Genetic analysis of vesicular stomatitis virus-New Jersey from the 1995 outbreak in the western United States

OBJECTIVE: To compare molecular associations between the vesicular stomatitis virus (VSV)-New Jersey isolates of the 1995 outbreak with those from previous outbreaks between 1982 and 1985 in the western United States. SAMPLE POPULATION: 23 virus isolates considered representative of the 1995 outbreak of vesicular stomatitis. PROCEDURE: Viral gene coding for surface-envelope protein G was evaluated by use of nucleotide sequencing and phylogenetic analysis. RESULTS: Changes in up to 0.77% of the nucleotide bases and 1.35% of the amino acids were detected among the 1995 viral isolates, whereas changes in up to 3.2 and 2.9% of the nucleotides and amino acids, respectively, were found, compared with the 1982 to 1985 viruses. Insertions or deletions were not found in the entire gene, which spanned 1,554 nucleotide bases. CONCLUSIONS AND CLINICAL RELEVANCE: Phylogenetic analysis indicated that the 1995 VSV-New Jersey belongs to a lineage distinct from that of the 1982 to 1985 viruses that caused previous outbreaks in the western United States. Furthermore, it also is distinct from strains from Central America and from the Georgian Hazelhurst strain.     

The influence of Mannheimia haemolytica A1 seed culture inoculum cell density on the production of leukotoxin in submerged culture supernatant

Mannheimia haemolytica leukotoxin is produced during the logarithmic growth phase in submerged culture in RPMI 1640 medium with and without the addition of foetal calf serum or albumin. In order to establish a pattern of optimal leukotoxin production in small volumes in submerged cultures and to define some parameters involved, two high leukotoxin producing Mannheimia haemolytica strains were grown in RPMI 1640 medium containing either FCS or BSA. The cell growth and leukotoxin production abilities of each strain were determined concomitantly every hour in RPMI 1640 medium containing each of the additives over a time period of 6 h. The growth performance of three dilutions of a standardized seed culture inoculum prepared with each of the cultures and additives were simultaneously compared with each other using the above parameters. The different seed culture inoculum dilutions had a definite effect on the time and quantity of leukotoxin production. Both strains demonstrated peak leukotoxin production after 4 h of active growth. The addition of albumin to both isolates gave slightly increased leukotoxin levels, and both showed that the peak leukotoxin was not associated with peak cell concentration. Obvious quantitative differences in the ability of different M. haemolytica strains to produce leukotoxin were noted. Strain 12296 produced optimal leukotoxin concentration from the medium (1/25) dilution of the seed culture inoculum after 4 h, whereas strain 1/10 produced the same concentration with the low (1/5) dilution seed culture inoculum, possibly reflecting the superior production ability of the first strain. However, each strain of M. haemolytica appeared to have its own specific logarithmic cell growth and leukotoxin production pattern. The peak cell density of M. haemolytica grown in submerged RPMI 1640 culture medium cannot be used as an indication of optimal leukotoxin levels.     

Hendra and Nipah virus infections.

The most important clinical and pathological manifestation of Hendra virus infection in horses and humans is that of severe interstitial pneumonia caused by viral infection of small blood vessels. The virus is also capable of causing nervous disease. Hendra virus is not contagious in horses and is spread by close contact with body fluids, such as froth from infected lungs. Diagnosis should be based on the laboratory examination of blood, lung, kidney, spleen, and, if nervous signs are present, also of the brain. Evidence of infection with the more recently identified and related Nipah virus was found in the brain of one horse in which there was inflammation of the meningeal blood vessels. Fruit bats, especially Pteropus s., have been incriminated as the natural and reservoir hosts of both Hendra and Nipah viruses.     

Immunoblot analysis of calpastatin degradation: evidence for cleavage by calpain in postmortem muscle.

A negative correlation exists between calpastatin activity and meat tenderness. Therefore, it is important to determine the mechanism of calpastatin inactivation in postmortem skeletal muscle. Western immunoblot analysis was performed to determine the protease(s) responsible for degradation of muscle calpastatin during postmortem storage. To accomplish this, purified calpastatin was digested with different proteases in vitro, and their pattern of calpastatin degradation was compared with that of calpastatin degradation in postmortem muscle. Polyclonal antibodies raised in mice against recombinant bovine skeletal muscle calpastatin were used to monitor calpastatin degradation. Lamb longissimus was stored at 4 degrees C and sampled at 0, 6, 12, 24, 72, 168, and 336 h postmortem. Postmortem storage produced a discrete pattern of calpastatin degradation products that included immunoreactive bands at approximately 100, 80, 65, 54, 32, and 29 kDa. Undegraded calpastatin (130 kDa) was barely detectable after 72 h of postmortem storage at 4 degrees C, and no immunoreactive calpastatin was observed by 336 h postmortem. For in vitro proteolysis, lamb longissimus calpastatin (0 h postmortem) was purified using Affi-Gel Blue chromatography. Calpastatin was digested with m-calpain, mu-calpain, cathepsin B, proteasome, trypsin, or chymotrypsin. Each of these enzymes degraded calpastatin. Immunoreactive fragments resulting from digestion of calpastatin with m- and mu-calpain were similar to each other and closely resembled those observed during postmortem aging of lamb longissimus at 4 degrees C. Digestion of calpastatin with mu-calpain reduced calpastatin activity. Degradation of calpastatin by other proteases resulted in unique patterns of immunoreactive fragments, distinct from that observed in longissimus. Thus, m- and(or) mu-calpain seem to be responsible for calpastatin degradation during postmortem storage of meat.     

Efficacy of fenbendazole granules and pyrantel pamoate suspension against Toxocara canis in greyhounds housed in contaminated runs.

The efficacy of fenbendazole granules against Toxocara canis in naturally infected greyhounds housed in contaminated environments was evaluated. Eight pens, each containing three to seven greyhounds, 3-12 months of age, were randomly allotted into two treatment groups. Greyhounds in Group 1 were treated with fenbendazole granules mixed in their feed at 50 mg/kg/day for 3 consecutive days once a month for 4 months. Greyhounds in Group 2 were treated with pyrantel pamoate suspension at 5.0 mg/kg per os once a month for 4 months. Quantitative fecal examinations were performed on days 0, 10 and then on the first day of each monthly treatment. Greyhounds administered fenbendazole had fecal egg count reductions (FECRs) of 95.8 and 99.8% at 10 and 31 days following initial treatment, respectively. Greyhounds administered pyrantel pamoate had FECRs of 85.8 and 88.3% at 10 and 31 days after the first treatment, respectively. T. canis fecal egg counts conducted from Day 31 through Day 128 were significant lower in those greyhounds administered fenbendazole as compared to greyhounds administered pyrantel pamoate. Fenbendazole produced FECRs in greyhounds from Day 31 through Day 128 by 96.8-99.8%. Pyrantel pamoate reduced fecal egg counts during the same time period 71.4-98.3%.     

A simulation model for studying the role of pre-slaughter factors on the exposure of beef carcasses to human microbial hazards.

A Monte Carlo simulation model was constructed for assessing the quantity of microbial hazards deposited on cattle carcasses under different pre-slaughter management regimens. The model permits comparison of industry-wide and abattoir-based mitigation strategies and is suitable for studying pathogens such as Escherichia coli O157:H7 and Salmonella spp. Simulations are based on a hierarchical model structure that mimics important aspects of the cattle population prior to slaughter. Stochastic inputs were included so that uncertainty about important input assumptions (such as prevalence of a human pathogen in the live cattle-population) would be reflected in model output. Control options were built into the model to assess the benefit of having prior knowledge of animal or herd-of-origin pathogen status (obtained from the use of a diagnostic test). Similarly, a facility was included for assessing the benefit of re-ordering the slaughter sequence based on the extent of external faecal contamination. Model outputs were designed to evaluate the performance of an abattoir in a 1-day period and included outcomes such as the proportion of carcasses contaminated with a pathogen, the daily mean and selected percentiles of pathogen counts per carcass, and the position of the first infected animal in the slaughter run. A measure of the time rate of introduction of pathogen into the abattoir was provided by assessing the median, 5th percentile, and 95th percentile cumulative pathogen counts at 10 equidistant points within the slaughter run. Outputs can be graphically displayed as frequency distributions, probability densities, cumulative distributions or x-y plots. The model shows promise as an inexpensive method for evaluating pathogen control strategies such as those forming part of a Hazard Analysis and Critical Control Point (HACCP) system.     

Antibodies produced by mice immunized with recombinant vaccinia viruses expressing two different types of a major Theileria sergenti surface antigen (p32) react with the native surface antigen.

A 32 kDa major surface antigen, p32, of Theileria sergenti at the piroplasm stage is the main target of the host immune response. The immunogenic property of the p32 varies in some strains among the population of Theileria sergenti in Japan where the Chitose type and the Ikeda type are the most common varieties. We have constructed vaccinia virus recombinants vv/p32C and vv/p32I which harbor the Chitose and Ikeda types of p32 gene, respectively. It was found that vv/p32C and vv/p32I produced type-specific p32 which did not cross react with the monoclonal antibodies (mAbs) against the other type of p32. When mice were immunized with vv/p32C and vv/p32I, antibodies against p32 were detectable 2 weeks after the immunization, and these antibodies reacted with the native surface antigen in purified T. sergenti merozoite.     

Sudden death of two horses associated with pulmonary aspergillosis.

The sudden death of two horses was attributed to the rapid and acute development of pulmonary aspergillosis. One horse was making excellent postoperative progress after a jejunal resection and anastomosis for intestinal adhesions. The other horse was being treated routinely for equine protozoal myeloencephalitis (EPM). Signs of fever and an increased respiratory rate were detected shortly before death in the first horse, but no premonitory clinical signs characteristic of pulmonary infection were detected in the horse being treated for EPM. Both horses developed rapidly debilitating, acute pulmonary mycosis and died unexpectedly.     

Long-term EKG in dogs: comparison between a computerized system and visual arrhythmia analysis]

Based on results of holter ecg recordings taken from 38 dogs the computerized analyses system turned out as method not exactly valuing the frequency and sensitivity of arrhythmias. Despite there was a significant correlation between the computerized and visual arrhythmia analysis for ventricular respectively supraventricular premature beats, in individual cases there was an important aberration between the particular values.