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Porcine reproductive and respiratory syndrome (PRRS) is considered one of the most relevant diseases of swine. The condition is caused by PRRS virus (PRRSV), an extremely variable virus of the Arteriviridae family. Its heterogeneity can be responsible, at least partially, of the poor cross-protection observed between PRRSV isolates. Neutralizing antibodies (NAs), known to play a role in protection, usually poorly recognize heterologous PRRSV isolates, indicating that most NAs are strain-specific. However, some pigs develop broadly reactive NAs able to recognize a wide range of heterologous isolates. The aim of this study was to determine whether PRRSV isolates that induce broadly reactive NAs as determined in vitro are able to confer a better protection in vivo. For this purpose two in vivo experiments were performed. Initially, 40 pigs were immunized with a PRRSV-1 isolate known to induce broadly reactive NAs and 24 additional pigs were used as controls. On day 70 after immunization, the pigs were divided into eight groups composed by five immunized and three control pigs and exposed to one of the eight different heterologous PRRSV isolates used for the challenge. In the second experiment, the same experimental design was followed but the pigs were immunized with a PRRSV-1 isolate, which is known to generate mostly strain-specific NAs. Virological parameters, specifically viremia and the presence of challenge virus in tonsils, were used to determine protection. In the first experiment, sterilizing immunity was obtained in three groups, prevention of viremia was observed in two additional groups, although the challenge virus was detected occasionally in the tonsils of immunized pigs, and partial protection, understood as a reduction in the frequency of viremia compared with controls, was recorded in the remaining three groups. On the contrary, only partial protection was observed in all groups in the second experiment. The results obtained in this study confirm that PRRSV-1 isolates differ in their ability to induce cross-reactive NAs and, although other components of the immune response might have contributed to protection, pigs with cross-reactive NAs at the time of challenge exhibited better protection, indicating that broadly reactive NAs might play a role in protection against heterologous reinfections. This study was supported by grants AGL2007-66695, AGL2008- 05708-C02 and CSD-2006-00007 from the Spanish Government and FEI 20/39 from the University Complutense of Madrid. FM-L and F-DF were funded by Project Consolider-Ingenio 2010 CDS2006-00007

Porcine reproductive and respiratory syndrome virus (PRRSV) isolates are classified in two different genotypes, based on genomic heterogeneity: type 1, which comprises European type isolates, and type 2, which includes North American type isolates. It is believed that members of both genotypes differ in some biological properties including pathogenicity, however extensive studies comparing isolates of both genotypes have never been carried out. The objective of the present study was to compare the pathogenic properties of six different PRRSV isolates, three of type 1 and three of type 2, in a young pig infection model. For this purpose, a total of 105 3-week-old piglets were divided in 7 groups of 15 animals that were exposed on day 0 of the experiment to one of the six isolates tested or were mock infected (negative control group). Clinical signs and rectal temperatures were recorded daily and blood samples were taken on days 3, 6, 9, 12, 15, 18 and 21 of the experiment. On days 7, 14 and 21 post-inoculation five animals per group were sacrificed, macroscopic lung lesions were evaluated and different tissue samples were collected to determine viral organic distribution. The results obtained indicate that type 2 isolates are more pneumovirulent than type 1 isolates, as demonstrated by the recording of respiratory clinical signs only in pigs exposed to type 2 viruses and by the severity of macroscopic and microscopic lung lesions in those pigs. However, no clear differences could be established between genotypes in systemic clinical signs or viral load and viral distribution after challenge. These results support the general idea that type 2 isolates induce more severe respiratory disease than type 1 isolates.

The aim of the present study was to establish the degree of diversity of porcine reproductive and respiratory virus (PRRSV) isolates that circulate in the same geographical area in different years. Nucleotide sequences of open reading frame (ORF) 5 were determined for 28 Spanish field PRRSV isolates from different years and three European-type modified live virus vaccines. Sequences were aligned using Clustal W software and a phylogenetic tree constructed using the neighbour joining method. The results of pairwise homology comparisons of nucleotide and deduced amino acid sequences of these PRRSV isolates indicate a tendency for heterogeneity to increase with time. The study of the phylogenetic tree revealed that Spanish PRRSV isolates constitute two well-defined clades and a group of unrelated sequences. The observed heterogeneity does not appear to be due to temporal evolution exclusively. Early and recent isolates group themselves into different clusters independently of the time of isolation, indicating the co-circulation of different variants and the maintenance of variants of the original isolates in the field.

The objective of this study was to determine the degree of protection conferred by a Lelystad-like modified live virus (MLV) vaccine against a heterologous wild-type porcine reproductive and respiratory syndrome virus (PRRSV) isolate of the same cluster. For this purpose, fourteen 3-week-old piglets were divided into three groups: Group A pigs were vaccinated with a modified live virus vaccine, Group B pigs were used as positive controls, and Group C pigs as negative controls. Twenty-eight days after the last dose of vaccine, all pigs in Groups A and B were inoculated with the Spanish PRRSV strain 5710. To evaluate efficacy, clinical signs were recorded and the presence of challenge virus was determined by virus isolation in blood samples and nasal swabs collected at various time points post-challenge (p.c.) and in tissue samples collected at necropsy 24 days p.c. After challenge, moderate clinical signs were observed in pigs from Groups A and B. In addition, all vaccinated pigs were viremic at least once, although viremia tended to be more sporadic in this group than in Group B pigs. PRRSV was detected in at least one tissue sample from four out of five pigs from Group A and in all pigs from Group B. The results indicate that the protection conferred by the MLV vaccine used in this study against a closely related virulent strain was only partial. The findings suggest that the degree of genetic homology of ORF5 between MLV vaccine and challenge isolate is not a good predictor of vaccine efficacy.

Porcine reproductive and respiratory syndrome virus (PRRSV) has been reported to be shed in the semen of infected boars. To determine whether the reproductive tissues could be a persistent source of virus and the possible origin of PRRSV found in semen of infected boars, 20 PRRSV-seronegative boars were intranasally inoculated with 5×10 6 median tissue culture infective doses (TCID 50 ) of PRRSV and necropsied at different times post-inoculation (p.i.) from Day 2 to Day 37 p.i. Blood samples were collected before experimental inoculation, at necropsy and at different times p.i. At necropsy, epididymal semen and reproductive tissues were collected and the presence of the virus determined by virus isolation. The infection of the boars was demonstrated by the isolation of the virus from the sera of all inoculated boars and by seroconversion. PRRSV was detected in serum samples from Day 2 to Day 23 p.i., although the viremic period was largely dependent on the individual response to infection. Viral replication was proven within different reproductive tissues from Day 2 to Day 23 p.i., being most consistently found in the epididymus. In addition, PRRSV was isolated in semen from Day 4 to Day 10 p.i. The correlation of a diminished viremia and the inability to isolate PRRSV from semen or reproductive tissues may be due to one of two possibilities. First, viremia is responsible for most of the virus isolated from reproductive tissues due to the movement of PRRSV-infected cells out of the blood and into the tissues. Second, viremia may initially seed the reproductive tissues with PRRSV, and then the virus is produced into the reproductive tract and shed into semen at low levels.

The objective of this study was to determine the protective properties of the Spanish MLV vaccine Amervac-PRRS® against porcine reproductive and respiratory syndrome virus (PRRSV) infection with the homologous wild-type strain5710. The experimental design comprised eighteen 3-week old pigs randomly divided into three groups. Animals fromgroup A received one dose of MLV vaccine Amervac-PRRS on days –42 and –21. Animals from groups B and C served as infected and uninfected controls, respectively. On day 0, pigs from groups A and B were intranasally challenged with homologous strain 5710 and 18 days after challenge all pigs were euthanized. The results reported in this study show that, although the titre of neutralizing antibodies elicited by vaccination was modest, MLV vaccine Amervac-PRRS® provided protection against homologous PRRSV infection and drastically reduced post-challenge viremia and completely block PRRSV dissemination to peripheral tissues.

We report a case of bacteremia in puppies caused by Streptococcus dysgalactiae subsp. dysgalactiae. Identification was achieved by phenotypic and molecular genetic methods. This is the first report of the recovery of S. dysgalactiae subsp. dysgalactiae from dogs.

Twenty-five gilts without measurable porcine reproductive and respiratory syndrome (PRRS) virus (PRRSV) serum antibody titres were used for this experiment. All of them were randomly assigned to one of the treatment groups at the time of artificial insemination. Twelve gilts were exposed to PRRSV, of these, six were slaughtered on day 10 after exposure and constituted group A. The remaining six were slaughtered on day 20 after infection and constituted group C. Thirteen gilts were used as controls, six of these were slaughtered on day 10 after treatment and constituted group B. The remaining seven were slaughtered on day 20 after treatment and constituted group D. The infected gilts were inoculated with PRRSV intranasally and intravenously in the ear vein. They were observed for clinical signs of infection and the effects on conception and fertilization rates were studied, while the gilts and their embryos were tested for PRRSV and homologous antibodies. The infected animals developed signs of PRRS associated with anorexia and slight pyrexia. Infection was verified by reisolation of the virus from serum and other tissue samples and also by seroconversion. Ten out of 12 infected gilts and 10 out of 13 controls were pregnant at the time of slaughter and the ratio of embryos to corpora lutea was the same in both, infected and control groups (0.75). Therefore, infection with PRRSV at the onset of gestation did not appear to interfere with conception and fertilization rates and subsequent pregnancy. The PRRSV was not isolated from any of the embryos collected at day 10 postexposure, but was present in 20-day-old embryos of group C gilts. In this group, 60% of litters were infected prenatally, with 16% of embryos infected. The proportion of dead embryos was three times greater than in a control group D (35.4% and 9.8%, respectively). The results of this report indicate that exposure of susceptible gilts to PRRSV at the onset of gestation has no significant effect on conception and fertilization rates. However, although infection does not appear to have any effect on the embryos before implantation, it can result in transplacental infection and embryo death.

Twenty-one gilts without measurable PRRSV serum antibody titres were identified for this experiment. Seven gilts were used as controls (Group C) and 14 as principals. Of these, 7 gilts were preimmunized to PRRSV and constituted Group B, while 7 gilts remained seronegative and constituted Group A. The principal gilts were inseminated with boar semen containing PRRSV and were killed 20 d later. The control gilts were treated similarly but were not exposed to PRRSV. Gilts were observed for clinical signs of infection. The effects on the conception rates were studied and gilts and embryos were tested for PRRSV and homologous antibodies. Group A and B gilts developed signs of PRRS associated with anorexia and slightly elevated body temperatures. Transmission of the infection was demonstrated by the isolation of PRRSV from serum and other tissue samples of principal gilts and also by seroconversion. The results show that early infection may have an insignificant effect or no effect on the conception and fertilization rates. However, exposure to PRRSV at the time of insemination can result in transplacental infection of embryos. In Group A gilts, 5 of 6 litters were infected prenatally with 7.6% of embryos infected. In Group B gilts, 1 of 5 litters and 1.3% of embryos were infected. Moreover, approximately 2 and 4 times more embryos were dead in litters of gilts from Group A and Group B than in gilts from control Group C. The isolation of PRRSV in 3 dead embryos suggests that the embryos may have died as a result of the direct effect of the virus. It can be concluded that the insemination of either seronegative or preimmunized gilts with boar semen containing PRRS V may have an insignificant effect or no effect on conception and fertilization rates, although it can result in transmission of the virus and embryonic infection and death.

The objective of this study was to compare the safety of all modified live virus vaccines commercially available in Europe against Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) under the same experimental conditions. For this purpose, one hundred and twenty three-week-old piglets, divided into five groups, were used. On day 0 of the experiment, nine pigs per group were removed and the remaining fifteen were vaccinated with the commercial vaccines Ingelvac PRRS MLV, Amervac PRRS, Pyrsvac-183 and Porcilis PRRS by the IM route or were mock vaccinated and used as controls. On day 3, the nine unvaccinated pigs were re-introduced into their respective groups and served as sentinel pigs. Clinical signs were recorded daily and lung lesions were determined on days 7, 14 and 21, when 5 vaccinated pigs per group were euthanized. Blood samples and swabs were taken every three days and different organs were collected at necropsy to determine the presence of PRRSV. None of the vaccines studied caused detectable clinical signs in vaccinated pigs although lung lesions were found. Altogether, these results indicate that all vaccines can be considered clinically safe. However, some differences were found in virological parameters. Thus, neither Pyrsvac-183 nor Porcilis PRRS could be detected in porcine alveolar macrophage (PAM) cultures or in lung sections used to determine PRRSV by immunohistochemistry, indicating that these viruses might have lost their ability to replicate in PAM. This inability to replicate in PAM might be related to the lower transmission rate and the delay in the onset of viremia observed in these groups

Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) is highly heterogenic. This heterogeneity has an effect on antigenic composition of PRRSV and might create differences in sensitivity to neutralization between isolates. The sensitivity to neutralization could be an important feature of PRRSV isolates because it is likely that isolates resistant to neutralization pose a significant challenge for the development of vaccines that elicit broad protective immunity. Nonetheless, little information is available for understanding or categorizing the viral neutralization phenotype of PRRSV isolates. Consequently, the main purpose of this study was to determine whether PRRSV isolates differ in their susceptibility to neutralization and if they can be classified in different categories based on their neutralization phenotype. For this purpose, a panel of 39 PRRSV isolates and a set of 30 hyperimmune monospecific sera were used in cross-neutralization assays. The results of this study indicate that PRRSV isolates differ in their sensitivity to neutralization and k-means clustering system allowed classifying the isolates in four different categories according to their neutralization phenotype: highly sensitive, sensitive, moderately sensitive and resistant to neutralization. Further analyses using two additional clustering systems that considered individual data for the classification of the isolates confirmed that classification obtained by k-means is accurate in most cases and that only in a few instances classification is less stringent. Sequences of GP3, GP4 and GP5 were analyzed but no correlation could be found between the sequence of previously identified neutralizing epitopes or the number of N-linked glycosylation sites in different proteins and the neutralization phenotype of the isolates. These data provide the first systematic assessment of overall neutralization sensitivities of a panel of diverse PRRSV isolates. The classification of the isolates provides a useful tool to facilitate the systematic characterization of neutralizing antibody production elicited by new vaccine candidates.

VP2 is the major component of canine parvovirus (CPV) capsids. The VP2-coding gene was engineered to be expressed by a recombinant baculovirus under the control of the polyhedrin promoter. A transfer vector that contains the lacZ gene under the control of the p10 promoter was used in order to facilitate the selection of recombinants. The expressed VP2 was found to be structurally and immunologically indistinguishable from authentic VP2. The recombinant VP2 shows also the capability to self-assemble, forming viruslike particles similar in size and appearance to CPV virions. These viruslike particles have been used to immunize dogs in different doses and combinations of adjuvants, and the anti-CPV responses have been measured by enzyme-linked immunosorbent assay, monolayer protection assays, and an assay for the inhibition of hemagglutination. A dose of ca. 10 micrograms of VP2 was able to elicit a good protective response, higher than that obtained with a commercially available, inactivated vaccine. The results indicate that these viruslike particles can be used to protect dogs from CPV infection.

Disease exacerbation was observed in pigs challenged with virulent porcine reproductive and respiratory syndrome virus (PRRSV) following immunisation with a recombinant GP5 sub-unit PRRSV vaccine (rGP5) produced in E. coli. Eighteen animals were divided into three experimental groups: group A were immunised twice IM with rGP5, 21 days apart; group B acted as positive controls (challenged but not immunised); and group C were negative controls. Pigs in groups A and B were challenged 21 days after the second immunisation of the group A animals. Following challenge, three pigs given rGP5 exhibited more severe clinical signs than the positive controls, including respiratory distress and progressive weight-loss. Although not statistically significant, the more severe disease exhibited by group A animals may suggest previous immunisation as a contributory factor. The mechanisms of these findings remain unclear and no association could be established between the severity of disease, non-neutralising antibody concentrations and tissue viral loads.

This study was designed to evaluate the efficacy of an inactivated vaccine based on a European-type strain of porcine reproductive and respiratory syndrome virus (PRRSV) against the reproductive form of the syndrome in breeding gilts, and any congenital disease in their piglets. Five gilts were vaccinated twice, following the manufacturer's instructions, before they were inseminated. Nine additional gilts remained unvaccinated and served as positive (five gilts) and negative (four gilts) controls. A European wild-type strain genetically divergent from the vaccine strain was used to challenge the five vaccinated and five unvaccinated positive control gilts at 90 days' gestation. The vaccination of the five seronegative gilts did not produce any clinical signs or adverse reactions. However, the vaccine failed to prevent the clinical signs associated with PRRSV infection, viraemia after the challenge and transplacental infection of their piglets. The reproductive performance of the vaccinated gilts was similar to that of the unvaccinated positive controls, and there were no statistically significant differences in most of the parameters tested. However, the preweaning mortality of the piglets born to the vaccinated gilts was significantly lower than that of the piglets born to the positive control gilts.

An unusual Gram-negative, catalase- and oxidase-positive, rod-shaped bacterium isolated from different clinical samples from two monkeys (Callithrix geoffroyi) was characterized by phenotypic and molecular genetic methods. The micro-organism was tentatively identified as a Pseudomonas species on the basis of the results of cellular morphological and biochemical tests. Fatty acid studies confirmed this generic placement and comparative 16S rRNA gene sequencing studies demonstrated that the unknown isolates were phylogenetically closely related to each other (100 % sequence similarity) and were part of the ‘Pseudomonas fluorescens intrageneric cluster’. The novel bacterium, however, was distinguished from other phylogenetically related species of Pseudomonas by DNA–DNA hybridization studies and biochemical tests. On the basis of both phenotypic and phylogenetic findings, it is proposed that the novel Pseudomonas isolates are classified as Pseudomonas simiae sp. nov. The type strain of P. simiae is OLiT (=CCUG 50988T=CECT 7078T).

The objective of this study was to evaluate the efficacy of two commercially available modified live virus vaccines for preventing the reproductive and early postnatal consequences of infecting (challenging) pregnant gilts with virulent porcine reproductive and respiratory syndrome virus (PRRSV). For this purpose 21 crossbred gilts were allocated to one or another of four groups (Groups A-D). Group A comprised four gilts neither vaccinated nor challenged; Group B comprised five gilts that were challenged but not vaccinated; Group C comprised seven gilts that were vaccinated (AmervacPRRS) and challenged; Group D comprised five gilts that were vaccinated (Pyrsvac-183) and challenged. Vaccination was 24 days before conception, and challenge was at 90 days of gestation. Both vaccine viruses and the challenge virus were European strains but differed in part from one another on the basis of their genetic (nucleotide) sequence. After challenge PRRSV was isolated from five (100%), four (57%), and two (40%) of the gilts of Groups B, C and D, respectively. Although vaccination failed to prevent a detectable viremia in all of the gilts of Groups C and D after they were challenged (or congenital infection of some of their pigs), it did provide a statistically significant level of protection in regard to the incidence of congenital infection, reproductive performance, and pig health and viability. Namely, for Groups C and D the numbers of liveborn pigs/litter and healthy pigs/litter throughout the early postnatal period were similar to those of Group A (nonvaccinated and nonchallenged) and far exceeded those of Group B (nonvaccinated and challenged).

The purpose of this study was to assess the effects of two currently available commercial European-type modified-live virus (MLV) vaccines against porcine reproductive and respiratory syndrome in a reproductive pig model. Sixteen 90-day pregnant gilts were divided into four groups and allocated to one of the following intranasal treatments: group A gilts served as negative controls; group B gilts were exposed to a virulent European field strain; group C gilts were exposed to vaccine strain VP046 Bis and group D gilts to vaccine strain All-183. The results indicated that MLV strains can replicate in breeding animals and have the ability to cross the placenta. In particular, viraemia was detected in all gilts in group C and 2/4 gilts in group D, at least at one time point. In addition, transplacental infection was demonstrated in 3/4 gilts in group C and 2/4 gilts in group D. However, congenital and early postnatal infection did not have a marked detrimental effect on piglet performance when compared to negative controls, and no statistically significant differences were observed in most cases. Conversely, the reproductive performance of gilts in group B was significantly worse than that of the other groups. Specifically, the number of born-alive piglets, the survival rate of piglets during lactation and the mean weight of weaned pigs were significantly lower. It was concluded that the two commercial European-type MLV vaccines tested had no marked detrimental effects in pregnant gilts, although the MLV strains can cross the placenta leading to the birth of congenitally infected piglets.

Twenty-five gilts without measurable serum antibody titres to porcine reproductive and respiratory syndrome virus (PRRSV) were identified and 16 were inoculated with PRRSV at seven, 14 or 21 days of gestation and killed 20 to 22 days later to determine the effect of the virus on their embryos. The remaining nine gilts were not exposed to PRRSV, but were killed at the same stages of gestation. The gilts were observed for clinical signs of infection and the gilts and their embryos were tested for PRRSV and homologous antibodies. The infection was demonstrated by the re-isolation of the virus and its detection by the reverse transcriptase polymerase chain reaction in serum and other tissue samples from the inoculated gilts, and also by seroconversion. However, the gilts remained healthy throughout the study, except for one which was depressed and anorexic for two days. Two of the litters from the gilts challenged with PRRSV on day 14 of gestation contained one and three infected live embryos; the other embryos from these two litters did not contain detectable virus, although most of the embryos in one of the litters were dead. The other nine litters from the gilts challenged with PRRSV and the control litters, showed no evidence of infection.

Fertilized pig ova, collected from 14 estrus-synchronized gilts 32 h after presumed ovulation were microinjected or cultured for 72 h in Beltsville Embryo Culture Medium-3 with or without Porcine Reproductive and Respiratory Syndrome Virus (PRRSV). To detect virus in the samples, virus isolation of swine alveolar macrophage, Reverse Transcriptase Polymerase Chain Reaction and Fluorescent Antibody techniques were employed. Microinjection or incubation of embryos with PRRSV did not significantly inhibit development of the porcine embryos in vitro when compared with that of controls (P = 0.75 and P = 0.14, respectively). Although either 10 to 20 TCID50 were microinjected or large concentrations of virus were used for embryo exposure by incubation, PRRSV was not detected in association with the embryos. It is concluded based on the experiments reported here that 4- to 16-cell stage pig embryos are not susceptible to productive infection with PRRSV in vitro.

Eleven boars seronegative to porcine reproductive and respiratory syndrome virus (PRRSV) were trained for semen collection: five boars were inoculated intranasally with 6 x 10(6)TCID(50)/ml of PRRSV (Group A); four boars were inoculated intranasally with 6 x 10(4)TCID(50)/ml (Group B); and two boars were used as uninfected control (Group C). Semen samples were collected at 7-d intervals from 49 d prior to experimental inoculation with PRRSV to 70 d after inoculation, and were examined for sperm volume, sperm concentration, sperm morphology, sperm motility and for the presence of PRRSV. The infection in boars was demonstrated by the reisolation of PRRSV from the serum of all inoculated boars. Rectal temperatures and general health of the boars were clinically normal throughout the trial. Differences were observed in the quality of semen collected from boars after experimental infection with PRRSV. This infection induced a significant decrease in sperm motility and in spermatozoa with normal acrosomes. Of the semen samples tested for virus isolation in swine alveolar macrophages PRRSV was only isolated in 1 boar from Group B. The virus was detected in an additional semen sample in Group A by the production of an antibody titer in a biological assay. All attempts to detect PRRSV by RT-PCR in semen samples were unsuccessful. Nevertheless, from our study it is possible to suggest that the PRRSV can occasionally be transmitted in the semen during the initial phase of the disease.

The use of modified live virus (MLV) vaccines is a common procedure to control porcine reproductive and respiratory syndrome virus (PRRSV) infection in the great majority of countries from America, Asia and Europe, including Spain. Current discriminatory techniques allow the detection of different MLV type-II vaccine strains. Herein we report a rapid and accurate technique aimed to discriminate between MLV type-I vaccine strains and Spanish field strains. This technique comprises a reverse transcription (RT) and nested polymerase chain reaction (nPCR) amplification of PRRSV ORF5 followed by a digestion of RT-nPCR products with two specific endonucleases, ItaI and AccI. Combined utilization of ItaI and AccI generates restriction fragments length polymorphisms (RFLP) patterns adequate for the differentiation of 30 Spanish field isolates, of which 12 were isolated between 1991 and 1995 and 18 between 2000 and 2003. These different RFLP patterns can be used to distinguish unequivocally between Spanish field strains of PRRSV and the three MLV type-I vaccines used in Spain: AmervacPRRS®, Pyrsvac-183® and PorcilisPRRS®., Para controlar la infección por el virus del síndrome reproductor y respiratorio porcino (PRRSV), en la gran mayoría de países de América, Asia y Europa, incluyendo España, se usan frecuentemente vacunas basadas en virus vivos modificados (MLV). En la actualidad existen técnicas discriminatorias que permiten detectar cepas vacunales del PRRSV de tipo II. El presente trabajo describe una técnica precisa y rápida para la diferenciación de cepas vacunales de tipo I del PRRSV y cepas de campo españolas. Esta técnica se basa en la transcripción reversa y posterior amplificación de la ORF5 del genoma del PRRSV utilizando la reacción en cadena de la polimerasa anidada, seguida de la digestión de los amplicones generados con dos endonucleasas específicas: ItaI y AccI. La utilización combinada de ambas enzimas genera patrones de polimorfismos en la longitud de fragmentos de restricción (RFLP), adecuados para la distinción de las 30 cepas de campo usadas, de las cuales 12 fueron aisladas entre 1991 y 1995 y 18 entre 2000 y 2003. Estos diferentes patrones pueden ser utilizados para distinguir entre cepas de campo españolas del PRRSV y las tres cepas vacunales de tipo I usadas en España: AmervacPRRS®, Pyrsvac-183® y PorcilisPRRS®.

Summary Serological response and reproductive performance were estimated in field trials of an inactivated virus vaccine against porcine parvovirus. Experiments were carried out in 10 selected pig breeding herds. A total of 277 seronegative gilts were used. Two hundred and twenty animals were vaccinated twice before mating, fourteen days apart and revaccinated after farrowing. Blood samples were obtained from both vaccinated and non-vaccinated (57 animal) control gilts, one week after the 2nd dose of vaccination, at farrowing time and one week after revaccination. Although there were considerable variations among the herds, the number of returns to oestrus in all herds was higher in vaccinated gilts (11.81%) than in the controls (10.52%). This difference, however, was not statistically significant. The reproductive performance results revealed the absence of an increase in the total born, as pooled values, in vaccinated gilts compared to controls. However, when these results are interpreted in relation to serological data, many control gilts were already seropositive before mating, or remained seronegative at farrowing. According to our results, the duration of immunity with this vaccine is apparently short, as there is a clear decrease in the titres between the 1st and the 2nd sampling times (2.35 ± 0.14 and 1.97 ± 0.08, respectively).

Summary 2,979 sera were collected from slaughtered swine in two geographic areas of Spain from 1987 to 1989. They were tested for antibodies against an H1N1- and H3N2-influenza virus by haemagglutination-inhibition tests (HI). The percentage of positive sera was higher in area I (78 %-69.2 %) than in area II (63.1 % -60.4%) for both viruses respectively. The coexistence of high titres to both H1N1- and H3N2-influenza virus became apparent in cold months simultaneously in each area, although influenza viruses circulated in the Spanish swine population for two years. Also this study suggests the possible circulation of A/Texas/1/77-like strains in Spain, results which have not been reported before. Resumen Se recogieron en matadero 2979 sueros a partir de porcino de cebo procedente de dos areas geograficas de Espana desde 1987 a 1989. Los sueros fueron enfrentados a un virus influenza H1N1 y a otro H3N2 mediante la tecnica de inhibicion de la hemoaglutinacion. Los porcentajes de sueros positivos fue mas elevado en el area I (78 % -69,2 %) que en el area II (63,1 %-60,4 %) para ambos virus, respectivamente. La coexistencia de cifras elevadas de anticuerpos frente a ambos virus se evidencio durante los meses frios y de forma paralela en las dos areas, aunque virus influenza circularon entre la poblacion porcina espanola a lo largo de los dos anos. Ademas en este estudio se sugiere la posible circulacion de virus analogos a la cepa A/Texas/1/77 en Espana, extremo no comunicado antes.

Eight boars 11 months of age that were seronegative to pseudorabies virus were trained for hand collection of semen. A genetically engineered pseudorabies virus vaccine was given to 7 of 8 boars, while the eighth was left unvaccinated to serve as a control. Semen samples were collected at 7 day intervals from 42 days prior to vaccination through 49 days after vaccination. Rectal temperatures and general health of the boars were clinically normal throughout the trial, and no clear differences were observed in the quality of semen collected from the boars before or after vaccination. The semen samples were tested for vaccine virus and none was detected. © 1992.

The presence of antibodies to two influenza viruses of the type A (H1N1 and H3N2) and to a porcine respiratory coronavirus was investigated in a study lasting a year. 735 blood serum samples were collected from 79 closed pig fattening farms in the province Segovia (Spain). Hemagglutination inhibition was used with influenza viruses. The percentage of positive results was 78.5% and 62.5% respectively for the serotypes H1N1 and H3N2. A clear reduction in the spread of antibodies was observed in the autumn. The ELISA technique was used with the porcine respiratory coronavirus. As antigen we used the antigenically related transmissible porcine gastroenteritis virus. Using this technique 87% of the sera were positive. Some of these sera with representative ELISA values were confirmed by means of serum neutralisation and radioimmune precipitation of the viral proteins. The incidence of these antibodies remained unchanged over the whole year of the investigation.