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An accelerated solid-phase enzyme-immunoassay has been developed which permits identification of antigens and antibodies to them within 30-40 min, even directly at the site of specimen collection. The method was tested on the models of foot-and-mouth disease virus, vesicular disease of swine, vesicular exanthema of swine, Aujeszky's disease, leukemia, and coronavirus infection of cattle.
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Blood samples collected from 120 wild swine (Sus scrofa) in thirteen Oklahoma (USA) counties during 1996 were tested for antibodies against six viral and two bacterial diseases. No antibodies to swine brucellosis, pseudorabies, transmissible gastroenteritis, and vesicular stomatitis were detected. Antibody titers to one or more leptospiral serovars were found in 44% of the samples, the two most frequent serovars being Leptospira interrogans serovars bratislava (29%) and pomona (27%). Antibody against porcine parvovirus and swine influenza virus was detected in 17% and 11% of the swine, respectively. Two samples (2%) were positive for antibody to the recently emerged porcine reproductive and respiratory syndrome virus.
To help policy makers determine the need for current regulations (which require cooking of swill prior to feeding to swine), an assessment of the likelihood of exposing domestic swine in the continental United States of America (USA) to selected foreign animal disease agents by feeding uncooked swill was carried out. The hazard was assumed to originate from contraband food items entering the USA and subsequently being discarded in household waste. Such food waste may be collected by licensed waste feeders and fed to swine. This study showed that, of the four diseases studied, the probability of exposure was highest for the classical swine fever (hog cholera) virus. The median annual likelihood of one or more contaminated loads of swill being fed to swine in the continental USA was estimated as follows: classical swine fever virus: 0.063, foot and mouth disease virus: 0.043, swine vesicular disease virus: 0.005, African swine fever virus: 0.005.
Human infectious diseases have been studied in pigs because the two species have common microbial, parasitic, and zoonotic organisms, but there has been no systematic evaluation of cytokine gene expression in response to infectious agents in porcine species. In this study, pigs were inoculated with two clinically and economically important parasites, Toxoplasma gondii and Ascaris suum, and gene expression in 11 different tissues for 20 different swine Th1/Th2-related cytokines, cytokine receptors, and markers of immune activation were evaluated by real-time PCR. A generalized Th1-like pattern of gene expression was evident in pigs infected with T. gondii, along with an increased anti-inflammatory gene expression pattern during the recovery phase of the infection. In contrast, an elevated Th2-like pattern was expressed during the period of expulsion of A. suum fourth-stage larvae from the small intestine of pigs, along with low-level Th1-like and anti-inflammatory cytokine gene expression. Prototypical immune and physiological markers of infection were observed in bronchial alveolar lavage cells, small intestinal smooth muscle, and epithelial cells. This study validated the use of a robust quantitative gene expression assay to detect immune and inflammatory markers at multiple host tissue sites, enhanced the definition of two important swine diseases, and supported the use of swine as an experimental model for the study of immunity to infectious agents relevant to humans.
The concept of fertility problems is defined in the introduction and their economic importance in pig breeding are pointed out. The infectious causes of the problem are reviewed. Infections with L. tarassovi and with Aujezky's virus regularly lead to practically endemic abortions; Brucella suis and swine-plague infections do so sporadically. The SMEDI syndrom is considered. The clinical symptoms, diagnosis, epizootiology, and the therapeutical and prophylactic measures of these infections are discussed. In the case of brucellosis, attention is paid to the differential diagnosis in the serological examination. The incidence of Yersinia enterocolitica among the Dutch pig population is high. This micro-organism has antigen components in common with brucellae. In Holland the significance of infectious agents as the causes of sterility is not considered to be high.
Vaccination programs to control Aujeszky's-disease virus (ADV) using gE-deleted vaccines are being considered in several European countries. Knowledge of factors influencing ADV-seropositivity for vaccinated herds might contribute to the success of these programs. A multivariable analysis of ADV-seropositivity in 1248 swine herds (332 farrow-to-finish, 260 farrow-to-feeder and 656 finishing herds) in the Emilia-Romagna region of Italy revealed that (1) high pig density (number of pigs in a 6-km radius), (2) gilt purchasing, and (3) increased number of fattening pigs were risk factors for farrow-to-finish herds. In farrow-to-feeder herds, ADV-seropositivity was related to (1) increased number of breeders, (2) heavy-gilt purchasing, and (3) increased pig density. In finishing herds, (1) increased herd size was related to ADV-seropositivity, whereas (2) periodic rearing suspension was protective.
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The results of a two-year study on swine stephanuriasis in Cuba indicate the high economic importance of this parasitosis in view of its pattern of local distribution and pathological changes occurring during the endogeneous phase of its life cycle. Under the present conditions of pig breeding in Cuba, it appears to be most economic to base control measures on a strict observance of general zoohygienic rules and on several specific measures suggested in the paper. The administration of expensive drugs is uneconomic because none of them is universally effective in any phase of the life cycle of S. dentatus. It is suggested to direct research work to suitable preventive measures and the choice of drugs to be administered during the most critical period in the spread of infection covering the time from birth to weaning.
A multiplex, real-time reverse transcription-polymerase chain reaction (RT-PCR) assay was developed that allowed simultaneous detection and rapid differentiation of vesicular stomatitis virus strains--New Jersey (VSV-NJ) and Indiana 1, 2, and 3 (VSV-IN1-3). This assay involves use of a set of VSV universal primers located in the L gene that amplify VSV-IN1-3 and VSV-NJ using probes that allow differentiation of the major serotypes Indiana and New Jersey. The assay was evaluated using reference VSV, foot-and-mouth disease virus, swine vesicular disease virus, and vesicular exanthema of swine virus. To estimate diagnostic sensitivity, 159 epithelial samples collected between 1996 and 2002 from naturally infected cattle in Colombia were used. The assay cut off was calculated by testing RNA extracted from 150 virus-negative bovine tissues consisting of tongue, soft palate, muzzle, coronary band, and lymph node. All infected cattle were test positive for VS by results of real-time RT-PCR analysis; results for 156 of 159 (98.1%) agreed with the serotype determination from the complement-fixation test. Amplification did not occur in any of the negative bovine epithelial samples, allowing the cut-off values for the assay to be set. The real-time RT-PCR assay was documented to be sensitive and specific for the detection of VSV-NJ and VSV-IN (1-3) strains from field samples in a single reaction, thereby supporting use of this assay in the differential diagnosis of vesicular virus diseases in cattle.
Antibodies to porcine circovirus (PCV) which is the smallest animal virus known so far were found in 77-95 per cent of sera from slaughter pigs gathered in Berlin and two districts of Northern Germany. About 60 per cent of these positive sera had relatively high titres similar to those in experimentally infected pigs 3-6 weeks after infection. This indicates that the animals might have become infected during the fattening period. Sera from 2-3 year old pigs from a laboratory animal breeding institution were also found positive (83 per cent) but titres were lower. Experimentally infected minipigs developed antibodies and virus was isolated from nasal swabs and from fecal samples. The animals neither showed any signs of illness nor were pathological changes noticable. The assumption that PCV is a common virus in all swine populations was strengthened by the finding of PCV antibodies in wild boars shot in the forests of the Berlin region.
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Erysipelothrix rhusiopathiae is pathogenic for both animals and humans, causing erysipelas in swine and erysipeloid in humans. In swine, disease may be either acute or chronic, resulting in the development of arthritis and endocarditis. In Japan, erysipelas remains an animal hygiene problem causing great economic loss as infected swine are disused. Human infection closely resembles that seen in swine, with both acute and chronic forms also. The most common presentation is erysipeloid, a localized cutaneous infection. In Western Australia, an erysipeloid-like infection referred to as "crayfish poisoning" occurs in lobster fishermen and handlers. A second type of presentation is a generalized cutaneous form involving lesions that progress from the initial site of infection or appear in remote areas. The third and most serious form of disease is a septicemia that is almost always linked to endocarditis. The mortality rate in Erysipelothrix endocarditis is still high (38%) and can be explained by the use of vancomycin (to which Erysipelothrix spp. are inherently resistant) as empirical therapy. Therefore, it is critical to have an early diagnosis of E. rhusiopathiae infection.Unfortunately, several problems exist with the diagnosis of E. rhusiopathiae infections by conventional cultural procedures, and these infections are often incorrectly diagnosed. First, because of their very small colony size and slow growth rates, it is difficult to isolate E. rhusiopathiae from heavily contaminated specimens. Various selective media have been described to improve the isolation of E. rhusiopathiae from contaminated specimens; however, not all contaminants are inhibited. The development of two polymerase chain reaction (PCR) methods has created an opportunity to greatly improve the efficiency with which these organisms are detected and identified. Makino et al. designed a PCR method that amplifies a 407-bp DNA fragment derived from the 16S rRNA coding sequence. The primers in this method are specific for the genus Erysipelothrix and do not differentiate between the species. A second set of primers designed by Shimoji et al. amplifies a 937-bp DNA fragment which is derived from a sequence associated with virulence of E. rhusiopathiae. These primers are specific for E. rhusiopathiae only. Shimoji et al. also utilized a selective enrichment medium based on tryptic soy broth containing ethidium bromide and sodium azide.
A plaque-purified preparation of foot-and-mouth disease virus (FMDV) of serotype C1 (C-S8c1-1), grown in cell culture, was used to infect nonimmunized pigs. No variant genomes were detected in the average populations of 50 viruses isolated from infected animals by direct RNA sequencing of the carboxy-terminal half of the VP1 gene. However, a mutant with altered phenotypic properties was present in low proportion in an infected animal. The frequency of mutants resistant to neutralization by SD6 monoclonal antibody (MAb) [SD6 epitope MAb-resistant mutants (MARMs)], directly estimated in virus from lesions of infected animals (without passage in cell culture), depended on the procedure used for its determination and ranged from 2.9 x 10(-6) (when the virus was incubated with the MAb prior to plating) to 2.6 x 10(-5) (when incubation with MAb was avoided and the MAb was maintained in the agar overlay of the titration assay). Such a difference was not found for C-S8c1-1, which consistently showed frequencies of about 4 x 10(-5). In addition, the repertoire of amino acid substitutions was similar among SD6 epitope MARMs isolated directly both from vesicles of infected animals and from C-S8c1-1. Thus, in spite of the conservation of the average sequence in the region of VP1 RNA analyzed, antigenic heterogeneity has been found in viral populations of FMDV upon replication in nonimmunized swine.