Showing papers in "Canadian Journal of Veterinary Research-revue Canadienne De Recherche Veterinaire in 2002"

Biofilm bacteria: formation and comparative susceptibility to antibiotics

Abstract: The Calgary Biofilm Device (CBD) was used to form bacterial biofilms of selected veterinary gram-negative and gram-positive pathogenic bacteria from cattle, sheep, pigs, chicken, and turkeys The minimum inhibitory concentration (MIC) and minimum biofilm eradication concentration (MBEC) of ampicillin, ceftiofur, cloxacillin, oxytetracycline, penicillin G, streptomycin, tetracycline, enrofloxacin, erythromycin, gentamicin, tilmicosin, and trimethoprim-sulfadoxine for gram-positive and -negative bacteria were determined Bacterial biofilms were readily formed on the CBD under selected conditions The biofilms consisted of microcolonies encased in extracellular polysaccharide material Biofilms composed of Arcanobacterium (Actinomyces) pyogenes, Staphylococcus aureus, Staphylococcus hyicus, Streptococcus agalactiae, Corynebacterium renale, or Corynebacterium pseudotuberculosis were not killed by the antibiotics tested but as planktonic bacteria they were sensitive at low concentrations Biofilm and planktonic Streptococcus dysgalactiae and Streptococcus suis were sensitive to penicillin, ceftiofur, cloxacillin, ampicillin, and oxytetracycline Planktonic Escherichia coli were sensitive to enrofloxacin, gentamicin, oxytetracycline and trimethoprim/ sulfadoxine Enrofloxacin and gentamicin were the most effective antibiotics against E coli growing as a biofilm Salmonella spp and Pseudomonas aeruginosa isolates growing as planktonic populations were sensitive to enrofloxacin, gentamicin, ampicillin, oxytetracycline, and trimethoprim/sulfadoxine, but as a biofilm, these bacteria were only sensitive to enrofloxacin Planktonic and biofilm Pasteurella multocida and Mannheimia haemolytica had similar antibiotic sensitivity profiles and were sensitive to most of the antibiotics tested The CBD provides a valuable new technology that can be used to select antibiotics that are able to kill bacteria growing as biofilms

Bovine viral diarrhea virus (BVDV) 1b: predominant BVDV subtype in calves with respiratory disease.

Abstract: The prevalence of bovine viral diarrhea virus (BVDV) infections was determined in 2 groups of stocker calves with acute respiratory disease. Both studies used calves assembled after purchase from auction markets by an order buyer and transported to feedyards, where they were held for approximately 30 d. In 1 study, the calves were mixed with fresh ranch calves from a single ranch. During the studies, at day 0 and at weekly intervals, blood was collected for viral antibody testing and virus isolation from peripheral blood leukocytes (PBLs), and nasal swabs were taken for virus isolation. Samples from sick calves were also collected. Serum was tested for antibodies to bovine herpesvirus-1 (BHV-1), BVDV1a, 1b, and 2, parainfluenza 3 virus (PI3V), and bovine respiratory syncytial virus (BRSV). The lungs from the calves that died during the studies were examined histopathologically, and viral and bacterial isolation was performed on lung homogenates. BVDV was isolated from calves in both studies; the predominant biotype was noncytopathic (NCP). Differential polymerase chain reaction (PCR) and nucleic acid sequencing showed the predominant subtype to be BVDV1b in both studies. In 1999, NCP BVDV1b was detected in numerous samples over time from 1 persistently infected calf; the calf did not seroconvert to BVDV1a or BVDV2. In both studies, BVDV was isolated from the serum, PBLs, and nasal swabs of the calves, and in the 1999 study, it was isolated from lung tissue at necropsy. BVDV was demonstrated serologically and by virus isolation to be a contributing factor in respiratory disease. It was isolated more frequently from sick calves than healthy calves, by both pen and total number of calves. BVDV1a and BVDV2 seroconversions were related to sickness in selected pens and total number of calves. In the 1999 study, BVDV-infected calves were treated longer than noninfected calves (5.643 vs 4.639 d; P = 0.0902). There was a limited number of BVDV1a isolates and, with BVDV1b used in the virus neutralization test for antibodies in seroconverting calves' serum, BVDV1b titers were higher than BVDV1a titers. This study indicates that BVDV1 strains are involved in acute respiratory disease of calves with pneumonic Mannheimia haemolytica and Pasteurella multocida disease. The BVDV2 antibodies may be due to cross-reactions, as typing of the BVDV strains revealed BVDV1b or 1a but not BVDV2. The BVDV1b subtype has considerable implications, as, with 1 exception, all vaccines licensed in the United States contain BVDV1a, a strain with different antigenic properties. BVDV1b potentially could infect BVDV1a-vaccinated calves.

Comparison of different methods for diagnosis of porcine proliferative enteropathy.

Abstract: The objectives of this study were: (1) to compare 2 methods of serology; (2) to compare 3 histologic techniques; and (3) to compare 2 methods of detecting shedding in pigs experimentally challenged with Lawsonia intracellularis. The sensitivities of these tests were determined by the detection of infection. Forty 5-week-old pigs were inoculated on day 0 with intestinal homogenate from pigs with proliferative enteropathy (PE). Clinical evaluation was done on day 7 and daily from day 14 to 28 postinoculation. Fecal shedding of L. intracellularis was monitored by use of polymerase chain reaction (PCR) analysis and immunoperoxidase staining at 7-day intervals. Serum was obtained on days 0 and 28 for serologic testing by glass slide and tissue culture indirect fluorescent antibody tests. At euthanasia on day 28, gross intestinal lesions were evaluated and ileum samples collected for histologic analyses. Ileal histologic sections from each animal were stained by hematoxylin and eosin, Warthin-Starry silver stain, and immunohistochemistry (IHC). Of the 40 pigs, 36 had gross lesions typical of PE at necropsy. The percentage of agreement between the 2 serologic methods was 94.4%. Immunoperoxidase stain of fecal smears was more sensitive than PCR for detecting fecal shedding, especially on day 21 (89.5% and 60.5%, respectively) and day 28 (59.4% and 37.5%, respectively) post-inoculation. The IHC stain was much more sensitive for detecting infection than the routinely used hematoxylin and eosin and Warthin-Starry silver stains. In conclusion, in experimentally infected pigs, both serologic methods were appropriate techniques for detecting infection. For fecal samples, PCR has low sensitivity. Immunohistochemistry is the best diagnostic tool for formalin-fixed samples.

Mechanical transmission of porcine reproductive and respiratory syndrome virus throughout a coordinated sequence of events during cold weather

Abstract: Using a field-based model, mechanical transmission of porcine reproductive and respiratory syndrome virus (PRRSV) was assessed throughout a coordinated sequence of events that replicated common farm worker behavior during cold weather (< 0°C). The model involved fomites (boots and containers), vehicle sanitation, transport, and the movement of personnel. A field strain of PRRSV was inoculated into carriers consisting of snow and water, and carriers were adhered to the undercarriage of a vehicle. The vehicle was driven approximately 50 km to a commercial truck washing facility where the driver's boots contacted the carriers during washing, introducing the virus to the vehicle interior. The vehicle was then driven 50 km to a simulated farm site, and the driver's boots mechanically spread virus into the farm anteroom. Types of containers frequently employed in swine farms (styrofoam semen cooler, metal toolbox, plastic lunch pail, and cardboard animal health product shipping parcel) contacted drippings from footwear on the anteroom floor. The truck wash floor, vehicle cab floor mats, boot soles, anteroom floor, and the ventral surface of containers were sampled to track the virus throughout the model. Ten replicates were conducted, along with sham-inoculated controls. At multiple sampling points PRRSV nucleic acid was detected in 8 of 10 replicates. In each of the 8 PCR-positive replicates, infectious PRRSV was detected on the surfaces of containers by virus isolation or swine bioassay. All sham-inoculated controls were negative. These results indicate that mechanical transmission of PRRSV can occur during coordinated sequence of events in cold weather.

Evaluation of health status of calves and the impact on feedlot performance: assessment of a retained ownership program for postweaning calves

Abstract: The objective of this study was to evaluate animal health status at entry to a feedlot against feedlot performance and carcass value. There were 24 herds represented by 417 calves in a retained ownership program. The health status at entry was represented by the levels of serum antibody to infectious bovine rhinotracheitis virus (IBRV), bovine viral diarrhea viruses 1 and 2 (BVDV1a, BVDV2), parainfluenza 3 virus (PI3V), bovine respiratory syncytial virus (BRSV), Mannheimia haemolytica, and Pasteurella multocida, as well as by the presence of virus in nasal swabs and blood leukocytes and the presence of bacteria in nasal swabs. The presence or absence of viruses or bacteria at entry did not predict subsequent illness. However, there were predictors of illness severity (number of treatments) and performance parameters of feedlot performance. Herds with a low morbidity rate had higher levels of BVDV1a antibodies than herds with a high morbidity rate. On both an individual-animal and a herd-average basis, calves with low levels of antibody to BVDV1a and BVDV2 had increased total treatment costs. Also, for individual animals and the herd as a whole, low levels of antibody to P. multocida, BVDV1a, and BVDV2 were related to decreased net value to owner (carcass value minus total feedlot cost). Calves treated twice or more had lower levels of antibody to BVDV1a than those treated once or not at all. Differences in herd morbidity rate and treatment costs were more related to appropriate timing of vaccine (last dose at or near delivery of calf) or lack of a 2nd dose of killed vaccine. This was best illustrated by the levels of antibody to BVDV1a. The results of this study were used to formulate recommendations for the subsequent year.

Presentation of postweaning Escherichia coli diarrhea in southern Ontario, prevalence of hemolytic E. coli serogroups involved, and their antimicrobial resistance patterns.

Abstract: Post-weaning Escherichia coli diarrhea (PWECD) in Ontario was investigated using a case-control study involving 50 Ontario nurseries. The clinical signs and the impact on productive parameters were determined by means of a producer survey. The hemolytic E. coli serogroups involved in PWECD (O149:K91:K88) were examined in this study. Based on a polymerase chain reaction test, the hemolytic E. coli from 82% of the case herds were positive for 3 enterotoxins (STa, STb, and LT), those from 12% of the case herds were positive for STb and LT only, and those from one herd (6%) were positive for 3 enterotoxins, as well as for verotoxin and F18 pili. The E. coli involved in disease were resistant to multiple antibiotics. Case farms commonly used a wide variety of antibiotics either in the feed or water, or as injectable drugs. The most common antibiotic used to treat PWECD on the study farms was apramycin, but evidence of resistance to this antibiotic was noted. The PWECD problem was commonly seen within a week of weaning but onset of diarrhea was reported as late as the grower-finisher stage. Growth rate was poorer in case herds and mortality was higher than in control herds, demonstrating that PWECD is an economically important disease in Ontario.

Occurrence and characterization of resistance to extended-spectrum cephalosporins mediated by β-lactamase CMY-2 in Salmonella isolated from food-producing animals in Canada

Abstract: Resistance of Salmonella to extended-spectrum cephalosporins (ESCs) is being reported with increasing frequency. In humans, infections with Salmonella resistant to ESCs threaten the efficacy of ceftriaxone, the drug of choice for treating salmonellosis in children. To determine the occurrence of resistance to ESCs, we examined 8426 strains isolated from food-producing animals in Canada in 1994–99 for reduced susceptibility or resistance to ceftriaxone. Of the 8 such strains identified (7 from turkeys and 1 from cattle), 5 had reduced susceptibility, and 3 were resistant; 2 were isolated in 1995, 1 was isolated in each of 1996 and 1997, and 4 were isolated in 1999. Isoelectric focusing showed that all 8 isolates produced a β-lactamase with a pI ≥ 9. The strains were resistant to cefoxitin and not inhibited by clavulanic acid. Primers specific for the Citrobacter freundii blaAmpC gene produced the expected product in the polymerase chain reaction. DNA sequencing showed that all isolates possessed the blaCMY-2 gene. Plasmid DNA from all 8 isolates transformed Escherichia coli DH10B, whereas only 1 isolate transferred blaCMY-2 conjugally. All transformants and the transconjugant were resistant to ampicillin, cefoxitin, ceftiofur, cephalothin, streptomycin, sulfisoxazole, and tetracycline. Southern blots of plasmids from the isolates, the transformants, and the transconjugant showed that blaCMY-2 was located on similar-sized plasmids (60 or 90 MDa) in the transformants and the transconjugant. In the S. Typhimurium DT104 and S. Ohio isolates, the floSt gene was found on the same plasmid. Class 1 integrons with the aadB gene cassette were detected in the S. Bredeney isolates but not in their transformants or the transconjugant. Pulsed-field gel electrophoresis and plasmid profiles indicated that both clonal dispersion and horizontal transfer of blaCMY-2 may have caused dissemination of the resistance determinant.

Neurohormonal and metabolic effects of medetomidine compared with xylazine in beagle dogs

Abstract: This study was aimed to investigate and compare the effects of medetomidine and xylazine on the blood level of some stress-related neurohormonal and metabolic variables in clinically normal dogs, especially focusing on time and dose relations of the effects. A total of 9 beagle dogs were used for 9 groups, which were treated with physiological saline solution (control), 10, 20, 40, and 80 microg/kg medetomidine, and 1, 2, 4, and 8 mg/kg xylazine, intramuscularly. Blood samples were taken at 10 times during 24 h from a central venous catheter. Plasma norepinephrine, epinephrine, cortisol, glucose, insulin, glucagon, and nonesterified fatty acid concentrations were determined. Both medetomidine and xylazine similarly and dose-dependently inhibited norepinephrine release and lipolysis. Medetomidine suppressed epinephrine release dose-dependently with greater potency than xylazine. Xylazine also tended to decrease epinephrine levels dose-dependently. The cortisol and glucagon levels did not change significantly in any treatment group. Both drugs suppressed insulin secretion with similar potency. Both medetomidine and xylazine increased glucose levels. The hyperglycemic effect of medetomidine, in contrast with xylazine, was not dose-dependent at the tested dosages. The results suggested that the effect of medetomidine on glucose metabolism may not be due only to alpha2-adrenoceptor-mediated actions.

Mechanical transmission of porcine reproductive and respiratory syndrome virus by mosquitoes, Aedes vexans (Meigen).

Abstract: The objective of this study was to determine whether porcine reproductive and respiratory syndrome virus (PRRSV) could be transmitted to naive pigs by mosquitoes following feeding on infected pigs. During each of 4 replicates, mosquito-to-pig contact took place on days 5, 6, and 7 after PRRSV infection of the donor pig. A total of 300 mosquitoes [Aedes vexans (Meigen)] were allowed to feed on each viremic donor pig, housed in an isolation room. After 30 to 60 s, feeding was interrupted, and the mosquitoes were manually transferred in small plastic vials and allowed to feed to repletion on a naive recipient pig housed in another isolation room. Prior to contact with the recipient pig, the mosquitoes were transferred to clean vials. Swabs were collected from the exterior surface of all vials, pooled, and tested for PRRSV. Separate personnel handled the donor pig, the recipient pig, and the vial-transfer procedure. Transmission of PRRSV from the donor to the recipient pig occurred in 2 out of 4 replicates. The PRRSV isolated from the infected recipient pigs was nucleic-acid-sequenced and found to be 100% homologous with the virus used to infect the donor pigs. Homogenates of mosquito tissues collected in all replicates were positive by either polymerase chain reaction or swine bioassay. All control pigs remained PRRSV negative, and PRRSV was not detected on the surface of the vials. This study indicates that mosquitoes (A. vexans) can serve as mechanical vectors of PRRSV.

Physiology and behavior of dogs during air transport.

Abstract: Twenty-four beagles were used to measure physiological and behavioral reactions to air transport. Each of 3 groups of 4 sedated (with 0.5 mg/kg body weight of acepromazine maleate) and 4 non-sedated (control) dogs was flown on a separate flight between Montreal, Quebec, and Toronto, Ontario, after being transported by road from Quebec City to Montreal. Saliva and blood samples were taken before ground and air transport and after air transport. The heart rate was monitored during the whole experiment except during ground transport, and behavior was monitored by video during air transport. Sedation did not affect any of the variables measured. The mean plasma cortisol concentration was significantly higher (P < 0.05) after ground transport than at baseline (225.3 vs 134.5 nmol/L); the mean salivary cortisol concentration was significantly higher (P < 0.05) after both ground and air transport than at baseline (16.2 and 14.8, respectively, vs 12.6 nmol/L). The mean neutrophil count was significantly higher (P < 0.05) after both ground and air transport than at baseline (80.6 and 81.4, respectively, vs 69.5 per 100 white blood cells), whereas the mean lymphocyte count was significantly lower (P < 0.05) (13.2 and 13.7, respectively, vs 22.4 per 100 white blood cells). Loading and unloading procedures caused the largest increase in heart rate. On average, the dogs spent more than 50% of the time lying down, and they remained inactive for approximately 75% of the time, except during take-off. These results suggest that transportation is stressful for dogs and that sedation with acepromazine, at the dosage and timing used, does not affect the physiological and behavioral stress responses of dogs to air transport.

Benign esophageal stricture in the dog and cat: a retrospective study of 20 cases.

Abstract: Twenty animals with benign esophageal strictures are presented. Most of the esophageal strictures were thought to be related to gastroesophageal reflux during ovariohysterectomy and were located at the distal portion of the thoracic esophagus (caudal to the base of the heart). For the dilation procedure, the endoscope tip or a balloon catheter was used and the outcome was generally considered to be good. The endoscope tip was an adequate instrument for dilation in some cases.

Serologic follow-up of a repopulated swine herd after an outbreak of proliferative hemorrhagic enteropathy

Abstract: Lawsonia intracellularis is an obligate intracellular organism that causes porcine proliferative enteropathy, a widespread infectious disease. Very little is known about the immune response and the epidemiologic features of the disease in the field. The aims of this study were to evaluate the duration and titers of antibody specific for L. intracellularis in gilts after an outbreak of proliferative hemorrhagic enteropathy (PHE), to evaluate maternal antibodies in piglets, and to evaluate seroconversion and fecal shedding in growing–finishing pigs. Thirty-six gilts in a herd that had recently experienced an outbreak of PHE, including 13 that had recovered, were bled 3 wk after the beginning of the outbreak and then every 3 wk until they became seronegative in 2 consecutive tests. Fourteen piglets from 5 gilts seropositive at farrowing and 5 piglets from 2 sows that remained seronegative were bled once or twice at the farrowing house and then every 3 wk until they reached market age. Fecal samples from these pigs were tested by polymerase chain reaction at 7 wk of age and then on the days of blood collection. After the PHE outbreak, the gilts had high serum antibody levels; the levels decreased over time, but antibody was still detectable for up to 3 mo in some animals. Four piglets from sows that were seropositive at farrowing had detectable passive antibodies up to 5 wk of age. Some nursery pigs started shedding L. intracellularis around 7 wk of age; peak shedding was observed between 13 and 16 wk. Antibody was not detected until 16 wk of age and was more often detected between 19 and 22 wk.

Pathogenic Shiga toxin-producing Escherichia coli in the intestine of calves.

Abstract: The purpose of this study was to compare the pathological effects of Shiga toxin-producing Escherichia coli (STEC) that vary in their association with bovine and human disease. Shiga toxin-producing E. coli of serotypes associated with both dysentery in calves and hemolytic uremic syndrome (HUS) in humans (O5:H-, O26:H11, O111:H-,O113:H21) were compared with O157:H7 STEC, which are associated with HUS in humans but not with disease in calves. The STEC were administered orally to 80 day-old chicks and into ligated loops in the ileum and colon of four 2- to 6-day-old calves. Examination of the ceca of the chickens 10 d postchallenge showed no adherence or tissue abnormality for any isolate. The calves were euthanized 8 to 10 h postinoculation, and sections of the intestinal loops were examined by light microscopy, transmission and scanning electron microscopy, and immunohistochemistry. All strains showed consistent focal adherence associated with mild lesions in the colon. Attaching and effacing lesions were observed with the eae-positive strains. Ileal lesions were similar to the colonic ones but were sometimes severe, with marked polymorphonuclear leukocyte proliferation in the lamina propria. It is concluded that chickens were unsuitable for studying interaction of STEC with the intestine and that there was no difference in the interaction of the ligated calf intestine with STEC of serotypes associated with disease in calves compared with O157:H7 STEC.

The use of negative binomial modelling in a longitudinal study of gastrointestinal parasite burdens in Canadian dairy cows

Abstract: The epidemiology of bovine gastrointestinal nematodes was investigated through a 1-year (October 1999 to September 2000) longitudinal study in 38 Canadian dairy herds from 4 different provinces (Prince Edward Island, Quebec, Ontario, Saskatchewan). For each herd, fecal egg counts from 8 randomly selected animals were performed on a monthly or quarterly basis. Larval cultures were performed once, to determine the species breakdown of the parasites. All producers were interviewed regarding herd management practices. The observed fecal egg counts were low in this study, with a range from 0 to 419 nematode eggs per 5 g of feces. The mean count was 9.8 and the median was 1. Standard transformations failed to normalize the data, which followed an over-dispersed Poisson distribution. A zero inflated negative binomial model was applied to assess factors that would influence the fecal egg counts. Identified associations were: egg counts were lowest in the winter and highest in the late spring; first-lactation cattle had higher counts than older cows; if manure was spread mechanically on pastures used by lactating cattle the egg counts were higher; and if manure was spread on heifer-pastures, the adult cows had lower counts. In herds where pasture use was more extensive, the cattle had higher fecal egg counts. The difference in pasture exposure was found to be a main contributor to an observed difference in fecal egg counts among herds in the 4 provinces.

Foot-and-mouth disease eradication efforts in the Republic of Korea.

Abstract: On March 20, 2000, a suspected vesicular disease in cattle was reported to the National Veterinary Research and Quarantine Service (NVRQS) of the Republic of Korea. This represented the index case of a foot-and-mouth disease (FMD) outbreak, which spread through several provinces. The Republic of Korea had been free of FMD for 66 years prior to the reintroduction of the virus and had recently suspended imports of pork and pork products from neighboring Japan owing to a reported FMD outbreak in that country. The Korean outbreak was ultimately controlled through the combination of preemptive slaughter, animal movement restrictions, and a strategy of ring vaccination. The purpose of this paper is to review the current FMD situation in Korea in the aftermath of its 2000 epizootic and how it may affect future efforts to eradicate or reduce risk of reintroduction of the disease into Korea.

Sequence analysis of old and new strains of porcine circovirus associated with congenital tremors in pigs and their comparison with strains involved with postweaning multisystemic wasting syndrome

Abstract: The entire genomes of 7 isolates of porcine circovirus (PCV) from pigs with congenital tremors (CT), type A2, or postweaning multisystemic wasting syndrome (PMWS) were cloned and sequenced. One isolate (CT-PCV-P7) originated from the late 1960s from a neonatal pig with CT, type A2. Two recent PCV isolates (CT-PCV-P5, CT-PCV-P6) were from 2 affected neonatal pigs, from different farms, with unrelated outbreaks of CT; type A2. Four isolates (PMWS-PCV-P1, PMWS-PCV-P2, PMWS-PCV-P3, PMWS-PCV-P4) originated from pigs with PMWS from 4 different farms. A comparative analysis of these PMWS and PCV isolates demonstrated 99% sequence identity with each other, and over 96% sequence identity with previously sequenced PCV2 isolates. The CT-PCV-P5 and CT-PCV-P6 isolates, however, shared 99% of the same identity with each other, and interestingly also with PMWS PCV isolates. There were no consistent genomic differences between PMWS and recent CT isolates. The CT-PCV-P7 showed 98% identity similarity to PK-15-derived PCV1 and demonstrated only 72% identity similarity to either CT-PCV-P5 or CT-PCV-P6. Phylogenetic analysis confirmed that the old isolate (CT-PCV-P7), and the new isolates (CT-PCV-P5, CT-PCV-P6, PMWS-PCV-P1, PMWS-PCV-P2, PMWS-PCV-P3, PMWS-PCV-P4) were correctly classified as PCV1 and PCV2, respectively.

Cliniconeuropathologic findings of familial frontal lobe epilepsy in Shetland sheepdogs

Abstract: We examined an epileptic focus by electroencephalography (EEG) by using an international 10-20 electrode system in 11 Shetland sheep dogs affected with familial idiopathic epilepsy. We also performed an evaluation of the amino acids in the cerebrospinal fluid (CSF) and a pathologic examination of the brains of 8 dogs that died from status epilepticus. Continuous electroencephalography demonstrated that an epileptic focus was initially detected in the frontal lobe, particularly the internal area, and that paroxysmal foci developed diffusely in other lobes of affected dogs with recurrent convulsions. The EEG analyses indicated spike and sharp wave complexes, which were considered to be paroxysmal discharges. An increased value for glutamate or aspartate was found in the CSF of some epileptic dogs. Histologically, acute neuronal necrosis and astrocytosis were distributed predominantly in the cingulate cortex and internal area of frontal cortex, less frequently in other areas of the cerebrum. The results of this study suggest that, initially, the dogs have an epileptic focus in the frontal lobe, and that the focus extends gradually to other areas of the cerebrum. Based on the distribution of neuronal necrosis and astrocytosis, acute neuronal damage may be related to the superexcitation of neurons following epilepsy.

Flaxseed (Linum usitatissimum) supplementation associated with reduced skin test lesional area in horses with Culicoides hypersensitivity

Abstract: The purpose of this study was to quantify the effect of flaxseed (Linum usitatissimum) supplementation on the skin test response of atopic horses. Six horses that displayed a positive skin test for allergy to extract from Culicoides sp. participated in the 42-day, placebo-controlled, double-blind, cross-over trial. Results showed that supplementation with flaxseed for 42 days in our experimental horses reduced the mean skin test response to Culicoides sp. This observation was concurrent with a significant decrease in the long-chain saturated fatty acids; behenic acid (22:0) and lignoceric acid (24:0), in the hair of horses receiving flaxseed. There was also a significant decrease in aspartate aminotransferase, and increase in serum glucose in the treatment animals at specific sampling points. It was concluded that; in this small pilot study, flaxseed was able to reduce the lesional area of the skin test response of atopic horses, alter the fatty acid profile of the hair, reduce inflammation, and did not elicit any negative side-effects in the experimental horses.

Improvements required for the detection of Salmonella Pullorum and Gallinarum.

Abstract: Detection of the specific Salmonella serovar Gallinarum, which is divided into the biovars Pullorum and Gallinarum, is compulsory under the national hygienic and sanitary control regulations of France for breeding flocks whose offspring are exported. Our aim was to examine the suitability of bacteriologic and serologic methods routinely used in France to screen serum samples and organs for S. Gallinarum. Since bacteriologic reference techniques are designed to isolate the commonly occurring non-typhoid serovars, such as S. Typhimurium, S. Enteritidis, and others that cause outbreaks of foodborne illness, they may not be particularly suitable for detecting S. Pullorum and S. Gallinarum. This hypothesis was confirmed by the inoculation of 10-wk-old chickens and 1-d-old chicks with various strains of S. Pullorum and S. Gallinarum. The most reliable enrichment media were selenite cystine and Rappaport-Vassiliadis broths. Moreover, on the usual plating media, colonies were small, grew more slowly than the common serovars (in 48 h instead of 24 h), and had an unusual appearance. Since the rapid slide agglutination (RSA) test is based only on antigens from standard and variant strains of S. Pullorum, it may not readily detect S. Gallinarum. In our study, it detected infection in all 10-wk-old chickens inoculated with S. Pullorum strains but did not detect any antibodies against S. Gallinarum. Therefore, S. Gallinarum antigens must be added to the S. Pullorum antigens used in the RSA test in order to detect antibodies produced by birds infected with either biovar.

Assessing the duration of persistence and shedding of porcine reproductive and respiratory syndrome virus in a large population of breeding-age gilts

Abstract: Porcine reproductive and respiratory syndrome virus (PRRSV) is an RNA virus in the order Nidovirales, family Arteriviridae, genus Arterivirus. The virus induces a prolonged viremia, replicates in macrophages, and produces persistent infection. The purpose of this study was to determine if PRRSV could persist for 90 d or more in a large population of breeding-age gilts housed under environmental conditions typical of commercial swine production and to determine if experimentally infected gilts could shed virus to naive sentinel gilts beyond 90 d postinfection. Using the intranasal route, we inoculated 120 PRRSV-naive gilts, 4 mo of age, with 5 mL of cell culture fluid containing a total dose of 10(2.4) TCID50 of a field isolate (MN-30100) of PRRSV. The index gilts were organized into 3 groups (A, B, and C), 40 gilts per group. To assess the dynamics of the experimental infection, a monitor group of 30 index gilts was blood-tested on days 0, 3, 7, 14, 30, 60, 90, 120, 150, and 180 postinfection. PRRSV viremia was detected with the polymerase chain reaction (PCR) on days 3, 7, and 14 and by virus isolation (VI) on days 7 and 14. PRRSV antibodies were detected from day 14 by enzyme-linked immunosorbent assay (ELISA). To assess shedding, 30 PRRSV-naive sentinel gilts were commingled with the index gilts on day 90 postinfection and tested by PCR, VI, and ELISA every 15 d until 180 d postinfection; all samples were negative. To assess persistence, 40 index and 10 sentinel gilts were slaughtered at 120 (group A), 150 (group B), or 180 (group C) d postinfection. Evidence of PRRSV was not detected by PCR or VI in any tissue samples from the 120 index gilts. These results indicate that persistence and shedding of PRRSV are of short duration in breeding-age gilts.

Validation of 2 commercial Neospora caninum antibody enzyme linked immunosorbent assays.

Abstract: This is a validation study of 2 commercially available enzyme linked immunosorbent assays (ELISA) for the detection of antibodies against Neospora caninum in bovine serum. The results of the reference sera (n = 30) and field sera from an infected beef herd (n = 150) were tested by both ELISAs and the results were compared statistically. When the immunoblotting results of the reference bovine sera were compared to the ELISA results, the same identity score (96.67%) and kappa values (K) (0.93) were obtained for both ELISAs. The sensitivity and specificity values for the IDEXX test were 100% and 93.33% respectively. For the Biovet test 93.33% and 100% were obtained. The corresponding positive (PV+) and negative predictive (PV−) values for the 2 assays were 93.75% and 100% (IDEXX), and 100% and 93.75% (Biovet). In the 2nd study, competitive inhibition ELISA (c-ELISA) results on bovine sera from an infected herd were compared to the 2 sets of ELISA results. The identity scores of the 2 ELISAs were 98% (IDEXX) and 97.33% (Biovet). The K values calculated were 0.96 (IDEXX) and 0.95 (Biovet). For the IDEXX test the sensitivity and specificity were 97.56% and 98.53%, whereas for the Biovet assay 95.12% and 100% were recorded, respectively. The corresponding PV+ and PV− values were 98.77% and 97.1% (IDEXX), and 100% and 94.44% (Biovet). Our validation results showed that the 2 ELISAs worked equally well and there was no statistically significant difference between the performance of the 2 tests. Both tests showed high reproducibility, repeatability and substantial agreement with results from 2 other laboratories. A quality assurance based on the requirement of the ISO/IEC 17025 standards has been adopted throughout this project for test validation procedures.

Recombinant equine interleukin-1β induces putative mediators of articular cartilage degradation in equine chondrocytes

Abstract: Interleukin-1 is considered a central mediator of cartilage loss in osteoarthritis in several species, however an equine recombinant form of this cytokine is not readily available for in vitro use in equine osteoarthritis research Equine recombinant interleukin-1β was cloned and expressed and its effects on the expression and activity of selected chondrocytic proteins implicated in cartilage matrix degradation were characterized Reverse transcriptase polymerase chain reaction methods were used to amplify the entire coding region of the equine IL-1β mRNA, which was cloned into an expression vector, expressed in E coli, and purified using a Ni2+ chromatographic method The effects of the recombinant peptide on chondrocyte gene expression were determined by Northern blotting using RNA from equine chondrocyte cultures hybridized to probes for matrix metalloproteinases (MMP 1, MMP 3, MMP 13), tissue inhibitor of matrix metalloproteinases 1 (TIMP 1) and cyclooxygenase 2 (COX 2) Effects on selected mediators of cartilage degradation (nitrite concentrations and MMP activity) were determined using conditioned medium from reIL-1β-treated equine cartilage explant cultures A recombinant peptide of approximately 21 kd was obtained Northern blotting analyses revealed a marked up-regulation of expression of all MMPs, TIMP 1, and COX 2 in mRNA from treated chondrocytes Furthermore, cartilage explants exposed to reIL-1β had augmented collagenase/gelatinase and stromelysin activities as well as increased concentration of nitrite in conditioned media The development of a biologically active, species-specific IL-1β provides a valuable tool in the study of osteoarthritis pathophysiology and its treatment in horses

Antibody response to an autogenous vaccine and serologic profile for Streptococcus suis capsular type 1/2

Abstract: An autogenous vaccine was developed, using sonicated bacteria, with a strain of Streptococcus suis capsular type 1/2. The objectives of this study were to evaluate the antibody response following vaccination and to assess the changes in antibody levels in pigs from a herd showing clinical signs of S. suis capsular type 1/2 infection in 6- to 8-week-old pigs. An enzyme-linked immunosorbent assay using the vaccine antigen was standardized. Results from a preliminary study involving 2 control and 4 vaccinated 4-week-old pigs indicated that all vaccinated pigs produced antibodies against 2 proteins of 34 and 43 kDa, respectively, and, in 3 out of 4 vaccinated pigs, against the 117-kDa muramidase-released protein. For the serologic profile, groups of 30 pigs from the infected herd were blood sampled at 2, 4, 6, 8, and 10 weeks of age. The lowest antibody level was observed between weeks 6 and 8, presumably corresponding to a decrease in maternal immunity. A marked increase was seen at 10 weeks of age, shortly after the onset of clinical signs in the herd. For the vaccination field trial, newly weaned, one-week-old piglets were divided into 2 groups of 200 piglets each (control and vaccinated); blood samples were collected from 36 piglets in each group at 2-week intervals for 12 weeks. A significant increase (P < 0.05) in antibody response was observed 4 weeks following vaccination and the level of antibodies stayed high until the end of the experiment. In the control group, the increase was only observed at 13 weeks of age, probably in response to a natural infection. The response to the vaccine varied considerably among pigs and was attributed, in part, to the levels of maternal antibodies at the time of vaccination. No outbreak of S. suis was observed in the control or vaccinated groups, so the protection conferred by the vaccine could not be evaluated.

Seroprevalence of porcine circovirus type 2 in swine populations in Canada and Costa Rica.

Abstract: Porcine circovirus (PCV) was recently divided into 2 antigenically distinct types that differ (65% amino acid identity) in the protein encoded by open reading frame 2 (ORF2). Porcine circovirus 1 is apparently non-pathogenic and, in contrast, PCV2 is associated with porcine multisystemic wasting syndrome (PMWS). Our objective was to determine the extent of exposure of normal pigs in Canada and Costa Rica to PCV2. Recombinant DNA techniques were used to produce an antigen from ORF2 of PCV2 that was suitable for the detection of antibody in swine sera. The presence of PCV2 nucleotide sequences was detected using polymerase chain reaction (PCR) techniques. Using these tests, specific antibody and nucleotide sequences were demonstrated in sera from a cohort of pigs during a PMWS outbreak. Antibody was detected in normal, healthy hogs slaughtered in Canada (82.4% of 386) and in Costa Rica (14.6% of 322). This is the first report indicating the presence of PCV2 in Latin America. More than 50% of these sera also contained PCV2 nucleotide sequence. Although these hogs were healthy when slaughtered, they were infected with PCV2 and may have previously been ill. The widespread occurrence of PCV2 in swine suggests that this virus is adapted to replication in porcine tissue.

Comparison of a central and a peripheral (cephalic vein) injection site for the measurement of cardiac output using the lithium-dilution cardiac output technique in anesthetized dogs.

Abstract: The objective of this study was to determine the agreement between cardiac output measured by central (cranial vena cava) versus peripheral (cephalic vein) venous injection of lithium chloride for lithium-dilution cardiac output (LiDCO) determination in the dog. Five dogs (2 males, 3 females), anesthetized with halothane, were used. With each dog, 12 alternating central and peripheral LiDCO measurements were made, resulting in 10 paired comparisons. A total of 50 comparisons were obtained, the cardiac output measurements ranging from 1.11 to 2.76 L/min. The LiDCO measurement from the cephalic vein was similar to that obtained from the recommended central venous site: the difference between the central and cephalic vein determinations for all measurements was 0.098 ± 0.336 L/min (mean ± 2 standard deviations). Linear regression analysis demonstrated a slope of 1.050 (95% confidence interval 0.904 to 1.196) and a y intercept of 0.005 (r = 0.902). Therefore, although the central venous site is recommended by the manufacturer, the cephalic vein can be used instead in the dog, eliminating the need for central venous catheterization and thus reducing time and expense.

Comparison of embryonated chicken eggs with MDCK cell culture for the isolation of swine influenza virus.

Abstract: Embryonated chicken eggs (ECE) and the Madin-Darby canine kidney (MDCK) cell line were compared for isolation of swine influenza virus (SIV) from nasal swabs and tissue samples. Samples originated from 30 pigs experimentally inoculated with 2 x 106 to 2 x 10(7) embryo infectious dose 50% (EID50)/mL of swine influenza strain A/Swine/Indiana/1726/88 (H1N1). The results were analyzed with McNemar's chi-squared test for symmetry. The results indicated that more samples were SIV-positive with ECE than with tissue culture (P < 0.001), suggesting that ECE remains the system of choice for isolation of SIV. It is recommend that routine use of both SIV isolation systems will increase the sensitivity of detection of virus shedding by considering the differences in growth and tropism of diverse SIV strains.

Field study of bovine coronavirus vaccine enriched with hemagglutinating antigen for winter dysentery in dairy cows.

Abstract: A field study of a vaccine; prepared by solubilizing cells infected with bovine coronavirus, Triton X-100, and mixing with an oil adjuvant, was performed at 9 farms over 4 prefectures. The cattle tested were Holstein dairy cows aged 2 to 10 years. A vaccination group consisted of 157 animals (including 132 pregnant cows) and a non-vaccinated control group consisted of 50 animals. The cows received 2 intramuscular injections of vaccine (2 mL) at 3-week intervals. Vaccinated cows did not develop abnormalities, such as a decrease in milk production volume, and all pregnant animals calved normally. The geometric mean of the hemagglutination inhibition antibody titer was 34.2 before vaccination in test cows. The titer had increased to 105.6, 3 weeks after the 1st injection and peaked at 755.6, 1 month after the 2nd injection. A high antibody titer persisted at 396.0; 241.0; and 201.5, at 3, 6, and 9 months after the 2nd injection, respectively. This confirms the safety and high antibody-response induced by this prototype vaccine. Therefore, this vaccine may be useful for the prevention of winter dysentery caused by bovine coronavirus infection.

Comparison of reverse transcription polymerase chain reaction, immunohistochemistry, and in situ hybridization for the detection of porcine epidemic diarrhea virus in pigs

Abstract: Reverse transcription polymerase chain reaction (RT-PCR), immunohistochemistry, and in situ hybridization were compared for the detection of porcine epidemic diarrhea virus (PEDV). Fifteen piglets experimentally infected with PEDV were used in the study. In addition, 94 diarrheic piglets submitted to the Department of Veterinary Pathology in Seoul National University for diagnosis of PEDV infection were used to compare the 3 methods. Antigen and nucleic acid of PEDV were detected in 15/15, 13/15, and 14/15 of the intestinal and fecal samples from the PEDV-inoculated pigs by RT-PCR, immunohistochemistry, and in situ hybridization, respectively. The virus was isolated from 15/15 of the jejunal samples from the PEDV-inoculated pigs. Neither PEDV antigen nor PEDV nucleic acid was detected in the intestinal and fecal samples from mock-infected control pigs. Of the 94 samples, 63 were positive for PEDV by all 3 techniques. Six samples were positive for PEDV by immunohistochemistry and in situ hybridization. Three samples were positive for PEDV by in situ hybridization and RT-PCR. Seven samples were positive for PEDV by RT-PCR. Although RT-PCR identified the presence of PEDV more frequently than the other methods, when only formalin-fixed tissues are submitted, immunohistochemistry and in situ hybridization would be useful methods for the detection of PEDV antigen and nucleic acid.

Changes in the prevalence of resistant Escherichia coil in cattle receiving subcutaneously injectable oxytetracycline in addition to in-feed chlortetracycline compared with cattle receiving only in-feed chlortetracycline.

Abstract: Information about the prevalence of antibiotic resistance in commensal enteric bacteria is of interest because these bacteria are potential indicators of selection pressure on enteric bacteria and represent a reservoir of resistance genes in potentially pathogenic bacteria. This study reports changes in the prevalence of resistance to antibiotics in commensal Escherichia coli from cattle receiving either subcutaneously injectable oxytetracycline in addition to in-feed chlortetracycline or only in-feed chlortetracycline. Resistance to 19 antibiotics was examined. The use of injectable oxytetracycline in addition to in-feed chlortetracycline was significantly associated (P < 0.05) with an increase in the prevalence of resistance only to chloramphenicol and sulfisoxazole.

Decreases in serum apolipoprotein B-100 and A-I concentrations in cows with milk fever and downer cows.

Abstract: Milk fever occurring during the peripartum period has been suggested to be caused by fatty liver developed during the non-lactating stage because diseased cows have increased serum concentrations of non-esterified fatty acids (NEFA) and show hepatic lipidosis. In cows with fatty liver and related diseases such as ketosis, serum concentrations of apolipoprotein (apo) B-100 and apoA-I are decreased. The purpose of the present study was to examine whether apoB-100 and apoA-I concentrations are similarly decreased in cows with milk fever. Apolipoprotein concentrations were also measured in cows with downer syndrome, which has been suggested to be related, at least in part, to milk fever. Compared with healthy cows during early lactation, apoB-100 and apoA-I concentrations were decreased in cows with milk fever and also in downer cows. In cows with milk fever, the decreases in apoB-100 and apoA-I concentrations were associated with increased NEFA and decreased cholesterol and phospholipid concentrations. However, in downer cows, serum lipid concentration changes were not as distinct as in cows with milk fever. These results, coupled with previous findings on the decreases in apoB-100 and apoA-I concentrations of cows with fatty liver-related diseases, suggest that fatty liver is involved in the development of milk fever and partly in that of downer cow syndrome.