFA tests was the same when either homologous or heterologous antigen was used in the IFA test. In general, the IFA test was more sensitive than the AGID test in diagnosing dourine, regardless of the antigen preparation used in the test or exposure group. Differences in test specificity were observed among both groups of exposed equids when either antigen was used (P < 0.05). The AGID test, using the American antigen, was more specific than the IFA test for sera from both groups of equids. When the Canadian antigen was used, the IFA test was a more specific test than the AGID test (P < 0.01).(ABSTRACT TRUNCATED AT 250 WORDS)

Evaluation of bacterins containing three predominant phage types of Salmonella enteritidis for prevention of infection in egg-laying chickens.

Six Salmonella enteritidis bacterin formulations differing in adjuvant content and whole-cell inactivation procedures were evaluated in egg-laying chickens. Chickens given S enteritidis bacterins containing modified Freund's incomplete adjuvant had greater humoral immune responses to S enteritidis than did birds given other bacterin formulations (P < 0.05). Better protection against infection by S enteritidis phage types 8, 13a, and 23 was obtained in birds vaccinated with bacterin 5. Bacterin 5 contained S enteritidis cells inactivated by 20% acetone and modified Freund's incomplete adjuvant.

Culture confirmation of the carrier status of Babesia caballi-infected horses.

Culture of horse blood for Babesia caballi identified four carrier horses among nine previously infected horses. Three of the carriers had no detectable parasitemias on stained blood smears, and sera from two carrier horses were complement fixation test negative. Three cultures were continuously cultivated. Cryopreserved fourth-passage B. caballi was successfully reestablished in vitro. Blood from a 10th horse previously subinoculated with blood from a suspected carrier was cultured, with negative results.

Serological evidence for the presence of Brucella antibodies in sheep and goats on Saint Croix, U.S. Virgin Islands.

A serological survey for Brucella antibodies in sheep and goats was completed on Saint Croix, United States Virgin Islands (USVI). Seroprevalence (at suspect or reactor titer levels) for B. melitensis antibodies was 11.3% for sheep and 2.5% for goats. This is the first report, of which we are aware, of B. melitensis antibodies in sheep or goats in the Caribbean islands.

Bacterial survival, lymph node changes, and immunologic responses of cattle vaccinated with standard and mutant strains of Brucella abortus.

Forty-eight cattle were used in 4 experiments; 6-week-old calves in experiments 1-3 (n = 24) and 10-month-old heifers in experiment 4 (n = 24). In experiments 1-3, 7 groups of 3 calves each were inoculated SC with 5 strains of Brucella abortus: virulent strain 2308 (2 groups), vaccine strain 19 (2 groups), and mutant strains RB51. 19 delta 31K, and 19 delta SOD. Sera and lymph node tissues were examined at 2-week intervals for evidence of infection. At postinoculation (PI) week 12, 2 calves in each group were given dexamethasone for 5 days. Calves were then euthanatized and lymphoid tissue, spleen, liver, and bone marrow were examined for evidence of B abortus. Calves given strain 2308 had large numbers of bacteria in their lymph nodes, marked granulomatous lymphadenitis in the deep cortex, and loss of lymphoid cells in superficial cortical areas. In addition, they had high serum antibody titers at PI week 16. Calves given strain 19, or genetic mutants derived from strain 19, cleared bacteria from lymph nodes more rapidly, had less lymphoid destruction, and developed antibody titers that did not persist for 16 weeks. The RB51 strain (rough) was cleared most rapidly from lymphoid tissues and induced serum antibody responses only to the core of the lipopolysaccharide molecule. Treatment of calves with dexamethasone did not cause B abortus to reappear in tissues of any calves, nor did serum antibody titers increase.(ABSTRACT TRUNCATED AT 250 WORDS)

Evaluation of a DNA-based probe for the detection of cattle experimentally infected with Babesia bigemina.

A digoxigenin-labeled probe was used for hybridization to various preparations of Babesia bigemina-infected erythrocyte extracts. Dot blot hybridization and immunological detection of DNA hybrids revealed that the probe was specific for B. bigemina DNA because it did not hybridize to the DNA of B. bovis, a closely related species. Studies of sensitivity showed that the probe would bind to as little as 1 ng of B. bigemina DNA, but not to 1 microgram of the B. bovis DNA. The probe reacted with equal intensity against seven B. bigemina isolates from different geographic areas. The lowest percentage of B. bigemina-infected erythrocytes detected was 0.001%, a level of parasitemia not usually detected with the light microscope. Six intact, mixed-breed steers, approximately 3 years old, were inoculated with a blood stabilate containing B. bigemina-infected erythrocytes. Blood samples collected from day -1 to day 86 postinoculation (PI) and prepared for DNA extraction were analyzed in a dot blot hybridization assay using a nonradioactive DNA probe. Hybridization reaction (HR) signals were compared to results obtained by light microscopy (LM) examination of Giemsa-stained blood smears and to antibody titers of serum samples assayed with the complement fixation test (CFT) and indirect fluorescent antibody test (IFAT). Four of six inoculated steers became infected with B. bigemina as assessed by LM. The parasitemias were low (less than 0.01-0.05) at day 10 PI. Only three steers were serologically positive by CFT (titer 1:40-1:160) and IFAT (1:1280). All four infected steers had positive HR signals in the dot blot assay. The HR signals were observed from day 10 to day 77 PI and were usually correlated with the presence of parasites in blood as observed by LM. The HR signals varied in intensity for different blood samples from the experimental animals and with day of blood sample collection. Although the signal intensity did not correlate with the parasitemia level estimated by LM, the nucleic acid hybridization assay was more sensitive than LM, CFT, or IFAT for the detection of B. bigemina-infected cattle.

Toward a population genetic analysis of Salmonella: genetic diversity and relationships among strains of serotypes S. choleraesuis, S. derby, S. dublin, S. enteritidis, S. heidelberg, S. infantis, S. newport, and S. typhimurium.

Variation in the chromosomal genomes of 1527 isolates of eight common serotypes (O and H antigen profiles) of Salmonella was assessed by analysis of electrophoretically demonstrable allelic polymorphism at 23 metabolic enzyme loci. Seventy-one distinctive electrophoretic types, representing multilocus genotypes, were identified. A basically clonal population structure was indicated by the presence of strong linkage disequilibrium among enzyme loci, the association of each serotype with a relatively small number of multilocus enzyme genotypes, and the global distribution of certain genotypes. For each of six of the serotypes, 83-96% of isolates were members of a single clone. The occurrence of each of four serotypes (S. derby, S. enteritidis, S. infantis, and S. newport) in isolates of clones belonging to several evolutionary lineages, some of which are distantly related, suggests that the horizontal transfer and recombination of chromosomal genes mediating expression of cell-surface antigens has been a significant process in the evolution of the salmonellae. Two divergent clone clusters of S. derby differ in the relative frequency with which they cause disease in birds versus mammals, and two major lineages of S. newport differ in the frequency with which their clones are associated with disease in humans versus animals.

Mastitis in Camelus dromedarius in Saudi Arabia.

The correlation between camels' milk samples collected from abnormal inflamed udders and samples positive in the California Mastitis Test (CMT) was +0.803 (P less than 0.01). The bacterial count ranges of milk samples differed significantly (P less than 0.05) for those with a negative CMT and those with a positive CMT. Infection with many but not all bacterial species was associated with positive CMT results. The highest percentage of camel milk samples was included in the bacterial count range of 3.0 x 10(2) to 3.0 x 10(3) cfu/ml rather than in the greater than 3.0 x 10(3) cfu/ml range for most of the bacterial species. The most predominant bacterial isolates were Micrococcus spp., Staphylococcus aureus, Streptococcus spp. and Corynebacterium spp. followed by eight other flora. Chloramphenicol was the most effective antimicrobial agent of six tested against 118 bacterial isolates. Preliminary observations are made on chemotherapy of mastitis cases in camels.

Incidence of brucellosis in domestic livestock in Saudi Arabia.

The results of an extensive serological survey for brucellosis antibodies using the standard plate agglutination procedure on 14,000 serum samples from native domestic animals and imported livestock over a five year period are reported. The incidence of brucellosis was highest (11.6%) in small ruminants reared intensively in breeding establishments, next highest (2.6%) in imported animals sacrificed during the Hajj season of 1977 and somewhat lower (1.5%) in local livestock sacrificed during the same Hajj season. The incidence was very low among the following groups of livestock: small ruminants raised on desert ranges (0.5%), small ruminants raised in small groups around individual homes (0.4%) and in commercial dairy herds (0.2%).

Comparative serologic study of equine piroplasmosis, with card and complement-fixation tests.

An agglutinating antigen and a rapid card test (CT) for equine piroplasmosis was developed. The antigen for the CT was prepared from lyophilized Babesia caballi complement-fixation (CF) antigen. Serum and plasma samples for testing were obtained from known B caballi-infected horses and clinically normal horses maintained at the laboratory. Serum samples also were obtained from horses outside the continental United States, in areas where piroplasmosis is endemic. Comparative CT and CF tests were done on all samples. The CT correctly identified 85% of 192 plasma samples from known infected and normal horses and 92% of 188 serum samples from these same horses. The CT results agreed closely with CF results. There was good agreement between CT and CF results. There was good agreement between CT and CF results on serum samples from horses outside the United States. Of 19 CF-positive samples, 90% were also CT-positive and 92% of 177 CF-negative samples were also CT-negative.

The identification of Babesia equi in Australia.

A Babesia parasite, isolated from the blood of a horse at Bowral, New South Wales, was identified on the basis of its morphological features, host specificity and serological reactions, as Babesia equi (Laveran 1901). The case was originally reported by Churchill and Best (1976, Aust. vet. J. 52: 487) and is the first record of equine babesiosis in Australia. In preliminary studies, the organism produced only a mild disease in an intact horse, but caused the typical clinical syndrome of acute babesiosis in a splenectomised horse, which died 19 days after the intravenous inoculation of the parasites.

Effect of imidocarb dipropionate and hemicastration on spermatogenesis in pony stallions.

The effect of imidocarb dipropionate [3, 3' bis-(2-imidazolin-2-yl) carbanilide dipropionate] on spermatogenesis in 3 pony stallions was studied. The drug alone had no deleterious effect on spermatogenesis when given intramuscularly 4 times at a dosage level of 4 mg/kg at 72-hour intervals. Hemicastration, with or without subsequent drug treatment, caused almost complete cessation of spermatogenesis 30 days after the operation.

Effect of antiparasitic medication in ponies on pasture.

Twenty Shetland ponies, 6 to 7 months old, were naturally infected with gastrointestinal nematodes and stomach bots. The ponies were allotted to 2 groups of 10 and were maintained on separate similar pastures that were free of infective larvae at the beginning of the study. The ponies in 1 group were treated monthly for 17 months with a therapeutic dose of a thiabendazole and piperazine mixture; those in the other group were not treated. During the 3rd and 5th months of the experiment, the ponies in the treated group were also given therapeutic doses of dichlorvos to remove bots. Various physical, hematologic, parasitologic, and blood chemical observations were made at weekly intervals. Each group of ponies was weighed at biweekly intervals. At the end of the experiment, the greatest differences between the treated and control ponies were in the mean number of worm eggs in fecal samples (0 vs 1,866 eggs/g), mean body weight (151.9 vs 117.0 kg), mean heart girth (126.5 vs 116.3 cm), mean packed cell volume (36.4 vs 30.8%), and mean serum protein content (8.47 vs 9.33 mg/100 ml), especially beta-globulin content (mean of 1.9 vs 3.4 mg/100 ml). The treated ponies remained clinically normal and were more spirited and more difficult to restrain for blood sampling and weighing than were the untreated controls. Parasitic nematode larvae were not recovered from grass samples from the pasture grazed by the treated ponies, but many such larvae (up to 500/300-g sample) were recovered from grass samples from the pasture grazed by the untreated ponies.