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Genetic relatedness of the nucleoprotein (NP) of recent swine, turkey, and human influenza A virus (H1N1) isolates.

The sequences of nucleoprotein (NP) genes of recent human and turkey isolates of influenza A viruses, which serologically could be correlated to contemporary swine viruses, were determined. These sequences were closely related to the NPs of these swine viruses and they formed a separate branch on the phylogenetic tree. While the early swine virus from 1931 resembled the avian strains in consensus amino acids of the NP and in its ability to rescue NP ts mutants of fowl plague virus in chicken embryo cells, the later strains on that branch were different: at 15 positions they have their own amino acids and they rescued the NP ts mutants only poorly. Of the NPs of the human New Jersey/76 isolates analysed, one clustered with the recent H1N1 swine viruses of the U.S.A., the other one with contemporary human strains. Since the NP is one of the main determinants of species specificity it is concluded that, although the H1N1 swine isolates from the U.S.A. form their own branch in the phylogenetic tree, they can be transmitted to humans and turkeys, but they do not spread further in these populations and so far have not contributed to human pandemics. It is not very likely that they will do so in future, since its branch in the phylogenetic tree develops further away from the human and avian branch.

Animals↗

Swine influenza virus and the recycling of influenza-A viruses in man.

Sera collected in 1967 and 1972 from people in the 0-100 age-group showed haemagglutination-inhibition (H.I.) antibody to swine virus A/Iowa/15/30 (Hsw1N1) in greatest number and with highest titre in people born before 1918. A slight decrease was observed from 1967 to 1972 in the number of sera with antibody to swine virus and in the height of the titres. The recently isolated A/New Jersey/10/76 (Hsw1N1) virus showed a result comparable to that of the Swine/1930 virus in sera of 1972. On the analogy of the findings in 1968, when the Hong Kong virus became epidemic in human populations and antibody to this virus was found in sera of people over 70 years, the suggestion is made that the recurrence of swine virus as an epidemic agent of human influenza may be expected around 1986. Fourfold or greater increase of antibody to Swine/1930 virus was observed in about 4--5% of people infected by or immunised with H3N2 viruses. This response occurred in people who had been in touch with the epidemic influenza-A viruses Hsw1N1, H0N1, and H1N1 during the swine era of 1918 to 1956. Following immunisation with H3N2 viruses of persons showing no response to H3N2 viruses in their serum 5% did show a fourfold or greater heterotypic H.I. antibody rise to swine virus. This finding is of consequence for the diagnostic serology of influenza.

Adolescent↗

Estrogen content of dairy and swine wastes.

Naturally occurring estrogens in animal wastes may cause negative environmental impacts, yet their abundance in animal waste treatment and storage structures is poorly documented. To better quantify estrogen concentrations in animal wastes, multiple waste samples were collected from treatment and storage structures at dairy and swine facilities and analyzed for concentrations of 17beta-estradiol (E2), estrone (E1), and 17alpha-estradiol by gas chromatography-mass spectroscopy and by enzyme linked immunosorbent assay (E2 only). Mass ratios of each estrogen to the macronutrients nitrogen, phosphorus, and potassium were also determined. Because manure application rates are typically macronutrient-based, estrogen to macronutrient ratios are proportional to areal mass application rates of estrogen to fields. Swine farrowing waste (from farrowing sows and piglets) had the highest ratios of E2 to macronutrients. Mean ratios in swine farrowing waste were roughly twice those in swine finishing waste (from growing male and nonpregnant female animals) and more than four times higher than those in dairy waste (from lactating cows in various stages of their reproductive cycles); these differences were statistically significant (alpha = 0.05). Estrone followed a similar trend. In contrast, ratios of 17alpha-estradiol to macronutrients were highest in dairy operations. These results can be used to better predict estrogen loading rates on fields receiving swine and dairy wastes.

Agriculture↗

Histopathologic and immunohistochemical comparison of human, rabbit, and swine aneurysms embolized with platinum coils.

BACKGROUND AND PURPOSE: The purpose of this study was to clarify the cellular mechanisms of aneurysmal healing by comparing histologic and immunohistochemical findings in experimental rabbit and swine aneurysms to a human aneurysm embolized with platinum coils. METHODS: Swine sidewall aneurysms (n = 5, harvested at 12 weeks) and elastase-induced rabbit aneurysms (n = 6, harvested at 24 weeks) were created and embolized. A single human aneurysm, embolized 6 years before death, was harvested following autopsy. All specimens were processed by using a modified paraffin embedding technique. Tissue was sectioned and stained with hematoxylin and eosin and Masson trichrome. Immunohistochemistry and immunofluorescence were performed with multiple antibodies, including alpha smooth muscle actin, myosin heavy chain, desmin, vimentin, and CD31. RESULTS: The human aneurysm's dome was filled with loose, hypocellular, amorphous tissue. The aneurysm's neck was completely covered with a thin layer of hypocellular tissue. Collagen and myofibroblasts were sparse in both the dome and neck. Rabbit aneurysms' domes were also filled with a loose, hypocellular tissue, amorphous matrix. In 5 of 6 aneurysms, a thin layer of hypocellular tissue ran along the neck. Collagen and myofibroblasts were sparse in the dome. Swine aneurysms were filled with densely infiltrated tissue, including chronic inflammatory tissue and extensive, attenuated collagen fiber bundles associated with myofibroblasts. Thick layers of myofibroblasts entirely bridged the necks. CONCLUSIONS: Absence of collagen deposition and scant myofibroblastic reaction to platinum coil embolization are seen in the rabbit model but not in swine aneurysms. The elastase-induced aneurysm model in rabbits is more suitable than sidewall swine aneurysms for testing of modified devices aimed at improving intra-aneurysmal fibrosis.

Actins↗

CDKN2A region polymorphism and genetic susceptibility to melanoma in the melim swine model of familial melanoma.

Some herds of miniature swine are genetically predisposed to cutaneous melanoma. To test if swine melanoma susceptibility could be linked to the CDKN2A gene, which is involved in a proportion of 9p21-linked human familial melanoma, we performed a genetic analysis of miniature pigs of the MeLiM strain. F(1) and backcross animals were generated by crossing 1 MeLiM boar with healthy Duroc sows. We isolated the swine CDKN2A gene and characterized a linked informative microsatellite marker, the S0644 marker. Using this marker and 2 flanking markers, we analyzed the segregation of the CDKN2A gene in a 3-generation pedigree. Allelic association, linkage analysis and haplotype analysis of these data led to exclusion of the CDKN2A gene as a candidate for melanoma susceptibility. Nonetheless, this analysis suggests an association with the swine 1q25 chromosomal region, which is homologous to the human 9p21 region.

Alleles↗

Comparative pathology of bacterial enteric diseases of swine.

Enteric bacterial infections are among the most common and economically significant diseases affecting swine production worldwide. Clinical signs of these infections include diarrhea, reduced growth rate, weight loss, and death of preweaned, weanling, grower-finisher, young and adult age breeding animals. The most common etiological agents include Escherichia coli, Clostridium perfringens, Lawsonia intracellularis, Salmonella enterica, and Brachyspira (Serpulina) spp. With the exception of Brachyspira (Serpulina) hyodysenteriae, the cause of swine dysentery, and Lawsonia intracellularis, the cause of proliferative enteropathy, the pathological changes seen with these agents closely resemble the diseases occurring in human beings. Histological changes in the intestines of swine with enteric bacterial infections include bacterial colonization without significant damage (e.g., certain enterotoxigenic E. coli and C. perfringens type A), attaching and effacing lesions with enteropathogenic E. coli and Brachyspira pilosicoli, the cause of colonic spirochetosis, inflammation with S. enterica, and necrotizing and hemorrhagic lesions with certain C. perfringens. Extraintestinal spread of bacteria and/or toxins occurs with some serotypes of E. coli and most serotypes of S. enterica. Enteric bacterial diseases of swine have been used as models to study the pathogenesis of similar diseases of human beings. Several of these pathogens are also important causes of food-borne disease in humans.

Animals↗

A six month dermal irritation test with anthralins in the Göttingen miniature swine.

Anthralins, especially dithranol, have been used for the treatment of chronic dermatoses for many years. Unfortunately dithranol stains the skin yellow or brown. It also irritates healthy skin more than psoriatic areas. A new anthralin derivative, butantrone may be tolerated better. Because the skin of the swine resembles human skin in many respects, the Göttingen miniature swine was selected as the test animal for the present study. 12 Göttingen miniature swines of the uncoloured line were randomized into three groups. The two anthralin derivatives were spread in three separate bases (ointment, stick and gel) and in three concentrations each on the allocated skin sites on the flanks of the animals. The responses of the skin were evaluated daily (except Sundays) for six months. Because there was a separate group for the control animals (bases only), for dithranol and butantrone, it was possible to make observations on systemic toxicity, too. Butantrone and dithranol turned out almost equally irritating to the skin of the miniature swine. The mean primary irritation scores in the butantrone group were slightly higher than in the dithranol group. On the other hand, the concentrations of butantrone were higher. It is possible that the release of the drugs from the gel formula was poorer than from the ointment and stick bases.

Animals↗

Relationship between fumonisin contamination of feed and mystery swine disease. A case-control study.

Fumonisin is a recently identified mycotoxin that has been shown to be the cause of pulmonary edema disease in swine and leukoencephalomalacia in horses. Mystery Swine Disease (MSD), is an economically devastating disease complex of unknown etiology that has been reported to have occurred in several swine producing states since 1988. To determine the relationship between MSD and fumonisin, a case-control study was carried out in Illinois in mid-1990. Feed samples collected from 12 case and 9 control farms were analyzed for fumonisin. Sera from swine on all farms was screened for titers against encephalomyocarditis (EMC) virus and concentrations of alpha-1 acid glycoprotein (an acute phase reactive protein). Fumonisin concentrations greater than or equal to 20 ppm were found on 1 control farm (1/9) and 8 case farms (8/12). Titers against EMC virus (greater than or equal to 1:16) were found on 5 control farms (5/9) and on 6 case farms (6/12). Farms with greater than or equal to 20 ppm fumonisin in the feed were at significantly increased risk (OR = 11.2, Fisher's exact test p = 0.037) for MSD. Furthermore, the pi2 test for trend was (p = 0.017), meaning that as the level of fumonisin in the feed increased, the risk of MSD also increased. The presence of EMC virus titers in the sow herd was not a significant risk for MSD (OR = 1.25, Fisher's exact test p = 0.75). Alpha-1 acid glycoprotein concentrations obtained from a 2-week old nursing pigs differed significantly (p = 0.0005) between MSD case and control herds.

Animal Feed↗

Transmission of Sarcocystis suihominis from humans to swine to nonhuman primates (Pan troglodytes, Macaca mulatta, Macaca irus).

Sporocysts of Sarcocystis suihominis obtained from human feces were used to infect swine. Heart, tongue, and skeletal muscle from experimentally infected and noninfected control swine were fed via stomach tube to nonhuman primates including chimpanzees (Pan troglodytes), rhesus monkeys (Macaca mulatta), and cynomolgus monkeys (Macaca irus). All primates fed infected swine tissues shed sporocysts beginning 13 to 15 days postinfection and were still shedding sporocysts at the conclusion of the experiment, 30 days postinfection. Rhesus and cynomolgus monkeys were fed infected swine tissues a second time and shed sporocysts. All primates remained in good health throughout both experiments and exhibited no unusual clinical signs as a result of infection.

Animals↗

Pseudorabies virus infectivity for swine skin characterized in vitro.

The infectivity of pseudorabies virus (PrV) was demonstrated in a cell substrate derived from swine skin explant cultures designated primary porcine skin cells (c/cSLA PPSC). c/cSLA PPSC infected with either wild type or TK- PrV strain Kaplan (Ka) developed typical cytopathologic changes (CPE) as early as 4 h post inoculation (p.i.). The CPE caused by PrV on c/cSLA PPSC was specifically neutralized by covalescent swine sera. Synthesis of late viral proteins was demonstrated in PrV-infected c/cSLA PPSC by indirect fluorescent antibody staining using monoclonal antibodies (mAbs) specific for PrV gIII. PrV induced protein synthesis was further confirmed by specific immunoprecipitation of 35S-methionine labeled viral polypeptides from PrV-infected c/cSLA PPSC with PrV convalescent swine serum, PrV immune mouse serum or mAb to PrV gIII. Moreover, the virus progeny derived from c/cSLA PPSC was shown to be infectious for MDBK cells and this infection was specifically neutralized by PrV convalescent swine serum. The capacity c/cSLA PPSC to support a complete growth cycle of PrV and the relative ease of deriving these cells from pigs can be applied in an autologous fashion in studies of cellular immunity where the MHC needs to be matched.

Animals↗

Virologic and serologic surveillance for human, swine and avian influenza virus infections among pigs in the north-central United States.

Influenza virus infection in pigs is both an animal health problem and a public health concern. As such, surveillance and characterization of influenza viruses in swine is important to the veterinary community and should be a part of human pandemic preparedness planning. Studies in 1976/1977 and 1988/1989 demonstrated that pigs in the U.S. were commonly infected with classical swine H1N1 viruses, whereas human H3 and avian influenza virus infections were very rare. In contrast, human H3 and avian H1 viruses have been isolated frequently from pigs in Europe and Asia over the last two decades. From September 1997 through August 1998, we isolated 26 influenza viruses from pigs in the north central United States at the point of slaughter. All 26 isolates were H1N1 viruses, and phylogenetic analyses of the hemagglutinin and nucleoprotein genes from 11 representative viruses demonstrated that these were classical swine H1 viruses. However, monoclonal antibody analyses revealed antigenic heterogeneity among the HA proteins of the 26 viruses. Serologically, 27.7% of 2,375 pigs tested had hemagglutination-inhibiting antibodies against classical swine H1 influenza virus. Of particular significance, however, the rates of seropositivity to avian H1 (7.6%) and human H3 (8.0%) viruses were substantially higher than in previous studies.

Amino Acid Sequence↗

Effect of levamisole on parenteral vaccines for swine dysentery.

Leucocyte migration-inhibition and humoral antibody responses (HAR) were demonstrated in swine immunized with particulate or soluble antigen of Treponema hyodysenteriae. Levamisole (at the recommended deworming level), given simultaneously with particulate vaccine did not significantly (P greater than 0.05) enhance nor suppress the leucocyte migration-inhibition response. However, an enhancing leucocyte migration-inhibition of the drug was seen in pigs given soluble vaccine in combination with levamisole compared with those receiving soluble antigen alone. Levamisole generally suppressed the HAR throughout the immunization schedule of pigs given soluble or particulate vaccine. There was no significant suppression (P greater than 0.05) of clinical signs of swine dysentery (SD) in animals given particulate vaccine nor in those receiving this vaccine plus levamisole. However, pigs receiving soluble vaccine plus levamisole had fewer clinical signs of SD as well as significantly (P less than 0.05) fewer shedding episodes of T. hyodysenteriae than those given soluble antigen alone. When compared with the control pigs, swine vaccinated with soluble antigen had fewer days of total diarrhoea and shedding episodes of T. hyodysenteriae. However, the diarrhoea of vaccinated pigs was not significantly (P greater than 0.05) delayed when compared to the unvaccinated swine.

Animals↗

Intimal cell masses in the abdominal aortas of swine fed a low-fat, low-cholesterol diet for up to twelve years of age.

The normal subendothelial intima of large arteries in man, swine and most other species is a variegated structure from birth onwards. In some regions it contains only a few scattered cells; in others there may be a continuous single layer of cells; and in still others the cells pile up to form what we have called intimal cell masses (ICM). The cells in the normal ICM are mostly smooth muscle cells although there is also a small resident population of monocyte-like cells. We have been studying the ICM in swine with emphasis on the abdominal aorta. We have found that atherosclerotic lesions in the abdominal aorta of swine induced by high-fat high-cholesterol diets begin by a hyperplastic reaction of the smooth muscle cells in the ICM and progress to form large lesions characterized by extensive regions of lipid-rich calcific necrotic debris similar to advanced lesions in man. Because of the putative key role of the ICM in atherogenesis we think that it is important to learn as much as possible about their natural history under conditions as normal as possible. In this report we present data on ICM in the abdominal aortas of 34 male and female Hormel miniature swine maintained on a low-fat low-cholesterol diet for up to 12 years of age. The ICM grow slowly with aging and in the distal portion of the aorta account for an average of 9% in the male and 15% in the female of the total cells in the aortic wall (intima + media).(ABSTRACT TRUNCATED AT 250 WORDS)

Age Factors↗

Role of the host cell nucleus in the replication of African swine fever virus DNA.

An examination by autoradiography of African swine fever virus-infected alveolar macrophages pulse labeled with [3H]thymidine showed that, at early times of viral DNA replication, the grains were localized exclusively in the nucleus in 20% of the cells, while in 45% the label was found in the cytoplasm. In the remaining 35%, newly synthesized DNA was detected in both the nucleus and the cytoplasm. At later times, the percentage of cells with grains in the nucleus decreased considerably. Pulse-chase experiments indicated that the DNA synthesized in the nucleus is then transported to the cytoplasm. The presence of virus-specific DNA sequences in the nucleus was confirmed by in situ hybridization of infected macrophages. Similar hybridization experiments with African swine fever virus-infected VERO cells followed by confocal microscopy also indicated the existence of a nuclear stage in the localization of the viral DNA. These results suggest a mechanism for African swine fever virus DNA replication with an initial stage in the nucleus followed by a cytoplasmic phase. Specific nuclear forms associated with the hybridization signal have been observed in African swine fever virus-infected macrophages and VERO cells. The nuclear forms seen in macrophages are consistent with a mechanism for the egress of the viral DNA from the nucleus that involves initial budding at the nuclear membrane.

African Swine Fever Virus↗

Immunodiagnosis of swine cysticercosis by indirect ELISA employing a heterologous antigen from Taenia crassiceps metacestode.

C. Larralde et al. (1990, Aoff. Pathol. Lab. Med., 114:926-928) demonstrated that heterologous antigen from the laboratory-adapted murine Taenia crassiceps metacestode may substitute those from Taenia solium in the immunodiagnosis of human cysticercosis by the indirect enzyme-linked immunosorbent assay (IE). This antigen is easily obtained at a laboratory level and solves the problem of T. solium cysticerci collection from naturally or experimentally infected swine. In this study an IE employing a heterologous antigen from the T. crassiceps metacestode was evaluated for the immunodiagnosis of swine cysticercosis. Sera from 300 swine free of T. solium cysticerci by post-mortem examination were employed to determine two IE cut-off values: 1) Mean ELISA values + 2 standard deviations (2 sigma cut-off) and 2) - Mean ELISA values + 3 standard deviations (3 sigma cut-off). The specificity of IE was 97% with the 2 sigma cut-off and 100% with the 3 sigma cut-off. When applied to ten sera from swine infected by cysticerci of T. solium by post-mortem examination, the sensitivity of IE was 100% independent of the cut off.

Animals↗

Detection of Eperythrozoon suis DNA from swine blood by whole organism DNA hybridizations.

A procedure is described for isolating Eperythrozoon suis DNA of sufficient quantity and purity to serve as a probe in whole-organism DNA hybridizations for detecting parasitized swine. The E. suis organisms were isolated from the blood of infected swine; the DNA was recovered and digested with restriction endonucleases and resolved on agarose gels. In DNA hybridizations using recovered E. suis DNA, blood samples from parasitized swine could be differentiated from uninfected, control samples. A high salt lysate recovery technique was used in sampling swine whole blood for E. suis DNA and found to offer many advantages in the collection and recovery process.

Animals↗

Monitoring experimental swine dysentery: rectal swab blood test and Serpulina (Treponema) hyodysenteriae detection.

In two experiments, ten postweaning piglets were inoculated intragastrically with cultures of Serpulina (Treponema) hyodysenteriae strain B204 and were individually monitored for swine dysentery. Eight control animals received sterile culture medium. Between the day of inoculation and 35 days later, rectal swab samples were assayed for fecal blood by means of a commercial test kit and for S. hyodysenteriae cells by direct (phase contrast) microscopy and selective culture methods. Optimal detection of S. hyodysenteriae required both microscopy and culturing. Of 41 rectal swab samples positive for fecal blood, 38 (93%) were positive by direct microscopy, by selective culture, or by both techniques. Of 118 samples negative for fecal blood, 115 (97%) were negative by direct microscopy, by selective culture, or by both techniques. Swab samples from control swine were negative for fecal blood. The fecal blood test was a convenient and reliable method for objectively monitoring individual animals with experimental swine dysentery. The occult blood test used in these studies might be generally useful for veterinary surgeons to monitor and diagnose swine intestinal disorders in which fecal blood is produced.

Animals↗

Characterization of transmissible gastroenteritis coronavirus S protein expression products in avirulent S. typhimurium delta cya delta crp: persistence, stability and immune response in swine.

The spike protein from transmissible gastroenteritis virus (TGEV) was expressed in attenuated S. typhimurium delta cya delta crp delta asd chi 3987. Three partially overlapping fragments of TGEV S gene, encoding the amino-terminal, intermediate, and carboxy-terminal end of the protein, as well as the full length gene were inserted into the asd+ plasmid pYA292 to generate recombinant plasmids pYATS-1, pYATS-2, pYATS-3, and pYATS-4, respectively, which were transformed into S. typhimurium chi 3987. Recombinant S. typhimurium chi 3987 (pYATS-1) and chi 3987 (pYATS-4) expressing constitutively a 53 kDa amino-terminal fragment of the S protein and the full length protein (144 kDa), respectively, showed high stability. After 50 generations in vitro 60% and 20% of the bacteria transformed with pYATS-1 and pYATS-4, respectively, expressed the S-protein antigen. Since S. typhimurium chi 3987 (pYATS-1) showed a better level of expression and stability in vitro, this recombinant strain was selected as a potential bivalent vector to induce both immunity to Salmonella and TGEV in swine. In order to study colonization of swine tissues by S. typhimurium delta cya delta crp, a gene conferring resistance to rifampicin was cloned into the chromosome of S. typhimurium chi 3987, generating chi 4509 strain. Both S. typhimurium chi 4509 (pYA292) and chi 4509 (pYATS-1) colonized the ileum of orally inoculated swine with clearance of bacteria between days 10-20 post-infection. The expression of the amino-terminal fragment of the S protein diminished the ability of S. typhimurium chi 4509 (pYATS-1) to colonize deep tissues. The recombinant strain S. typhimurium chi 3987 (pYATS-1) induced TGEV specific antibodies in both serum and saliva of orally inoculated swine.

Animals↗