Persistence of porcine parvovirus in swine infected in utero and followed through maturity

The potential of porcine parvovirus (PPV) to persistently infect swine exposed in utero was studied. Forty eight 80- to 95-day-old fetuses from 5 PPV seropositive sows were inoculated intramusculary with a virulent strain of PPV or with cell culture medium (controls). Blood samples were collected at birth prior to nursing and at monthly intervals thereafter and tested for antibodies to PPV. Virus-inoculated and control pigs were euthanized at either 1 week before birth (-1), at birth (0) and at weeks 2, 4, 6, 8, 10, 22, and 28 after birth. Presence of viral DNA and antigen was evaluated using slot blot DNA hybridization and indirect FA techniques, respectively. All inoculated fetuses (n = 26) and 7 control fetuses (n = 22) seroconverted in utero, and these pigs maintained antibody titers greater than log10 2 for the period of testing (0-38 weeks after birth). After passive antibody titers had reached subdetectable levels in control animals, animals remained seronegative through an additional 14 weeks of testing in spite of close contact with infected pigs. Virus antigen was not detected in any tissues examined from pigs euthanized at term. In contrast, PPV DNA was detected consistently from pigs at birth from various tissues, and from the lung of one pig at 6 weeks of age and from the lymph nodes of one pig euthanized at 28 weeks of age. The results indicate that pigs infected with PPV in utero may be persistently infected, however the likelihood of shedding to contact animals is minimal.     

Seroepidemiologic study of natural transmission of Mycoplasma hyopneumoniae in a swine herd

A cohort of 57 pigs in a farrow-to-finish swine herd with mild clinical mycoplasmal disease was followed to determine patterns of seroconversion to Mycoplasma hyopneumoniae (MH), detected with an enzyme-linked immunosorbent assay (ELISA). Survival analysis was used to evaluate the relationship between time to seroconversion and possible risk factors for MH infection (or enzootic pneumonia).

Pigs were housed in outdoor pens at approximately 9 weeks of age, when passively acquired MH antibodies had decayed. From 9 to 11 weeks of age and during a 5 week period, pigs were exposed by direct (nose-to-nose) or indirect contact to older seropositive gilts. Blood samples were collected from each pig at 3 week intervals until market age, when they were either slaughtered or selected for breeding. Antibody concentration was measured as the ratio of optical densities of the serum sample to the positive control (S/P). Based on the sample distribution of S/P ratios from pigs in an MH-free herd, pigs were considered positive when S/P ratios were greater than 0.34. At the beginning of the study, all pigs were seronegative to MH. Seroconversion was first detected after 21 days, and was most frequent about 11 weeks after exposure to older seropositive gilts. By the end of the study, 11 pigs (19%) had seroconverted, with S/P ratios ranging from 0.40 to 1.11. The presence of gross lung lesions showed a moderate to good agreement with ELISA results (K = 0.62). Histologic lesions were evident in virtually all slaughtered pigs, ranging from mild, non MH-specific lesions to severe lesions typical of MH infection. No secondary respiratory pathogens were isolated. Clinical signs were mild and there was no significant difference (P > 0.4) in weight gain between seropositive and seronegative pigs, or between pigs with and without lung lesions. A Cox regression model was fitted to the seroconversion data, and opportunity of contact (direct or indirect) was the only significant variable. After adjustment for breed and antibody S/P ratio prior to exposure, pigs in direct contact with seropositive gilts were seven times more likely to seroconvert than those in only indirect contact.     

Longitudinal study of hepatitis E virus infection in Spanish farrow-to-finish swine herds

Hepatitis E is a zoonotic disease and is highly prevalent in European swine livestock. There is a need to compare the infection dynamics of hepatitis E virus (HEV) between herds with the same production system and determine the percentage of animals that could arrive infected at slaughter age. Therefore, a longitudinal study was performed in six Spanish farrow-to-finish affected farms. Twenty piglets per farm were monitored from nursery to slaughter. RT-PCR and serology techniques were applied to analyze longitudinally collected sera and/or faecal samples. Liver and bile samples were also taken at the abattoir. Anti-HEV IgM were firstly detected at 7 weeks of age in 5 farms whereas at 13 weeks of age in 1 farm (farm 2). At slaughter age 50–100% of pigs had seroconverted to anti-HEV IgG in the former 5 farms whereas in the other herd only 5% of pigs were IgG seropositive (farm 2). Six out of 96 livers and 5 out of 80 biles analyzed were HEV positive at the abattoir (total percentage of infected animals: 11.5%). All these positive animals had already seroconverted except 2 pigs of farm 2. Hence, pigs can be seronegative at slaughter age being infected during the latest fattening period. Manipulation of HEV-infected livers or other organs from pigs could be considered a possible route of transmission in Spanish abattoirs. This study represents the first longitudinal survey on swine HEV infection dynamics conducted in different herds.     

Indication of transmission of BVDV in the absence of persistently infected (PI) animals

In a closed dairy herd all animals were tested serologically for BVD antibodies twice a year during a 6-year period. Seroconversions were detected every year. At the start of the 6-year monitoring period blood samples from all animals were examined by virus isolation. No persistently infected animals were identified. Entire-herd culturing for BVDV was repeated at the end of the third year. Samples from all newborn female calves were examined for BVDV at approximately 2 months of age and older. During the entire monitoring period BVDV was isolated in one newborn calf twice with an interval of 3 weeks. The mother had seroconverted during pregnancy. Five congenitally infected non-PI calves were identified, the mothers of which had seroconverted during late pregnancy; repeated sampling proved the calves to remain seropositive in a seronegative age cohort. Although direct and indirect introduction of BVDV from outside the herd can never be excluded it seems highly unlikely in this closed herd. The findings indicate that transmission of BVDV can take place over a long period of time in the absence of PI animals. This observation may have serious consequences for control programmes.     

Neospora caninum: Serological follow-up in dairy cows during pregnancy

We conducted a longitudinal study to follow-up the anti-Neospora caninum serologic status in 30 initially seropositive and 83 initially seronegative cows during their pregnancy. Study cows were blood-sampled every other month during pregnancy until parturition. Blood serum samples were screened for anti-N. caninum antibodies by ELISA. Cows that seroconverted were re-tested by immunoblot as a confirmation test. Among 30 seropositive cows, 28 cows remained seropositive during the whole pregnancy, whereas 2 cows transiently tested negative at least once during pregnancy. Among 83 seronegative cows, 82 cows remained seronegative and 1 cow tested positive three times during the sixth, eighth and last month of pregnancy. As only 2 out of 30 seropositive animals and 1 out of 83 animals changed their serologic status during pregnancy, the study results indicate that there is only a minor temporal instability of anti-N. caninum antibody reactivity in adult cattle.     

Pleuropneumonia in swine caused by Haemophilus parahaemolyticus. A study of the epidemiology of the infection

Haemophilus parahaemolyticus infection was studied in a herd with continuous production, i.e., continuous introduction of stock to replace animals delivered for slaughter. None of 30 seronegative pigs contracted the infection when exposed to contact with two pigs that were seropositive after inoculation with H. Parahaemolyticus three weeks earlier. After aerosol infection had been applied in the building an acute outbreak with a morbidity rate of 100 per cent developed in less than 24 hours. Following recovery the majority of the 16 pigs present became seropositive, and when 30 seronegative pigs were introduced 7 weeks later, antibody response occurred in three of them. The persistence of H. parahaemolyticus in pigs that had been infected during the acute outbreak was confirmed at slaughter, in that the organism was re-isolated from the tonsils of 2 of these pigs. Most serum titres persisted for several months, but some animals showed just a transient antibody response.     

Role of birds in transmission of classical swine fever virus

Active transmission of classical swine fever virus (CSFV) was studied in six birds (five ravens, one hooded crow) and two laying hens. Cloacal swabs, blood and organs of birds and hens as well as blood and organ samples of pigs which had been fed with faeces derived from CSFV infected birds or which had come in contact with faeces of infected hens were negative for CSFV. None of the animals seroconverted during the study. This result demonstrates that active virus transmission by these animals is unlikely. Dissemination of CSFV from wild boar to domestic pigs is discussed.     

Natural postnatal Neospora caninum infection in cattle can persist and lead to endogenous transplacental infection

A serological follow-up study of 3.5 years duration was done of a dairy herd that had experienced a mass seroconversion to Neospora caninum following a point source exposure shortly before the 17th of January 2000. A total of 913 blood samples of 244 animals at seven sampling dates were used to investigate the seroprevalence dynamics in the herd. Most postnatally infected cattle remained seropositive during the period of investigation but 11 animals became seronegative after 6-27 months indicating transient infection. Six animals seroconverted later than the main group of 45 animals and 5 animals became seronegative after at least two seropositive records possibly due to a low infection dose or difference in the haplotypes of the infected animals. In total 58% (14/24) of the offspring of postnatally infected dams was seropositive. Nine of 16 (56%) daughters originating from inseminations after the postnatal infection of their dams were seropositive indicating endogenous transplacental infection.     

Confirmation of pseudorabies virus infection, using virus recrudescence by dexamethasone treatment and in vitro lymphocyte stimulation

A corticosteroid treatment regimen, together with peripheral blood lymphocyte transformation and virus isolation, were used as epizootiologic tools to study pigs seropositive to pseudorabies (PR) virus, but of unknown PR viral infection status. After treatment with 160 mg of dexamethasone daily for 5 days, PR virus was isolated from 2 of 3 adult littermate boars that carried antibodies to PR virus. These pigs also transferred infection to a specific-pathogen-free sentinel penmate . After dexamethasone treatment, increased lymphocyte stimulation indices were detected in all PR virus seropositive pigs. Pseudorabies virus was not isolated from a seronegative littermate gilt of the seropositive boars. The gilt also remained PR virus seronegative and negative for lymphocyte transformation. Before dexamethasone treatment, the 3 boars and 1 gilt were seronegative to the PR viral serum neutralization (SN) test at 11 weeks of age. At 13 weeks of age while still SN negative, these pigs were moved to a building and randomly penned with 76 other SN-negative pigs. At 18 weeks of age, all pigs in the building were PR virus seronegative, but at 25 weeks of age, the 3 boars were seropositive and the gilt was seronegative. There were no other pigs in the building that had antibodies to PR virus before the addition or after the removal of these pigs. Seemingly, the 3 boars were infected with PR virus before 13 weeks of age indicating that PR virus infection of swine may occur in the absence of detectable SN antibodies.     

Light and electron microscopic findings in the intestine of spontaneous and experimentally-produced African swine fever]

Light and electron microscopical studies were carried out after experimental induced and spontaneous infection with African swine fever virus. The experimental infection was performed in 18 pigs divided into two groups consisting of 9 animals. The pigs of group I were inoculated with virulent isolate E 70, those of group II with attenuated isolate E 75. Two infected pigs of group I and one control animal were killed on days 3, 5 or 7 p.i., two pigs of group II and one control animal were killed on days 9, 11 or 13 p.i. Additionally 19 spontaneous infected and seropositive animals and two seronegative pigs without clinical signs were examined. Infection with African swine fever virus resulted in slight morphological alterations of the intestine. The pathological findings showed a more intense involvement of the large intestine, especially the caecum and colon, than the small intestine, where the ileum was mostly affected. In all three groups edema and vacuolisation could be observed in endothelial cells. In spite of beginning degenerative signs, especially after spontaneous infection, the fenestrated structure of the endothelium was conserved in most of the cases. In animals infected with virulent isolate the vascular lumina contained aggregations of fibrin, which was severe pronounced in the pigs of the other groups. In the area of these alterations disturbance of permeability with extravasation could be found. In all groups single virions or virus aggregates could be identified in concave depressions of the erythrocyte membrane or free within the blood plasma, in some cases enveloped by a plasma-like material.(ABSTRACT TRUNCATED AT 250 WORDS)     

Skin test as herd diagnosis for Aujeszky's disease (Pseudorabies) in swine

Summary The value of a skin test for the diagnosis of Aujeszky's disease (pseudorabies) in swine was examined. Semipurified and concentrated antigens, obtained by ether/tween 80 inactivation of Aujeszky's disease virus, were inoculated intradermally into seronegative, experimentally infected and vaccinated swine and into pigs with maternal antibodies. A specific skin reaction, characterized by a visible indurated swelling, was observed within 24 hours in animals with active immunity, No reaction was seen in pigs with maternal immunity or in seronegative uninfected and unvaccinated animals. In the infected and vaccinated groups, 89% and 58% respectively of the animals with seroneutralizing antibodies were positive by the skin test response. Positive reactions were observed as early as 8 days after the infection. All but one seronegative animals remained free of antibodies after 2 consecutive skin tests and the course of decline of maternal antibodies was not changed. An earlier skin test did not lead to sensitization for a later application. The present results, together with field experience on 8 farms, revealed that the skin test could be considered a reliable method for diagnosis of Aujeszky's disease in swine on a herd basis.     

Infection of pigs by aerosols of Aujeszky's disease virus and their shedding of the virus

On three consecutive days, six pigs were exposed for 15 minutes to aerosols of Aujeszky's disease virus. The total estimated dose was 4·5 log₁₀ 50. Within each isolation room, a sentinel pig was placed on a deck two feet away from the infected pig. The breath of the pigs that had inhaled the aerosols was collected on days 3, 7 and 13. The respiratory and other clinical signs of the infected pigs resembled those in field cases of Aujeszky's disease. All the pigs infected with Aujeszky's disease virus seroconverted within seven to 10 days after infection. Among the sentinel pigs, clinical signs were minimal and only three seroconverted.     

Pigs orally inoculated with swine hepatitis E virus are able to infect contact sentinels

The purpose of the present study was to explore the most likely natural route of infection of swine hepatitis E virus (HEV) by oral inoculation of pigs and to investigate the potential infection by direct contact exposure. A preliminary experiment was performed to assess the infectiousness of the bile used as source of virus. Once confirmed, 16 pigs were inoculated via oral drop with an HEV positive bile suspension containing 2 × 10⁵ genome equivalents per pig. Nine animals were kept as contact sentinels and 12 more pigs were used as negative controls. A number of pigs from the three groups were euthanized at 16, 32 and 64 days post-inoculation. From the HEV inoculated group, three pigs shed virus in faeces, two had virus RNA in bile at necropsy and two seroconverted. In the contact group, two animals showed presence of HEV RNA in bile. This study demonstrates that pigs orally inoculated with a single HEV dose got infection, although few animals had evidence of infection. Moreover, the virus was successfully transmitted to direct contact exposed pigs.     

Seroprevalence of Actinobacillus (Haemophilus) pleuropneumoniae serotype-1 infection in swine herds in Quebec

Seroprevalence of Actinobacillus (Haemophilus) pleuropneumoniae serotype-1 infection was evaluated in pigs on 7 farms in Quebec. Commercial cross-bred herds A to G, ranging from 110 to 235 sows and infected with A pleuropneumoniae serotype-1 were selected. Five pigs/litter were selected at random and were identified (group 1). Blood samples were obtained from group-1 pigs at 2 to 4, 14, 28, 42, and 56 days of age. Blood also was obtained from group-1 pigs remaining in the postweaning unit at 70 days of age, and from 20 to 40 sows 1 to 3 times. To determine prevalence of seropositive pigs in all age groups for the entire study period in herds C to G, blood samples were obtained from 20 pigs/age group (group 2) selected at random at 28, 42, and 56 days of age at each visit. Group-1 pigs were included when they reached 28, 42, and 56 days of age. Pigs were serologically monitored in herds A and B for 3 months and in herds C to G for 5 to 6 months. Serologic status of pigs at 2 to 4 days of age was not statistically associated with status at 42 days (P = 0.6293) and at 56 days (P = 0.3098) of age for the same pigs. Therefore, seronegative pigs 2 to 4 days old did not seroconvert earlier than did those with detectable maternal antibodies at 2 to 4 days old. Only about 50% of the 70-day-old pigs were seropositive at 56 days. Seemingly, pigs seroconverted late in the postweaning period.(ABSTRACT TRUNCATED AT 250 WORDS)     

Correlation of tissue infection and serologic findings in pigs fed Toxoplasma gondii oocysts

Each of thirteen 6-week-old pigs was inoculated per os with 10,000 sporulated oocysts of Toxoplasma gondii. By postinoculation day (PID) 13, pigs were seropositive by the indirect fluorescent antibody test. Beginning on PID 13 and every 7 days thereafter through PID 97, 1 pig was killed and 6 tissues were examined for T gondii. Of the 13 pigs, 11 were infected, including the 1st pig killed on PID 13, although none of the pigs had gross lesions of toxoplasmosis. Tissues harboring T gondii most frequently were the heart and brain; organisms were detected less frequently in the longissimus muscles, diaphragm, and liver. Toxoplasma gondii was not detected in the bronchial lymph nodes. There was good correlation between antibody and presence of T gondii in these pigs. One additional pig, maintained as a noninfected control, remained seronegative and had no evidence of infection when killed on PID 97.     

Sero-epidemiological screening of pig sera collected at the slaughterhouse to detect herds infected with Aujeszky's disease virus, porcine influenza virus and Actinobacillus (Haemophilus) pleuropneumoniae in the framework of an integrated quality control (IQC) system

Over a period of six months, approximately 4700 blood samples were collected from 97 pig-finishing farms in the provinces of Noord-Brabant and Gelderland and screened for antibodies with respect to Aujeszky's disease virus (ADV), porcine influenza virus (PI) and Actinobacillus (Haemophilus) pleuropneumoniae (App). There were significant differences in the percentages of seropositive pigs between the two provinces, which may be related to the difference in the density of the pig population in the two provinces. In practice, it was possible to perform a reliable sera collecting procedure at the slaughterhouse. No farms remained seronegative with respect to most of the disease agents during the sampling period. There was a high degree of variation in the percentages of seropositive pigs per farm as to most of the disease agents. Evidence was found that animals that were seropositive with respect to ADV were significantly more susceptible to becoming seropositive with respect to App. serotype 2, and vice versa. The same connection was observed for PI serotype H1N1 and PI serotype H3N2. Furthermore, evidence was found that pigs seropositive with respect to PI serotype H1Ni only, or to PI serotype H1N1 and ADV or PI serotype H3N2 show a significant decrease in average daily weight gain compared to pigs that were seronegative.     

A congenital persistent swine fever infection. II. Immune response to swine fever virus and unrelated antigens

After the disappearance of maternal antibodies, no antibodies were detected in five pigs that were congenitally infected with the Bergen strain of swine fever virus. The pigs lived for 2–11 months. One pig showed low levels of precipitating antibody during the last month of its life. The pigs had a normal immune response to sheep red blood cells and to pig parvovirus. These observations suggest the presence of immunological tolerance to swine fever virus in these pigs.Cell mediated immunity, measured by phytohaemagglutinin lymphocyte stimulation, appeared to be normal.     

Serologic status of pseudorabies virus in growing-finishing pigs in quarantined herds

It has been reported that pseudorabies virus (PRV) stops spreading within growing-finishing sections of a large percentage of infected farrow-to-finish herds. This study was designed to follow the PRV status of growing-finishing pigs in a sample of infected herds. Fifteen infected herds were selected, of which 11 had seropositive finishing pigs and 4 had seronegative finishing pigs. These herds were visited quarterly for one year, and a cross section of growing-finishing pigs was tested for the presence of anti-PRV antibodies. The 4 herds that initially were seronegative remained seronegative, whereas of the 11 herds initially seropositive, 4 remained seropositive, 4 became seronegative, and 3 became temporarily seronegative before becoming seropositive again. Three characteristics serologic profiles were observed: one indicating continued viral spread; one indicating no spread for at least the preceding 3 months; and one indicating that PRV spread had recently ceased in this section of the herd. Results of our study indicated that periodic monitoring of a cross section of the growing-finishing pigs for their PRV serologic status was valuable for determining whether PRV was actively spreading in this section of the herd.     

Modelling Salmonella spread within a farrow-to-finish pig herd

Delivery of infected pigs to the slaughterhouse is a major source of pork meat contamination by bacterial hazards to humans. We propose a model of Salmonella spread within a farrow-to-finish pig herd, assuming the prevalence in infected delivered pigs depends on the whole pig life-time and growing process. This stochastic discrete-time model represents both the population dynamics in a farrow-to-finish pig herd using batch management, and Salmonella spread. Four mutually exclusive individual health states were considered: Salmonella-free, seronegative shedder, seropositive shedder and seropositive not shedding carrier, making the distinction between seropositive animals and shedders. Since indirect transmission is the main route of transmission, the probability of infection depends on the quantity of Salmonella in the pigs' environment (Q). A dose effect function is used with two thresholds, assuming saturation in exposure for high Q vs. a minimum exposure for low Q. Salmonella is introduced in an initially Salmonella-free 150-sow herd. Prevalence of shedders and seroprevalence are calculated over time in batches of sows and pigs, and in groups of delivered pigs, composed of pigs from different batches. The model shows very variable seroprevalence over time within a herd among delivered groups, as well as among replications. The mean seroprevalence and the mean shedding prevalence are 19.3% and 13.8% respectively. A sensitivity analysis shows that the Salmonella quantity shed and the maternal protective factor are the most influential parameters on Salmonella prevalence in delivered pigs.     

Evaluation of transmission of swine influenza type A subtype H1N2 virus in seropositive pigs

OBJECTIVE: To examine clinical signs, virus infection and shedding, and transmission of swine influenza virus (SIV) subtype H1N2 among seropositive pigs. ANIMALS: Eighteen 3-week-old pigs with maternal antibodies against SIV subtypes H1N1, H3N2, and H1N2. PROCEDURE: Ten pigs (principal) were inoculated intranasally with subtype H1N2 and 2 groups of contact pigs (n = 4) each were mixed with principal pigs on day 7 (group 1) or 28 (group 2). Two principal pigs each were necropsied on days 4, 14, 21, 28, and 42 days after inoculation. Four pigs in each contact group were necropsied 35 and 14 days after contact. Virus excretion was evaluated after inoculation or contact. Lung lesions and the presence of SIV in various tissues were examined. RESULTS: Mild coughing and increased rectal temperature were observed in principal pigs but not in contact pigs. Nasal virus shedding was detected in all principal pigs from day 2 for 3 to 5 days, in group 1 pigs from day 2 for 4 to 9 days after contact, and in group 2 pigs from day 4 for 2 to 6 days after contact. Trachea, lung, and lymph node specimens from infected pigs contained virus. Antibody titers against all 3 subtypes in all pigs gradually decreased. CONCLUSIONS AND CLINICAL RELEVANCE: Protection from viral infection and shedding was not observed in pigs with maternal antibodies, but clinical disease did not develop. Vaccination programs and good management practices should be considered for control of SIV subtype H1N2 infection on swine farms.