L.A.M.G. van Leengoed

High volume hemofiltration improves right ventricular function in endotoxin-induced shock in the pig

This study assessed the influence of continuous high volume hemofiltration on right ventricular function of pigs with endotoxin induced shock. Eighteen anesthetized and ventilated pigs were studied for 240 min after the start of infusion of 0.5 mg/kg endotoxin over 30 min. Right ventricular ejection fraction (RVEF) was measured by rapid response thermodilution technique. After endotoxin infusion, the pigs were randomly divided into 3 groups: group 1 as a control group, receiving endotoxin only, group 2 to observe the effects of zero balance high volume veno-venous hemofiltration with removal of ultrafiltrate at a rate of 6000 ml/h, and group 3 to evaluate the effect of the extracorporeal circuit itself on RVEF. The decline of RVEF in group 2 was less than in group 1 (0.04±0.02 vs 0.21±0.03 (mean±SEM);p<0.001). The decline of RVEF in group 3 (0.24±0.02) was more pronounced than that in group 1 (p<0.05). The differences in the course of RVEF between group 1 and group 2 could not be explained by differences in heart rate, preload or afterload. Cardiac output and mean arterial pressure were significantly higher in group 2 than in group 1 (p<0.01). It is concluded that in this model, high volume hemofiltration improves RVEF and cardiac performance by removal of vasoactive mediators, responsible for myocardial depression.

Acquisition of Clostridium difficile by piglets

Clostridium difficile is recognized as an important cause of nosocomial diarrhoea in humans especially in association with administration of antibiotics. In pigs, C. difficile can cause neonatal enteritis and can be isolated from faeces from both diseased and healthy animals. The presented prospective study describes how soon C. difficile can be isolated from newborn piglets after normal parturition and how C. difficile spreads within a pig farm. Six sows, their farrowing crates and their litters at one farm were sampled until C. difficile was found in all piglets. Within 48 h after birth, all 71 piglets became positive for C. difficile (two piglets were already positive within 1h post partum), all sows became positive within 113 h after parturition and the farrowing crates were found intermittently positive. C. difficile could also be detected in air samples and in samples of teats of the sows. All isolates belonged to PCR ribotype 078. Twenty-one C. difficile ribotype 078 isolates, found at the farm, were further analyzed by MLVA (multiple-locus variable-number tandem repeat analysis) and belonged to one clonal complex, except one isolate. To be sure that piglets were not born already infected with C. difficile ribotype 078, 38 caesarean derived piglets were sampled immediately after surgery. All piglets tested negative at delivery and stayed negative for C. difficile ribotype 078 during the 21 days in which they were kept in sterile incubators. This study shows that C. difficile ribotype 078 spreads easily between sows, piglets and the environment. Vertical transmission of C. difficile ribotype 078 was not found and is very unlikely to occur.

A quantitative assessment of the effectiveness of PRRSV vaccination in pigs under experimental conditions.

This paper presents a quantitative approach to evaluate effectiveness of vaccination under experimental conditions. We used two consecutive experimental designs to investigate whether PRRSV transmission among vaccinated pigs was reduced compared to control pigs and to estimate the reproduction parameter R. Based upon data analysis and power calculations the series of small-scale vaccination-challenge experiments ended with multiple one-to-one experiments. This new experimental design has considerable power to detect the effect of vaccination on transmission if R is close to but still above one in vaccinated pigs. The last experiment showed that transmission was not significantly reduced and the R for vaccinated pigs was estimated to be larger than 4.9. This is remarkable because duration and level of viremia were significantly reduced by vaccination.

Streptococcus suis infections in pigs in the Netherlands (Part I)

Data are presented on the incidence of various streptococcal infections in pigs in the Netherlands. 314 Strains isolated in the course of routine post-mortem diagnosis were examined. The most frequently occurring streptococcus was S. subacidus (bio) type II which was isolated in 31.2% of the cases. S. suis type 2 (Serogroup R) and S. equisimilis (Serogroup C) constituted 16.2% and 13.7% of the isolates respectively. Besides meningitis, endocarditis and polyserositis S. suis type 2 infections may frequently be associated with pneumonia (42%). The biochemical profiles of the various S. suis and S. subacidus (bio) types are presented. The profile of both species is almost identical. It seems justified to use the name S. suis for strains with this characteristic profile and to abandon the name S. subacidus. Haemolysis does not appear to be a suitable characteristic to screen for S. subacidus/S. suis types. In comparing three serological methods for typing S. suis type 2, gel precipitation using Fullers extract and slide agglutination give an almost 100% correlation. These two methods are recommended for serotyping.

Dual infections of PRRSV/influenza or PRRSV/Actinobacillus pleuropneumoniae in the respiratory tract.

To study the effect of a previous porcine respiratory and reproductive syndrome-infection (PRRS) of the respiratory tract on influenza virus and Actinobacillus pleuropneumoniae (App) infections, 3-week-old specific-pathogen-free (spf) piglets were intranasally infected with PRRS virus. One week later, when the lung alveolar macrophages of PRRSV infected pigs were lowest in number, a second infection was applied by intranasal aerosol of influenza virus H3N2 or by endobronchial instillation of a mildly virulent App. The first experiment consisted of two groups (only influenza infection or dual PRRSV/influenza infection). A second experiment consisted of 4 groups (only influenza infection, only PRRSV infection, dual PRRSV/influenza infection and uninfected controls). At day 2, 4, 14 and 21 after influenza infection, two pigs were killed and sampled for virological and histopathological examination. Influenza H3N2 virus caused only a mild inflammation of the smaller bronchioli. Previous PRRSV infection did not influence clinical signs during influenza infection. Next, we studied in two experiments the effect of dual PRRSV/App infection during the acute stage at two days after App infection. In a third experiment, the influence of PRRSV on more chronic stages of App infection was studied at two weeks after the App infection. At the end of the experiments, the pigs were killed. Lungs were ranked according to size and kind of the lesions. Lesions were cut and measured, samples were taken for virological and histopathological examination. Statistical analysis of the ranked lung-lesions in the first experiment showed a distinct but small effect of previous PRRSV infection on the development of App-lesions. In PRRSV infected pigs. App produced a more severe disease. The second and third experiment however failed to show any influence of the previous PRRSV infection on the App infection. We conclude that previous PRRSV infection of the respiratory tract of spf pigs does not necessarily enhance the severity of secondary infections of the respiratory tract.

Introduction, persistence and fade-out of porcine reproductive and respiratory syndrome virus in a Dutch breeding herd: a mathematical analysis

The objective of this study was to investigate the dynamics of PRRSV infection and to quantify transmission within a breeding herd, and its impact on herd performance. For this purpose a longitudinal study was performed in a closed breeding herd of 115 sows. Statistical methods and Monte Carlo simulations based on stochastic SIR models were used to analyse the observational data. Moreover, a case-control study was performed to determine whether seroconversion of sows during gestation was associated with aberrant litters. The transmission parameter R was estimated to be 3.0 (95% confidence interval 1.5-6.0) for the model version based on the most plausible assumptions that the infectious period lasts 56 days and no lifelong immunity exists after infection. Based on simulations using a breeding herd of equal size the average time-to-extinction was estimated to be 6 years; using a herd of twice the size, it was 80 years. Furthermore, in contrast to the epidemic phase of the disease, the endemic phase was not detrimental to herd performance.

Estimating the incidence of influenza-virus infections in Dutch weaned piglets using blood samples from a cross-sectional study.

A cross-sectional study was carried out on 32 Dutch breeding herds to estimate the incidence of influenza-virus infections in piglets before the start of the finishing period, at the age of approximately 10 weeks. Longitudinal studies on two herds (8 and 10 litters, respectively) were done to obtain an average decay function for maternal antibodies.Each participating farm in the cross-sectional study was visited twice within 5 months; each time, blood samples were taken randomly from one compartment (a separate room with separate air flow) of 4-5-week-old piglets and one compartment of 8-9-week-old piglets. These blood samples (a total of 2598; 16-23 per compartment, depending on its size) were tested in a haemagglutination inhibition test for antibodies against influenza-virus subtypes H1 and H3. Samples from 8-9-week-old piglets from the first sampling period (n=660) were also tested in an IgM ELISA. For each individual herd and each influenza-virus subtype separately, the decay function derived from the longitudinal studies was used to calculate an expected seroprevalence in 8-9-week-old piglets, which was then compared to the observed seroprevalence. Depending on subtype and sampling period, between 10 and 15 of the 32 herds were suspected of virus circulation during the weaning period because the observed seroprevalence was significantly higher than the expected seroprevalence (P<0.05). In the first sampling period the IgM ELISA confirmed six of these outbreaks. However, due to the small window of detection of the IgM ELISA (compared to the length of the weaning period), it will always underestimate the number of infections. Infections in the first half of the weaning period will no longer be detectable because IgM antibodies have already disappeared. In individual pigs, an incidence of 16-17% was estimated for each subtype over a 4-week period between the age of 4-5 and 8-9 weeks. For each influenza subtype, 80% of the piglets will enter the finishing facilities without antibodies or with decaying maternal antibodies. These piglets may be susceptible to an infection with influenza virus.

RELEASE OF PROINFLAMMATORY CYTOKINES RELATED TO LUTEOLYSIS AND THE PERIPARTURIENT ACUTE PHASE RESPONSE IN PROSTAGLANDIN-INDUCED PARTURITION IN COWS

An acute phase response was previously found in cows at parturition, which might be associated with uterine cytokine release. Five late pregnant cows were implanted with vascular catheters in both the maternal aorta and uterine vein. Blood samples were taken to study temporal relationships between changing plasma levels of proinflammatory cytokines and the periparturient acute phase response following prostaglandin (PG)-induced luteolysis at Day 275 of gestation. The plasma levels of three proinflammatory cytokines, tumor necrosis factor-alpha (TNF-alpha), interleukin-1 (IL-1) and interleukin-6 (IL-6), as well as progesterone (P4), PGFM and serum amyloid A (SAA) were measured every 4 h between PG induction and expulsion of the calf. In the arterial plasma, progesterone levels dropped to baseline levels within 10 h following PG treatment, indicative of complete luteolysis. Contrary to expectations, the uterine vein samples showed lower proinflammatory cytokine levels compared with the maternal aorta values. A classical acute phase response, as assessed by SAA, was observed during the expulsive stage, but not during luteolysis.

Seroprevalence of porcine reproductive and respiratory syndrome virus in Dutch weaning pigs

To determine whether under Dutch field conditions PRRSV infection occurs in weaning pigs before the finishing period, a cross-sectional study was performed on 32 breeding farms to estimate the seroprevalence of antibodies directed against PRRSV in 4- to 5-week-old and 8- to 9-week-old pigs. Farms were visited twice within 5 months, and during each sampling an average of 20 sera were randomly collected from a unit of 4- to 5-week-old and a unit of 8- to 9-week-old pigs. The sera (n = 2568) were tested in the IDEXX-ELISA for the presence of antibodies directed against PRRSV. The seroprevalence of PRRSV in 4- to 5-week-old pigs and 8- to 9-week-old pigs varied between both samplings for each farm. The seroprevalence in the younger pigs was significantly higher than in the older pigs for both samplings (p < 0.05), suggesting the presence of maternal antibodies. In addition, a longitudinal study was performed to evaluate the IDEXX-ELISA in detecting maternal antibodies directed against PRRSV and to determine the rate of decline of these antibodies in field sera. From serological results of eight litters, an average decay function was computed to quantify the maternal immunity to PRRSV. A seroprevalence in 8- to 9-weeks-old pigs of > or = 0.20 was calculated to indicate an active immune response to PRRSV. In the cross-sectional study in the pigs twenty-three percent of the units with 8- to 9-week-old pigs were considered to have an active serological response against PRRSV. We conclude that most Dutch pigs are seronegative for PRRSV at the start of the finishing period, since the results of this study showed that 77% of the units with 8- to 9-week-old pigs had a seroprevalence < 0.20.

Evaluation of Four Different Diagnostic Tests To Detect Clostridium difficile in Piglets

ABSTRACT Clostridium difficile is emerging as pathogen in both humans and animals. In 2000 it was described as one of the causes of neonatal enteritis in piglets, and it is now the most common cause of neonatal diarrhea in the United States. In Europe, C. difficile infection (CDI) in both neonatal piglets and adult sows has also been reported. Diagnosis of this infection is based on detection of the bacterium C. difficile or its toxins A and B. Most detection methods, however, are only validated for diagnosing human infections. In this study three commercially available enzyme immunoassays (EIAs) and a commercial real-time-PCR (Becton, Dickinson, and Company) were evaluated by testing 172 pig fecal specimens (139 diarrheic and 33 nondiarrheic piglets). The results of each test were compared to those of cytotoxicity assays (CTAs) and toxigenic culture as the “gold standards.” Compared to CTAs, the sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) were, respectively, as follows: for real-time PCR, 91.6, 37.1, 57.6, and 82.5%; for Premier Toxins A&B (Meridian), 83.1, 31.5, 53.1, and 66.7%; for ImmunoCard Toxins A&B kit (ICTAB; Meridian), 86.6, 56.8, 66.9, and 80.7%; and for VIDAS (bioMérieux), 54.8, 92.6, 85.0, and 72.8%. Compared to toxigenic culture, the sensitivity, specificity, PPV, and NPV were, respectively, as follows: for real-time PCR, 93.0, 34.7, 50.0, and 87.5%; for Premier Toxins A&B, 80.3, 27.7, 43.8, and 66.7%; and for ICTAB, 80.0, 46.2, 52.8, and 75.4%; and for VIDAS, 56.4, 89.8, 77.5, and 76.7%. We conclude that all tests had an unacceptably low performance as a single test for the detection of C. difficile in pig herds and that a two-step algorithm is necessary, similar to that in cases of human CDI. Of all of the assays, the real-time PCR had the highest NPV compared to both reference methods and is therefore the most appropriate test to screen for the absence of C. difficile in pigs as a first step in the algorithm. The second step would be a confirmation of the positive results by toxigenic culture.