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Inactivated whole-virus vaccines against porcine parvovirus (PPV) can prevent disease but not infection and virus shedding after heterologous virus challenge. Here, we showed that the same is true for a homologous challenge. Pregnant sows were vaccinated with an experimental inactivated vaccine based on PPV strain 27a. They were challenged on day 40 of gestation with the virulent porcine parvovirus PPV-27a from which the vaccine was prepared (homologous challenge). On day 90 of gestation, the fetuses from vaccinated sows were protected against disease, while the fetuses of the non-vaccinated sows (control group) exhibited signs of parvovirus disease. All gilts, whether vaccinated or not vaccinated, showed a boost of PPV-specific antibodies indicative of virus infection and replication. Low DNA copy numbers, but not infectious virus, could be demonstrated in nasal or rectal swabs of immunized sows, but high copy numbers of challenge virus DNA as well as infectious virus could both be demonstrated in non-vaccinated sows.

The importance of foodborne viruses is increasingly recognized. Thus, the effect of commonly used food preservation methods on the infectivity of viruses is questioned. In this context, we investigated the antiviral properties of d , l -lactic acid, sodium chloride and sodium nitrite by in vitro studies. Two model viruses, Feline Calicivirus (FCV) and Enteric Cytophatic Human Orphan (ECHO) virus, were chosen for this study simulating important foodborne viruses (human noroviruses (NoV) and human enteroviruses, resp.). The model viruses were exposed to different solutions of d , l -lactic acid (0.1–0.4% w/w, pH 6.0–3.2), of sodium chloride (2–20%, w/v) and of sodium nitrite (100, 150 and 200 ppm) at 4 and 20 °C for a maximum of 7 days. Different results were obtained for the two viruses. ECHO virus was highly stable against d , l -lactic acid and sodium chloride when tested under all conditions. On the contrary, FCV showed less stability but was not effectively inactivated when exposed to low acid and high salt conditions at refrigeration temperatures (4 °C). FCV titers decreased more markedly at 20 °C than 4 °C in all experiments. Sodium nitrite did not show any effect on the inactivation of both viruses. The results indicate that acidification, salting or curing maybe insufficient for effective inactivation of foodborne viruses such as NoV or human enteroviruses during food processing. Thus, application of higher temperature during fermentation and ripening processes maybe more effective toward the inactivation kinetics of less stable viruses. Nevertheless, more studies are needed to examine the antiviral properties of these preserving agents on virus survival and inactivation kinetics in the complex food matrix.

In recent years, it has been shown that some parvoviruses exhibit high substitution rates, close to those of RNA viruses. In order to monitor and determine new mutations in porcine parvovirus (PPV), recent PPV field isolates from Austria, Brazil, Germany and Switzerland were sequenced and analysed. These samples, together with sequences retrieved from GenBank, were included in three datasets, consisting of the complete NS1 and VP1 genes and a partial VP1 gene. For each dataset, the nucleotide substitution rate and the molecular clock were determined. Analysis of the PPV field isolates revealed that a recently described amino acid substitution, S436T, appeared to be common in the VP2 protein in the Austrian, Brazilian and German virus populations. Furthermore, new amino acid substitutions were identified, located mainly in the viral capsid loops. By inferring the evolutionary dynamics of the PPV sequences, nucleotide substitution rates of approximately 10−5 substitutions per site per year for the non-structural protein gene and 10−4 substitutions per site per year for the capsid protein gene (for both viral protein datasets) were found. The latter rate is similar to those commonly found in RNA viruses. An association of the phylogenetic tree with the molecular clock analysis revealed that the mutations on which the divergence for both capsid proteins was based occurred in the past 30 years. Based on these findings, it was concluded that PPV variants are continuously evolving and that vaccines, which are based mainly on strains isolated about 30 years ago, should perhaps be updated.

Since highly pathogenic avian influenza virus H5N1 emerged in 1997, avian influenza is considered one of the most important infectious diseases globally. In respect of virus transmission to humans, the consumption of raw poultry products remains of serious concern. In this study, data about survival time and inactivation kinetics of two low pathogenic avian influenza virus (AIV) strains (H3N8, H5N6) in short fermented raw sausage were obtained. In addition, the impact of the preserving factors d,l-lactic acid and sodium chloride on virus infectivity was evaluated through in vitro studies. Virus infectivity was confirmed in embryonated chicken eggs. Inactivation of H3N8 was seen in d,l-lactic acid solutions (0.15 and 0.20%, pH 4.40–4.70 and pH 3.80–3.91) at both temperatures (20 vs. 4°C) during 3 days of exposure. However, infectious virus particles could still be detected after exposure to 0.1% d,l-lactic acid (pH 5.80–5.99). In all NaCl solutions (2, 6 and 12% w/v), infectivity of the H3N8 strain decreased steadily but reduction of the virus titre increased significantly with higher temperature. In raw sausages, decline in virus titre was observed for both strains during ripening and storage. Thereby, decline of virus infectivity was dependent on time and temperature with a more marked effect at higher temperatures (22 vs. 7°C). At refrigeration (7°C), both viruses maintained infectivity over 14 days. Results indicate that appropriate processing of short fermented raw poultry sausage is likely to reduce risk of virus exposure due to adequate inactivation of AIV during ripening and storage.

Overview Feline panleukopenia virus (FPV) infects all felids as well as raccoons, mink and foxes. This pathogen may survive in the environment for several months and is highly resistant to some disinfectants. Infection Transmission occurs via the faecal–oral route. Indirect contact is the most common route of infection, and FPV may be carried by fomites (shoes, clothing), which means indoor cats are also at risk. Intrauterine virus transmission and infection of neonates can occur. Disease signs Cats of all ages may be affected by FPV, but kittens are most susceptible. Mortality rates are high – over 90% in kittens. Signs of disease include diarrhoea, lymphopenia and neutropenia, followed by thrombocytopenia and anaemia, immunosuppression (transient in adult cats), cerebellar ataxia (in kittens only) and abortion. Diagnosis Feline panleukopenia virus antigen is detected in faeces using commercially available test kits. Specialised laboratories carry out PCR testing on whole blood or faeces. Serological tests are not recommended, as they do not distinguish between infection and vaccination. Disease management Supportive therapy and good nursing significantly decrease mortality rates. In cases of enteritis, parenteral administration of a broad-spectrum antibiotic is recommended. Disinfectants containing sodium hypochlorite (bleach), peracetic acid, formaldehyde or sodium hydroxide are effective. Vaccination recommendations All cats – including indoor cats – should be vaccinated. Two injections, at 8–9 weeks of age and 3–4 weeks later, are recommended, and a first booster 1 year later. A third vaccination at 16–20 weeks of age is recommended for kittens from environments with a high infection pressure (cat shelters) or from queens with high vaccine-induced antibody levels (breeding catteries). Subsequent booster vaccinations should be administered at intervals of 3 years or more. Modified-live virus vaccines should not be used in pregnant queens or in kittens less than 4 weeks of age.

Feline calicivirus (FCV) is a highly variable virus. More severe, systemic forms of FCV infection have been observed recently.Sick, acutely infected or carrier cats shed FCV in oronasal and conjunctival secretions. Infection occurs mainly through direct contact.The main clinical signs are oral ulcers, upper respiratory signs and a high fever. Feline calicivirus may be isolated from nearly all cats with chronic stomatitis or gingivitis. Cats with 'virulent systemic FCV disease' variably show pyrexia, cutaneous oedema, ulcerative lesions on the head and limbs, and jaundice. Mortality is high and the disease is more severe in adult cats.Diagnosis of FCV can be achieved by virus isolation or reverse-transcriptase PCR. Viral RNA can be detected in conjunctival and oral swabs, blood, skin scrapings or lung tissue using PCR. Positive PCR results should be interpreted with caution, as these may be a consequence of low-level shedding by persistently infected carriers. The diagnosis of virulent systemic FCV disease relies on clinical signs and isolation of the same strain from the blood of several diseased cats.Supportive therapy (including fluid therapy) and good nursing care are essential. Anorexic cats should be fed highly palatable, blended or warmed food. Mucolytic drugs (eg, bromhexine) or nebulisation with saline may offer relief. Broad-spectrum antibiotics may be administered to prevent secondary bacterial infections. Feline calicivirus can persist in the environment for about 1 month and is resistant to many common disinfectants.Two injections, at 9 and 12 weeks of age, are recommended, followed by a first booster 1 year later. In high-risk situations, a third vaccination at 16 weeks is recommended. Boosters should be given every 3 years. However, cats in high-risk situations should be revaccinated annually. Cats that have recovered from caliciviral disease are probably not protected for life, particularly if infected with different strains. Vaccination of these cats is still recommended.

The banning of specified risk materials (SRMs) from food chain is one of the most important measures to protect the consumer against any exposition with the agent of transmissible spongiform encephalopathies (TSEs). In order to control the SRM-ban, suitable methods for the detection of SRMs have to be developed. In this study homogenized samples of central nervous system (CNS) tissue from cattle (n=38), sheep (n=38) and pig (n=40) were analyzed by gas chromatography-mass spectrometry (GC-MS). The analysis was focused on the identification of fatty acid ratios for species differentiation and age estimation. We found a novel fatty acid ratio (C24:1(n-9)/(n-7))/(2OH-C24:1(n-9)/(n-7) for species differentiation. For the first time, we used multivariate analysis for species differentiation, based on fatty acid ratios. It showed an excellent predictive ability (Q_(cum)(2))Q_(cum)(2)=0.842. For age estimation in cattle CNS we discovered a novel fatty acid ratio 2OH-C25:0/2OH-C24:1(n-7). The ratio is strongly correlated with age (coefficient of correlation (r)=0.935, Spearman-Rho). The corresponding regression analysis which allows the prediction of age by this ratio was acceptable (coefficient of determination (R(2))=0.831).

The present study was conducted to study the impact of the virulence factors invC and sseD of the two type III secretion systems of Salmonella enterica serovar Typhimurium (S. Typhimurium) on the pathogenesis of the porcine S. Typhimurium DT104 infection. For this purpose, two S. Typhimurium mutant strains with a disrupted invC gene of the Salmonella pathogenicity island 1 or with a disrupted sseD gene of the Salmonella pathogenicity island 2 have been studied in experimental infection of pigs. Twenty-two 7-week-old male hybrid pigs were either infected with one of the mutants or the wild-type S. Typhimurium DT104 strain. Each group was examined for clinical signs, Salmonella shedding rate and the specific antibody response. Survival and replication were evaluated by qualitative and quantitative determination of the colonization rate. The humoral and cellular immune responses were examined using isotype-specific ELISA and quantitative real-time PCR of IL-2, IL-4, IL-10, IL-12 and IFN-gamma. The results proved that both mutants had a lower virulence (with marked differences between both mutants) than the wild-type and that both virulence factors have importance in porcine salmonellosis. Only pigs infected with the wild-type S. Typhimurium DT104 exhibited typical clinical symptoms of salmonellosis like anorexia, vomiting, disturbed demeanour, fever and diarrhoea. Deletion of the invC gene resulted in a significantly reduced colonization rate. Interestingly, the mRNA expression of both type-1 and type-2 cytokines were significantly decreased in pigs infected with either the invC-mutant and the sseD-mutant strain.

The aim of this study was to investigate the subcutaneous tissue response to administration of a single dose of multi-component vaccine in the cat. Three groups of 15 cats were injected with one of three vaccine products with saline as a negative control. Cats in group A received non-adjuvanted vaccine; cats in group B received vaccine with a lipid-based adjuvant; whilst those in group C were vaccinated with a product adjuvanted with an alum–Quil A mixture. The vaccine and saline injection sites were sampled on days 7, 21 and 62 post-vaccination. Biopsies of these vaccine sites were examined qualitatively and scored semi-quantitatively for a series of parameters related to aspects of the inflammatory and tissue repair responses. These data were analysed statistically, including by principal component analysis. At all three time points of the experiment, there was significantly less inflammation associated with administration of non-adjuvanted vaccine ( p = 0.000). Although there was evidence of tissue repair by day 62 in all groups, those cats receiving adjuvanted vaccines had evidence of residual adjuvant material accumulated within macrophages at this late time point. The severity of tissue reactions may vary significantly in response to vaccines which include adjuvants or are non-adjuvanted.

Tierhalter und Zuchter informieren sich zunehmend im Internet uber Impfungen und deren Notwendigkeit, auch uber Impfschaden, und dem Tierarzt werden Fragen gestellt. Diese als Laienansuchen abzulehnen ware eine verpasste Gelegenheit zu einem Impfgesprach, das einer moglichen - aber nicht zwangslaufigen - Wiederimpfung vorausgehen sollte.

In order to clarify the epidemiology of bovine protothecal mastitis, 30 Prototheca zopfii mastitis isolates were genetically investigated. Based on the 18S rDNA, which allows a differentiation of the former species P. zopfii in two distinct P. zopfii genotypes and Prototheca blaschkeae sp. nov., newly developed genotype-specific PCR-assays as well as RFLP-assays were applied. All mastitis isolates investigated could be assigned to P. zopfii genotype 2 suggesting that this genotype is the aetiological agent of bovine Prototheca mastitis.

The pathogenicity of two recent German field isolates of Porcine parvovirus (PPV-27a and PPV-143a) and two vaccine viruses [PPV-NADL-2 and PPV-IDT (MSV)], which are used for the production of inactivated vaccines, was investigated by inoculation of pregnant sows at day 40 of gestation. Post-infection sera of these sows as well as antisera prepared in rabbits by immunization with the four above-mentioned PPV isolates and with the virulent strain PPV-Challenge (Engl.) were tested for their homologous and heterologous neutralization activities. All antisera had high neutralization activity against the vaccine viruses, the PPV-Challenge (Engl.) virus and PPV-143a, but much lower activity against PPV-27a. These results suggest that PPV-27a represents a new antigenic variant or type of PPV and vaccines based on the established vaccine viruses may not be fully protective against this field isolate. PPV-27a has been characterized based on the amino acid sequences of the capsid protein as a member of a new and distinct PPV cluster ( Zimmermann et al., 2006 ). Interestingly, the homologous neutralizing antibody titres of the sera of all three pigs and both rabbits inoculated or immunized with PPV-27a were 100- to 1000-fold lower than the heterologous titres against any of the other viruses. The low homologous neutralizing antibody titres suggest a possible, yet undefined, immune escape mechanism of this PPV isolate.

Ferlaviruses are important pathogens of snakes. However, factors influencing the pathogenicity of individual isolates as well as optimal protocols for virus detection in tissues of infected snakes have been insufficiently studied. The objectives of this study were to compare virus detection using previously described PCR and cell culture protocols following infection with three genetically distinct ferlaviruses in corn snakes (Pantherophis guttatus) as a model species. Groups of 12 corn snakes were each inoculated intratracheally with a genogroup A, B, or C ferlavirus. Tracheal washes and cloacal swabs were tested for virus shedding on days 16 and 28. Three animals were each euthanized on days 4, 16, 28, and 49. Beside immunohistochemistry of lung tissue, several organs (lung, intestine, pancreas, kidney, brain) were tested for the presence of ferlavirus. Distinct differences were noted in the pathogenicity of the three viruses, with a genotype B isolate causing the greatest pathology. PCR was more sensitive in comparison to cell culture, but results varied depending on the tissues. Ferlaviruses spread rapidly into the tissues, including the brain. Overall average detection rate was 72%, and was highest on day 16. There were differences between the groups, with the most virulent strain causing 100% positive samples at the end of the study. Some snakes were able to clear the infection. Shedding via cloaca was seen only on day 28. For ante-mortem sampling, a tracheal wash sample is recommended, for post mortem diagnosis, a pooled organ sample should be tested.

Overview: Borna disease virus (BDV) has a broad host range, affecting primarily horses and sheep, but also cattle, ostriches, cats and dogs. In cats, BDV may cause a non-suppurative meningoencephalomyelitis (‘staggering disease’). Infection: The mode of transmission is not completely elucidated. Direct and indirect virus transmission is postulated, but BDV is not readily transmitted between cats. Vectors such as ticks may play a role and shrews have been identified as a potential reservoir host. Access to forested areas has been reported to be an important risk factor for staggering disease. Disease signs: It is postulated that BDV may infect nerve endings in the oropharynx and spread via olfactory nerve cells to the central nervous sytem. A strong T-cell response may contribute to the development of clinical disease. Affected cats develop gait disturbances, ataxia, pain in the lower back and behavioural changes. Diagnosis: For diagnostic purposes, detection of viral RNA by reverse transcription PCR in samples collected from cats with clinical signs of Borna disease can be considered diagnostic. Serology is of little value; cats without signs of Borna disease may be seropositive and yet not every cat with BDV infection has detectable levels of antibodies. Human infection: A hypothesis that BDV infection may be involved in the development of selected neurological disorders in man could not be confirmed. A research group within the German Robert Koch Institute studied the potential health threat of BDV to humans and concluded that BDV was not involved in the aetiology of human psychiatric diseases.

Overview: Cytauxzoon species are apicomplexan haemoparasites, which may cause severe disease in domestic cats, as well as lions and tigers. For many years, cytauxzoonosis in domestic cats was only reported in North and South America, but in recent years the infection has also been seen in Europe (Spain, France and Italy). Infection: Cytauxzoon felis is the main species; it occurs as numerous different strains or genotypes and is transmitted via ticks. Therefore, the disease shows a seasonal incidence from spring to early autumn and affects primarily cats with outdoor access in areas where tick vectors are prevalent. Domestic cats may experience subclinical infection and may also act as reservoirs. Clinical signs: Cytauxzoonosis caused by C felis in the USA is an acute or peracute severe febrile disease with non-specific signs. Haemolytic anaemia occurs frequently; in some cats neurological signs may occur in late stages. The Cytauxzoon species identified in Europe differ from C felis that causes disease in the USA and are probably less virulent. The majority of infected cats have been healthy; in some cases anaemia was found, but disease as it occurs in the USA has not been reported to date. Diagnosis: Diagnosis is usually obtained by Cytauxzoon detection in blood smears and/or fine-needle aspirates from the liver, spleen and lymph nodes. PCR assays are able to detect low levels of parasitaemia and may be used for confirmation. Treatment: Currently a combination of the antiprotozoal drugs atovaquone and azithromycin is the treatment of choice. Concurrent supportive and critical care treatment is extremely important to improve the prognosis. Cats that survive the infection may become chronic carriers for life. Prevention: Cats with outdoor access in endemic areas should receive effective tick treatment.