Effects of heat stress on the antimicrobial drug resistance of Escherichia coli of the intestinal flora of swine.

The effects of heat stress on the antimicrobial drug resistance of Escherichia coli of the intestinal tract of swine were studied in animals from a farm that had not been supplementing antimicrobials in feed for the past 10 years. In one study, 10 finisher hogs were heat stressed (34 degrees C) for 24 h. Antimicrobial resistance levels after stress were significantly higher (P < 0.05) when compared with pre-stress levels for amikacin, ampicillin, cephalothin, neomycin and tetracycline from faecal samples. This high level of resistance persisted to slaughter that occurred at 10 days post-stress for most of the antimicrobials mentioned. In a second study, samples of different sections of the gastrointestinal tract were collected after heat stress and compared with control, non-stressed animals. Results indicated that E. coli which colonized the ileum and caecum had a higher level of resistance to ampicillin and tetracycline than the E. coli which colonized the colon and rectum. When animals were exposed to heat stress, resistance to ampicillin and tetracycline of E. coli in the lower digestive tract increased (P < 0.05) to a level similar to that observed in the ileum and caecum. Based on these findings, an investigation was made to test the hypothesis that (a) an increase in intestinal motility increases shedding of resistant E. coli and (b) heat stress induces a reduction in intestinal transit time in swine. For each study, two groups of three, randomly selected finisher hogs each were formed (treated and control groups). In study (a), induction of increased motility and peristalsis was obtained using an intramuscular injection of the cholinergic drug neostigmine methylsulphate. Escherichia coli isolates were obtained from the ileum, caecum, colon and rectum after animals were slaughtered. A higher level of ampicillin-resistant E. coli was found in the caecum (40%) than in other segments of the intestinal tract. In treated animals, level of resistance increased for organisms from the colon and rectum. Similar results were obtained for tetracycline resistance. In study (b), intestinal transit time was measured using chromium-EDTA as a marker. Swine were euthanized and samples were collected throughout the intestinal tract (duodenum to rectum) 8 h after administration of the marker to control and heat-stressed animals. Results indicated a reduced transit time for the stressed group. These findings corroborate the initial hypothesis that an outflow of resistant organisms moves from the upper tract (ileum and caecum) to the lower tract (colon and rectum).

Effects of cold stress on the antimicrobial drug resistance of Escherichia coli of the intestinal flora of swine.

Faecal samples were initially collected from pigs of different age groups, over periods considered to be seasonally normal and stable (baseline), and during times in which drastic drops in environmental temperature (cold stress) occurred. Baseline bacterial resistance to ampicillin and tetracycline were significantly higher (P < 0.05) in younger than in older pigs. Also, when animals were exposed to excessively cold conditions, there was a significant (P < 0.05) increase in ampicillin and tetracycline resistance in Escherichia coli for animals of all age groups. These results may indicate that factors other than feeding or use of antibiotics may play a role in establishing or maintaining the antibiotic resistance microflora of pigs, especially in those operations where animals are maintained outdoors, with minimal protection against extreme weather conditions.

Stability of porcine reproductive and respiratory syndrome virus in the presence of fomites commonly found on farms.

OBJECTIVE: To determine the survival of porcine reproductive and respiratory syndrome virus (PRRSV) on nonliving substances (fomites) at 25 to 27 C. DESIGN: Prospective controlled study. SAMPLE POPULATION: 3 solid, 6 porous, and 7 liquid fomites. PROCEDURE: The fomites were contaminated with known concentrations of PRRSV. Samples for virus isolation were obtained on day 0 through day 11, assayed in cell cultures, and stained with fluorescent antibody conjugate. RESULTS: The virus was recovered only on day-0 samples of alfalfa, wood shavings, straw, plastic, boot rubber, and stainless steel. Virus was isolated from city water through day 11, from well water through day 9, and from 2 buffer solutions for 4 and 6 days. The virus was isolated only on day 0 from swine saliva, urine, and fecal slurry. CLINICAL IMPLICATIONS: Results indicated that PRRSV is a fairly labile virus, but because of its duration of viability in water, contamination of drinking water and lagoons by PRRSV-shedding swine would serve as sources of virus to infect susceptible swine.

Zoonoses in practice.

Twenty-five zoonoses of importance in the United States are focused upon in this article. Each is presented in outline form. Items are presented in a convenient format that can be of use in explaining zoonoses to owners of animals in which a specific zoonoses has been diagnosed. Also described are specific actions to be taken in diagnosing suspected zoonotic diseases, in preventing transmissions of zoonoses from infected animals to humans in contact or from infected humans to susceptible animals, in notifying physicians of potential human exposure to zoonotic disease, and in reporting to health authorities of those diseases that require notification.

Further studies on the susceptibility of raccoons (Procyon lotor) to a rabies virus of skunk origin and comparative susceptibility of striped skunks (Mephitis mephitis).

Two raccoons (Procyon lotor) were inoculated in the masseter muscles with 10(5.9) mouse intracerebral lethal dose50 (MICD50) of a rabies virus isolated from a naturally infected Iowa (USA) striped skunk (Mephitis mephitis). Five striped skunks were inoculated with either 10(0.7) or 10(2.1) MICLD50 of the same isolate. All five skunks died within 35 days following inoculation. Both raccoons survived 273 days without adverse effects, and virus was not isolated from saliva samples taken at between 25 and 273 days following inoculation.

A serosurvey of swine and free-living species on Iowa farms for antibodies against encephalomyocarditis virus.

Swine and free-living nonporcine species from 20 Iowa swine farms were surveyed for antibodies against encephalomyocarditis virus (EMCV). The microtitration serum neutralization test was used, and antibody titers >/= 1:8 were considered positive. The overall prevalence in 267 sows in various groups was 37.8% (range 20-86%). The prevalence in sows maintained in total confinement was significantly lower than in sows not maintained in total confinement (p = 0.01). Prevalence in sows was not associated with other herd characteristics or with abundance estimates of rats and/or mice.Free-living animals tested included 74 domestic cats (Felis domestica), 203 house mice (Mus musculus), 15 mice of the genus Peromyscus, nine Norway rats (Rattus norvegicus), 34 opossums (Didelphis virginiana), 14 raccoons (Procyon lotor), and seven striped skunks (Mephitis mephitis). Of these, the only seropositive animals were two domestic cats.The results of this study failed to implicate the free-living species surveyed as important reservoirs of EMCV for swine, and suggested that swine themselves and/or some species not included in this survey are the main reservoir of EMCV for swine in Iowa.

Demonstration of rabies virus-specific antibody in the sera of free-ranging Iowa raccoons (Procyon lotor).

Between 1984 and 1988, a study was conducted to evaluate the frequency of rabies virus neutralizing antibodies in raccoons (Procyon lotor) in two counties in Iowa. Nine hundred eighty five raccoons were trapped and tagged in Guthrie and Cerro Gordo counties during the spring, summer and fall of each year. Sex, age and weight were recorded for each animal and a blood sample was collected. Serum samples were tested for the presence of serum neutralizing antibodies (SNA) by the rapid fluorescent focus inhibition test (RFFIT), mouse serum neutralization test (MSN), and an indirect fluorescent antibody (IFA) technique for detecting immunoglobulin G. Fifty-one raccoons (5%) were found to have SNA by the RFFIT. Thirty-six serum samples (24 with RFFIT antibody titer greater than 3.0, and 12 less than 3.0) were also tested by the MSN, with results correlating well with the RFFIT results (r = 0.86, P less than 0.01, Kappa = 0.93). In 35 raccoons with SNA by the RFFIT, six individuals had immunoglobulin G binding activity by the IFA test. These results provided serologic evidence of exposure of raccoons to rabies virus in an area free of enzootic raccoon rabies.

The epidemiology of toxoplasmosis on Iowa swine farms with an emphasis on the roles of free-living mammals.

Multiparous sows from 19 central Iowa swine farms were tested for antibodies against Toxoplasma gondii by the modified direct agglutination test. Antibody titers of 1:32 or greater were considered positive. Rodents, domestic cats, opossums (Didelphis virginiana), raccoons (Procyon lotor) and striped skunks (Mephitis mephitis) were live-trapped on each farm and similarly tested for antibodies. The overall prevalence of T. gondii antibodies in the species tested was 39/273 (14.3%) swine, 31/74 (41.9%) cats, 2/588 (0.3%) house mice (Mus musculus), 0/21 mice of the genus Peromyscus, 0/9 Norway rats (Rattus norvegicus), 1/34 (2.9%) opossums, 4/14 (28.6%) raccoons and 2/7 (28.6%) striped skunks. The overall prevalence was significantly greater in adult cats versus juvenile cats, adult male cats versus adult female cats, and adult raccoons versus juvenile raccoons. The prevalence of T. gondii antibodies in sows was compared with the prevalence in each non-swine species on a farm basis in order to identify existing associations. The prevalence in sows (and each of the non-swine species) was also analyzed on a farm basis for association with farm characteristics or swine management practices, including the degree of confinement of swine, population size and average parity of breeding female swine, estimated cat population size, and estimated mouse and rat abundance. Average titers of seropositive animals were compared on a species basis. The prevalence in sows which were totally and continuously confined was lower than that in sows which were not totally and continuously confined. The prevalence in sows from farms with an average parity of less than 2.0 was significantly lower than that in sows from farms with an average parity of 2.0 or greater. These results suggested that the prevalence of T. gondii antibodies in swine increased with age and that prevalence in swine could be reduced through total confinement. No associations could be established between prevalence in sows and prevalence in non-swine species or other farm characteristics/swine management practices. However, the high prevalence of T. gondii antibodies in cats suggested that fecal contamination of the environment by cats may be the most significant source of toxoplasmosis for swine. The extremely low prevalence of T. gondii antibodies in house mice suggested that this species was not an important source of T. gondii for swine in Iowa.

Experimental inoculation of raccoons (Procyon lotor) with rabies virus of skunk origin.

To determine raccoon (Procyon lotor) susceptibility and serum neutralizing antibody response to a skunk salivary gland rabies virus, raccoons were inoculated with a rabies virus isolated from a naturally-infected striped skunk (Mephitis mephitis). Raccoons were divided into four groups of three animals each. A dilution of the rabies virus suspension, 10(2.4), 10(3.4), or 10(4.8), mouse intracerebral lethal dose50 (MICLD50), was administered into the masseter muscles of each animal. Three negative control animals received only diluent. Saliva and sera were collected on post-inoculation days 35, 63 and 92 for virus isolation and determination of serum neutralizing antibody titer. All animals survived the 92 day observation period and none exhibited the behavioral changes classically associated with clinical rabies virus infections. Rabies virus was not detected in the saliva of any raccoon and two of the three animals receiving the highest inoculum developed serum neutralizing antibodies (SNA). On day 92, a challenge suspension of New York City/Georgia (NYC/GA) strain rabies virus in fox salivary glands (10(3.2) MICLD50) was administered to all 12 raccoons. All animals succumbed to rabies virus except the two animals that had earlier developed SNA. The results of this study provided evidence about the susceptibility of raccoons to a skunk rabies virus and demonstrated that exposed raccoons could survive for at least 92 days following exposure. Furthermore, animals developing SNA under such circumstances were capable of withstanding challenge with rabies virus that was fatal for seronegative raccoons.

Seroprevalence of antibodies against encephalomyocarditis virus in swine of Iowa.

A total of 2,614 swine from 104 herds located throughout Iowa were tested for antibodies against encephalomyocarditis virus (EMCV) by use of the microtitration serum neutralization test. The sample was composed of 587 sows and gilts and 2,027 finishing swine. A statistically significant (P less than 0.002) difference was observed between prevalence in sows and gilts (17.2%) and that in finishing swine (12.2%). Breeding swine maintained in total confinement (20.5%) had significantly (P = 0.04) higher prevalence than did breeders maintained in other types of housing (12.1%), whereas prevalence in finishing swine raised in total confinement (6.4%) was significantly (P = 0.02) lower that in finishers not raised in total confinement (13.6%). Association was not detected between prevalence and herd size or between prevalence and season of the year. Adjusting for test specificity and sensitivity, the true prevalence of EMCV infection in swine in Iowa was estimated to be 13.8% in breeding stock and 8.5% in finishing swine. On a herd basis, 89.4% (93/104) of the herds had one or more EMCV-positive swine.

Survey to estimate prevalence of Leptospira interrogans infection in mature cattle in the United States.

A total of 5,142 kidney tissue samples and 5,111 serum samples from mature cattle in 49 states and Puerto Rico were collected at slaughter. Age of cattle ranged from 1 to 16 years (mean, 6.6 years). Leptospires were isolated from 88 (1.7%) kidney tissues, and 2,493 (49%) sera contained antibodies against 1 or more of 12 Leptospira interrogans serovars. Leptospires were observed by immunofluorescence in 41 (0.8%) kidney tissues. Using agglutinin-absorption tests, 73 (83%) isolates were identified as serovar hardjo, 11 (12.5%) as serovar pomona, and 4 (4.5%) as serovar grippotyphosa. By use of restriction endonuclease analysis studies of chromosomal DNA, all isolates differed from reference serovars but were identical to strains previously isolated from cattle or swine in the United States. Of the serovar hardjo isolates, 85% were identical to restriction endonuclease analysis type (genotype) hardjo-bovis A and 11 (15%) were identical to genotype hardjo-bovis B. Serovar pomona isolates were identical to genotypes kennewicki A (64%) or kennewicki B (36%), and serovar grippotyphosa isolates were identical to the RM 52 strain. Isolation rates were significantly (P less than 0.001) higher for beef cattle than for dairy cattle and were higher (P less than 0.001) for bulls than for cows. Combined culture and immunofluorescence results indicated that 2% of mature cattle were renal carriers of leptospires.

Relationships between prevalence of Leptospira interrogans in cattle, and regional, climatic, and seasonal factors.

On the basis of serologic test results and isolation of leptospires from mature cattle, distribution and prevalence of Leptospira interrogans serovars and genotypes were compared by state and region of the United States. Relationships between isolation rate and month of sample collection, mean regional temperature, and mean regional precipitation were examined. Isolation rate and seroprevalence were significantly (P less than 0.001) higher for southeastern, south central, and Pacific coastal regions than for other regions of the United States. Isolates of genotypes hardjo-bovis A and kennewicki A and B, and of serovar grippotyphosa appeared to be randomly distributed. Genotype hardjo-bovis B isolates came from a southern area of the country that extends from Georgia to New Mexico. To the authors' knowledge, this is the first recorded isolation of serovar hardjo from Hawaii. Although significant relationship was not documented between isolation rate and month or season of the year, seroprevalence for summer, fall, and winter was significantly (P less than 0.001) higher than that for spring. Regional isolation rate was related more to mean temperature (r = 0.83; P less than 0.05) than to mean precipitation amount (r = 0.34; P greater than 0.50).

Virus survival in the environment.

Viruses pass into the environment from clinically ill or carrier hosts; although they do not replicate outside living animals or people, they are maintained and transported to susceptible hosts. Population concentrations and movement, both animal and human, have been steadily increasing in this century, enhancing transmission of respiratory and enteric viruses and compounding the difficulty of preventing environmental transmission. Studies on environmental survival factors of viruses have been most definitive for polioviruses, foot and mouth disease viruses and Aujeszky's disease virus. In addition, heat resistance studies have been reported on adenoviruses, African swine fever virus and the Norwalk virus. Resistance to disinfectants has been studied for many viruses, including picornaviruses, papovaviruses, reoviruses and retroviruses. Survival of viruses in and on a variety of fomites has been studied for influenza viruses, paramyxoviruses, poxviruses and retroviruses. The subacute spongiform encephalopathy agents, under extensive current studies, are being found to have incredible stability in the environment.

Survival of pseudorabies virus in the presence of selected diluents and fomites.

Because of the importance of environmental survival of pseudorabies virus to proposals to eradicate the virus from swine in the United States, survival of the virus was studied in various diluents and on combinations of diluents and solid fomites at 25 C. Suspensions of the virus in phosphate-buffered saline and saline G solutions remained infectious for at least 10 days. Infectivity of other virus/diluent suspensions decreased to less than 10 plaque-forming units/ml in 14 days (swine urine), 7 days (well water), 4 days (swine saliva), 2 days (lagoon water and swine nasal washings), and 1 day (swine pit effluent, chlorinated water, and bile). Suspensions of pseudorabies virus in saline G solution and on the solid fomites, whole corn, and steel remained infectious for at least 7 days. Infectivity of other virus/diluent/fomite combinations decreased to less than 10 plaque-forming units/ml in 7 days. The role of the fomites as vehicles for transmission of infection is discussed.

A current assessment of the role of porcine parvovirus as a cause of fetal porcine death.

One hundred one litters containing 1 or more dead porcine fetuses were collected at an Iowa abattoir during a 2-month interval and examined for evidence of viral infection. Each of 1,137 fetuses (302 dead, 835 alive) of these litters was tested for porcine parvovirus (PPV) antigens by direct immunofluorescence microscopy (FA) of fetal lung. Antigens of PPV were detected in the lungs of most of the fetuses of 11 of the litters. The 11 FA-positive litters contained 105 dead (100 FA-positive) and 14 live (12 FA-positive) fetuses. Infectious PPV was isolated from 10 of the 11 FA-positive litters and from 3 of the 90 FA-negative litters. No cytopathogenic agents other than PPV were isolated from any of the litters. Eleven of 101 (11%) litters examined and 100 of 302 (33%) dead fetuses examined were FA positive for viral antigen, indicating that PPV remains as a major cause of porcine fetal death.

Serological evidence of infection of dogs and man in Nigeria by lyssaviruses (family Rhabdoviridae).

A survey was conducted for serum neutralizing antibodies against 3 members of the Lyssavirus group (rabies, Lagos bat and Mokola viruses) in Nigerian dogs and humans. Of 463 unvaccinated dogs sampled, 142 (30.7%) had antibodies against rabies; 39.2% of the stray dogs were positive. Of 241 dogs tested, 17.4% had antibodies against Mokola virus and 5.8% against Lagos bat virus. 28.6% of human samples tested had antibodies against rabies and, of 158 human samples tested, 12 (7.5%) had antibodies against Mokola virus and 4 (2.5%) had antibodies against Lagos bat virus.

Porcine leptospirosis in Iowa.

The epidemiology of leptospirosis in Iowa swine was examined on the basis of serologic results and herd data from 55 herds in the National Animal Health Monitoring System (NAHMS) program and culture results and histories from 578 cases of reproductive failure submitted to the Iowa Veterinary Diagnostic Laboratory during a 3-year period. Thirty-eight percent of sera from NAHMS herds contained antibodies against 1 or more of 12 leptospira antigens. Leptospires were isolated from 9 (1.6%) of 578 cases of reproductive failure. Seven (78%) of the isolates were identified as Leptospira interrogans serovar kennewicki and 2 (22%) as serovar grippotyphosa. In 7 herds from which leptospires were isolated, attack rates ranged from 1% to 84%. Clinical leptospirosis, characterized by reproductive failure and confirmed by isolation of leptospires, was sporadic. No significant differences in farrowing averages and reproductive problems were observed between vaccinated and nonvaccinated NAHMS herds or between herds with higher (43-63%) or lower (14-40%) percentages of animals that were serologically positive against serovar bratislava.