Laboratory model to evaluate the role of aerosols in the transport of porcine reproductive and respiratory syndrome virus.

The aim of this study was to develop a model to evaluate the aerosol transmission of porcine reproductive and respiratory disease virus (PRRSV). PRRSV (MN 30-100 strain, total dose 3 x 10(6) virus particles) was aerosolised and transported up to 150 m and a portable air sampler was used to collect air samples at 1, 30, 60, 90, 120 and 150 m (five replicates at each distance) and the air samples were tested by TaqMan PCR and virus isolation. The infectivity of the aerosolised PRRSV was tested by exposing six PRRSV-naive pigs for three hours to aerosolised virus that had been transported 150 m. PRRSV RNA was detected in all five replicate air samples collected at 1, 30, 60 and 90 m, in four of the five collected at 120 m, and in three of the five collected at 150 m. Infectious PRRSV was detected by virus isolation at 1 and 30 m (all five replicates), 60, 90 and 120 m (three of the five) and 150 m (two of the five). There was a 50 per cent reduction in the log concentration of PRRSV RNA every 33 m. Three of the six pigs exposed to PRRSV-positive aerosols became infected, and PRRSV RNA was detected in air samples and on swab samples collected from the interior of the chambers that housed the infected pigs while they were being exposed.

Studies on the carriage and transmission of porcine reproductive and respiratory syndrome virus by individual houseflies (Musca domestica).

The objectives of the study were to determine the site of porcine reproductive and respiratory syndrome virus (PRRSV) in individual houseflies, to assess whether an individual housefly could transmit PRRSV to a susceptible pig, and to compare the ability of PCR, virus isolation and a pig bioassay to detect PRRSV in houseflies. In the first experiment 26 houseflies were fed on a pig infected experimentally with PRRSV; 13 were processed as a whole fly homogenate, while an exterior surface wash and a gut homogenate were collected from the other 13. Infectious PRRSV was recovered from nine of the whole fly homogenates, 12 of the gut homogenates and one of the exterior surface washes. In the second experiment, two of 10 individual houseflies, which had fed on an infected pig, transmitted PRRSV to a susceptible pig in a controlled manual transmission protocol. In the third experiment, single flies or pools of 30 flies were immersed in different concentrations of a PRRSV inoculum, then tested by PCR, virus isolation and bioassay. The virus was detected at a concentration of 10(1) TCID50/ml by PCR, 10(2) TCID50/ml by the bioassay and 10(3) TCID50/ml by virus isolation.

Transmission of porcine reproductive and respiratory syndrome virus by houseflies (Musca domestica).

Three hundred houseflies were allowed to feed on donor pigs viraemic with porcine reproductive and respiratory syndrome virus (PRRSV) on the fifth, sixth and seventh days after the pigs had been inoculated with the virus. After 60 seconds, the flies' feeding was interrupted, and they were transferred manually to feed to repletion on a naive recipient pig housed in a separate room. To enhance the chance of the flies obtaining the pigs' blood, the back of each pig was scarified with sandpaper until a slight haemorrhage was visible. The PRRSV was transmitted from the donor to the recipient pigs, and PRRSV RNA was detected by reverse transcriptase-PCR from homogenates of the flies. In a second experiment, 210 houseflies were allowed to feed to repletion on a PRRSV-infected pig on the sixth day after it had been inoculated, and were then maintained under laboratory conditions. Groups of 30 flies were collected immediately after they had fed and six, 12, 24, 48, 72 and 96 hours later, and were tested for PRRSV. Homogenates of the flies collected up to six hours after feeding were PCR- and pig bioassay-positive, but the others were negative by both tests.

Evaluation of aerosol transmission of porcine reproductive and respiratory syndrome virus under controlled field conditions.

The aim of this study was to determine whether porcine reproductive and respiratory syndrome virus (PRRSV) could be transmitted by aerosol under field conditions. A total of 210 five-month-old PRRSV-negative pigs were housed in a mechanically ventilated finishing facility containing 11 pens. Pen 1 contained 10 pigs (indirect contact controls) and pen 2 remained empty, providing a barrier of 2.5 m from the remaining pigs in pens 3 to 11. Fifteen or 16 of the pigs in each of pens 3 to 11 were infected experimentally with a field isolate of PRRSV and the other six or seven pigs served as direct contact controls. Five days after the pigs were infected, two trailers containing 10 five-week-old PRRSV-naive sentinel pigs were placed along each side of the building; one was placed 1 m from the exhaust fans on one side of the building, and the other was placed 30 m from the fans on the other side, and the sentinel pigs remained in the trailers for 72 hours. They were then moved to separate buildings on the same site, 30 and 80 m, respectively, from the infected barn, and their PRRSV status was monitored for 21 days. The direct and indirect contact control pigs became infected with PRRSV but the sentinel pigs did not.

Diagnostic investigation of chronic porcine reproductive and respiratory syndrome virus in a breeding herd of pigs.

Forty-five sows and 15 boars were selected at random from a breeding herd known to be chronically infected with porcine reproductive and respiratory syndrome virus (PRRSV) and lymphoid, immune-privileged, and non-lymphoid/non-immune-privileged tissues were tested for the presence of the virus by PCR, virus isolation, and immunohistochemistry. The virus was isolated from the lateral retropharyngeal lymph node of one sow; the isolate was nucleic acid sequenced and determined to be of field origin, and it was inoculated into two PRRSV-naive pregnant sows (A and B) at 95 days of gestation. They were necropsied 14 days later and samples of maternal and fetal tissue and blood samples were collected. Sow A had 10 fresh, six partially autolysed, and two mummified fetuses, and sow B had six fresh and viable fetuses. Viral nucleic acid was detected by PCR in tissue pools from each sow and also from pooled fetal tissues, and the virus was isolated from fetal pools from sow A.

An evaluation of test and removal for the elimination of porcine reproductive and respiratory syndrome virus from 5 swine farms.

The objective of this field study was to evaluate the protocol of test and removal (T&R) for the elimination of porcine reproductive and respiratory syndrome virus (PRRSV) from 5 chronically infected breeding herds. The T&R protocol involved sampling the entire breeding herd in one day, testing sera by polymerase chain reaction and ELISA to detect previously exposed and/or infected animals, and subsequently removing them from the herd. Following completion of T&R, breeding herds were monitored for 12 consecutive months, using ELISA, for the presence of antibodies to PRRSV. In order to be classified as a PRRSV-negative herd, all samples collected over the 12-month monitoring period were required to be negative by ELISA (s/p ratio < 0.4). At the conclusion of the monitoring period, all 5 farms were PRRSV-negative, according to the defined testing criteria. Approximately 2.2% (74/3408) ELISA false positive samples were detected across all 5 farms during the monitoring period. The diagnostic cost required during the T&R protocol was approximately US $10.66 per animal tested. Limitations of the study were a lack of herds with large (> 2000 sows) breeding herd inventories, and herds with a history of PRRSV vaccination.

Identification of genetically diverse sequences (ORF 5) of porcine reproductive and respiratory syndrome virus in a swine herd.

The ability of genetically diverse strains of porcine reproductive and respiratory syndrome virus (PRRSV) to coexist in a 1750-sow farm was assessed through the case study describing a chronically infected farm, and also by an animal experiment involving the use of swine bioassay. The case study employed a program of monitoring sera from suckling, nursery, and finishing pigs for the presence of PRRSV by polymerase chain reaction (PCR) and virus isolation (VI). The swine bioassay tested homogenates, consisting of lymphoid and pulmonary tissues, collected from 60 breeding animals from the same farm. The open reading frame (ORF) 5 portion of selected positive PRRSV detected from sera or tissues were nucleic acid sequenced and their phylogenies compared. The results indicated the presence of 3 genetically diverse groups, designated PRRSV-A, -B, and -C. Sequence heterology ranged from 5.8 to 11% between groups. Sequence homology ranged from 98.7 to 99.8% within groups. Swine bioassay verified the presence of PRRSV-A in 1 of 60 animals, and no evidence of strains B or C were detected. This paper indicates that based on the evaluation of ORF 5, genetically diverse strains of PRRSV appear to coexist, although the frequency and significance of this observation is not understood.

Transmission of porcine reproductive and respiratory syndrome virus from persistently infected sows to contact controls.

The objective of this study was to determine if porcine reproductive and respiratory syndrome virus (PRRSV) could persist in non-pregnant sows and if persistently infected sows could transmit virus to naive contact controls. Twelve PRRSV-naive, non-pregnant sows (index sows) were infected with a field isolate of PRRSV and housed in individual isolation rooms for 42 to 56 days postinfection. Following this period, 1 naive contact sow was placed in each room divided by a gate allowing nose-to-nose contact with a single index sow. Index sows were not viremic at the time of contact sow entry. Virus nucleic acid was detected by polymerase chain reaction, and infectious virus was detected by virus isolation in sera from 3 of the 12 contact sows at 49, 56, and 86 days postinfection. All 3 infected contacts developed PRRSV antibodies. Virus nucleic acid was detected in tissues of all of the 12 index sows at 72 or 86 days postinfection. Nucleic acid sequencing indicated that representative samples from index and infected contacts were homologous (> 99%) to the PRRSV used to infect index sows at the onset of the study. This study demonstrates that PRRSV can persist in sows and that persistently infected sows can transmit virus to naive contact animals.

Epidemiological and diagnostic observations following the elimination of porcine reproductive and respiratory syndrome virus from a breeding herd of pigs by the test and removal protocol.

The protocol of test and removal for the elimination of porcine reproductive and respiratory syndrome (PRRS) virus was applied to an 825-sow breeding herd. All the adult animals were tested and serum samples analysed by ELISA and polymerase chain reaction (PCR). Eighty-eight animals (10 x 7 per cent) were removed from the herd and, of these, three were ELISA-pOSitive and PCR-positive, and 85 were ELISA-positive and PCR-negative. They tended to be either individual sows, or groups of four to six animals housed in adjacent gestation stalls. Four of the ELISA-positive, PCR-negative sows were slaughtered and PRRS virus nucleic acid was detected in a sample of sternal lymph node from one of them. After the completion of the test and removal protocol, the breeding and finishing populations were monitored for 12 consecutive months by ELISA. The 960 samples taken were negative for PRRS virus antibodies.

Attempted elimination of porcine reproductive and respiratory syndrome virus from a seedstock farm by vaccination of the breeding herd and nursery depopulation.

An attempt was made to eliminate the virus of porcine reproductive and respiratory syndrome from a seedstock farm by using the combined strategies of vaccination and nursery depopulation. The breeding herd was vaccinated with a modified-live virus vaccine; all breeding and lactating adult animals were vaccinated twice, with a 30-day interval between vaccinations. All the sows were vaccinated in this way except for those in the third trimester of gestation (66 to 114 days) which were vaccinated on day 7 of lactation and 30 days later. A serological profiling system was developed to assess when the piglets became infected. Pigs from vaccinated sows were profiled at weekly intervals after weaning, using immunofluorescence tests for the detection of IgM and IgG, a serum neutralising antibody test, and virus isolation. After completion of the protocol, the nursery and finishing sites were monitored for 15 months. Evidence of reinfection in the finishing stage was detected 16 months after depopulation, but not in the nursery or the breeding herd. The source of the virus was not determined, but suspected origins included a lack of biosecurity, aerosol transmission from another infected farm or a persistently infected pig.

Evaluation of the effects of nursery depopulation of the profitability of 34 pig farms.

The financial impact of nursery depopulation was assessed on 34 pig farms by constructing a partial budget model to measure the profitability of the nursery production. The model measured margin over variable cost and used production data generated from a previous study; it assumed that fixed costs remained constant throughout the study and that feed cost, weaned pig cost and market price per nursery pig also remained fixed. The mean margin over variable cost per sow on the 34 farm after nursery depopulation was Pounds 116. Thirty-two of the farms showed reductions in this cost, ranging from Pounds 20 to Pounds 408 per sow, in the 12 months after nursery depopulation compared with the previous 12 months. Of the two farms which did not show an increase in profitability, one showed no change and the other showed a net loss of Pounds 8 per sow. The sows' serostatus for porcine reproductive and respiratory syndrome virus infection was monitored but there was no significant difference between the margin over variable cost per sow of the seropositive (Pounds 130) and seronegative (Pounds 170) herds.

Indirect fluorescent IgM antibody response of pigs infected with porcine reproductive and respiratory syndrome syndrome virus.

IgG and IgM antibody responses were examined by an indirect fluorescent antibody method in pigs following inoculation with different porcine reproductive and respiratory syndrome virus (PRRSV) isolates or a vaccine virus. Viremia was also examined in the pigs. The IgG antibody was first detected between 9 and 14 days post inoculation (PI) and maintained high titers for at least 7 weeks PI. No change in IgG antibody titers was observed when the pigs were reinoculated with PRRSV 35 days PI. IgM antibody was detected between 5 and 28 days PI in the pigs. Reinoculation at 35 days PI caused a short term rise of IgM antibody. Virus was isolated from sera collected between 2 and 21 days PI. The IgM antibody was detected regularly in sera collected during viremia and up to 1-2 weeks after the viremic periods. These results suggest that pigs with detectable IgM antibody are probably pigs with recent infection and that routine testing of IgM antibody in purchased breeding pigs from seropositive farms may be useful in identification of pigs with recent infection.

Strategies to control PRRS: a summary of field and research experiences.

Various methods for the control of PRRS virus have been published. The technology of nursery depopulation (ND) appears to effectively control the spread of virus between members of endemically infected populations. ND consists of a strategic adjustment in pigflow based on the presence of specific serologic patterns as detected by the indirect fluorescent antibody test. This pattern indicates a low seroprevalence of antibodies detected in the breeding herd and recently weaned piglets (< or = 10%), in contrast to a high (> 50%) seroprevalence in 8 to 10 week old piglets. ND has been carried out on swine farms in the US and results indicate improvements in nursery piglet growth rate and mortality levels. Three examples are provided in the following text. Recently a modified live virus vaccine (RespPRRS, NOBL Laboratories/Boerhinger Ingleheim) has become commercially available. It is currently approved for use in piglets from 3 to 18 weeks of age; however, potential for the use in adult animals is currently under investigation.