Infectivity and pathobiology of H7N1 and H5N8 high pathogenicity avian influenza viruses for pigeons (Columba livia var. domestica).

Avian influenza (AI) is one of the most important viral diseases in poultry, wildlife and humans. Available data indicate that pigeons play a minimum role in the epidemiology of AI. However, a degree of variation exists in the susceptibility of pigeons to highly pathogenic AI viruses (HPAIVs), especially since the emergence of the goose/Guangdong H5 lineage. Here, the pathogenesis of H5N8 HPAIV in comparison with a H7N1 HPAIV and the role of pigeons in the epidemiology of these viruses were evaluated. Local and urban pigeons (Columba livia var. domestica) were intranasally inoculated with 105 ELD50 of A/goose/Spain/IA17CR02699/2017 (H5N8) or A/Chicken/Italy/5093/1999 (H7N1) and monitored during 14 days. Several pigeons inoculated with H5N8 or H7N1 seroconverted. However, clinical signs, mortality, microscopic lesions and viral antigen were only detected in a local pigeon inoculated with H5N8 HPAIV. This pigeon presented prostration and neurological signs that correlated with the presence of large areas of necrosis and widespread AIV antigen in the central nervous system, indicating that the fatal outcome was associated with neurological dysfunction. Viral RNA in swabs was detected in some pigeons inoculated with H7N1 and H5N8, but it was inconsistent, short-term and at low titres. The present study demonstrates that the majority of pigeons were resistant to H5N8 and H7N1 HPAIVs, despite several pigeons developing asymptomatic infections. The limited viral shedding indicates a minimum role of pigeons as amplifiers of HPAIVs, regardless of the viral lineage, and suggests that this species may represent a low risk for environmental contamination. RESEARCH HIGHLIGHTS H7N1 and H5N8 HPAIVs can produce subclinical infections in pigeons. The mortality caused by H5N8 HPAIV in one pigeon was associated with neurological dysfunction. Pigeons represent a low risk for environmental contamination by HPAIVs.

Experimental infection of domestic geese (Anser anser var. domesticus) with H5N8 Gs/GD and H7N1 highly pathogenic avian influenza viruses.

Prior to the emergence of the Asian-origin H5 Goose/Guangdong/1/96 (Gs/GD) lineage, highly pathogenic avian influenza viruses (HPAIV) had rarely caused high mortalities in domestic geese. In 2016/2017 European epidemics, H5N8 Gs/GD clade 2.3.4.4 Group B produced an unprecedented number of outbreaks in waterfowl holdings. In this study, the pathogenesis of H5N8 HPAIV in comparison with H7N1 HPAIV, and the role of domestic geese in the epidemiology of these viruses, were evaluated. Local and commercial geese (Anser anser var. domesticus) were intranasally inoculated with 105 ELD50 of A/goose/Spain/IA17CR02699/2017 (H5N8) or A/Chicken/Italy/5093/1999 (H7N1) and monitored daily during 15 days. H5N8 was highly virulent to domestic geese, reaching 100% mortality by 10 days post-infection. Systemic microscopic necrotizing lesions associated with widespread AIV-antigen were detected by IHC techniques, the central nervous system being the most severely affected. High viral loads, measured by qRT-PCR, were present in all samples collected: oral and cloacal swabs, plasma tissues, and moderate levels in pool water. Domestic geese were also susceptible to H7N1 infection, as demonstrated by seroconversion and detection of viral RNA in tissues and plasma in some geese, but all lacked clinical signs. Viral shedding was confirmed in only some geese and was restricted to the oral route, but levels were high and still detected at the end of the study. Overall, H7N1 presents a lower lethality and shedding than H5N8 in geese; however, the viral shedding indicates that these species could play a role in the epidemiology of Gs/GD and other lineages of HPAIVs. RESEARCH HIGHLIGHTS H5N8 Gs/GD clade 2.3.4.4 Group B is highly virulent to domestic geese. The severity of H5N8 is associated with multisystemic replication. H7N1 can infect domestic geese but is avirulent to this species. Domestic geese could play a role in the epidemiology of Gs/GD HPAIVs.

Non-linear quantization error reduction for the temperature measurement subsystem on-board LISA Pathfinder.

Laser Interferometer Space Antenna (LISA) Pathfinder is a mission to test the technology enabling gravitational wave detection in space and to demonstrate that sub-femto-g free fall levels are possible. To do so, the distance between two free falling test masses is measured to unprecedented sensitivity by means of laser interferometry. Temperature fluctuations are one of the noise sources limiting the free fall accuracy and the interferometer performance and need to be known at the ∼10 µK Hz-1/2 level in the sub-millihertz frequency range in order to validate the noise models for the future space-based gravitational wave detector LISA. The temperature measurement subsystem on LISA Pathfinder is in charge of monitoring the thermal environment at key locations with noise levels of 7.5 µK Hz-1/2 at the sub-millihertz. However, its performance worsens by one to two orders of magnitude when slowly changing temperatures are measured due to errors introduced by analog-to-digital converter non-linearities. In this paper, we present a method to reduce this effect by data post-processing. The method is applied to experimental data available from on-ground validation tests to demonstrate its performance and the potential benefit for in-flight data. The analog-to-digital converter effects are reduced by a factor between three and six in the frequencies where the errors play an important role. An average 2.7 fold noise reduction is demonstrated in the 0.3 mHz-2 mHz band.

Transmission of Porcine reproductive and respiratory syndrome virus 1 to and from vaccinated pigs in a one-to-one model.

The present study examined transmission by contact of Porcine reproductive and respiratory syndrome virus (PRRSV) 1 in a one-to-one model to vaccinated and unvaccinated pigs and from vaccinated infected pigs to other vaccinated pigs. The experiment started by randomly assigning weaned pigs to groups V (n=24) and U (n=26). V pigs were vaccinated with a commercial live attenuated PRRSV vaccine and the U animals were kept as unvaccinated controls. Twenty-eight days later, 6U pigs were separated and allocated in individual boxes. The remaining 20U pigs were intranasally inoculated with PRRSV isolate 3267 (from now on designated as seeder (S) pigs) and 48h later were distributed in boxes where they were commingled with either V or U pigs in 1:1 groups (first contact phase), resulting in 6S:U and 14S:V pairs. As soon as a V pig was detected to be viremic because of contact with a S, the infected V (from now on designated as Vinf) was transferred (<24h after detection) to a new pen where it was comingled with a new V pig (designated as V2) in a second contact phase. For the first contact phase, pigs were maintained 21days at maximum and for the second contact phase the maximum exposure period was 14days. Two V pigs tested positive for the vaccine virus (>99.5% similarity) when they were relocated with the corresponding V2 pigs and they were removed; thus, only 12Vinf were finally considered. All V pigs (12/12) exposed to S animals became infected although the first detection of viremia occurred at 13.6±3.6days, one week later than in U (p<0.05). Also, duration of viremia was shorter for Vinf compared to U, (5.5±4.3days versus 12.5±2.7days). The Vinf group showed remarkable individual variability: eight animals had a viremic period of 5 or less days (3.0±1.4) while the remaining four had a longer viremic period of more than one week (10.8±2.9). This situation was not observed in U. In the second contact phase, transmission from Vinf to V2 pigs occurred in 7/8 cases (87.5%). The mean duration of viremia for V2 was 4.8±3.4 and two different patterns were again observed: two animals had viremias of 9-10days and the rest averaged 3.0±1.4days (range: 2-5days). Vaccinated groups Vinf and V2 had a significantly lower PRRSV shedding in oral fluids for at least the first 9days after the onset of the viremia compared to U, and shedding for V2 was even significantly lower (p<0.05) than shedding for Vinf. Our experimental design reproduced the worst-case scenario for evaluating the effect of vaccination and, under such conditions; it was still efficacious in slowering PRRSV transmission and decreasing the global viral load and particularly oral shedding.

A novel variant of the infectious bronchitis virus resulting from recombination events in Italy and Spain.

Infectious bronchitis is considered to be one of the most devastating diseases in poultry. Control of its spread is typically attempted through biosecurity measures and extensive vaccination. However, the remarkable genetic and antigenic variability of the virus, which originate from both mutations and recombination events, represents an unsolved challenge for this disease. The present study reports on the emergence and spread of recombinant clusters detected in Italy and Spain between 2012 and 2014. A total of 36 Spanish and Italian infectious bronchitis virus (IBV) field strains were investigated and genetically characterized using phylogenetic, molecular, recombination and selection pressure analyses of the complete S1 gene. Based on the partial S1 sequencing, 27 IBV strains originating from Spain and nine from Italy were initially classified as being closely related to the Guandong/Xindadi (XDN) genotype. Phylogenetic analysis of the complete S1 gene revealed that the XDN strains formed a homogeneous clade with the Spanish IBV isolates within the QX genotype, whereas there was higher variability within the Italian strains. Recombination analysis determined that these strains belonged to four groups, which originated from independent recombination events between the QX and 793B IBV genotypes. Our data support the hypothesis of two different scenarios: firstly, in Spain, the large and homogeneous clade probably originated from a single offspring of the recombinant founder, which became dominant and spread throughout the country. Secondly, the nine Italian recombinants, which are characterized by three different recombination patterns, probably represent less fitted strains, because they were less viable with respect to their recombinant parents.

Six-Year Follow-up of Slaughterhouse Surveillance (2008-2013): The Catalan Slaughterhouse Support Network (SESC).

Meat inspection has the ultimate objective of declaring the meat and offal obtained from carcasses of slaughtered animals fit or unfit for human consumption. This safeguards the health of consumers by ensuring that the food coming from these establishments poses no risk to public health. Concomitantly, it contributes to animal disease surveillance. The Catalan Public Health Protection Agency (Generalitat de Catalunya) identified the need to provide its meat inspectors with a support structure to improve diagnostic capacity: the Slaughterhouse Support Network (SESC). The main goal of the SESC was to offer continuing education to meat inspectors to improve the diagnostic capacity for lesions observed in slaughterhouses. With this aim, a web-based application was designed that allowed meat inspectors to submit their inquiries, images of the lesions, and samples for laboratory analysis. This commentary reviews the cases from the first 6 years of SESC operation (2008-2013). The program not only provides continuing education to inspectors but also contributes to the collection of useful information on animal health and welfare. Therefore, SESC complements animal disease surveillance programs, such as those for tuberculosis, bovine cysticercosis, and porcine trichinellosis, and is a powerful tool for early detection of emerging animal diseases and zoonoses.

Virological and serological characterization of vaccinated and non-vaccinated piglet subpopulations coming from vaccinated and non-vaccinated sows.

The present study describes the virological and serological profiles of PCV2 vaccinated (V) and non-vaccinated (NV) piglet subpopulations coming from V and NV sows in a PCV2 subclinically infected farm. Four hundred seventy-six piglets born from V or NV sows were further subdivided in a total of four groups: NV sows-NV pigs (NV-NV), NV sows-V pigs (NV-V); V sows-NV pigs (V-NV) and V sows-V pigs (V-V). Seventy-five pigs were randomly selected at the beginning of the trial from each group and they were bled at 4, 8, 12, 16, 21 and 25 weeks of age. All animals included in the trial were weighed at 4 and 25 weeks of age and their average daily weight gain (ADWG) was calculated. Serum samples obtained at different time points were used to assess PCV2 infection (viremia) and the level of antibodies by means of immunoperoxidase monolayer assay (IPMA) against this pathogen. IPMA titers (classified in high, medium or low) and PCR results (positive or negative) were analyzed using a multiple correspondence and K-means cluster analysis. According to these tests, animals included in the study were classified into the following four clusters: (1) 93 piglets that were viremic mainly from 12 to 25 weeks of age and with PCV2 antibody titers increasing over time; (2) 75 piglets with late PCV2 infection and seroconversion (later than 16 weeks of age); (3) 26 piglets with high but decreasing PCV2 antibody titers and low percentages of PCV2 PCR positive serum samples; and (4) 105 piglets with medium and high IPMA titers throughout the trial and sporadic PCR positive samples. The defined subpopulations of piglets were observed in all experimental groups (NV-NV, NV-V, V-NV and V-V) although in variable percentages. Thus, animals from clusters 1 and 2 belonged mainly to the NV-NV and V-NV groups and animals from clusters 3 and 4 were distributed mainly into the NV-V and V-V groups. Finally, the ADWG of pigs belonging to clusters 3 and 4 was significantly higher (p=0.02) than that of pigs belonging to clusters 1 and 2. Within each cluster, no statistically significant differences were found in ADWG between treatment groups.

Effect of porcine circovirus type 2 (PCV2) load in serum on average daily weight gain during the postweaning period.

Porcine circovirus type 2 (PCV2) is a ubiquitous virus that mainly affects nursery and fattening pigs causing systemic disease (PCV2-SD) or subclinical infection. A characteristic sign in both presentations is reduction of average daily weight gain (ADWG). The present study aimed to assess the relationship between PCV2 load in serum and ADWG from 3 (weaning) to 21 weeks of age (slaughter) (ADWG 3-21). Thus, three different boar lines were used to inseminate sows from two PCV2-SD affected farms. One or two pigs per sow were selected (60, 61 and 51 piglets from Pietrain, Pietrain×Large White and Duroc×Large White boar lines, respectively). Pigs were bled at 3, 9, 15 and 21 weeks of age and weighted at 3 and 21 weeks. Area under the curve of the viral load at all sampling times (AUCqPCR 3-21) was calculated for each animal according to standard and real time quantitative PCR results; this variable was categorized as "negative or low" (<10(4.3) PCV2 genome copies/ml of serum), "medium" (≥10(4.3) to ≤10(5.3)) and "high" (>10(5.3)). Data regarding sex, PCV2 antibody titre at weaning and sow parity was also collected. A generalized linear model was performed, obtaining that paternal genetic line and AUCqPCR 3-21 were related to ADWG 3-21. ADWG 3-21 (mean±typical error) for "negative or low", "medium" and "high" AUCqPCR 3-21 was 672±9, 650±12 and 603±16 g/day, respectively, showing significant differences among them. This study describes different ADWG performances in 3 pig populations that suffered from different degrees of PCV2 viraemia.

Humoral response and colostral antibody transfer following 'one-dose' pre-mating vaccination of sows against porcine circovirus type-2.

The objective of this study was to investigate the presence of porcine circovirus type 2 (PCV2) DNA and antibody to the virus in the serum and colostrum of sows vaccinated prior to mating and in their offspring. Seventy-seven sows were randomly distributed into vaccinated (V, n=36) and non-vaccinated (NV, n=41) groups. One week before mating, sows were given a PCV2 vaccine (V group) or PBS (NV group) IM. Blood samples were taken from the sows at fixed time-points and colostrum samples were taken at farrowing. Blood samples were also taken from the piglets of the sows at 4 weeks of age. The results indicated that vaccination prior to mating elicited a strong, homogeneous humoral response and, in consequence, more homogeneous colostral PCV2 antibody concentrations.

Simultaneous porcine circovirus type 2 and Mycoplasma hyopneumoniae co-inoculation does not potentiate disease in conventional pigs.

The aim of this study was to assess the effect of simultaneous experimental inoculation of porcine circovirus type 2 (PCV2; intranasal delivery) and Mycoplasma hyopneumoniae (Mhyo; transtracheal delivery) into conventional, seropositive 6-week-old piglets. Thirty-six male piglets were assigned randomly to four groups: control (n=6), PCV2 (n=6), Mhyo (n=12) and PCV2+Mhyo (n=12). Blood samples and faecal and nasal swabs were collected at 0, 7, 14 and 21 days post inoculation (dpi). No significant clinical signs attributable to PCV2 infection were observed during the experiment. Coughing was recorded in three pigs from the Mhyo group and six from the PCV2+Mhyo group. No significant differences in mean body weight and rectal temperature were observed between the groups. Mild microscopical lesions similar to those reported for post-weaning multisystemic wasting syndrome were observed in two PCV2 pigs and in one PCV2+Mhyo animal. Mhyo-compatible lung lesions were observed in 21/24 pigs inoculated with Mhyo (10 from the Mhyo group and 11 from the PCV2+Mhyo group). PCV2 was detected by in-situ hybridization in 3/12 PCV2 and in 4/12 PCV2+Mhyo animals. No significant differences in PCV2 load (serum and nasal and faecal swabs), duration of viraemia or antibody titre were detected between PCV2-inoculated groups. No significant differences in Mhyo load in nasal swabs, percentage of Mhyo-seropositive pigs and mean lung score was detected between Mhyo-inoculated groups. Under the conditions of the present study, concurrent inoculation of PCV2 and Mhyo did not result in potentiation of clinical signs and lesions attributed to either infection.

Enhancing DNA immunization by targeting ASFV antigens to SLA-II bearing cells.

One of the main criticisms to DNA vaccines is the poor immunogenicity that they confer on occasions, at least in large animals. Confirming this theory, immunization with plasmid DNA encoding two African swine fever virus genes in frame (pCMV-PQ), failed in inducing detectable immune responses in pigs, while it was successful in mice. Aiming to improve the immune responses induced in swine, a new plasmid was constructed, encoding the viral genes fused in frame with a single chain variable fragment of an antibody specific for a swine leukocyte antigen II (pCMV-APCH1PQ). Our results clearly demonstrate that targeting antigens to antigen professional cells exponentially enhanced the immune response induced in pigs, albeit that the DNA vaccine was not able to confer protection against lethal viral challenge. Indeed, a viremia exacerbation was observed in each of the pigs that received the pCMV-APCH1PQ plasmid, this correlating with the presence of non-neutralizing antibodies and antigen-specific SLA II-restricted T-cells. The implications of our discoveries for the development of future vaccines against African swine fever virus and other swine pathogens are discussed.

Selected Swine viral pathogens in indoor pigs in Spain. Seroprevalence and farm-level characteristics.

A serosurvey on porcine reproductive and respiratory syndrome virus (PRRSV), swine influenza virus (SIV), Aujeszky's disease virus gE protein (ADV gE), porcine parvovirus (PPV) and porcine circovirus type 2 (PCV2) was carried out in Spanish pig herds. The serosurvey consisted of two studies. First, a retrospective study assessed the proportion of seropositive boar, sow and fattening pig herds and their seroprevalences to PRRSV, SIV, ADV gE and PPV from 2003 to 2005 and to PCV2 from 2000 to 2005. Such information was obtained from routine serologic analyses from two veterinary diagnostic laboratory services. Second, a cross-sectional study in sow and fattening pig herds from 44 farms (without vaccination interferences on serologic analyses) was performed to provide information on seroprevalences and co-seropositivity to PRRSV, SIV, ADV gE and PCV2 (PPV was excluded because of widespread vaccination) and to elucidate their relationships with farm characteristics, management and productive parameters. Similar seroprevalences were observed in both studies, although some variations were obtained, probably because of vaccination schedules, number of tested sera, sampling age and regional variations. Percentage of PRRSV and SIV seropositive herds was over 85% for sows, around 80% for fatteners and around 50% for boar studs. The proportion of ADV gE seropositive sow herds decreased from 41% to 30% between 2003 and 2005, whereas such decrease was from 41% to 33% in fattening pig herds and from 13% to 4% in boar studs PCV2 antibodies were widespread as well as those against PPV; in the latter case, if antibodies were elicited by infection and/or vaccination was not assessed. Concurrent presence of PCV2, PRRSV and SIV antibodies was found in 89% and 66% sow and fattening herds, respectively. No statistical associations were obtained between seroprevalences or co-seropositivity and farm characteristics, management or productive parameters.

Sow vaccination modulates the colonization of piglets by Haemophilus parasuis.

Haemophilus parasuis is the etiologic agent of Glässer's disease in pigs and a colonizer of the upper respiratory tract of healthy pigs. A good balance between colonization and immunity is important to avoid a disease outbreak. This work studied the colonization of H. parasuis in healthy piglets coming from vaccinated and non-vaccinated sows. Piglets from vaccinated sows had higher IgG levels at early time points and subsequently were colonized later and to a lower degree than piglets from non-vaccinated ones. The variability of H. parasuis isolates was investigated by 2 genotyping methods: enterobacterial repetitive intergenic consensus (ERIC)-PCR and multilocus sequence typing (MLST). A high turnover of strains was found in both groups of piglets, with few strains found on more than one sampling occasion. We found a higher number of H. parasuis strains (16 strains) within a given farm than previously thought. Overall, more H. parasuis diversity was found in piglets from non-vaccinated sows than in those from vaccinated sows. These results indicate that vaccination of sows in a farm delays the colonization of piglets and reduces the carriage and heterogeneity of H. parasuis strains.

Risk factors associated with pleuritis and cranio-ventral pulmonary consolidation in slaughter-aged pigs.

Examination of lung lesions at the slaughterhouse is a useful tool to estimate the importance of respiratory disease at farm, regional or national level. The objective of the present work was to describe the prevalence of gross lung lesions at slaughter, with a special focus on pleuritis and cranio-ventral pulmonary consolidation, and to identify major risk factors for these lesions. Data from 107 farms involving approximately 11,000 pigs enabled gross lung lesions to be correlated with serology to different swine respiratory pathogens as well as with production system characteristics and vaccination schedules. Pleuritis and cranio-ventral pulmonary consolidation lesions were recorded in 26.8% and 55.7% of slaughter-aged pigs, respectively. Among lungs with pleuritis, 50.1% had lesions compatible with Actinobacillus pleuropneumoniae (App) infection. Antibodies to porcine reproductive and respiratory syndrome (PRRSV), three subtypes (H1N1, H1N2 and H3N2) of swine influenza virus (SIV), App and Mycoplasma hyopneumoniae (Mhyo) were highly prevalent (>82%) in most of the farms. In a multivariable analysis, it was estimated (R(2)=0.40) that the percentage of animals with pleuritis compatible with App infection depended on the existence of an all in-all out by room management system and App and PRRSV herd seroprevalence. Moreover, it was possible to foresee (R(2)=0.59) that cranio-ventral pulmonary consolidation lesions (EP-like lesions) were affected by the type of farm ventilation, the presence of respiratory symptoms during the fattening period and Mhyo and SIV H1N2 herd seroprevalence.

Interferon-gamma induction correlates with protection by DNA vaccine expressing E2 glycoprotein against classical swine fever virus infection in domestic pigs.

Classical swine fever (CSF) is a highly contagious viral infection affecting domestic and wild pigs. For classical swine fever virus (CSFV), immunization with plasmids expressing different versions of glycoprotein E2 has proven an effective way to induce protection. Previously, we have also shown that immunization with DNA vaccine expressing glycoprotein E2 (DNA-E2) induced specific T helper cell responses in the absence of neutralizing antibodies. However, the role of T cell responses in protection against CSFV is largely unknown. Here we have extended these studies to deeply characterize the role of T cell responses by a DNA-E2 and their correlation with protection against CSFV infection. Thus, pigs vaccinated with the DNA vaccine induced a strong cellular immune response, characterized by the specific induction IFN-gamma expressing T cells after vaccination without any detectable levels of CSFV neutralizing antibodies. Constant levels of CSFV-specific IFN-gamma producing cells observed from the beginning of the infection until 7 days after challenge in vaccinated animals might contribute to early control of CSFV replication, at least until neutralizing antibodies are developed. Severe clinical signs of the disease, including high titers of viremia, pyrexia and virus spread to different organs, were recorded in the non-vaccinated challenged animals, in comparison to the vaccinated animals where only one animal showed mild clinical signs and a short peak of viremia. Lack of complete protection in this animal correlated with a delay on the induction of neutralizing antibodies, detectable only from day 11 post-CSFV challenge. Conversely, the rest of the pigs within the group developed neutralizing antibodies as early as at day two post-challenge, correlating with sterile protection. Finally, an inverse correlation seemed to exist between early induction of IFN-alpha and the protection observed, while IL-10 seemed to be differentially regulated in vaccinated and non-vaccinated animals. Our results support the relevance of the induction of a strong T cellular response to confer a solid protection upon DNA vaccination against CSFV. Further experiments are needed to be done in order to clarify the key cytokines playing a role in CSFV-protection and to obtain emergency vaccines capable to confer robust and fast protection.

A genetically engineered chimeric vaccine against porcine circovirus type 2 (PCV2) improves clinical, pathological and virological outcomes in postweaning multisystemic wasting syndrome affected farms.

The present study describes the effects of a commercially available genetically engineered chimeric vaccine against porcine circovirus type 2 (PCV2) on clinical, pathological and virological features in three multi-site farms suffering from postweaning multisystemic wasting syndrome (PMWS). The vaccine product was able to reduce clinical signs, PCV2 viral load in lymphoid organs and/or sera, and overall mortality in nurseries and fattening units. This is the first time in which is shown that a PCV2 vaccine is able to decrease specifically PMWS-associated mortality. Another novelty of this study is the assessment of PMWS-like histological lesions in a large number of vaccinated and non-vaccinated pigs under field conditions.

Effect of dietary level of protein and fiber on the productive performance and health status of piglets.

To study the interaction between the levels of protein and fiber on the productive performance and health status of piglets, ninety-six 35-d-old piglets (9.11 +/- 0.60 kg of BW) were placed in 32 pens of 3 animals each and allotted to 4 dietary treatments for 21 d. The 4 diets were based on rice, dairy products, and soybean meal in a 2 x 2 factorial arrangement of treatments, with 2 levels of CP (15.4 vs. 19.4%, as-fed basis) and 2 levels of dietary fiber [DF; low fiber (LF) 5.3% NDF and high fiber (HF) 7.15% NDF, as-fed basis]. The HF diet was developed by supplementing the basal diet with 40 g/kg of wheat bran and 20 g/kg of sugar beet pulp. Animal performance was obtained weekly with samples of feces collected for microbiology on the first and the last experimental day and scored from 1 (liquid) to 4 (hard). On the last day, 1 pig from each pen was sampled for blood analyses of the acute-phase protein, major acute-phase protein of pigs (PigMap) and subsequently killed to register the digestive tract weight (including contents) and colon histology. Pigs fed the HF diets had greater ADG (390 vs. 457 g; P < or = 0.001) and large intestine weight (4.4 vs. 5.4% of BW; P < or = 0.05). This coincided with a greater (P < or = 0.05) short-chain fatty acid concentration (especially of acetic and butyric acids), a decrease in Escherichia coli counts (7.77 vs. 6.86 log of cfu/g of feces, P < or = 0.05), and an increase in the ratio of lactobacilli:enterobacteria (0.76 vs. 1.37, P < or = 0.05). However, CP level did not modify the productive performance, but 20% CP increased P < or = 0.05) the relative weight (% of BW) of the small (6.5 vs. 7.7) and large intestine (3.8 vs. 4.3). In the large bowel, the 20% CP diet increased the numbers of goblet cells (4.6 vs. 5.4/100 microm; P < or = 0.05) and reduced the numbers of intraepithelial lymphocytes (1.8 vs. 1.3/100 microm; P < or = 0.05). In relation to health status, increasing DF was dependent of the dietary CP content. Supplementing the 16% CP diet with DF reduced the fecal score and increased the antibiotics interventions, whereas the opposite was the case in the 20% CP diet. Pigs fed the 20% CP diet showed decreased (P < or = 0.05) PigMap concentrations than pigs fed 16% CP diets. As a whole, CP showed major effects on the gastrointestinal weight and gut barrier integrity, whereas DF increased the productive performance and promoted major changes in the microbial colonization and fermentation variables.

Effectiveness and safety of a treatment regimen based on isoniazid plus vaccination with Mycobacterium tuberculosis cells' fragments: field-study with naturally Mycobacterium caprae-infected goats.

The identification of a herd of goats with tuberculosis let us test a new treatment regimen against latent tuberculosis infection (LTBI). Using large animal experimental models allows a better approach to understanding human tuberculosis according to immunopathological parameters. Based on an initial study showing a correlation between the ESAT-6-specific interferon (IFN)-gamma secretion and the severity of pulmonary lesions, this parameter was used in combination with an X-ray examination to screen the animals to be included in the efficacy and safety studies. All the animals proved to be infected with Mycobacterium caprae. The efficacy study was run in animals distributed in three experimental groups according to treatment: untreated (CT), treated with isoniazid (INH), and treated with INH + RUTI (a vaccine based on M. tuberculosis cell fragments) inoculated twice. RUTI temporarily increased the IFN-gamma production after stimulating the peripheral blood with ESAT-6, purified protein derivative and RUTI in vitro. The INH chemotherapy reduced both pulmonary and extra pulmonary affectation, but not disease in pulmonary lymph nodes. The addition of RUTI may have decreased extrapulmonary disease further but had no benefit to lung or lung lymph-nodes itself. Safety studies showed that inoculation of RUTI caused a temporary increase of rectal temperature (1-2 degrees C) and local swelling, both adverse effects being well tolerated. Neither systemic toxicity nor mortality was induced by the vaccination. The control of goats' infection by the therapeutic regimen consisting in INH chemotherapy + RUTI as well as its safety, represented a further step towards testing its effects in human LTBI in a future.

One dose of a porcine circovirus 2 (PCV2) sub-unit vaccine administered to 3-week-old conventional piglets elicits cell-mediated immunity and significantly reduces PCV2 viremia in an experimental model.

The immunogenicity and efficacy generated by one dose of a PCV2 sub-unit vaccine (Porcilis PCV) were evaluated in 3-week-old conventional piglets. Vaccination induced both humoral and cell-mediated responses against PCV2, which were increased after the challenge with a PCV2 genotype "b" isolate. High levels of maternally derived antibodies (IPMA >or= 10 log(2)) at the time of vaccination were found to interfere with the active seroconversion, whereas titres below 8 log(2) allowed the development of a proper antibody response. Nevertheless, the immunity induced by one dose of the product was partly protective against PCV2 infection, since viremia, shedding and viral load in tissues were significantly reduced in vaccinated pigs compared to controls.