bluuebunny/crossref_metadata_2025 huggingface dataset:
Comparison of Diagnostic Tests for Bacterial Kidney Disease in Juvenile Steelhead Trout (Oncorhynchus Mykiss)
Authors: White Melvin Randall , Wu Ching Ching , Albregts Sharon R.
Published: May 2011

In order to accurately diagnose bacterial kidney disease caused by Renibacterium salmoninarum in steelhead trout, kidney tissue from experimentally infected fish was evaluated using a commercially available enzyme-linked immunosorbent assay (ELISA) test kit, fluorescent antibody (FA) testing, bacteriologic culture, and histopathology.

Seventy-five steelhead trout were randomly assigned to 1 of 4 groups and intraperitoneally inoculated with 0.15 ml saline ( n = 20), 1 × 1010 organisms/ml ( n = 18), 1 × 108 organisms/ml ( n = 18), or 1 × 106 organisms/ml ( n = 19) of R. salmoninarum.

ELISA, FA, and bacteriologic culture were positive for R. salmoninarum from the kidney tissue of the 2 groups infected with the highest doses. Although the ELISA and FA tests were accurate when compared to the bacteriologic culture from the 2 groups infected with the higher doses of the organism, they were less sensitive at the lowest level of inoculum.

Histopathology was not specific for this disease; however, all infected fish had a marked proliferative histiocytic interstitial nephritis, characterized by marked expansion of the renal hematopoietic tissue by histiocytes without tissue necrosis. Other microscopic findings included splenitis and myositis (at the injection site) of some fish.
Evaluation of a Commercial Automated System and Software for the Identification of Veterinary Bacterial Isolates
Authors: Patten V. H. , Shin S. J. , Cole J. , Watson C. W. , Fales W. H.
Published: May 2011

A commercial gram-negative bacterial autoidentification plate was originally developed using bacterial isolates of human origin. Three veterinary diagnostic laboratories conducted a 2-phase trial to enhance the database for veterinary use. The first phase consisted of testing the plate with 447 bacterial isolates of veterinary origin and incorporating that data into the existing database. Emphasis was placed on the Actinobacillus, Bordetella, Pasteurella and Enterobacteriaceae groups, since the Pseudomonas taxon was quite complete. The second phase of the trial consisted of evaluating the enhanced database using 270 clinical veterinary isolates normally encountered in veterinary laboratories. For the Actinobacillus, Bordetella, Pasteurella and Enterobacteriaceae groups, 72% of the bacterial isolates were identified correctly to genus and 61% to species after 5 hours incubation. Eighty-nine percent of the bacterial isolates were identified correctly to genus and 85% to species after 18 hours incubation. All identifications in phase 1 and phase 2 were confirmed using conventional methods.
Role of Sodium Azide in Reducing Nonspecific Color Development in Enzyme Immunoassays
Authors: Saini Parmesh K. , Webert Donald W. , Judkins Joanne C.
Published: May 2011

Improved enzyme immunoassay (EIA) procedures achieved by incorporating sodium azide during predilution of serum samples in a solid-phase EIA for the detection of anti- Toxoplasma antibody in swine using a peroxidase conjugate and in all washes of a bovine brucellosis rapid card test EIA using alkaline phosphatase conjugate are reported. Without this modification, substantial background interference was encountered that showed direct correlation with the degree of hemolysis of the serum samples. Anti- Toxoplasma gondii antibody-negative samples, separated by subjective groupings based on degree of hemolysis, into “clear,” “slight,” and “gross/total” samples, had a mean ± standard deviation of 0.150 ± 0.072, 0.187 ± 0.105, and 0.232 ± 0.108, respectively. The incorporation of sodium azide during the initial step of serum dilution dramatically eliminated the background, giving a mean ± standard deviation of 0.079 ± 0.029, 0.076 ± 0.022, and 0.081 ± 0.029, respectively. The level of endogenous peroxidase activity, a possible factor for this nonspecific interference, was considerably elevated in some of the swine sera. The clear, slight, and gross/total categories had relative levels of 1%, 2%, and 51% peroxidase activity compared to the conjugate peroxidase activity of 100%. Whereas sodium azide could be used only in sample predilution in the swine toxoplasmosis peroxidase-conjugate test, in the bovine brucellosis alkaline phosphatase-conjugate card test it could be used in all wash cycles. Many brucellosis card test results were visually uninterpretable because of significant background color when the manufacturer's wash reagent was used. The substitution of a wash reagent containing sodium azide eliminated background color, giving a visually unambiguous test.
Comparative Pathogenicity of Nine US Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) Isolates in a Five-Week-Old Cesarean-Derived, Colostrum-Deprived Pig Model
Authors: Halbur Patrick G. , Paul Prem S. , Meng Xiang-Jin , Lum Melissa A. , Andrews John J. , Rathje John A.
Published: May 2011

One hundred forty-six 5-week-old cesarean-derived, colostrum-deprived (CDCD) pigs were inoculated intranasally with 1 of 9 US porcine reproductive and respiratory syndrome virus (PRRSV) isolates. Differences were found in severity of clinical respiratory disease, rectal temperatures ( P ≤0.001), gross lung lesions ( P ≤ 0.001), and microscopic lung lesions ( P ≤ 0.05). Gross lung lesions were generally most severe 10 days postinoculation and were distributed primarily in the cranial, middle, and accessory lobes and ventromedial portion of the caudal lung lobes. Mean gross lung lesion scores estimating the percentage of lung affected by pneumonia at 10 days postinoculation ranged from 16.7% ± 2.8% (x X ± SEM, n = 10) for isolate ISU-51 to 62.4% ± 5.7% ( n = 10) for isolate ISU-28. Microscopic lung lesions were characterized by hyperplastic and hypertrophied type 2 pneumocytes, septal infiltration by mononuclear cells, and accumulation of necrotic alveolar exudate. Lymph node follicular hyperplasia and focal necrosis was seen with all 9 isolates. This CDCD pig model was useful for demonstration of significant differences in pathogenicity among US PRRSV isolates. This difference in pathogenicity may help explain the variation in severity of clinical disease observed in field outbreaks of porcine reproductive and respiratory syndrome and should provide for meaningful comparison of PRRSV genotypes.
Diagnostic Serologic Testing of Cage and Aviary Birds for Chlamydiosis and Suggested Confirmatory Testing
Authors: Grimes James E. , Arizmendi Francisco , Carter Craig N. , Sneed Loyd
Published: May 2011

A 2 × 2 contingency table was constructed to demonstrate the relationships between detectable Chlamydial antibody activity and clinical health status of tested birds. The table revealed that 65.5% of clinically ill birds were antibody positive by elementary body agglutination (EBA) (≥10 titers) and 59.0% were antibody positive by latex agglutination (LA). Thus, EBA was slightly more sensitive than LA in detecting antibody activity. Of the clinically normal birds, 96.7% were antibody negative (< 10 titers) by EBA and 98.3% were antibody negative by LA. Individual serum or plasma samples from a group of mixed types of psittacine birds and cockatiels were tested as a separate group, and relationships between EBA-detectable antibody activity and health status were obtained from a 2 × 2 contingency table. Sixty-six percnt of birds clinically ill with signs of chlamydiosis in the mixed-type group were antibody positive, whereas only 32.3% of clinically ill cockatiels were antibody positive. Statistical analysis of the contingency table using a chi-square test demonstrated that the EBA test differentiates between individual birds on the basis of health status ( P < 0.001). When testing paired serum or plasma samples by EBA, LA, and direct complement fixation (DCF), the highest percentage of significant (≥ 4–fold change) titer decreases was detected by LA, and the highest percentage of significant titer increases was detected by DCF. Examples of EBA, LA, and DCF titers in paired and multiple serum or plasma samples are presented to show the variety of responses that can occur. Results reflected variations seen in individual testing of birds with titer variability seen in the first sample tested. Additional types of testing believed necessary for confirming or ruling out an infectious process in birds are outlined. The current interpretations of serologic results are given.
Lung and Nasal Lesions Caused by a Swine Chlamydial Isolate in Gnotobiotic Pigs
Authors: Rogers Douglas G. , Andersen Arthur A. , Hunsaker Breck D.
Published: May 2011

The objective of this study was to determine whether a chlamydial isolate recovered from nasal swabs from swine with pneumonia could cause pneumonia and rhinitis in gnotobiotic pigs. The identity of the isolate currently is unknown, but it shares characteristics with Chlamydia trachomatis. After propagation in Vero cells and preparation of the inoculum (2.5 × 1010 inclusion-forming units/ml), Chlamydiae were instilled into nostrils (1.0 ml/nostril) and lungs (2.0 ml intralaryngeally) of 15 anesthetized 3-day-old gnotobiotic piglets. Five age-matched gnotobiotic piglets were anesthetized and sham infected with uninfected cell culture lysates. Two treated piglets were moribund and 2 were severely dyspneic prior to necropsy 7 days postinfection (DPI), whereas remaining treated piglets showed mild dyspnea upon exertion throughout the study. All treated piglets developed diarrhea. All treated piglets necropsied 7–21 DPI had extensive consolidation in cranial, middle, and accessory lung lobes; a majority of these piglets also had extensive consolidation in the caudal lobes. Treated piglets necropsied 28 and 35 DPI had a lobular pattern of consolidation in all lung lobes. Histologically, lesions in lungs from treated piglets necropsied 7, 14, and 21 DPI were characterized by bronchointerstitial pneumonia with foci of type II pneumocyte hypertrophy and hyperplasia; pneumocytes and bronchial and bronchiolar epithelial cells were markedly vacuolated. Alveolar macrophages, peribronchitis, peribronchiolitis, and peri-vasculitis were seen in lungs from treated piglets necropsied 28 and 35 DPI; those necropsied 28 DPI also had foci of lymphohistiocytic and plasmacytic infiltrates. Turbinate lesions in all treated piglets were characterized by mild multifocal lymphoplasmacytic and occasionally neutrophilic rhinitis. Immunohistochemistry detected chlamydial antigen in bronchial and bronchiolar epithelial cells, pneumocytes, and inflammatory cells in treated piglets necropsied 7, 14, and 21 DPI. Positive staining was limited to alveolar macrophages in treated piglets necropsied 28 and 35 DPI. Chlamydial antigen was detected in turbinate epithelial cells at all necropsy intervals. Ultrastructurally, chlamydiae were seen with glycogen particles in vacuoles or free in the cytoplasm of bronchial and bronchiolar epithelial cells and pneumocytes. The results indicated that the chlamydial isolate used in this
Development of Murine Monoclonal Antibodies for the Immunohistochemical Diagnosis of Systemic Bovine Aspergillosis
Authors: Jensen H. E. , Aalbaek B. , Lind P. , Krogh H. V. , Frandsen P. L.
Published: May 2011

Murine monoclonal antibodies (MAbs) against water-soluble somatic antigens (WSSA) and the wall fraction (WF) from Aspergillus fumigatus were produced by fusion of splenocytes from immunized BALB/c mice with mouse myeloma X63-Ag 8.653 cells. The supernatants of in vitro cultured hybridomas were initially screened for reactivity with the WSSA and the WF from A. fumigatus and WSSA of other fungi in an enzyme-linked immunosorbent assay (ELISA). Supernatants reacting only with A. fumigatus antigens were subsequently screened for homologous and heterologous reactivity with immunohistochemical techniques using formalin-fixed, paraffin-embedded tissues from experimentally infected mice. Because of a high immunohistochemical reactivity with homologous fungi, 4 MAbs raised against A. fumigatus WSSA and WF were selected for a further evaluation of cross-reactivity (diagnostic specificity) in immunohistochemical and immunoblotting assays. In immunohistochemical assays, all MAbs raised against WSSA cross-reacted heavily with a number of other fungal species. All 4 MAbs (MAb-WF-AF-1-4) raised against the WF reacted strongly with hyphae of Aspergillus spp.; hyphae of Scedosporium apiospermum were also strongly labeled by MAb-WF-AF-3 and-4. The 2 specifically reacting MAbs (MAb-WF-AF-1 and-2) were of the IgM biotype and were precipitating, and in immunoblotting experiments both bound to a 106-kD antigen of the WF, whereas they did not bind to WSSA of A. fumigatus. One of the 2 aspergillosis-specific MAbs (MAb-WF-AF-1) was used to screen 145 mycotic lesions of cattle. The diagnoses on bovine lesions obtained by MAb-WF-AF-1 were compared with results based on reactivity with heterologously absorbed polyclonal antibodies and, for some lesions, to culture results. In the vast majority of lesions ( n = 133), the MAb-WF-AF-1 and the polyclonal anti-Aspergillus antibodies reacted in a similar pattern, i.e., positively in 41 aspergillosis lesions and negatively in 92 zygomycotic lesions. Hyphae in 3 of 12 lesions that were not stained by the polyclonal antibodies reacted with the specific MAb-WF-AF-1; i.e., aspergillosis was diagnosed. The characteristics of the 2 MAbs (MAb-WF-AF-1 and-2) raised against the WF of A. fumigatus in ELISA and immunoblotting and immunohistochemical assays justify their application for the in situ diagnosis of systemic aspergillosis of cattle.
Chromosomal Aneuploidy Associated with Spontaneous Abortions and Neonatal Losses in Cattle
Authors: Schmutz S. M. , Moker J. S. , Clark E. G. , Orr J. P.
Published: May 2011

Pericardial sac samples from 77 bovine aborted fetuses and stillborn calves were submitted for tissue culture; cells from 55 of these samples were grown successfully in culture. Six of the 55 karyotyped fetuses (10.7%) had an abnormal chromosome complement, in 3 of which (5.5%) the abnormality was probably the cause of death. This level of abnormality is relatively high when one considers that most fetuses were gt; 8 months gestational age. Approximately 5–7% of human stillbirths and 50% of first-trimester aborted fetuses have chromosome anomalies. If a similar situation exists in cattle, as suggested by these data, chromosome abnormalities may be a major cause of early fetal loss in cattle. Most chromosomally abnormal fetuses had multiple malformations, which suggests that the diagnostic use of chromosome analysis is most cost effective for malformed fetuses and newborns. Twins were present in a higher proportion of these fetuses than expected based on their incidence among liveborn cattle.
Detection of Antibodies against Foot-and-Mouth Disease Virus Using a Liquid-Phase Blocking Sandwich ELISA (LPBE) with a Bioengineered 3D Protein
Authors: O'Donnell Vivian K. , Boyle David B. , Sproat Kathy , Fondevila Norberto A. , Forman Anthony , Schudel Alejandro A. , Smitsaart Eliana N.
Published: May 2011

A liquid-phase blocking sandwich enzyme-linked immunosorbent assay (ELISA-3D) was developed to detect specific antibodies to the 3D protein in sera from foot-and-mouth disease (FMD) virus (FMDV)-infected animals. The assay uses a nonstructural 3D recombinant protein and two polyclonal antisera, one for capture (bovine) and the other for detector (guinea pig). The specificity of the assay was demonstrated by negative results with 101 sera of cattle from the FMD-free zone in Argentina and with bovine and porcine sera raised against various RNA and DNA viruses. The ELISA-3D was able to detect antibodies in cattle after natural or experimental infection with FMDV of A, O, or C types as early as 5 days postinfection and at later stages in persistently infected animals. Comparison of the results with those obtained with the routinely used agar gel immunodiffussion test and a previously described ELISA, both employing a partially purified virus-infection-associated antigen, shows that the ELISA-3D is highly sensitive and specific and gives reproducible results. Its use as a tool for monitoring viral activity and for certification of FMDV-free animals is recommended.
Application of a Polymerase Chain Reaction Assay for Detection of Proliferative Enteritis-Affected Swine Herds
Authors: Holyoake Patricia K. , Jones Gary F. , Davies Peter R. , Foss Dennis L. , Murtaugh Michael P.
Published: May 2011

A polymerase chain reaction (PCR) assay was used to confirm the presence of ileal symbiont (IS) intracellularis in 3 swine herds with a history of proliferative enteritis (PE). Two pooled fecal specimens, each comprising 5 individual stool samples, were collected from pen floors to screen for the presence of IS intracellularis and determine the age range of pigs shedding the organism. IS intracellularis was detected in the feces of clinically normal 10–25week-old grower/finisher pigs, indicating that this age range of pigs was the main source of infection for younger nursery pigs. Shedding continued without clinical disease when 10–100 g/ton of tylosin or 10 g/ton of chlortetracycline was added to the feed. PCR testing of pooled fecal samples can be used to identify groups of pigs affected with PE. The results of this study indicate that this PCR assay has the potential to accurately assess the IS intracellularis infection status of swine herds and the association of IS intracellular-is with PE and growth performance.
Phenotypic and Ribosomal RNA Characterization of Arcobacter Species Isolated from Porcine Aborted Fetuses
Authors: Schroeder-Tucker Linda , Wesley Irene V. , Kiehlbauch Julia A. , Larson David J. , Thomas Lee Ann , Erickson Gene A.
Published: May 2011

Aerotolerant organisms resembling Campylobacter, now designated as Arcobacter, have been described from aborted farm animals and from cases of human enteritis worldwide. The goals of this study were 1) to attempt to recover Arcobacter spp. from cases of porcine abortion, 2) to characterize these isolates by phenotype and ribotype, and 3) to compare the usefulness of ribotype and phenotype patterns for identifying Arcobacter butzleri and the DNA hybridization groups 1A and 1B of A. cryaerophilus. Isolates of Arcobacter spp. from North Carolina and Iowa were recovered from porcine tissues. In Iowa, Arcobacter spp. were recovered from 43% (13/30) of porcine abortion cases evaluated. Isolations were made from placenta (44%), kidney (44%), and stomach contents (12%), which were the only tissues examined. The most reliable biochemical tests for A. butzleri included growth in 1% glycine and in 1.5% NaCl, weak catalase activity, and resistance to cadmium chloride. Arcobacter cryaerophilus strains were characterized by strong catalase activity and sensitivity to cadmium chloride. The DNA hybridization groups 1A and 1B of A. cryaerophilus could not be distinguished by biochemical tests. This represents the first description of A. cryaerophilus DNA group 1A in animals within the United States.
Novel Indirect Fluorescent Antibody Test for Lyme Disease
Authors: Chambers Margaret A. , Swango Larry J. , Wright James C.
Published: May 2011

An indirect fluorescent antibody (IFA) test was developed using a novel format of Borrelia burgdorferi organisms adhered to a monolayer of cultured endothelial cells derived from an equine tumor. Sensitivity and specificity of the new IFA test for detecting anti- B. burgdorferi antibodies were evaluated using sera from dogs inoculated with live B. burgdorferi or vaccinated with B. burgdorferi bacterin or leptobacterins and from unvaccinated specific-pathogen-free (SPF) dogs. To compare the new IFA test with existing tests, serum samples were submitted to independent laboratories to be tested by enzyme-linked immunosorbent assay (ELISA) and a traditional IFA test. Samples were also tested with 2 commercially available membrane-bound ELISA kits. Both Borrelia-inoculated dogs and dogs vaccinated with B. burgdorferi bacterin developed levels of antibody detectable by the new IFA test. Dogs vaccinated with a combination canine vaccine or leptobacterin for food animal use developed detectable levels of antibody against Leptospira but remained seronegative for Borrelia by the new IFA test, as did the unvaccinated SPF dogs. The new IFA test was sensitive, detecting antibodies against B. burgdorferi as early as 7 days postinoculation. It was also specific, showing no cross-reactivity with anti- Leptospira antibodies induced by vaccination with leptobacterins. The new IFA test compared favorably with both the standardized traditional IFA test and ELISA. Results from both membrane-bound ELISA kits were not consistent when compared with each other or with the new IFA test. The new IFA test had low nonspecific fluorescence, which made it easier to evaluate and reduced the human error and variability of test results.
Effects of Environmental Heat and Intake of Tall Fescue Seed Infested with Acremonium Coenophialum on the Acid-Base Status of Young Bulls
Authors: Wallace L. L. Mills , Vogt D. W. , Lipsey R. J. , Gamer G. B. , Cornell C. N.
Published: May 2011

Effects of high environmental temperature and dietary intake of tall fescue ( Festuca arundinacea) seed containing the endophyte Acremonium coenophialum on bovine acid-base status were studied using 3 groups of bull calves (2 Simmental, 1 Angus). Experimental animals were housed in controlled-climate chambers and subjected to gradual increases in environmental temperature, first while being fed an endophyte-free diet and then while being fed a diet containing 17% endophyte-infested fescue seed. Marked acid-base disturbances were not observed in any animals. In general, Pco2, HCO3-, base excess, and arterial blood pH values were reduced in response to heat stress, both with endophyte-free and endophyte-containing diets. In most individuals anion gap increased. These results reflected metabolic compensation for mild chronic alveolar hyperventilation and retention of organic acids. These findings suggest that, under conditions similar to those found during the summer in central Missouri, normal cattle should not be at great risk of developing respiratory alkalosis or other severe acid-base disturbances as a result of heat stress and/or intake of tall fescue endophyte.
Immunoblot and Ultrastructural Analysis of Antigens Extracted from Dichelobacter Nodosus with Potassium Thiocyanate
Authors: Whittington Richard
Published: May 2011

The antigens extracted from Dichelobacter nodosus with potassium thiocyanate were analyzed by western blotting with sera from footrot-free sheep and from sheep infected with D. nodosus to identify components specific for infection. Several components with molecular mass < 33 kD were associated with infection, particularly bands of 32.5 kD, 30.5 kD, and 28 kD. Components with molecular mass > 56 kD may be responsible for false-positive reactions observed when sera of footrot-free sheep react with the potassium thiocyanate extract in solid phase enzyme-linked immunosorbent assays (ELISAs). Bands of 62.5 kD, 56 kD, 40 kD, and 19 kD were recognized for most sheep regardless of their disease status or ELISA reactivity and therefore do not appear to be detected by the ELISA. No components of the potassium thiocyanate extract were completely specific for infection. Antigens in the extract were identified primarily on the cell envelope by immunogold electron microscopy. Labeling was concentrated in the periplasm, with minor labeling of the cell surface. The extract consisted of tangled strands of particles with electron-dense cores, and few vesicular structures were observed.
Distribution of Lesions in Cattle Infected with Mycobacterium Bovis
Authors: Whipple Diana L. , Bolin Carole A. , Miller Janice M.
Published: May 2011

Detailed postmortem examinations were conducted on 30 cattle from a dairy herd with bovine tuberculosis to determine the distribution of lesions in Mycobacterium bovid-infected cattle. Twenty-four different tissue specimens from each animal were examined for gross lesions and collected for bacteriologic culturing and histologic examination. Tuberculosis was confirmed in 15 cattle with evidence of infection in 1 or more of the following tissues: medial retropharyngeal, parotid, tracheobronchial, mediastinal, caudal deep cervical, and subiliac lymph nodes; palatine tonsil; and lung. Gross and histologic lesions were present most frequently in lymph nodes of the thoracic region. Mycobacterium bovis was isolated from 3 cattle that had no gross lesions of tuberculosis. One animal had lesions only in the subiliac lymph node, which is not routinely examined during slaughter surveillance. Results of this study indicate that not all cattle infected with M. bovis have visible lesions of tuberculosis in sites that are routinely inspected. These findings are important because detection of gross lesions of tuberculosis during inspection of carcasses at slaughter is the primary method for detection of tuberculous cattle and herds in the United States.
Evidence Suggesting a Point Source Exposure in an Outbreak of Bovine Abortion Due to Neosporosis
Authors: McAllister Milton M. , Huffman E. M. , Hietala Sharon K. , Conrad Patricia A. , Anderson Mark L. , Salman Mo D.
Published: May 2011

A Holstein dairy farm suffered an abortion outbreak due to neosporosis. Abortion losses were > 18%. Cows with the highest Neospora antibody titers were at the greatest risk of aborting. Mummified fetuses were found after the 43rd day of the outbreak. The epidemic curve was suggestive of a point source exposure, which is consistent with the hypothesis that Neospora can be spread by a definitive host.
A Sequential Study of Experimental Porcine Paramyxovirus (LPMV) Infection of Pigs: Immunostaining of Cryostat Sections and Virus Isolation
Authors: Allan G. M. , McNeilly F. , Walker I. , Linne T. , Moreno-Lopez J. , Hernandez P. , Kennedy S. , Carroll B. P. , Herron B. , Foster J. C. , Adair B.
Published: May 2011

La Piedad Michoacan Paramyxovirus (LPMV) is a recently recognized paramyxovirus infecting pigs throughout Mexico. Disease syndromes observed in field cases associated with LPMV infection include neurologic, respiratory, and reproductive disorders. Clinical signs and the distribution of LPMV virus and antigen in tissue samples from pigs experimentally infected with LPMV by natural routes were studied. Severe neurologic disease and death occurred following experimental inoculation of 3- and 17-day-old pigs. All of the pigs inoculated at 3 days of age were either dead or moribund by 8 days after inoculation, whereas 30% of the pigs inoculated at 17 days of age were affected. Virus was consistently recovered from or demonstrated in tissues from the respiratory tract of both groups of pigs. LPMV and antigen were also demonstrated in central nervous system (CNS) tissues from these pigs; however, differences in virus distribution within the CNS were demonstrated in the 2 groups. In the pigs inoculated at 17 days of age, isolation of LPMV was restricted to the olfactory bulb and midbrain. In contrast, in the pigs inoculated at 3 days of age, isolation of LPMV was more widespread throughout the CNS tissue examined. Virus excretion studies indicated that nasal spread of LPMV was more important than fecal spread. Comparatively large quantities of infectious LPMV were consistently recovered from urine samples of experimentally infected pigs.
PCR Detection of Acute Ehrlichia Canis Infection in Dogs
Authors: McBride Jere W. , Corstvet Richard E. , Gaunt Stephen D. , Chinsangaram Jarasvech , Akita Geoffrey Y. , Osburn Bennie I.
Published: May 2011

A polymerase chain reaction (PCR)-based detection assay that specifically detected Ehrlichia canis in dogs with acute infections was developed. A region of the 16S ribosomal RNA gene of E. canis was targeted for PCR amplification and chemiluminescent hybridization (CH) with a complementary internal 287-base pair (bp) oligonucleotide probe. The CH improved the PCR assay sensitivity 1,000-fold as compared with visualization on ethidium bromide-stained agarose gels. The PCR assay with CH (PCRKH) detected as little as 30 fg of E. canis genomic DNA, the equivalent of approximately 150 E. canis organisms. The 495-bp product defined by the specific primers was not detected when genomic DNA from E. platys, E. chaffeensis, E. risticii, and E. equi were used in the PCR/CH assay. The PCR/CH assay was tested with unfractionated blood samples collected from 9 dogs experimentally infected with E. canis. The PCR/CH assay had greater detection sensitivity than did cell culture isolation (CCI) from infected blood. PCR/CH detected E. canis 7 days prior to CCI in 4 of 6 experimentally infected dogs. The results obtained with the PCR/CH assay otherwise consistently matched the results obtained by CCI. This PCR/CH assay is a rapid, sensitive, and specific method for E. canis detection with sensitivity comparable to or exceeding that of CCI. A diagnosis of E. canis using this PCR/CH assay can be made in 2 days as compared with 14 weeks for CCI. The PCR/CH assay appears to be an acceptable alternative or complement to current diagnostic techniques.
Water Hemlock (Cicuta Douglasii) Toxicoses in Sheep: Pathologic Description and Prevention of Lesions and Death
Authors: Panter Kip E. , Baker Dale C. , Kechele Phil O.
Published: May 2011

Water hemlock causes numerous livestock losses in North America every year. Description of pathologic and serum biochemical changes has been lacking in the literature. Tubers of western water hemlock ( Cicuta douglasii) induced excessive salivation, tremors, grand mal seizures, skeletal and cardiac myodegeneration, and death in sheep given 1.2–2.7 g fresh tuber/kg body weight by gavage. Seizures were intermittent with periods of relaxation until death occurred from anoxia during seizure activity. In sheep given 1.5–2.5 times the lethal dose of water hemlock by gavage, intravenous administration of sodium pentobarbital at the onset of the first seizure prevented further seizure activity and skeletal and cardiac myodegeneration and resulted in rapid and complete recovery.
Immunohistochemical Detection of Hog Cholera Viral Glycoprotein 55 in Paraffin-Embedded Tissues
Authors: de las Mulas Juana Martín , Ruiz-Villamor Eduardo , Donoso Sergio , Quezada Manuel , Lecocq Claudio , Sierra Miguel Angel
Published: May 2011

Formalin-fixed, paraffin embedded tissues obtained from 40 pigs inoculated with a field isolate of hog cholera virus were examined for the presence of Gp55, a major structural protein of the virus envelope, using a monoclonal antibody-based immunohistochemical test with the avidin-biotin-peroxidase complex method. Immunoreactivity was detected in hog cholera virus-infected tissues but not in control pigs tissues, African swine fever virus-infected tissues, or bovine viral diarrhea virus-infected porcine or bovine tissues. The first positive reactions were seen in lymphatic tissues, digestive tract and skin on postinoculation day (pid) 4, respiratory and urinary tissues on pid 5, nervous tissues on pid 6, and endocrine tissues on pid 7. These staining reactions persisted until the last observation on pid 18. Hog cholera virus antigen was not detected in heart tissue at any time. The highest levels of antigen detection were found in tonsils, spleen, and pancreas, although the esophageal mucosa and skin epithelial cells were also intensely and widely stained. The cellular staining pattern of Gp55 had a ubiquitous distribution. It was found in epithelial cells, macrophages and circulating monocytes, endothelial cells, lymphoid cells, and glial cells. The results showed a high specificity and high sensitivity for detecting hog cholera Gp55 in formalin-fixed, paraffin embedded tissue samples. This method allows precise association of Gp55 with specific cells, tissues, and histologic lesions, making the technique suitable for use in routine diagnosis of hog cholera.
Ergotism and Photosensitization in Swine Produced by the Combined Ingestion of Claviceps Purpurea Sclerotia and Ammi Majus Seeds
Authors: López Tomas A. , Campero Carlos M. , Chayer Ricardo , de Hoyos Miguel
Published: May 2011

Poisoning of domestic animals happens frequently in the southeast of Buenos Aires Province (Argentina). Intoxications are produced mainly by the ingestion of plants and mycotoxins, but animals are rarely affected simultaneously by both types of agents. One herd of pigs suffered simultaneous intoxications by ergot alkaloids from Claviceps purpurea sclerotia and furocoumarins from Ammi majus seeds. Pigs were fed a diet composed of wheat (poor quality) or corn and protein and vitamin supplements. This diet was completed with forage sorghum. Nervous signs were first observed 5–7 days after the initiation of feeding the suspect ration. These signs were followed by cutaneous irritation. Snout ulcers, eyelid edema, and conjunctivitis were observed in several piglets. Ten days after the start of feeding the incriminated ration, 8 abortions were observed. Many of the sows that were nursing piglets developed udder edema and teat cracking. Dermal lesions were observed in most of the animals with unpigmented areas in the skin but not in a Duroc-Jersey boar. Removal of the incriminated diet and feeding of another diet prepared with good-quality wheat allowed all the animals to recover in 15 days. The herd experienced normal pregnancies and parturitions, litter sizes, and piglet weights when fed a cleaned portion of the poor-quality wheat. No photosensitization lesions were observed. Examination of impurities in the suspected wheat indicated the presence of 2.2% of A. majus seeds and 0.14% of C. purpurea sclerotia. The quantitative analysis indicated the presence of 3.2 g xanthotoxin and 0.65 g bergaptene/100 g A. majus seeds and 0.73 g ergot alkaloids (expressed as ergonovine) per 100 g of C. purpurea sclerotia. Qualitative analysis demonstrated the presence of ergotamine, ergocristine, and ergonovine. These results indicate that clinical signs and lesions were caused by the ingestion of large quantities of these biologically active compounds.
In Vivo Indicators of Pathologic Ruminal Sulfide Production in Steers with Diet-Induced Polioencephalomalacia
Authors: Gould Daniel H. , Cummings Bruce A. , Hamar Dwayne W.
Published: May 2011

Two groups of 3 120–160-kg Holstein steers were fed a diet high in carbohydrate and low in long fiber and either with or without added sodium sulfate. Prior to and during the course of feeding the experimental diet, the concentrations of rumen hydrogen sulfide gas and rumen fluid sulfide were determined by a simple sulfide detector tube method and by sulfide-selective electrode, respectively. Other measurements included rumen fluid pH, blood creatine kinase, and blood sulfhemoglobin. Two of the 3 steers fed the high-sulfate diet developed signs and lesions of polioencephalomalacia. Clinical signs included episodic ataxia and blunted or absent menace reaction. Increased ruminal H2S gas concentrations occurred in all 3 steers consuming the diet with added sulfate. The onset of clinical signs coincided with the onset of elevated H2S concentrations. These increases were 40–60 times the values measured in the steers consuming the diet without added sulfate. In contrast, increases in rumen fluid sulfide concentrations usually rose to 4 times that of control steers. The steers fed an identical diet but without added sulfate exhibited no signs or lesions of polioencephalomalacia and no elevations of sulfide in rumen gas or fluid. All steers had a modest decrease in rumen fluid pH associated with the transition to the concentrate diet. No significant changes were observed in any of the blood measurements of any of the steers. An additional pair of steers was fed the experimental diet with or without added sulfate to compare the ruminal H2S gas concentrations estimated by H2S detector tubes with those estimated by a different method of analysis utilizing charcoal trapping of H2S, conversion to sulfate, and measurement of the sulfate. Both methods yielded comparable estimates of H2S concentration. Overall, these data indicate that changes in rumen gas cap H2S concentrations are larger than changes in rumen fluid sulfide concentration and that estimation of rumen gas cap H2S concentration may be a practical approach to detecting pathologic increases in ruminal H2S gas. This simple, rapid, minimally invasive method should be useful for estimating the H2S content of ruminal gas under field conditions.
Laboratory Identification and Enteropathogenicity Testing of Serpulina Pilosicoli Associated with Porcine Colonic Spirochetosis
Authors: Muniappa Nagaraja , Mathiesen Michelle R. , Duhamel Gerald E.
Published: May 2011

Pathogenic intestinal spirochetes of swine include Serpulina hyodysenteriae, a strongly β-hemolytic spirochete that causes swine dysentery, and S. pilosicoli, a weakly B-hemolytic intestinal spirochete (WBHIS) that causes porcine colonic spirochetosis. Because of the existence of nonpathogenic WBHIS in the normal swine colon, it is important to develop laboratory procedures for accurate identification of S. pilosicoli. The purpose of the present study was to assess hippurate hydrolysis and polymerase chain reaction (PCR) amplification of specific 16S ribosomal RNA (rRNA) sequences for identification of porcine S. pilosicoli. Additionally, the enteropathogenicity of 8 field isolates of porcine S. pilosicoli was determined by challenge exposure of 1-day-old chicks and sequential histologic examination of the cecal mucosa. The field isolates of porcine S. pilosicoli hydrolyzed hippurate and yielded S. pilosicoli-specific products by PCR amplification of 16S rRNA sequences. Although all of the field isolates of porcine S. pilosicoli attached to the cecal epithelium of challenge-exposed chicks by day 21 postinoculation, some isolates had locally invasive phenotypes. We concluded that identification of porcine S. pilosicoli could be made on the basis of results of hippurate hydrolysis and 16S rRNA PCR amplification. Challenge inoculation of 1-day-old chicks followed by histologic examination of the cecal mucosa demonstrated the enteropathogenicity of porcine S. pilosicoli.
Investigation of the Selenium Status of Aborted Calves with Cardiac Failure and Myocardial Necrosis
Authors: Orr James P. , Blakley Barry R.
Published: May 2011

Lesions of heart failure, specifically cardiac dilation or hypertrophy along with a nodular liver (chronic passive congestion) and ascites, have been found in 4-5% of aborted bovine fetuses. In this study, a group of 22 such fetuses was compared with groups of aborted fetuses without lesions of heart failure and with nonaborted fetuses obtained from a slaughterhouse. The fetuses were necropsied, tissues were taken for histopathology, and samples were collected for routine bacteriologic and virologic examinations. Liver and kidney tissue was saved for selenium analysis. Histopathologic examinations of myocardium of fetuses with cardiac failure revealed myocardial necrosis and mineralization in 7 fetuses, lymphocytic myocarditis in 5 fetuses, myocardial fibrosis in 5 fetuses, or no microscopic lesions in 5 fetuses. Mean liver selenium levels were 5.5 (μmol/kg in the fetuses with heart lesions, 6.5 μmol/kg in the fetuses without heart lesions and 7.5 μmol/kg in fetuses from the slaughterhouse; these differences were statistically significant. The results suggest that selenium deficiency in bovine fetuses may cause myocardial necrosis and heart failure. This study also provides data on normal liver and kidney selenium levels in bovine fetuses from the analyses of 19 nonaborted fetuses.
Polymerase Chain Reaction for Detection of Guinea Pig Adenovirus
Authors: Pring-Åkerblom Patricia , Blažek Karel , Schramlová Jana , Kunstýr Ivo
Published: May 2011

Lack of in vitro cultivation methods has inhibited the development of rapid, reliable diagnostic procedures for adenovirus-associated necrotizing bronchopneumonia in guinea pigs. Because polymerase chain reaction (PCR) techniques are well established for human adenoviruses, primers for the amplification of guinea pig adenovirus DNA were evaluated. The DNA for PCR was purified from the lung tissue of spontaneously infected and healthy guinea pigs. Adenovirus DNA could only be detected in the lungs of the infected animals. Subsequent sequence analysis of PCR products revealed that the guinea pig adenovirus is a distinct adenovirus.
Sodium Toxicity and Pathology Associated with Exposure of Waterfowl to Hypersaline Playa Lakes of Southeast New Mexico
Authors: Meteyer Carol U. , Dubielzig Richard R. , Dein F. Joshua , Baeten Laurie A. , Moore Melody IS. , Jehl Joseph R. , Wesenberg Katherine
Published: May 2011

Cause of mortality was studied in waterfowl in hypersaline playa lakes of southeast New Mexico during spring and fall migration. Mortality was not common in wild ducks resting on the playas during good weather. However, when birds remained on the lakes for prolonged periods of time, such as during experimental trials and stormy weather, a heavy layer of salt precipitated on their feathers. Sodium toxicity was the cause of death for all experimental mallards housed on playa water and for 50% of the wild waterfowl found moribund or dead during the spring of 1995. Gross lesions included heavy salt precipitation on the feathers, ocular lens opacities, deeply congested brains, and dilated, thin-walled, fluid-filled cloacae. Microscopic lesions in the more severely affected birds included liquefaction of ocular lens cortex with lens fiber swelling and multifocal to diffuse ulcerative conjunctivitis with severe granulocytic inflammation, edema, and granulocytic vasculitis resulting in thrombosis. Inflammation similar to that seen in the conjunctiva occasionally involved the mucosa of the mouth, pharynx, nasal turbinates, cloaca, and bursa. Transcorneal movement of water in response to the hypersaline conditions on the playa lakes or direct contact with salt crystals could induce anterior segment dehydration of the aqueous humor and increased osmotic pressure on the lens, leading to cataract formation.
Experimental use of a Dot-Blot Assay to Measure Serologic Responses of Cattle Vaccinated with Brucella Abortus Strain RB51
Authors: Olsen S. C. , Stevens M. G. , Cheville N. F. , Schurig G.
Published: May 2011

Brucella abortus strain RB51 was recently approved as an official brucellosis calfhood vaccine for cattle by the Animal and Plant Health Inspection Service branch of the United States Department of Agriculture. Currently available serologic surveillance tests for B. abortus do not detect seroconversion following SRB51 vaccination. The purpose of this study was to evaluate a dot-blot assay using γ-irradiated strain RB51 bacteria for its specificity and sensitivity to detect antibody responses of cattle vaccinated with strain RB51. Dot-blot titers of sera at a recommended dosage (1010 colony-forming units) were similar to those of sera from cattle vaccinated with similar numbers of B. abortus strain 19 and greater ( P < 0.05) than titers of nonvaccinated cattle. In the first 12 weeks after vaccination with 1010 colony-forming units of strain RB51, the RB51 dot-blot assay had 100% specificity for titers of 80 or less and a 53% sensitivity for titers of 160 or greater. Sensitivity of the RB51 dot-blot assay peaked at 4 weeks after vaccination with 1010 colony-forming units of strain RB51. Dot-blot responses of sera from cattle vaccinated with a reduced dosage of strain RB51 (109 colony-forming units) did not differ ( P > 0.05) from titers of sera from nonvaccinated cattle. Following intraconjunctival challenge with B. abortus strain 2308, titers on the RB51 dot-blot assay did not differ ( P > 0.05) between nonvaccinated cattle and cattle vaccinated at calfhood with strain 19 or strain RB51.
An Improved Method for Cultivation of Mycobacterium Paratuberculosis from Bovine Fecal Samples and Comparison to Three Other Methods
Authors: Stabel Judith R.
Published: May 2011

A new method (NADC) for isolation of Mycobacterium paratuberculosis from fecal samples is described and evaluated using fecal samples from a known paratuberculosis-infected herd of cattle. The NADC method includes centrifugation of the total fecal sample supernatant and use of a 2-step decontamination protocol. The growth rate of M. paratuberculosis and contamination rate of cultures when using this method are compared to 3 other published methods: sedimentation, centrifugation, and Cornell. Sensitivity was lowest for the Cornell method precluding detection of some low shedders; however, contamination was not observed for this method. Contamination was the most severe in samples processed by the centrifugation method but was also high for the sedimentation method, resulting in unreadable culture tubes for some fecal samples. The NADC method was 10-fold more sensitive for detection of M. paratuberculosis colonies and contamination was significantly reduced compared to other 3 methods.
Restriction Fragment Length Polymorphism Analysis of Mycobacterium Bovis Isolates from Captive and Free-Ranging Animals
Authors: Whipple Diana L. , Clarke P. Ryan , Jarnagin Jerald L. , Payeur Janet B.
Published: May 2011

Mycobacterium bovis isolates from cattle, captive elk, and free-ranging mule deer and coyotes were examined by restriction fragment length polymorphism (RFLP) analysis. DNA extracted from each isolate was digested with restriction endonucleases AluI and PvuII. DNA probes used for Southern hybridizations were a 37-base oligonucleotide and a 123-base–pair sequence specific for the insertion sequence IS 6110 and a plasmid, pTBN12, which contains a polymorphic GC-rich repetitive sequence present in several species of mycobacteria. Generally, M. bovis isolates originating from a single herd of either cattle or captive elk had identical RFLP patterns, whereas isolates from unrelated sources had distinct patterns. The RFLP patterns for M. bovis isolates from free-ranging mule deer and coyotes were identical to patterns observed for isolates from a captive elk herd that was located in the area where the free-ranging animals were found. These results indicate that the captive elk herd may have been the source of M. bovis that infected the free-ranging animals. Results of this study show that RFLP analysis is a useful tool for differentiation of M. bovis isolates and for molecular epidemiology studies to determine possible sources of infection in outbreaks of tuberculosis in animals.
Immunohistochemical Detection of Brucella Abortus Antigens in Tissues from Aborted Bovine Fetuses Using a Commercially Available Polyclonal Antibody
Authors: Pérez José , Quezada Manuel , López Juana , Casquet Olga , Sierra Miguel A. , de las Mulas Juana Martin
Published: May 2011

A commercially available polyclonal antibody and an avidin-biotin-peroxidase immunohisto-chemical technique were used to detect Brucella abortus antigens in formalin-fixed, paraffin-embedded tissues of lung and liver from 20 aborted bovine fetuses. Thirteen fetuses were obtained from farms with a previous history of brucellosis, and 7 were collected from farms without a history of brucellosis. Among the 13 aborted bovine fetuses obtained from farms with a history of brucellosis, immunoreactivity to B. abortus was detected in lung (9 fetuses) and in liver (1 fetus), whereas Brucella was cultured from abomasal contents in 9 fetuses (8 were immunohistochemically positive). In addition, 11 dams of these 13 aborted bovine fetuses had antibodies to Brucella. Brucella abortus was not detected by immunohistochemistry in the 7 aborted bovine fetuses collected from farms without a history of brucellosis. Bacteriologic culture and serologic tests were also negative for Brucella. The results of this study revealed that the immunohistochemical technique was sufficiently sensitive for detecting B. abortus antigens in formalin-fixed lung tissues from naturally aborted bovine fetuses. Although additional studies are necessary to rule out cross-reaction of the polyclonal antibody with other microorganisms that cause bovine abortion, this immunohistochemical technique could be a complementary tool to serology and bacteriology for the diagnosis of brucellosis.