Influenza surveillance in pigs: balancing act between broad diagnostic coverage and specific virus characterization.

BACKGROUND: Monitoring of infectious diseases on swine farms requires a high diagnostic sensitivity and specificity of the test system. Moreover, particularly in cases of swine influenza A virus (swIAV) it is desirable to include characterization of the virus as precisely as possible. This is indispensable for strategies concerning prophylaxis of swIAV and furthermore, to meet the requirements of a purposeful monitoring of newly emerging swIAV strains in terms of vaccine design and public health. Within the present cross-sectional study, we compared the diagnostic value of group samples (wipes of surfaces with direct contact to mouth/nose, dust wipes, udder skin wipes, oral fluids) to individual samples (nasal swabs, tracheobronchial swabs) for both swIAV identification and characterization. Sampling included different stages of pig production on 25 sow farms with attached nursery considered as enzootically infected with swIAV. Firstly, samples were analyzed for IAV genome and subsequently samples with Ct-values < 32 were subtyped by multiplex RT-qPCR. RESULTS: Nasal swabs of suckling piglets and nursery pigs resulted in a higher odds to detect swIAV (p < 0.001) and to identify swIAV subtypes by RT-qPCR (p < 0.05) compared to nasal swabs of sows. In suckling piglets, significant higher rates of swIAV detection could be observed for nasal swabs (p = 0.007) and sow udder skin wipes (p = 0.036) compared to contact wipes. In the nursery, group sampling specimens were significantly more often swIAV positive compared to individual samples (p < 0.01), with exception of the comparison between contact wipes and nasal swabs (p = 0.181). However, in general nasal swabs were more likely to have Ct-value < 32 and thus, to be suitable for subtyping by RT-qPCR compared to dust wipes, contact wipes, udder skin wipes and tracheobronchial swabs (p < 0.05). Interestingly, different subtypes were found in different age groups as well as in different specimens in the same holding. CONCLUSION: Although population-based specimens are highly effective for swIAV monitoring, nasal swabs are still the preferable sampling material for the surveillance of on-farm circulating strains due to significantly higher virus loads. Remarkably, sampling strategies should incorporate suckling piglets and different age groups within the nursery to cover as many as possible of the on-farm circulating strains.

Examination on the Occurrence of Coinfections in Diagnostic Transmittals in Cases of Stillbirth, Mummification, Embryonic Death, and Infertility (SMEDI) Syndrome in Germany.

The stillbirth, mummification, embryonic death, and infertility (SMEDI) syndrome is most commonly associated with porcine parvovirus 1 (PPV1) infections. Little is known about the occurrence of coinfections with SMEDI-associated pathogens and the associations among these pathogens. In our study, we included 40 SMEDI-affected litters from 18 different farms. In total, 158 out of 358 available fetuses from diagnostic transmittals were selected by systematic random sampling and examined for PCV2, PCV3, PPV1, and Leptospira spp. by q-PCR. Results from diagnostic materials showed the following results: in eleven farms, PCV2 was present; in nine farms, PPV1 was present; in five farms, PCV3 was present; and in two farms, Leptospira spp. was present. The detection of Leptospira spp. was significantly associated with a PCV2 coinfection (OR: 26.3; p < 0.001). PCV3 positivity resulted in a reduced probability of detecting PCV2 in the corresponding fetus (OR: 0.078; p = 0.008). Fetal maceration was associated with Leptospira spp. detection (OR: 8.6; p = 0.003), whereas mummification (p = 0.047), reduced crown-rump length (p < 0.001), and bodyweight (p = 0.001) of fetuses were significantly associated with PPV1 and PCV2 coinfection and thus, presumably, a shorter time to death after infection, indicating an enhanced negative effect on the development of fetuses with PCV2 + PPV1 coinfection.

Explorative Field Study on the Use of Oral Fluids for the Surveillance of Actinobacillus pleuropneumoniae Infections in Fattening Farms by an Apx-Real-Time PCR.

Oral fluids (OFs) represent a cost effective and reliable tool for surveillance purposes, mostly regarding viruses. In the present study, we evaluated the suitability of OFs for surveillance purposes concerning Actinobacillus (A.) pleuropneumoniae infections in fattening pigs under field conditions. OFs were examined with an Apx-toxin real-time PCR that detects the genes encoding for Apx I-, Apx III-, and Apx IV-toxin. For this purpose, we conducted a pen-wise collection of OFs over one fattening period from fattening pigs of two farms (farm A and B) with a known history of A. pleuropneumoniae infection. Lung lesions were determined at slaughter to estimate the extend of pulmonary lesions and pleural affection. Apx III- and Apx IV-toxin DNA were present in the OFs of both farms whereas Apx I-toxin DNA was present on farm A only. We were able to detect Apx I-, Apx III-, and Apx IV-toxin DNA in different patterns directly after introduction of the new pigs in the farms and over the entire study period. In summary, or results indicate the suitability of OFS for the early detection and surveillance of A. pleuropneumoniae in fattening farms.

Cross-Sectional Study on the Prevalence of PCV Types 2 and 3 DNA in Suckling Piglets Compared to Grow-Finish Pigs in Downstream Production.

Vertical transmission is a consistently discussed pathway of porcine circovirus type 2 (PCV2) and porcine circovirus type 3 (PCV3) transmission in pigs. To evaluate the presence of PCV2 and PCV3 in piglets, we collected tissue samples from 185 piglets that were crushed within the first week of life from 16 farms located in Germany and Austria. Pooled samples consisting of thymus, inguinal lymph node, myocardium, lung and spleen were examined for PCV2 and PCV3 by qPCR. Furthermore, oral fluid samples (OFS) from grow−finish pigs were collected and examined the same way. In piglets, PCV2 was highly prevalent (litters: 69.4%; piglets: 61.6%), whereas PCV3 prevalence was low (litters: 13.4%; piglets: 13.0%). In total, 72.6% and 67.2% of all collected OFS were PCV2 or PCV3 positive, respectively. Sow vaccination against PCV2 was identified as a protective factor concerning PCV2 in piglets (OR: 0.279; CI: 0.134−0.578; p < 0.001), whereas the porcine reproductive and respiratory syndrome virus (PRRSV) vaccination of sows was identified as a protective factor concerning PCV3 in piglets (OR: 0.252 CI: 0.104−0.610; p = 0.002). Our results show that PCV2, but not PCV3, is ubiquitous in suckling piglets and that early PCV3 infections might be modulated by PRRSV−PCV3 interaction. However, the ubiquitous nature of both viruses in older pigs could be confirmed.

[Hind limb paralysis in fattening pigs due to a new strain of porcine Teschovirus A11]. / Hinterhandlähmungen bei Mastschweinen im Zusammenhang mit einem neuen Stamm des porzinen Teschovirus A11.

In a fattening farm in southern Germany, paralysis of the hind limbs was observed in 2 age groups (50 kg as well as 60 kg) during a 4 week period. Despite a low morbidity of 3.3 % the majority of the affected animals needed to be euthanized in consequence to the progression of their hind limb paralysis. During pathomorphological examinations of 2 affected fattening pigs severe lymphohistiocytic meningoencephalomyelitis and vasculitis were detected. Immunhistochemistry revealed the presence of Porcine Teschovirus antigen in all parts of the central nervous system as well as in several cell types (neurons, glia cells, endothelial cells, mononuclear cells). Porcine Teschovirus was detected by PCR in spinal cord samples. The subsequently performed phylogenetic analysis PCR revealed a close relation (88 % full genome sequence) to porcine Teschovirus A11 strain "Dresden". Other swine relevant pathogens were excluded by PCR, bacteriologic examination and sequencing. Following a period of 4 weeks no additional cases of hind limb paralysis were observed in the fattening farm.

Cross-sectional study on the in-herd prevalence of Mycoplasma hyopneumoniae at different stages of pig production.

BACKGROUND: A cross-sectional study was carried out to assess the prevalence of Mycoplasma hyopneumoniae infections before vaccination in 3-week-old piglets and to gain information about infection dynamics. METHODS: In 13 German and three Austrian farms with a known history of enzootic pneumonia, 790 piglets and 158 sows were sampled (blood samples, tracheobronchial swabs [TBS] [piglets], laryngeal swabs [LS] [sows]), and 525 pen-based oral fluids (OFs) were collected in growing and fattening pigs. Laboratory diagnostics included enzyme-linked immunosorbent assay (ELISA) and real-time polymerase chain reaction (PCR) analyses. RESULTS: Antibodies to M. hyopneumoniae were present in 87.5 per cent of all herds. Seroprevalence ranged from 0.0 to 100.0 per cent and 0.0 to 88.0 per cent in sows and piglets, respectively. M. hyopneumoniae-deoxyribonucleic acid (DNA) was present in 3.8 and 0.4 per cent of LS and TBS, respectively. Gilts had a 10.9 times higher chance being M. hyopneumoniae PCR-positive than older sows. In 75.0 per cent of all farms, M. hyopneumoniae-DNA was present in OFs. Detection rate was significantly higher in OFs of 20-week-old than in younger pigs (p < 0.001). CONCLUSION: Results indicate that M. hyopneumoniae infections of the lower respiratory tract in piglets are rare but highlight the role of gilts in maintaining infection in the herd. Collecting OFs seems promising for surveillance, if coughing occurs simultaneously.

Detection of porcine circovirus type 3 DNA in serum and semen samples of boars from a German boar stud.

Porcine circovirus type 3 (PCV3) is regularly reported in association with various clinical presentations, including porcine dermatitis and nephropathy syndrome (PDNS)-like lesions, respiratory signs, congenital tremor, and reproductive disorders. To investigate the epidemiology of PCV3 in a boar stud, we analysed fresh boar semen and matching sera from 181 boars from a German stud supplying semen for artificial insemination (AI) to approximately 740 breeder farms for PCV3 DNA. PCV3 DNA was detected in 1.7% semen samples and 24.3% sera. Spearman rho correlation demonstrated a significant positive correlation between boar age and quantitative DNA (by PCR quantification cycles [Cq] values) in serum samples (r = 0.636; P < 0.001). Sera from boars up to 12 months of age had higher viral loads (P < 0.001) and were PCV3-positive significantly more often (P < 0.01) than older boars. Detection of PCV3 DNA was not associated with breed (P> 0.05). PCV3 DNA was detected sporadically in fresh boar semen. Based on the assumption that processing fresh semen reduces viral load in semen used for AI, it is likely that the risk of sexual transmission of PCV3 during AI in is low. However, young boars may contribute to the maintenance of PCV3 infection in boar studs.

Update on shedding and transmission routes of porcine haemotrophic mycoplasmas in naturally and experimentally infected pigs.

Horizontal transmission of Mycoplasma suis via parenteral exposure during standard practices or through bites during fightings have been identified as key epidemiological routes. However, as knowledge gaps on other potential shedding and transmission routes exist, the present study combines both laboratory experiments and field surveys to gain new insights into the epidemiology of porcine haemotrophic mycoplasmas. Splenectomised pigs were orally inoculated with a M. suis field strain and investigated for clinical signs related to infectious anaemia of pigs (IAP) and the presence of M. suis in blood, urine and saliva samples by qPCR. All blood samples were negative for M. suis and animals did not show obvious clinical signs of IAP throughout the entire study period. Additionally, urine, nasal and saliva samples from sows of conventional piglet producing farms and semen samples from a boar stud revealed no detection of M. suis and 'Candidatus Mycoplasma haemosuis' by qPCR. Thus, the results indicate that blood-independent transmission routes might be of minor relevance under field conditions.

[The porcine respiratory disease complex (PRDC) - a clinical review]. / Der Porcine Respiratory Disease Complex (PRDC) ­ eine klinische Übersicht.

The porcine respiratory disease complex describes a clinical condition that often manifests as treatment-resistant respiratory disease of growing to finishing pigs. Its multifactorial etiology includes infectious and non-infectious factors. Besides management and hygiene conditions, particularly viral and bacterial pathogens contribute to the development and course of PRDC. The porcine reproductive and respiratory syndrome virus (PRRSV), porcine circovirus type 2 (PCV2), influenza A virus (IAV) and Mycoplasma (M.) hyopneunoniae are considered as the major pathogens involved in PRDC. The clinical outcome and necropsy findings may differ depending on the involvement of the different pathogens. The complex nature of the PRDC impedes the diagnostic and preventive measures on affected farms. The present review provides insight into the pathomorphology, pathogenesis and inter-pathogen-interactions and aims to support practitioners in implementing purposeful diagnostic and preventive measures.

Discriminating the eight genotypes of the porcine circovirus type 2 with TaqMan-based real-time PCR.

BACKGROUND: The porcine circovirus type 2 (PCV2) is divided into eight genotypes including the previously described genotypes PCV2a to PCV2f and the two new genotypes PCV2g and PCV2h. PCV2 genotyping has become an important task in molecular epidemiology and to advance research on the prophylaxis and pathogenesis of PCV2 associated diseases. Standard genotyping of PCV2 is based on the sequencing of the viral genome or at least of the open reading frame 2. Although, the circovirus genome is small, classical sequencing is time consuming, expensive, less sensitive and less compatible with mass testing compared with modern real-time PCR assays. Here we report about a new PCV2 genotyping method using qPCR. METHODS: Based on the analysis of several hundred PCV2 full genome sequences, we identified PCV2 genotype specific sequences or single-nucleotide polymorphisms. We designed six TaqMan PCR assays that are specific for single genotypes PCV2a to PCV2f and two qPCRs targeting two genotypes simultaneously (PCV2g/PCV2d and PCV2h/PCV2c). To improve specific binding of oligonucleotide primers and TaqMan probes, we used locked nucleic acid technology. We evaluated amplification efficiency, diagnostic sensitivity and tested assay specificity for the respective genotypes. RESULTS: All eight PCV2 genotype specific qPCRs demonstrated appropriate amplification efficiencies between 91 and 97%. Testing samples from an epidemiological field study demonstrated a diagnostic sensitivity of the respective genotype specific qPCR that was comparable to a highly sensitive pan-PCV2 qPCR system. Genotype specificity of most qPCRs was excellent. Limited unspecific signals were obtained when a high viral load of PCV2b was tested with qPCRs targeting PCV2d or PCV2g. The same was true for the PCV2a specific qPCR when high copy numbers of PCV2d were tested. The qPCR targeting PCV2h/PCV2c showed some minor cross-reaction with PCV2d, PCV2f and PCV2g. CONCLUSION: Genotyping of PCV2 is important for routine diagnosis as well as for epidemiological studies. The introduced genotyping qPCR system is ideal for mass testing and should be a valuable complement to PCV2 sequencing, especially in the case of simultaneous infections with multiple PCV2 genotypes, subclinically infected animals or research studies that require large sample numbers.

Bone regeneration of minipig mandibular defect by adipose derived mesenchymal stem cells seeded tri-calcium phosphate- poly(D,L-lactide-co-glycolide) scaffolds.

Reconstruction of bone defects represents a serious issue for orthopaedic and maxillofacial surgeons, especially in extensive bone loss. Adipose-derived mesenchymal stem cells (ADSCs) with tri-calcium phosphates (TCP) are widely used for bone regeneration facilitating the formation of bone extracellular matrix to promote reparative osteogenesis. The present study assessed the potential of cell-scaffold constructs for the regeneration of extensive mandibular bone defects in a minipig model. Sixteen skeletally mature miniature pigs were divided into two groups: Control group and scaffolds seeded with osteogenic differentiated pADSCs (n = 8/group). TCP-PLGA scaffolds with or without cells were integrated in the mandibular critical size defects and fixed by titanium osteosynthesis plates. After 12 weeks, ADSCs seeded scaffolds (n = 7) demonstrated significantly higher bone volume (34.8% ± 4.80%) than scaffolds implanted without cells (n = 6, 22.4% ± 9.85%) in the micro-CT (p < 0.05). Moreover, an increased amount of osteocalcin deposition was found in the test group in comparison to the control group (27.98 ± 2.81% vs 17.10 ± 3.57%, p < 0.001). In conclusion, ADSCs seeding on ceramic/polymer scaffolds improves bone regeneration in large mandibular defects. However, further improvement with regard to the osteogenic capacity is necessary to transfer this concept into clinical use.

Detection of Mycoplasma suis in pre-suckling piglets indicates a vertical transmission.

BACKGROUND: Transmission of Mycoplasma (M.) suis mainly occurs via iatrogenic or zootechnical manipulations or due to ranking fights. Other transmission routes including ingestion of secretes/excretes; blood-sucking arthropods and intra-uterine transmission have thought to play an epidemiological role without being experimentally proven. To investigate a vertical transmission of M. suis under field conditions blood samples from pre-suckling piglets and their corresponding dam were examined for M. suis by quantitative polymerase chain reaction (qPCR) in 21 farms in Southern Germany. RESULTS: A total of 14.35% of the 474 blood samples from pre-suckling piglets reacted qPCR positive. Additionally, M. suis was detected in 65 (31.25%) of the 208 sows at farrowing. On farm level, 16 (76.2%) of the 21 farms had at least one M. suis positive animal. M. suis positive farms had an average of 0.41 more stillborn piglets per litter than M. suis negative farms (p = 0.007). CONCLUSION: The present study provides further insights into M. suis infection dynamics as it is the first detection of M. suis in piglets immediately after birth prior to colostrum intake and the first large scale investigation of M. suis in sows at farrowing.

Cross-sectional study on viraemia and shedding of porcine circovirus type 2 in a subclinically infected multiplier sow herd.

Vertical and horizontal transmission of porcine circovirus type 2 (PCV2) plays an important role for the spread of PCV2 within piglet-producing farms and following production steps. Further information is crucial to learn about the principles of PCV2 circulation among sows in piglet-producing farms to improve preventive healthcare concerning porcine circovirus diseases (PCVD) in downstream production steps. The present study was conducted as a cross-sectional study in a 400 sow multiplier herd in Germany with no PCV2 vaccination. Blood, faeces and saliva of the sows in all stages of production were tested for PCV2-DNA by real-time PCR. Results were analysed under respect of the parity and stage of production of the sows. PCV2-DNA in faeces or saliva was observed especially in young sows. Highest rates of viraemia in productive sows were found in the early stages of pregnancy. The results revealed that particularly gilts from the quarantine and rearing area and sows up to the second parity play a major role for the spread of PCV2 and thus for the maintenance of PCV2 infection in sow herds. Furthermore, the stage of production had a significant influence on the detection rate of PCV2-DNA in serum, saliva or faeces of the sows.

Full genome characterization of porcine circovirus type 3 isolates reveals the existence of two distinct groups of virus strains.

BACKGROUND: The occurrence of the novel porcine circovirus type 3 (PCV3) was reported from the Americas, Asia and Europe. Although this virus was detected in association with various clinical syndromes in pigs, its role as possible swine pathogen remains unclear. PCV3 was detected with high prevalence in Polish farms, but to date no genome sequences were available from European PCV3 strains. METHODS: We collected 1060 serum samples from piglets at the age of 20-24 weeks from 53 farms distributed all over Germany. PCV3 DNA was detected using a real-time PCR and subsequently complete PCV3 genome sequences were obtained after multiply primed rolling circle amplification and sequencing of overlapping PCR products. Phylogenetic analysis was performed by neighbor-joining method and maximum likelihood method. RESULTS: We obtained 15 complete PCV3 genome sequences as well as nine partial sequences including the putative ORFs 1, 2 and 3 from PCV3 viremic animals in German pig farms. Phylogenetic analysis of these German as well as 30 full genome sequences received from GenBank divided the PCV3 strains into two main groups and several subclusters. Furthermore, we were able to define group specific amino acid patterns in open reading frame 1 and 2. CONCLUSION: PCV3 is distributed with high prevalence in German pig industry. Phylogenetic analysis revealed two clearly separated groups of PCV3 strains, which might be considered as PCV3 genotypes. Specific nucleotide and amino acid marker positions may serve for easy and fast intraspecies classification and genotyping of PCV3 strains. No correlation between PCV3 variants with their geographical origin was evident. We found the same diversity of PCV3 strains in Germany as in other countries. We hypothesize that PCV3 is not a newly emerging virus in the German pig population. Future studies will have to show, if PCV3 genotype specific biological properties are evident.

Field study on the safety and efficacy of intradermal versus intramuscular vaccination against Mycoplasma hyopneumoniae.

The present study compares the safety and efficacy of a needle-free, intradermal Mycoplasma hyopneumoniae vaccine to an intramuscular one. 420 piglets (21+3 days of age) were randomly assigned to two vaccination groups (intradermal vaccination V1 (n=138), intramuscular vaccination V2 (n=144)) and one unvaccinated control group (CG, n=138). As safety parameters clinical observations, local injection site reactions (ISR) and rectal temperatures were assessed. Average daily weight gain (ADWG) and pneumonic lung lesions (LL) were measured as efficacy parameters. ISRs were minor in V1. After both vaccinations, no adverse impact on appetite was observed and mean rectal temperatures remained within physiological range. ADWG during the fattening period was significantly higher in vaccinated groups (V1: 913.4 g, V2: 924.5 g) compared with CG (875.6 g). No differences in ADWG were observed between V1 and V2. Vaccinated pigs had a significantly reduced mean extent of LL compared with CG. V1 was superior in reducing the extent and prevalence of LL compared with V2. These results reveal that a needle-free intradermal vaccination is safe and efficacious in reducing both the prevalence and extent of lung lesions, as well as in improving performance parameters, in a farrow-to-finish farm with a late onset of M hyopneumoniae infection.

[Occurrence of genotypes of porcine circovirus (PCV2) in pig farms using different vaccination strategies against PCV2]. / Vorkommen von Genotypen des porzinen Circovirus Typ 2 (PCV2) in Schweinebeständen mit unterschiedlichen Impfstrategien gegen PCV2.

OBJECTIVE: Since 2004/2005 a worldwide shift of the detection rate of porcine circovirus (PCV) has been observed from PCV2a towards PCV2b. Currently commercially available vaccines are based on genotype PCV2a. The study was conducted as a pilot study to evaluate the occurrence of PCV2a and PCV2b in farms with different vaccination strategies against PCV2. MATERIAL AND METHODS: For this purpose a total of 405 piglets originating from nine farms (three farms with sow vaccination [SI], piglet vaccination [FI] and no vaccination [NI] against PCV2, respectively) were enrolled and followed from day 3 of life until slaughter. Serum of the piglets was examined for PCV2-DNA by quantitative PCR, genotype differentiating duplex PCR, and after sequencing of the total genome, PCV2 isolates were phylogenetically assigned. The evaluation included the data from 383 animals. RESULTS: In eight farms PCV2 could be detected (1x PCV2a; 6x PCV2b; 1x PCV2a and PCV2b). PCV2b was found in SI-, NI- and FI-farms, whereas PCV2a was only detected in SI- and NI-farms. A proportion of 55.4% was PCV2-positive at least once during the entire study period (FI: 7.8%, SI: 65.4%, NI: 93.7%). Of these samples 4.7% were PCV2a-, 92.2% PCV2b- and 2.4% PCV2a- and PCV2b-positive. The mean content of PCV2-DNA in the serum of PCV2b positive animals was significantly higher than from PCV2a positive animals. PCV2 isolates were identified as PCV2b-1A (5/9 farms), PCV2b-1B (1/9 farms) und PCV2a-2D (2/9 farms). CONCLUSION AND CLINICAL RELEVANCE: The increased detection rate of PCV2b in comparison to PCV2a could be confirmed. The present study gives hint that the vaccination of piglets using PCV2a-based vaccines may lead to a further shift of the detection rate from PCV2a to PCV2b. To assess the clinical relevance of this observation, extensive comparative studies should be taken into account, which also evaluate the efficacy of PCV2a-based vaccines in PCV2a- and PCV2b-positive farms.

[Detection of Salmonella Choleraesuis var. Kunzendorf in a fattening pig with septicaemic salmonellosis. A case report]. / Nachweis von Salmonella Choleraesuis var. Kunzendorf bei einem Mastschwein mit septikämischer Salmonellose. Ein Fallbericht.

The present case reports the detection of Salmonella (S.) Choleraesuis var. Kunzendorf and porcine circovirus type 2 in an organic fattening pig suffering from septicaemic salmonellosis and porcine dermatitis and nephropathy syndrome. Six weeks after pigs had been housed in an organic fattening farm, an increase in mortality, diarrhea and coughing was observed. In recent years, S. choleraesuis var. Kunzendorf has been frequently detected in wild boars in Germany, whereas the same serovar did not play an important role in the Western European domestic pig population. A direct transmission of this serovar from wild boars to domestic pigs in this case could not be proven. However, because wild boars are important reservoirs for the spread of epizootic diseases to the domestic pig population, this case emphasises the importance of taking epidemiological relationships under consideration and to comply with biosecurity measures according to German law (Schweinehaltungshygieneverordnung).

Assessment of safety and reproductive performance after vaccination with a modified live-virus PRRS genotype 1 vaccine in pregnant sows at various stages of gestation.

The objective of the present study was to assess safety and efficacy of a new modified live-virus porcine reproductive and respiratory syndrome (PRRS) genotype 1 vaccine in pregnant sows at various stages of gestation under field conditions. A total of 505 sows and gilts were allocated to two treatment groups and maintained in separate facilities. Animals of group 1 were vaccinated with a commercial modified live genotype 1 PRRSV vaccine (control product, CP), while animals of group 2 were immunized with a new modified live genotype 1 PRRSV vaccine (investigational veterinary product, IVP) (ReproCyc® PRRS EU, Boehringer Ingelheim Vetmedica GmbH). Injection site reactions were noted to be significantly less frequent in the IVP group compared to the CP group for pain (p=0.039), redness (p=0.030), heat (p=0.016) and swelling (p=0.002). The mean total number of piglets alive at weaning did not differ significantly between both study groups (10.6 vs. 11.0, p=0.375). However, pre-weaning mortality was significantly higher (p=0.005) in piglets from the CP group (14.1% vs. 10.9%). Analyses of reproductive performance data for both groups did not result in statistically significant differences between CP group and IVP group for number of piglets alive (12.7 and 12.6, respectively), healthy live (11.9 and 11.8), weak (0.7 and 0.5), stillborn (1.0 and 0.8) and mummified piglets (0.3 and 0.2) per litter. No differences were detected between both groups for piglet birth weights, while body weights at weaning (7.2kg vs. 6.6kg, p=0.026) and average daily gain (0.2445kg vs. 0.2211kg, p=0.037) were significantly higher in piglets from the IVP group. In conclusion, the administration of a single dose of ReproCyc® PRRS EU to sows and gilts at various stages of gestation confirmed non-inferiority to a commercial PRRS vaccine regarding safety and efficacy parameters under field conditions.

Effects of the glucagon-like peptide-1 receptor agonist liraglutide in juvenile transgenic pigs modeling a pre-diabetic condition.

BACKGROUND: The glucagon-like peptide-1 receptor (GLP1R) agonist liraglutide improves glycemic control and reduces body weight of adult type 2 diabetic patients. However, efficacy and safety of liraglutide in adolescents has not been systematically investigated. Furthermore, possible pro-proliferative effects of GLP1R agonists on the endocrine and exocrine pancreas need to be further evaluated. We studied effects of liraglutide in adolescent pigs expressing a dominant-negative glucose-dependent insulinotropic polypeptide receptor (GIPR(dn)) in the beta-cells, leading to a pre-diabetic condition including disturbed glucose tolerance, reduced insulin secretion and progressive reduction of functional beta-cell mass. METHODS: Two-month-old GIPR(dn) transgenic pigs were treated daily with liraglutide (0.6-1.2 mg per day) or placebo for 90 days. Glucose homeostasis was evaluated prior to and at the end of the treatment period by performing mixed meal and intravenous glucose tolerance tests (MMGTT and IVGTT). Finally animals were subjected to necropsy and quantitative-stereological analyses were performed for evaluation of alpha- and beta-cell mass, beta-cell proliferation as well as acinus-cell proliferation. RESULTS: MMGTT at the end of the study revealed 23% smaller area under the curve (AUC) for glucose, a 36% smaller AUC insulin, and improved insulin sensitivity, while IVGTT showed a 15% smaller AUC glucose but unchanged AUC insulin in liraglutide- vs. placebo-treated animals. Liraglutide led to marked reductions in body weight gain (-31%) and food intake (-30%) compared to placebo treatment, associated with reduced phosphorylation of insulin receptor beta (INSRB)/insulin-like growth factor-1 receptor beta (IGF1RB) and protein kinase B (AKT) in skeletal muscle. Absolute alpha- and beta-cell mass was reduced in liraglutide-treated animals, but alpha- and beta-cell mass-to-body weight ratios were unchanged. Liraglutide neither stimulated beta-cell proliferation in the endocrine pancreas nor acinus-cell proliferation in the exocrine pancreas, excluding both beneficial and detrimental effects on the pig pancreas. CONCLUSIONS: Although plasma liraglutide levels of adolescent transgenic pigs treated in our study were higher compared to human trials, pro-proliferative effects on the endocrine or exocrine pancreas or other liraglutide-related side-effects were not observed.

Field evaluation of the efficacy, compatibility and serologic profiling of a combined vaccine against porcine reproductive and respiratory syndrome and Haemophilus parasuis in nursery pigs.

The aim of the study was to compare the efficacy and compatibility of a separate or combined vaccination against the porcine reproductive and respiratory syndrome (PRRS) and Haemophilus (H.) parasuis. The study was conducted in a 1200 head nursery farm. A total of 360 piglets at an age of 26 days were randomized into three groups. Group A was vaccinated separately against H. parasuis (Porcilis(®)Glässer) and PRRS (Porcilis(®)PRRS), group B was vaccinated with a combined vaccine of both vaccines and group C remained unvaccinated as control group. The compatibility was evaluated by measurement of the body temperature and a palpation score of the injection site 0, 4, 24 and 72 h after vaccination. During the nursery and the fattening period the average daily weight gain (ADWG), the number of runts and the mortality was evaluated. Additionally blood samples were taken every 2 weeks during the nursery period to perform an OppA-ELISA and a PCR for PRRS virus. No significant difference could be seen regarding the body temperature between group A and group C. Piglets which were vaccinated with the combined vaccine showed a significantly higher body temperature 4 and 72 h post vaccination than piglets from group A. The palpation score was significantly higher in group A 4 and 24h post vaccination compared to the control group, whereas no significant difference was observed between group A and B. No significant differences between groups were seen in the ADWG during the nursery period. The mortality rate during the nursery period was significantly higher in group C than in group A. The ADWG during fattening was significantly higher in the vaccinated groups than in group C. A PRRS genotype1 field virus was detected at the end of the nursery period. No significant differences were observed in the number of OppA-ELISA positive animals, but vaccinated pigs seemed to react earlier. All pigs of the vaccinated groups that were positive in the OppA-ELISA did not develop Glässer's disease and remained in the study until slaughter. The combined administration had no negative influence on efficacy but showed a slightly worse compatibility than the separate administration of both vaccines. The results of the present study indicate that vaccination against Glässer's disease using Porcilis(®)Glässer might influence the results of the OppA-ELISA.